Assessment of milk metabolites as biomarkers for predicting feed efficiency in dairy sheep.

Estimating feed efficiency (FE) in dairy sheep is challenging due to the high cost of systems that measure individual feed intake. Identifying proxies that can serve as effective predictors of FE could make it possible to introduce FE into breeding programs. Here, 39 Assaf ewes in first lactation were evaluated regarding their FE by 2 metrics, residual feed intake (RFI) and feed conversion ratio (FCR). The ewes were classified into high, medium and low groups for each metric. Milk samples of the 39 ewes were subjected to untargeted metabolomics analysis. The complete milk metabolomic signature was used to discriminate the FE groups using partial least squares discriminant analysis. A total of 41 and 26 features were selected as the most relevant features for the discrimination of RFI and FCR groups, respectively. The predictive ability when utilizing the complete milk metabolomic signature and the reduced data sets were investigated using 4 machine-learning algorithms and a multivariate regression method. The Orthogonal Partial Least Square algorithm outperformed other ML algorithms for the FCR prediction in the scenarios using the complete milk metabolite signature (r2 = 0.62 ± 0.06) and the 26 selected features (0.62 ± 0.15). Regarding RFI predictions, the scenarios using the 41 selected features outperformed the scenario with the complete milk metabolite signature, where the Multilayer feedforward artificial neural network (r2 = 0.18 ± 0.14) and extreme gradient boosting (r2 = 0.17 ± 0.15) outperformed other algorithms. The functionality of the selected metabolites implied that the metabolism of glucose, galactose, fructose, sphingolipids, amino acids, insulin, and thyroid hormones was at play. Compared with the use of traditional methods, practical applications of these biomarkers might simplify and reduce costs in selecting feed-efficient ewes.

Relationship between feed efficiency and resilience in dairy ewes subjected to acute underfeeding.

Selection of dairy sheep based on production levels has caused a loss of rusticity, which might compromise their future resilience to nutritional challenges. Although refocusing breeding programs toward improved feed efficiency (FE) is expected, more-efficient ewes also seem to be more productive. As a first step to examine the relationship between FE and resilience in dairy sheep, in this study we explored the variation in the response to and the recovery from an acute nutritional challenge in high-yielding Assaf ewes phenotypically divergent for FE. First, feed intake, milk yield and composition, and body weight changes were recorded individually over a 3-wk period in a total of 40 sheep fed a total mixed ration (TMR) ad libitum. Data were used to calculate their FE index (FEI, defined as the difference between the actual and predicted intake estimated through net energy requirements for maintenance, production, and weight change). The highest and lowest FE ewes (H-FE and L-FE groups, respectively; 10 animals/group) were selected and then subjected to the nutritional challenge (i.e., withdrawing the TMR and limiting their diet only to the consumption of straw for 3 d). Afterward, sheep were fed again the TMR ad libitum. Temporal patterns of variation in performance traits, and ruminal fermentation and blood parameters were examined. A good consistency between FEI, residual feed intake, and feed conversion ratio was observed. Results supported that H-FE were more productive than L-FE sheep at similar intake level. Average time trends of milk yield generated by a piecewise model suggest that temporal patterns of variation in this trait would be related to prechallenge production level (i.e., H-FE presented quicker response and recovery than L-FE). Considering all studied traits, the overall response to and recovery from underfeeding was apparently similar or even better in H-FE than in L-FE. This would refute the initial hypothesis of a poorer resilience of more-efficient sheep to an acute underfeeding. However, the question remains whether a longer term feed restriction might impair the ability of H-FE ewes to maintain or revert to a high-production status, which would require further research.

Effect of lipid supplementation on the endogenous synthesis of milk cis-9,trans-11 conjugated linoleic acid in dairy sheep and goats: A tracer assay with 13C-vaccenic acid.

A major proportion of milk rumenic acid (RA; cis-9,trans-11 CLA) is synthesized through mammary Δ9-desaturation of vaccenic acid (VA; trans-11 18:1). Diet composition may determine the relative contribution of this endogenous synthesis to milk RA content, with effects that might differ between ruminant species. However, this hypothesis is mostly based on estimated values, proxies of stearoyl-CoA desaturase (SCD) activity, and indirect comparisons between publications in the literature. With the aim of providing new insights into this issue, in vivo Δ9-desaturation of 13C-labeled VA (measured via milk 13C-VA and -RA secretion) was directly compared in sheep and goats fed a diet without lipid supplementation or including 2% of linseed oil. Four Assaf sheep and 4 Murciano-Granadina goats were used in a replicated 2 × 2 crossover design to test the effects of the 2 dietary treatments during 2 consecutive 25-d periods. On d 22 of each period, 500 mg of 13C-VA were i.v. injected to each animal. Dairy performance, milk fatty acid profile, including isotope analysis, and mammary mRNA abundance of genes coding for SCD were examined on d 21 to 25 of each period. Supplementation with linseed oil improved milk fat concentration and increased the content of milk VA and RA. However, the isotopic tracer assay suggested no variation in the relative proportion of VA desaturated to milk RA, and the percentage of this CLA isomer deriving from SCD activity would remain constant regardless of dietary treatment. These results put into question a major effect of lipid supplementation on the endogenous synthesis of milk RA and support that mammary Δ9-desaturation capacity would not represent a limiting factor when designing feeding strategies to increase milk RA content. The lack of diet-induced effects was common to caprines and ovines, but inherent interspecies differences in mammary lipogenesis were found. Thus, the higher proportions of VA desaturation and endogenous synthesis of milk RA in sheep supported a greater SCD activity compared with goats, a finding that was not associated with the similar mRNA abundance of SCD1 in the 2 species. On the other hand, transfer efficiency of the isotopic tracer to milk was 37% higher in caprine than in ovine, suggesting a greater efficiency in mammary fatty acid uptake from plasma in caprine.

Rumen biohydrogenation and milk fatty acid profile in dairy ewes divergent for feed efficiency.

A sustainable increase in livestock production would require selection for improved feed efficiency, but the mechanisms underlying this trait and explaining its large individual variation in dairy ruminants remain unclear. This study was conducted in lactating ewes to test the hypothesis that rumen biohydrogenation (BH) would differ between high- and low-efficiency animals, and these differences would be reflected in rumen fatty acid (FA) profile and affect milk FA composition. A second aim was to identify differences in FA that may serve as biomarkers of feed efficiency. Data of daily feed intake and milk yield and composition, as well as body weight, were collected individually over a 3-wk period in 40 ewes. The difference between the mean actual and predicted feed intake (estimated through metabolizable energy requirements for maintenance, production, and body weight change) over the period was used as the feed efficiency index (FEI) to select 8 of the highest feed efficiency (H-FE) and 8 of the lowest feed efficiency (L-FE) animals. In addition, residual feed intake (RFI) was estimated as the residual term from the regression of feed intake on various energy sinks. Rumen and milk FA composition were characterized by using gas chromatography, and results were analyzed using a statistical model that included the fixed effect of the group (H-FE vs. L-FE). The FEI averaged -0.29 ± 0.046 and 0.81 ± 0.084 in H-FE and L-FE, respectively, whereas RFI averaged -0.16 ± 0.084 and 0.18 ± 0.082, respectively. The correlation coefficient between both metrics was 0.69. Feed intake was similar in both groups, but H-FE showed greater milk yield, with increases in lactose content and yield, and in milk protein and fat production. Results from rumen FA profiles included a lower proportion of 18:2n-6, cis-9 18:1, and of several of their BH metabolites, and a greater concentration of 18:0, which may indicate that the apparent BH would be more complete in more efficient sheep. Milk FA analysis suggested that the greater fat yield in the H-FE group was mostly explained by increased de novo FA synthesis, whereas their milk would have lower proportions of cis-9 18:1 and C20 to 22n-6 polyunsaturated FA than L-FE. Stepwise multiple linear regression suggested that milk C20 to 22n-6 PUFA might be convenient biomarkers to discriminate more efficient dairy sheep. Further research is needed to validate these findings (e.g., under different dietary conditions).

Effect of dietary lipids and other nutrients on milk odd- and branched-chain fatty acid composition in dairy ewes.

Milk odd- and branched-chain fatty acids (OBCFA) are largely derived from bacteria leaving the rumen, which has encouraged research on their use as biomarkers of rumen function. Targeted research has examined relationships between these fatty acids (FA) and dietary components, but interactions between the effects of lipids and other nutrients on milk OBCFA are not well characterized yet. Furthermore, factors controlling milk OBCFA in sheep are largely unknown. Thus, the present meta-analysis examined relationships between diet composition and milk OBCFA using a database compiled with lot observations from 14 trials in dairy ewes fed lipid supplements. A total of 47 lots received lipid supplements, whereas their respective controls (27 lots) were fed the same basal diets without lipid supplementation. Relationships between milk OBCFA and dietary components were first assessed through a principal component analysis (PCA) and a correlation analysis. Then, responses of milk OBCFA to variations in specific dietary components (selected on the basis of the PCA) were examined in more detail by regression analysis. According to the loading plot, dietary unsaturated C18 FA loaded opposite to major milk OBCFA (e.g., 15:0, 15:0 anteiso, and 17:0) and were strongly correlated with principal component 1, which described 46% of variability. Overall, regression equations supported this negative, and generally linear, relationship between unsaturated C18 FA levels and milk OBCFA. However, the influence of C20-22 n-3 polyunsaturated FA and saturated FA was more limited. The PCA also suggested that dietary crude protein is not a determinant of milk OBCFA profile in dairy ewes, but significant relationships were observed between some OBCFA and dietary fiber or starch, consistent with a potential role of these FA as biomarkers of rumen cellulolytic and amylolytic bacteria. In this regard, regression equations indicated that iso FA would show opposite responses to increasing levels of acid detergent fiber (positive linear coefficients) and starch (negative linear coefficients). Lipid supplementation would not largely affect these associations, supporting the potential of OBCFA as noninvasive markers of rumen function under different feeding conditions (i.e., with or without lipid supplementation). Because consumption of these FA may have nutritional benefits for humans, the use of high-fiber/low-starch rations might be recommended to maintain the highest possible content of milk OBCFA in dairy sheep.

Endogenous synthesis of milk cis-9,trans-11 conjugated linoleic acid in dairy ewes: Quantification using 13C-labeled vaccenic acid and comparison with estimates based on cobalt administration.

Isotopic tracers are used to directly quantify the effect of mammary Δ9-desaturation on milk fatty acid (FA) composition, but very few studies have applied this method to measuring the endogenous synthesis of rumenic acid (RA; cis-9,trans-11 conjugated linoleic acid) in cows and goats, and no publications exist in ewes. In sheep, knowledge about the contribution of stearoyl-coenzyme A desaturase (SCD) to milk FA secretion is derived mostly from indirect estimates based on inhibition of the enzyme by oral administration of cobalt, a cost-effective method that has not been validated to date. To fill both gaps, we conducted an isotopic tracer assay in sheep to quantify the proportion of endogenous RA in milk for the first time in this species. We then compared the results with estimates derived from a Co administration assay performed on the same animals. First, 5 lactating ewes received an intravenous injection of 200 mg of [1-13C]trans-11 18:1 (vaccenic acid, VA), the precursor for RA production by SCD activity. At -24, -15, 0, 9, 24, 33, 48, 57, 72, 81, and 96 h post-injection, we recorded milk yield and collected milk samples to examine fat concentration and FA profile. We conducted compound-specific isotope analysis of VA and RA by gas chromatography-combustion isotope ratio mass spectrometry. Afterward, in the Co administration assay, ewes received a daily dose of 7 mg of Co/kg of body weight for 5 d. We analyzed milk samples for composition before and on the last days of cobalt dosing. On average, 17% of the injected amount of [1-13C]VA was transferred to milk within 96 h post-injection, and up to 29% of the VA taken up by the mammary gland was desaturated to milk RA. Under our conditions, the mean proportion of this conjugated linoleic acid isomer deriving from Δ9-desaturation represented 82 to 90% of the amount secreted in milk. However, the proportion estimated in the Co assay with calculations based on individual FA concentrations was lower (on average, 46%). When we calculated the same estimates based on changes in Δ9-desaturation ratios after Co dosing, the higher values of endogenous RA (75%) did not differ from the results of the isotopic tracer assay. Nevertheless, correlation analysis between the results obtained through [1-13C]VA or Co administration revealed no significant relationship, which would prevent acceptance of the latter as a reliable alternative to isotopic labeling to examine mammary Δ9-desaturation in dairy ewes.

Analysis of the management of antipsychotics in a group of prisons.

Objective: To assess the use of prescribed antipsychotic treatments in the Spanish prison population in order to determine whether there are differences in efficacy and cost between the different long-acting antipsychotic injectables (LAIs). Material and method: An observational, retrospective study was carried out in twelve prisons and in two prison psychiatric hospitals. To assess efficacy, all the clinical histories of patients with some kind of LAI were reviewed and only those who were in a situation of therapeutic stability were selected, defined as those treatments that had not undergone any change in the three last months, both in doses and in the association of another antipsychotic. Results: Of the 11,953 inmates included in the study, 1,851 (15.5%) received prescriptions for an antipsychotic, 415 in penitentiary psychiatric hospitals (87%) and 1,436 in prisons (12%), which is a much higher prevalence than that found in the population. Regarding the prescription pattern of LAIs, paliperidone is the most widely prescribed in prison psychiatric hospitals and prisons, followed by aripiprazole, zuclopenthixol and risperidone. Finally flufenazine, olanzapine and paliperidone quarterly, which are scarcely represented. Of the 292 patients with LAI, 41% (121 patients) are with monotherapy and 59% (171 patients) are with polytherapy, which is similar to the data found in the bibliography. Discussion: The use of prescribed LAIs amongst inmates is widespread. Polytherapy is also a common phenomenon, although there is a downward trend due to the use of more recently marketed drugs. Since the use of LAI is long-term, with numerous side effects and a wide range of prices, it is essential to consider the patient's risk factors and the cost of treatment alongside the therapeutic efficacy.

Rumen bacterial community responses to DPA, EPA and DHA in cattle and sheep: A comparative in vitro study.

The role of marine lipids as modulators of ruminal biohydrogenation of dietary unsaturated fatty acids may be explained by the effects of their n-3 polyunsaturated fatty acids (PUFA) on the bacterial community. However, the impact of individual PUFA has barely been examined, and it is uncertain which bacteria are truly involved in biohydrogenation. In addition, despite interspecies differences in rumen bacterial composition, we are not aware of any direct comparison of bovine and ovine responses to dietary PUFA. Therefore, rumen fluid from cannulated cattle and sheep were used as inocula to examine in vitro the effect of 20:5n-3 (EPA), 22:5n-3 (DPA), and 22:6n-3 (DHA) on the bacterial community. Amplicon 16 S rRNA sequencing suggested that EPA and DHA had a greater contribution to the action of marine lipids than DPA both in cattle and sheep. Certain effects were exclusive to each ruminant species, which underlines the complexity of rumen microbial responses to dietary fatty acids. Based on changes in bacterial abundance, Barnesiella, Prevotella, Paraprevotella, Hallela, Anaerovorax, Succiniclasticum, Ruminococcus and Ruminobacter may be involved in the ruminal response in biohydrogenation to the addition of marine lipids, but further research is necessary to confirm their actual role in ruminal lipid metabolism.

Objectively and subjectively measured physical activity and its relation to cardiovascular risk factors in older people-Which is most appropriate?

OBJECTIVES: To determine the strength of the association between objectively and subjectively measured physical activity (PA) and cardiovascular risk factors (CVRF) in a sample of older adults to assess which measure of PA is most appropriate for use in this population. STUDY DESIGN AND OUTCOME MEASURES: The study was a cross-sectional analysis of a community-dwelling sample aged 60 years or more (61 women and 34 men). Data collection included anthropometric measurements, blood analytics, and objective and subjective measurements of PA from accelerometers and Yale PA Survey questionnaires, respectively. RESULTS: Although the questionnaire indicated a higher summary index (total units) of PA in men than in women, objective measurements of total PA (mean counts per minute and steps per day) did not differ by sex. However, we obtained different results between women and men when we analyzed the relation between objective and subjective PA parameters and CVRF. In women, parameters that indicate cardiovascular risk, such as body mass index (BMI), waist circumference (WC), glucose, and total cholesterol, negatively correlated with objective PA parameters, while HDLc positively correlated with objective PA parameters and negatively with subjective PA parameters. In men, BMI and WC were negatively correlated with objective PA parameters, while blood cholesterol, LDLc, and triglycerides negatively correlated with subjective PA parameters. CONCLUSIONS: In women, it seems more appropriate to use objective methods to measure PA, while in men, objective and subjective methods seem to be complementary. Both methods of measurement, accelerometers and questionnaires, should be used simultaneously in research studies in older populations to establish precise relationships between PA and cardiovascular risk.

Serum adiponectin is associated with body composition and cognitive and psychological status in older adults living in long-term nursing homes.

Frailty is a geriatric syndrome characterized by decreasing functional reserves and increasing vulnerability to falls, injuries and declining health, leading to dependence upon caregivers. Frailty is associated not only with physical decline, but also with cognitive and psychological impairments in aging people. Higher serum adiponectin concentration has been linked to better performance on frailty measures but also to worse cognitive status. Nonetheless, several studies have proposed adiponectin as a frailty biomarker. To further delineate the relationship between adiponectin serum levels and frailty-related parameters, we studied a population of 112 long-term nursing home residents (aged 84.9 ±â€¯6.7) and analyzed their serum adiponectin levels in conjunction with frailty-related parameters including body composition, physical fitness, cognitive function, psychological parameters and quality of life. Frailty was assessed following the Fried Frailty Criteria, the Clinical Frailty Scale and the Tilburg Frailty Indicator. In women, higher serum adiponectin levels were associated with lower body weight, body mass index, body fat mass, fat mass/height2, lean mass, lean mass/height2 and smaller waist circumference and hip circumference (p < 0.05). In men, the association was positive (p < 0.05) between serum adiponectin and percentage of fat mass and negative between serum adiponectin and percentage of lean mass. Interestingly, in men, better cognitive function was inversely related to adiponectin (p < 0.05) while decreased anxiety was linked to a higher concentration of adiponectin in women (p < 0.05). According to the Tilburg Frailty Indicator, frail men had lower levels of adiponectin than those who were not frail (p < 0.05). Variables that predicted adiponectin concentration in multiple regression models were different for women and men. In women, lean mass and anxiety were independent negative predictors of blood adiponectin (ß = -0.363, p = 0.002; ß = -0.204, p = 0.067, respectively). In men, the Montreal Cognitive Assessment (MOCA) test was the only parameter to remain significant in the regression model (ß = -439, p = 0.015). The results of our study show that adiponectin is linked to body composition, cognitive function and anxiety in long-term nursing home residents with differential relationships by sex. Further studies should be conducted to determine whether adiponectin is a valid and reliable frailty biomarker.

In vitro biohydrogenation of 13C-labeled α-linolenic acid in response to ruminal alterations associated with diet-induced milk fat depression in ewes.

The basis for marine lipid-induced milk fat depression (MFD) has not been established yet, but recent reports suggest the putative contribution of shifts in the ruminal metabolism of α-linolenic acid (ALA). To test this hypothesis, an isotopic tracer approach was used in batch cultures of rumen microorganisms with inoculum collected from cannulated ewes fed either a total mixed ration without lipid supplementation (control inoculum) or the same diet supplemented with 2% of fish oil, which is known to cause MFD in lactating sheep (FO-MFD inoculum). The [1-13C]ALA was added at a dose of 1% of incubated dry matter and the proportions of 13C-labeled fatty acids (FA) were examined after 24 h of incubation, using complementary gas chromatography and gas chromatography-combustion isotope ratio mass spectrometry (GC-C-IRMS) analyses. Expected differences in FA profiles were confirmed between control and FO-MFD inocula (e.g., large decreases in 18:0 and increases in most 18:1 and 18:2 intermediates, particularly trans isomers, to fish oil supply). The biohydrogenation of 13ALA was extensive and yielded multiple metabolites, with a total of 48 chromatographic peaks showing 13C enrichment, regardless of the inoculum type. However, although ALA was biohydrogenated through common pathways under standard or MFD conditions, large changes in the accumulation of 13C-labeled FA suggest important differences in the relative contribution of each specific route. First, increased accumulation of trans-11-containing FA in FO-MFD incubations was accompanied by a general repression of the trans-13/14 pathway (supported by lower trans-13+14 18:1 or trans-11,trans-13 18:2 proportions), together with a lower production of cis FA (e.g., cis-9, -12, and -15 18:1 and some cis,cis 18:2). Results also downplayed the relevance of the trans-11 to trans-10 shift as an effective marker of diet-induced MFD in sheep, and challenged the involvement of some trans-10 intermediates (e.g., trans-10 18:1 and trans-10,cis-15 18:2) in the low-fat milk syndrome in this species. Conversely, increased abundance of most 18:3 intermediates (including some unidentified isomers) might be representative of ruminal alterations related to fish oil supplementation in ewes. Further research is necessary to examine the potential association between these findings and MFD in lactating animals.

Effects of shearing 2 breeds of dairy ewes during lactation under mild winter conditions.

The lactational effects of shearing (CO, control unshorn; SH, shorn) were investigated in 48 dairy ewes of 2 breeds (Lacaune, LC, n = 24; Manchega, MN, n = 24) having a similar stage of lactation (120 ± 6 d in milk) and body frame (65.1 ± 1.5 kg of body weight and 2.4 ± 0.1 body condition score), but differing in fleece and milk production. Ewes were penned indoors, adapted to the diet (alfalfa hay ad libitum and fixed amount of concentrate), and allocated for 30 d in 8 balanced groups to which the experimental treatments were applied. All ewes were sheared on the same day. Feed intake by pen and milk yield by ewe were recorded daily. Individual samples of milk (d -3, 3, 5, 7, and 15) and blood (d -7, 3, 7, and 15) were collected, as well as body weight and body condition score measured (d -15, 0, and 15), related to shearing. Pooled milk samples per pen were also collected before and after shearing for milk fatty acid analysis (d -3 and 15). Average temperatures in the barn before (12.6 ± 0.7°C) and after (13.7 ± 0.4°C) shearing were mild. Fleece was heavier in MN than in LC (1.04 ± 0.10 vs. 0.75 ± 0.09 kg/ewe) and tended to cover more body surface in MN than in LC ewes. Responses to shearing varied according to breed, the rectal temperature after shearing only decreasing significantly in the MN (-0.36 ± 0.09°C). Feed intake increased in the LC-SH (5%), when compared with LC-CO, but did not vary in the MN ewes. Ingestibility of the alfalfa hay, expressed as filling units for sheep and monitored in 2 groups of 6 dry and unshorn ewes of each breed (73.0 ± 2.5 kg of body weight and 3.1 ± 0.2 body condition score), was constant throughout the experiment (0.99 ± 0.03 filling units for sheep/kg of dry matter). Regarding milk production, LC-SH ewes yielded 10% more milk (1.38 ± 0.06 vs. 1.52 ± 0.05 kg/d) than LC-CO ewes, but no differences were detected in MN ewes (0.74 ± 0.03 kg/d, on average). No differences in the concentration of major milk components by effect of the shearing treatment were detected in either breed, but LC-SH ewes yielded 9% more milk protein than did LC-CO ewes. No relevant effects of shearing were detected on milk fatty acid profiles, although MN ewes showed lower C4:0, C6:0, C14:0, trans-11 and trans-12 C18:1 contents than did LC ewes. Moreover, no changes by effect of shearing were detected in plasma glucose, nonesterified fatty acids, cortisol, or insulin values in either breed, or in body weight or body condition score. In conclusion, shearing dairy ewes during lactation under mild winter conditions is a suitable management option that may increase feed intake and milk production, without deleterious effects on milk composition.

Análisis del manejo de antipsicóticos inyectables de larga duración en varios centros penitenciarios / Analysis of the management of antipsychotics in a group of prisons

Objetivos: Evaluar el uso de los tratamientos antipsicóticos prescritos en la población reclusa española y determinar si hay diferencias, en cuanto a eficacia y coste, entre los diferentes antipsicóticos inyectables de larga duración (ALD). Material y método: Estudio observacional, retrospectivo, efectuado en doce centros penitenciarios (CCPP) y en dos hospitales psiquiátricos penitenciarios. Para evaluar la eficacia, se revisaron todas las historias clínicas de pacientes con algún ALD y se seleccionaron los que estaban en situación de estabilidad terapéutica, definida como aquellos tratamientos que no habían experimentado ningún cambio en los tres últimos meses, tanto en dosis como en la asociación de otro antipsicótico. Resultados: De 11.953 internos incluidos, 1.851 (el 15,5%) tenían prescrito algún antipsicótico; 415 (el 87%) en los hospitales psiquiátricos penitenciarios y 1.436 (el 12%) en los CCPP. La paliperidona es el ALD más prescrito en hospitales psiquiátricos penitenciarios y en CCPP, seguido del aripiprazol, el zuclopentixol y la risperidona. La flufenazina, la olanzapina y la paliperidona trimestral están poco prescritos. El 41% de los pacientes eran tratados con monoterapia y el 59% con politerapia. Con los ALD de más reciente incorporación, se ha observado una tendencia a la disminución de la politerapia. Discusión: La prevalencia de prescripción de ALD en población reclusa es muy superior a la de la población no presa. La politerapia está muy extendida, aunque muestra una tendencia descendente con el uso de los fármacos más recientemente comercializados. Se recomienda que además de la eficacia terapéutica se valore a la hora de prescribir el riesgo de interacciones farmacológicas y el coste del tratamiento

Objective: To assess the use of prescribed antipsychotic treatments in the Spanish prison population in order to determine whether there are differences in efficacy and cost between the different long-acting antipsychotic injectables (LAIs). Material and method: An observational, retrospective study was carried out in twelve prisons and in two prison psychiatric hospitals. To assess efficacy, all the clinical histories of patients with some kind of LAI were reviewed and only those who were in a situation of therapeutic stability were selected, defined as those treatments that had not undergone any change in the three last months, both in doses and in the association of another antipsychotic. Results: Of the 11,953 inmates included in the study, 1,851 (15.5%) received prescriptions for an antipsychotic, 415 in penitentiary psychiatric hospitals (87%) and 1,436 in prisons (12%), which is a much higher prevalence than that found in the population. Regarding the prescription pattern of LAIs, paliperidone is the most widely prescribed in prison psychiatric hospitals and prisons, followed by aripiprazole, zuclopenthixol and risperidone. Finally flufenazine, olanzapine and paliperidone quarterly, which are scarcely represented. Of the 292 patients with LAI, 41% (121 patients) are with monotherapy and 59% (171 patients) are with polytherapy, which is similar to the data found in the bibliography. Discussion: The use of prescribed LAIs amongst inmates is widespread. Polytherapy is also a common phenomenon, although there is a downward trend due to the use of more recently marketed drugs. Since the use of LAI is long-term, with numerous side effects and a wide range of prices, it is essential to consider the patient's risk factors and the cost of treatment alongside the therapeutic efficacy

Review: Modulating ruminal lipid metabolism to improve the fatty acid composition of meat and milk. Challenges and opportunities.

Growth in demand for foods with potentially beneficial effects on consumer health has motivated increased interest in developing strategies for improving the nutritional quality of ruminant-derived products. Manipulation of the rumen environment offers the opportunity to modify the lipid composition of milk and meat by changing the availability of fatty acids (FA) for mammary and intramuscular lipid uptake. Dietary supplementation with marine lipids, plant secondary compounds and direct-fed microbials has shown promising results. In this review, we have compiled information about their effects on the concentration of putative desirable FA (e.g. c9t11-CLA and vaccenic, oleic, linoleic and linolenic acids) in ruminal digesta, milk and intramuscular fat. Marine lipids rich in very long-chain n-3 polyunsaturated fatty acids (PUFA) efficiently inhibit the last step of C18 FA biohydrogenation (BH) in the bovine, ovine and caprine, increasing the outflow of t11-18:1 from the rumen and improving the concentration of c9t11-CLA in the final products, but increments in t10-18:1 are also often found due to shifts toward alternative BH pathways. Direct-fed microbials appear to favourably modify rumen lipid metabolism but information is still very limited, whereas a wide variety of plant secondary compounds, including tannins, polyphenol oxidase, essential oils, oxygenated FA and saponins, has been examined with varying success. For example, the effectiveness of tannins and essential oils is as yet controversial, with some studies showing no effects and others a positive impact on inhibiting the first step of BH of PUFA or, less commonly, the final step. Further investigation is required to unravel the causes of inconsistent results, which may be due to the diversity in active components, ruminant species, dosage, basal diet composition and time on treatments. Likewise, research must continue to address ways to mitigate negative side-effects of some supplements on animal performance (particularly, milk fat depression) and product quality (e.g. altered oxidative stability and shelf-life).

Milk fat depression in dairy ewes fed fish oil: Might differences in rumen biohydrogenation, fermentation, or bacterial community explain the individual variation?

Dairy ewes show large individual variation in the extent of diet-induced milk fat depression (MFD) but reasons behind this variability remain uncertain. Previous results offered no convincing support for these differences being related to relevant changes in the milk fatty acid (FA) profile, including potentially antilipogenic FA, or in the transcript abundance of candidate genes involved in mammary lipogenesis. Therefore, we hypothesized that alterations in the processes of rumen biohydrogenation and fermentation, as well as in the bacterial community structure, might account for individual variation in fish oil-induced MFD severity. To test this explanation, 15 ewes received a total mixed ration without lipid supplementation (control; n = 5) or supplemented with 20 g of fish oil/kg of dry matter [10 animals divided into those showing a strong (RESPON+; -25.4%; n = 5) or a mild (RESPON-; -7.7%; n = 5) decrease in milk fat concentration] for 5 wk. Rumen fermentation parameters, biohydrogenation metabolites, and bacterial structure and diversity were analyzed in rumen samples collected before and after treatments. Although the fish oil supplementation increased the concentration of demonstrated or putative antilipogenic FA (e.g., cis-9 16:1, cis-11 18:1, or trans-10,cis-12 CLA), surprisingly, none of them differed significantly in relation to the extent of MFD (i.e., between RESPON- and RESPON+), and this was the case only for a few minor FA (e.g., cis-6+7 16:1 or 17:0 anteiso). Changes in total volatile FA, acetate, and propionate concentrations were associated with MFD severity, with higher decreases in more susceptible animals. Individual responses were not related to shifts in rumen bacterial structure but some terminal restriction fragments compatible with Clostridiales, Ruminococcaceae, Lachnospiraceae, and Succiniclasticum showed greater abundances in RESPON-, whereas some others that may correspond to Prevotella, Mogibacterium, and Quinella-related spp. were more abundant in RESPON+. Overall, the results suggest that individual variation in MFD severity in dairy ewes fed fish oil cannot be fully explained by differences in the processes of rumen biohydrogenation and fermentation or in the bacterial community, and further research would be necessary to elucidate the large variability in the responsiveness to MFD-inducing marine lipids.

In vitro ruminal biohydrogenation of eicosapentaenoic (EPA), docosapentaenoic (DPA), and docosahexaenoic acid (DHA) in cows and ewes: Intermediate metabolites and pathways.

A great deal of uncertainty still exists about intermediate metabolites and pathways explaining the biohydrogenation (BH) of 20- and 22-carbon polyunsaturated fatty acids (PUFA). Therefore, this study was conducted to provide further insight into the ruminal metabolism of 20:5 n-3 (EPA), 22:5 n-3 (DPA), and 22:6 n-3 (DHA), the main n-3 PUFA present in the marine lipids used in dairy ruminant feeding, and to examine potential differences between bovine and ovine. To meet this aim, we investigated the 20- and 22-carbon metabolites accumulated during in vitro incubation of EPA, DPA, and DHA with rumen inocula from cows and ewes. The PUFA were added at a dose of 2% incubated dry matter and digesta samples were analyzed after 24 h of incubation using complementary gas-liquid chromatography of fatty acid methyl esters and gas chromatography-mass spectrometry of 4,4-dimethyloxazoline derivatives. Results suggested that the main BH pathway of EPA and DPA would proceed via the reduction of the double bond closest to the carboxyl group (cis-5 in EPA and cis-7 in DPA); curiously, this mechanism seemed of much lower importance for DHA. Thus, DPA would not be a major intermediate product of DHA and their BH might actually follow separate pathways, with the accumulation of numerous unique metabolites in each case. A principal component analysis supported this hypothesis, with a clear separation between PUFA treatments in the score and loading plots. Within EPA and DPA groups, cow and ewe samples loaded separately from each other but not distant. No conjugated 20:5, 22:5, or 22:6 isomer compatible with the initial product of EPA, DPA, or DHA metabolism, respectively, was identified in the ruminal digesta, although this would not unequivocally exclude their transient formation. In this regard, results from DPA incubations provided the first indication that the metabolism of this very long chain PUFA may involve the formation of conjugated double bond structures. The BH of EPA, DPA, and DHA resulted in the appearance of several tentative trans-10-containing metabolites, showing a general trend to be more abundant in the digesta of ewes than in that of cows. This finding was speculated to have some relationship with the susceptibility of dairy sheep to marine lipid-induced milk fat depression. Differences in the relative proportion of intermediate products would also suggest an influence of ruminant species on BH kinetics, with a process that would likely be slower and less complete in cows than in ewes.

Individual variation of the extent of milk fat depression in dairy ewes fed fish oil: Milk fatty acid profile and mRNA abundance of candidate genes involved in mammary lipogenesis.

Dairy ewes are less prone than cows to milk fat depression (MFD) but suffer from this syndrome when marine lipids are added to their diet to modulate milk fatty acid (FA) profile. However, there are large individual differences in MFD extent, and the reasons behind this variability are uncertain. On this basis, a study was conducted in lactating sheep to test the hypotheses that individual susceptibility to the low-fat milk condition may be explained by differences in (1) the milk concentration of some FA, particularly antilipogenic FA, or (2) the transcriptional regulation of mammary lipogenesis. For 5 wk, 15 ewes received a total mixed ration supplemented with 0 (control; n = 5) or 20 g of fish oil/kg of dry matter [10 animals selected out of 22 and divided into those showing marked (RESPON+; n = 5) or mild (RESPON-; n = 5) MFD]. Milk production and composition, including a comprehensive FA profile, were examined on 3 consecutive days before and after treatments. Candidate gene expression was also analyzed before the start of the trial and at its end using RNA isolated from milk somatic cells. According to the experimental design, the fish-oil-induced decrease in milk fat concentration was much stronger in RESPON+ (-25.4%) than in RESPON- (-7.7%). Milk from all ewes fed the supplemented diet showed rather uniform changes in the proportion of potentially healthy FA (such as cis-9,trans-11 18:2, trans-11 18:1, or 20:5n-3) and of those with confirmed or putative antilipogenic effects (e.g., cis-9 16:1, trans-10 and cis-11 18:1, trans-9,cis-11 18:2, and 10-oxo-18:0), without significant variation between RESPON+ and RESPON-. It was not possible to relate the very few exceptions to this general trend (e.g., in cis-7 16:1 and 22:6n-3) to responsiveness. Major mechanisms involved in mammary lipogenesis, specifically the uptake and de novo synthesis of FA, appeared to be unequally inhibited in ewes displaying different degrees of MFD, with molar yields of >16C FA being unaffected in RESPON-. However, this was not reflected in candidate gene expression. Supplementation with fish oil showed a tendency to lower the mRNA abundance of lipogenic genes such as ACSS2, FASN, LPIN1, FADS2, and INSIG1, but only SCD and GPAT4 tended to differ between RESPON- and RESPON+. Overall, these results offer no convincing support for the initial hypotheses, so further research must be pursued to explain the individual variation in MFD severity.

In vitro response to EPA, DPA, and DHA: Comparison of effects on ruminal fermentation and biohydrogenation of 18-carbon fatty acids in cows and ewes.

The modulation of milk fat nutritional quality through fish oil supplementation seems to be largely explained by the action of n-3 very long chain polyunsaturated fatty acids (PUFA) on ruminal biohydrogenation (BH) of C18 fatty acids (FA). However, relationships among this action, disappearance of those PUFA in the rumen, and potential detrimental consequences on ruminal fermentation remain uncertain. This study compared the effect of 20:5n-3 (eicosapentaenoic acid; EPA), 22:5n-3 (docosapentaenoic acid; DPA), and 22:6n-3 (docosahexaenoic acid; DHA) on rumen fermentation and BH of C18 FA and was conducted simultaneously in cows and sheep to provide novel insights into interspecies differences. The trial was performed in vitro using batch cultures of rumen microorganisms with inocula collected from cannulated cows and ewes. The PUFA were added at a dose of 2% incubated dry matter, and treatment effects on ruminal C18 FA concentrations, PUFA disappearances, and fermentation parameters (gas production, ammonia and volatile FA concentrations, and dry matter and neutral detergent fiber disappearances) were examined after 24 h of incubation. A principal component analysis suggested that responses to PUFA treatments explained most of the variability; those of ruminant species were of lower relevance. Overall, EPA and DHA were equally effective for inhibiting the saturation of trans-11 18:1 to 18:0 and had a similar influence on ruminal fermentation in cows and sheep (e.g., reductions in gas production and acetate:propionate ratio). Nevertheless, DHA further promoted alternative BH pathways that lead to trans-10 18:1 accumulation, and EPA seemed to have specific effects on 18:3n-3 metabolism. Only minor variations attributable to DPA were observed in the studied parameters, suggesting a low contribution of this FA to the action of marine lipids. Although most changes due to the added PUFA were comparable in bovine and ovine, there were also relevant specificities, such as a stronger inhibition of 18:0 formation in cows and a greater increase in 18:3n-3 metabolites in sheep. No direct relationship between in vitro disappearance of the incubated PUFA and effect on BH (in particular, inhibition of the last step) was found in either cows or ewes, calling into question a putative link between extent of disappearance and toxicity for microbiota. Conversely, an association between the influence of these PUFA on ruminal lipid metabolism and fermentation may exist in both species. In vivo verification of these findings would be advisable.

Endogenous synthesis of milk oleic acid in dairy ewes: In vivo measurement using 13C-labeled stearic acid.

The use of stable isotopes is a reliable and risk-free alternative to radioactive tracers for directly examining in vivo fatty acid (FA) metabolism. However, very limited information is available in ruminants, and none is available in sheep. Therefore, we conducted an experiment in dairy ewes to determine, for the first time in this species, the uptake, Δ9-desaturation, and secretion of 13C-labeled stearic acid (SA) into milk with the aim of measuring in vivo endogenous synthesis of milk oleic acid (OA) and stearoyl-CoA desaturase activity. Six lactating Assaf ewes fed a total mixed ration (forage:concentrate ratio = 30:70) received an intravenous injection of 2 g of 13C-labeled SA. At -24, -15, 0, 4, 8, 12, 16, 20, 24, 36, 48, 60, and 72 h postinjection (p.i.), milk yield was recorded and milk samples were collected to examine fat concentration and FA composition, including compound-specific isotope analysis of SA and OA by gas chromatography-combustion isotope ratio mass spectrometry. Over the p.i. period, the SA proportion ranged from 7.6 to 8.3% of total FA, with a maximum 13C enrichment of 1.9%, whereas OA was more abundant (14.3-15.4% of total FA) and had lower 13C enrichments (up to 0.69%). On average, 15% of the isotopic tracer was transferred to milk within 72 h p.i., and 47 to 50% of the SA taken up by the mammary gland would have been desaturated to OA. The proportion of oleic acid being synthesized endogenously was estimated to represent between 48 and 57% of the amount secreted in milk. Further research under different dietary conditions is recommended.

mRNA abundance of genes involved in mammary lipogenesis during fish oil- or trans-10,cis-12 CLA-induced milk fat depression in dairy ewes.

Milk fat depression (MFD) caused by trans-10,cis-12 18:2 is known to be mediated in cows and ewes by downregulation of mammary lipogenic genes. However, transcriptional mechanisms underlying marine lipid-induced MFD have not been well defined yet and the few available studies in ovine are not consistent. This trial was conducted to directly compare changes in animal performance, milk fatty acid composition, and particularly mammary mRNA abundance of candidate lipogenic genes and transcription factors in response to the inclusion of fish oil or trans-10,cis-12 18:2 in the dairy sheep diet. To meet this objective, 12 lactating Assaf ewes (on average, 64 days in milk, producing 1.72 kg of milk/d with 5.17% of fat) were divided into 3 groups and offered a total mixed ration without supplementation (control) or supplemented with 2.4% dry matter of fish oil (FO treatment) or 1% dry matter of a commercial product rich in trans-10,cis-12 18:2 (CLA treatment) for 39 d. Measurements and samplings were conducted before starting the treatments and at the end of the trial. Milk samples were used for RNA extraction from somatic cells. Feed intake was not affected by lipid supplements, and as designed, reductions in milk fat concentration (-31%) were similar in the 2 treatments, although the unpredicted increase in milk production with FO counteracted the anticipated reduction in milk fat yield. Nevertheless, this did not preclude the detection of FO-induced decreases in the mRNA abundance of candidate lipogenic genes [e.g., acyl-CoA synthetase short-chain family member 2 (ACSS2), fatty acid synthase (FASN), and lipin 1 (LPIN1)], thus supporting the hypothesis that transcriptional regulation would be a relevant component of this type of MFD in sheep. Expected CLA-induced downregulation of some genes, such as FASN or sterol regulatory element binding transcription factor 1 (SREBF1), could not be detected in our samples, which might be related, at least in part, to high inter-individual variation and relatively advanced lactation stage (on average 102-103 d in milk on d 38 and 39). Overall, direct comparison of the effects of dietary FO and CLA on transcript abundance of candidate lipogenic genes and transcription factors suggest that there might be relevant differences in the transcriptional control mechanisms underlying the MFD induced by each kind of supplement (i.e., fish oil or CLA).