Oral chondroitin sulfate and prebiotics for the treatment of canine Inflammatory Bowel Disease: a randomized, controlled clinical trial

BACKGROUND Canine inflammatory bowel disease (IBD) is a chronic enteropathy of unknown etiology, although microbiome dysbiosis, genetic susceptibility, and dietary and/or environmental factors are hypothesized to be involved in its pathogenesis. Since some of the current therapies are associated with severe side effects, novel therapeutic modalities are needed. A new oral supplement for long-term management of canine IBD containing chondroitin sulfate (CS) and prebiotics (resistant starch, β-glucans and mannaoligosaccharides) was developed to target intestinal inflammation and oxidative stress, and restore normobiosis, without exhibiting any side effects. This double-blinded, randomized, placebo-controlled trial in dogs with IBD aims to evaluate the effects of 180 days administration of this supplement together with a hydrolyzed diet on clinical signs, intestinal histology, gut microbiota, and serum biomarkers of inflammation and oxidative stress. RESULTS Twenty-seven client-owned biopsy-confirmed IBD dogs were included in the study, switched to the same hydrolyzed diet and classified into one of two groups: supplement and placebo. Initially, there were no significant differences between groups (p > 0.05) for any of the studied parameters. Final data analysis (supplement: n = 9; placebo: n = 10) showed a significant decrease in canine IBD activity index (CIBDAI) score in both groups after treatment (p < 0.001). After treatment, a significant decrease (1.53-fold; p < 0.01) in histologic score was seen only in the supplement group. When groups were compared, the supplement group showed significantly higher serum cholesterol (p < 0.05) and paraoxonase-1 (PON1) levels after 60 days of treatment (p < 0.01), and the placebo group showed significantly reduced serum total antioxidant capacity (TAC) levels after 120 days (p < 0.05). No significant differences were found between groups at any time point for CIBDAI, WSAVA histologic score and fecal microbiota evaluated by PCR-restriction fragment length polymorphism (PCR-RFLP). No side effects were reported in any group. CONCLUSIONS The combined administration of the supplement with hydrolyzed diet over 180 days was safe and induced improvements in selected serum biomarkers, possibly suggesting a reduction in disease activity. This study was likely underpowered, therefore larger studies are warranted in order to demonstrate a supplemental effect to dietary treatment of this supplement on intestinal histology and CIBDAI

Validation of an automated assay for the measurement of cupric reducing antioxidant capacity in serum of dogs

BACKGROUND: The objective of the present study was to optimize and validate an automated method to assess the total antioxidant capacity (TAC) in serum of dogs using the cupric reducing antioxidant capacity (CUPRAC) methodology (TAC(c)) with bathocuproinedisulfonic acid disodium salt as chelating agent, evaluating also possible variations due to the use of two different automated analyzers. The method is based on the reduction of Cu(2+) into Cu(1+) by the action of the non-enzymatic antioxidants that are present in the sample. RESULTS: Imprecision was low in both apparatus utilized, and the results were linear across serial Trolox and canine serum samples dilutions. Lipids did not interfere with the assay; however, hemolysis increased the TAC(c) concentrations. When TAC(c) concentrations were determined in ten healthy (control) dogs and in twelve dogs with inflammatory bowel disease (IBD), dogs with IBD had lower TAC(c) concentrations when compared with the healthy dogs. CONCLUSIONS: The method validated in this paper is precise, simple, and fast and can be easily adapted to automated analyzers

Causes, consequences and biomarkers of stress in swine: an update

BACKGROUND: In recent decades there has been a growing concern about animal stress on intensive pig farms due to the undesirable consequences that stress produces in the normal physiology of pigs and its effects on their welfare and general productive performance. This review analyses the most important types of stress (social, environmental, metabolic, immunological and due to human handling), and their biological consequences for pigs. The physio-pathological changes associated with stress are described, as well as the negative effects of stress on pig production. In addition an update of the different biomarkers used for the evaluation of stress is provided. These biomarkers can be classified into four groups according to the physiological system or axis evaluated: sympathetic nervous system, hypothalamic-pituitary-adrenal axis, hypothalamic-pituitary-gonadal axis and immune system. CONCLUSIONS: Stress it is a process with multifactorial causes and produces an organic response that generates negative effects on animal health and production. Ideally, a panel of various biomarkers should be used to assess and evaluate the stress resulting from diverse causes and the different physiological systems involved in the stress response. We hope that this review will increase the understanding of the stress process, contribute to a better control and reduction of potential stressful stimuli in pigs and, finally, encourage future studies and developments to better monitor, detect and manage stress on pig farms

INFLUENCIA DE DIVERSAS CONDICIONES ANALÍTICAS EN LA DETERMINACIÓN DE COLINESTERASA EN SANGRE ENTERA MEDIANTE EL MÉTODO DE ELLMAN

The main sources of variation that can produce variability on whole blood cholinesterase determinations were studied and evaluated. For this purpose, dog whole blood aliquots were analyzed under different analytical conditions. An increase on cholinesterase activity was observed as reaction temperature was raised. The highest cholinesterase activity was obtained at pH range between 8.0 and 8.5; however, non-enzimatic substrate hydrolysis was also increased when pH exceeded 7.5, so the use of pH 7.5 would be recommended. Although acetylthiocholine iodide concentration of 5x10-3 M and butyrylthiocholine iodide concentration of 10x10-3 M yielded the highest cholinesterase activity, an increase of non-enzymatic substrate hydrolysis was also observed at these substrate concentrations, so the use of both substrates at 1x10-3 M concentration was preferred. 5,5’- dithiobis-2-nitrobenzoic acid as chromophore, and acetylthiocholine iodide and butyrylthiocholine iodide as substrates, were stable for at least three months if frozen. Based on these results, this work in contributing to a better knowledge of the sources of variation affecting whole blood cholinesterase determination, and the standardization of analytical conditions.En este trabajo se pretende estudiar y evaluar las principales fuentes de variación que pueden provocar variabilidad en la determinación de colinesterasa en sangre entera. Para este fin se analizaron alícuotas de sangre entera de perro bajo distintas condiciones analíticas. Se observó un aumento de la actividad colinesterasa conforme se incrementó la temperatura de reacción. La mayor actividad colinesterasa se encontró en un intervalo de pH entre 8.0 y 8.5; sin embargo, una fuerte elevación de la hidrólisis no enzimática del sustrato tiene lugar cuando el pH excede de 7.5, lo que haría recomendable el empleo de pH 7.5. Aunque los valores más elevados de actividad se alcanzaron con una concentración del sustrato acetiltiocolina iodada de 5x10-3 M y de butiriltiocolina iodada de 10x10-3 M, se describió un fuerte aumento de la hidrólisis no enzimática de los sustratos a estas concentraciones, por lo que se prefirió emplear una concentración de ambos sustratos de 1x10-3 M. El cromóforo ácido 5,5’-ditiobis-2-nitrobenzoico, y los sustratos acetiltiocolina iodada y butiriltiocolina iodada, permanecieron estables al menos durante tres meses si eran conservados a una temperatura de –20ºC. Los datos aportados permitirán contribuir a un mejor conocimiento de las fuentes de variación que afectan a la determinación de colinesterasa en sangre entera, y a la estandarización de las condiciones analíticas utilizadas

FLUOROMETRÍA EN TIEMPO RETARDADO: CONCEPTOS GENERALES Y APLICACIONES EN MEDICINA VETERINARIA

Time-resolved fluorometry is a novel technology that emerges to replace radioimunoassay and appears as a feasible alternative to ELISA. This detection system has led to the development of highly sensitive immunoassays in which antigens or antibodies are labelled with lanthanide chelates, which emit fluorescence capable of being quantified. The high sensitivity makes this technology very useful in the measurement of substances present in small quantities in different organic fluids, such as urine, blood or saliva. Recent advances in fluorometry have provided ultrarapid and ultrasensitive assays for the quantification of acute phase proteins and myocardial infarction markers in human medicine, whereas in veterinary medicine one of its main applications focuses on hormones determination.La fluorometría en tiempo retardado es una tecnología novedosa que surge con la intención de reemplazar al radioinmunoanálisis y se presenta como una posible alternativa al ELISA. Este sistema de detección ha permitido el desarrollo de inmunoensayos altamente sensibles en los que antígenos o anticuerpos son marcados con quelatos de lantánidos, emisores de fluorescencia susceptible de ser cuantificada. Su gran sensibilidad hace que sea una herramienta eficaz en el análisis de compuestos que se encuentran en pequeñas concentraciones en diferentes fluidos orgánicos, tales como orina, sangre o saliva. Los crecientes avances en esta metodología han proporcionado ensayos ultrarápidos y ultrasensibles para la determinación de proteínas de fase aguda y marcadores de infarto de miocardio en medicina humana, mientras que en medicina veterinaria una de sus principales aplicaciones se centra en la determinación de hormonas

PRINCIPALES APLICACIONES DE LAS PROTEÍNAS DE FASE AGUDA EN LA CLÍNICA CANINA

The term acute phase proteins reflects all the changes that occur in the organism after a tissue injury with the purpose of restoring its homeostasis. During this response there is a change in the concentration of some plasma proteins collectively known as acute phase proteins. Some of these proteins are haptoglobin, C reactive protein, serum amyloid A and ceruloplasmin. Healthy dogs have negligible or low levels of these proteins, however their concentrations increase quickly in response to infection or inflammatory conditions. Therefore the appearance of a high result of some of these proteins in a dog would be indicative that this animal it is not healthy and that a secondary detailed diagnostic study should be made. In addition acute phase proteins have an important application in the monitoring of the response to treatment, and they could be an aid to determine the effectiveness of the therapy and to predict possible relapses.El término Respuesta de Fase Aguda refleja todos los cambios que se producen en el organismo de un animal tras sufrir una lesión tisular, y que van encaminados a restablecer su homeostasis. Durante el desarrollo de esta respuesta se producen, entre otros cambios, variaciones en los niveles de algunas proteínas plasmáticas, denominándose a éstas de forma general, Proteínas de fase Aguda, entre las que se encuentran la haptoglobina, proteína C reactiva, amiloide A sérico y ceruloplasmina. En perros sanos estas proteínas se encuentran en niveles no detectables o muy bajos; sin embargo, sus concentraciones plasmáticas aumentan rápidamente en respuesta a diversos procesos inflamatorios e infecciosos. Por tanto la aparición de un resultado elevado de alguna de estas proteínas en un animal, sería indicativo de que dicho animal no está sano, y constituiría un aviso para la realización de un estudio clínico-diagnóstico mas detallado. Además van a tener una importante aplicación en la monitorización de la respuesta a los tratamientos empleados, ayudando a determinar la eficacia de los mismos e incluso a predecir la posible existencia de recidivas

MÉTODOS PARA MEDIR EL GRADO DE LA OBESIDAD EN PERROS: ENTRE LA FÍSICA Y LA BIOQUÍMICA

Obesity is defined as an exesive accumulation of adipose tissue in the body. Frequently the main cause of obesity in dogs is an execive ingest or an inadequate metabolism, that causes positive energy balance. Besides of the high number of associated diseases that obesity can cause, longevity of obese animals is two years shorter than normal weight animals. For these reasons the correct determination, valoration and treatment of obesity in dogs is of great importance nowadays in routine practice. In this review we will deal with several methods for obesity grade measurement that can be divided into two groups: physical and biochemical. In physical we will include body weight, morphometric methods, tape measurements, dual X-ray absorptiometry (DXA), deuterium isotope dilution (D2O). Regarding the biochemical methods, we will focus on two proteins: leptin and adiponectin that in the future could be used as biomarkers of obesity in canine species.La obesidad se define como un acumulo excesivo de tejido adiposo en el cuerpo. La causa de obesidad normalmente es una excesiva ingesta o un metabolismo inadecuado, que ocasiona un balance positivo de energía. Además de las múltiples enfermedades que se asocian con la obesidad en el perro, se ha visto que los animales obesos viven hasta dos años menos en comparación con animales sanos. Esto hace que, hoy en día, sea de gran importancia la correcta determinación, valoración y tratamiento de los perros obesos. Para medir grado de obesidad se han descrito muchos métodos que pueden ser divididos en dos grupos: físicos y bioquímicos. En este trabajo se van a analizar los métodos mas utilizados en veterinaria para medir grado de obesidad en el perro; así dentro de los métodos físicos se estudiarán el peso corporal, las medidas antropométricas, escalas morfológicas, absorciometría de rayos X de doble energía (DXA) y dilución de isótopos de oxido de deuterio (D2O). Por otra parte dentro de los métodos bioquímicos, nos centraremos en dos proteínas que en el futuro podrán ser utilizados como biomarcadores de la obesidad: la leptina y adiponectina

HAPTOGLOBINA COMO INDICADOR DE ESTRÉS EN VACAS DE LIDIA

The influence of physical and psychological stress on the concentration in plasma of the acute phase protein haptoglobin (Hp) was studied in 20 fi ghting cows aged 3 to 6 years. Four groups were prepared, the first one or control group comprised livestock with no apparent stimulation. The other groups corresponded to animals of traditional bull running that were sacrifi ced at 30, 54 and 78 h after the end of the spectacle. The mean serum Hp concentration from control fighting cows was 0,093 g/l, while it was 0,13 g/l at 30h, 0,19 g/l at 54 h and 0,5 g/l at 78 h after the end of the spectacle. These results show that in the traditional bull running, Hp concentration is elevated and signifi cant differences are observed at 54 h, although the maximum levels appeared at 78 h. In conclusion, in fighting cows the serum hp concentration was more elevated due to a situation of physical and psychological stress and may be a useful indicator of stress levels in the animal.Se ha estudiado la infl uencia del estrés físico y psicológico sobre las concentraciones plasmáticas de la proteína de fase aguda haptoglobina (Hp) en 20 vacas de raza bovina de lidia de 3 a 6 años de edad. Los animales se dividieron en 4 grupos, el primero, utilizado como control, corresponde a vacas en la ganadería sin estimulación aparente, los tres grupos restantes corresponden a vacas que se utilizaron en encierros tradicionales y que fueron sacrificados a las 30, 54 y 78 horas de la celebración del espectáculo. Las concentraciones medias de Hp en suero para las vacas control fueron 0,093 g/l y 0,13 g/l a las 30h, 0,19 g/l a las 54 h y 0,5 g/l a las 78 h de la celebración del encierro. Estos resultados indican que en los encierros tradicionales se estimula la producción de Hp en vacas de lidia, presentando valores estadísticamente significativos a partir de las 54 horas de la realización del mismo y situándose los valores máximos a las 78 horas. Como conclusión, hemos de considerar que en situaciones de estrés en vacas de lidia los niveles de Hp en plasma son superiores a los normales y estos parámetros podrían ser utilizados como un indicador del nivel de estrés sufrido por el animal

Haptoglobin as an indicator of stress in lidia breeding cows

Se ha estudiado la influencia del estrés físico y psicológico sobre las concentraciones plasmáticas de la proteína de fase aguda haptoglobina (Hp) en 20 vacas de raza bovina de lidia de 3 a 6 años de edad. Los animales se dividieron en 4 grupos, el primero, utilizado como control, corresponde a vacas en la ganadería sin estimulación aparente, los tres grupos restantes corresponden a vacas que se utilizaron en encierros tradicionales y que fueron sacrifi cados a las 30, 54 y 78 horas de la celebración del espectáculo. Las concentraciones medias de Hp en suero para las vacas control fueron 0,093 g/l y 0,13 g/l a las 30h, 0,19 g/l a las 54 h y 0,5 g/l a las 78 h de la celebración del encierro. Estos resultados indican que en los encierros tradicionales se estimula la producción de Hp en vacas de lidia, presentando valores estadísticamente signifi cativos a partir de las 54 horas de la realización del mismo y situándose los valores máximos a las 78 horas. Como conclusión, hemos de considerar que en situaciones de estrés en vacas de lidia los niveles de Hp en plasma son superiores a los normales y estos parámetros podrían ser utilizados como un indicador del nivel de estrés sufrido por el animal