RESUMEN
Isoprenoids are vital for all organisms, in which they maintain membrane stability and support core functions such as respiration1. IspH, an enzyme in the methyl erythritol phosphate pathway of isoprenoid synthesis, is essential for Gram-negative bacteria, mycobacteria and apicomplexans2,3. Its substrate, (E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate (HMBPP), is not produced in metazoans, and in humans and other primates it activates cytotoxic Vγ9Vδ2 T cells at extremely low concentrations4-6. Here we describe a class of IspH inhibitors and refine their potency to nanomolar levels through structure-guided analogue design. After modification of these compounds into prodrugs for delivery into bacteria, we show that they kill clinical isolates of several multidrug-resistant bacteria-including those from the genera Acinetobacter, Pseudomonas, Klebsiella, Enterobacter, Vibrio, Shigella, Salmonella, Yersinia, Mycobacterium and Bacillus-yet are relatively non-toxic to mammalian cells. Proteomic analysis reveals that bacteria treated with these prodrugs resemble those after conditional IspH knockdown. Notably, these prodrugs also induce the expansion and activation of human Vγ9Vδ2 T cells in a humanized mouse model of bacterial infection. The prodrugs we describe here synergize the direct killing of bacteria with a simultaneous rapid immune response by cytotoxic γδ T cells, which may limit the increase of antibiotic-resistant bacterial populations.
Asunto(s)
Diseño de Fármacos , Inhibidores Enzimáticos/farmacología , Proteínas de Escherichia coli/antagonistas & inhibidores , Bacterias Gramnegativas/efectos de los fármacos , Bacterias Gramnegativas/inmunología , Activación de Linfocitos/efectos de los fármacos , Viabilidad Microbiana/efectos de los fármacos , Oxidorreductasas/antagonistas & inhibidores , Linfocitos T Citotóxicos/efectos de los fármacos , Animales , Farmacorresistencia Microbiana , Resistencia a Múltiples Medicamentos , Inhibidores Enzimáticos/química , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Femenino , Semivida , Humanos , Leucocitos Mononucleares/efectos de los fármacos , Leucocitos Mononucleares/inmunología , Leucocitos Mononucleares/microbiología , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Pruebas de Sensibilidad Microbiana , Simulación del Acoplamiento Molecular , Oxidorreductasas/deficiencia , Oxidorreductasas/genética , Oxidorreductasas/metabolismo , Profármacos/farmacocinética , Profármacos/farmacología , Especificidad por Sustrato , Porcinos/sangre , Linfocitos T Citotóxicos/inmunologíaRESUMEN
The brains of humans and other mammals are highly vulnerable to interruptions in blood flow and decreases in oxygen levels. Here we describe the restoration and maintenance of microcirculation and molecular and cellular functions of the intact pig brain under ex vivo normothermic conditions up to four hours post-mortem. We have developed an extracorporeal pulsatile-perfusion system and a haemoglobin-based, acellular, non-coagulative, echogenic, and cytoprotective perfusate that promotes recovery from anoxia, reduces reperfusion injury, prevents oedema, and metabolically supports the energy requirements of the brain. With this system, we observed preservation of cytoarchitecture; attenuation of cell death; and restoration of vascular dilatory and glial inflammatory responses, spontaneous synaptic activity, and active cerebral metabolism in the absence of global electrocorticographic activity. These findings demonstrate that under appropriate conditions the isolated, intact large mammalian brain possesses an underappreciated capacity for restoration of microcirculation and molecular and cellular activity after a prolonged post-mortem interval.
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Autopsia , Encéfalo/irrigación sanguínea , Encéfalo/citología , Circulación Cerebrovascular , Microcirculación , Porcinos , Animales , Encéfalo/metabolismo , Encéfalo/patología , Isquemia Encefálica/metabolismo , Isquemia Encefálica/patología , Caspasa 3/metabolismo , Supervivencia Celular , Arterias Cerebrales/fisiología , Modelos Animales de Enfermedad , Hipoxia Encefálica/metabolismo , Hipoxia Encefálica/patología , Inflamación/metabolismo , Inflamación/patología , Neuroglía/citología , Neuronas/citología , Neuronas/metabolismo , Neuronas/patología , Perfusión , Daño por Reperfusión/prevención & control , Porcinos/sangre , Sinapsis/metabolismo , Sinapsis/patología , Factores de Tiempo , VasodilataciónRESUMEN
BACKGROUND: Blood sampling from neonatal piglets is related to multiple disadvantages. Therefore, a new, alternative matrix is required to assess piglets' early immune status efficiently. The present study aimed to assess the usefulness of processing fluid for determining selected piglets' immune parameters. 264 pigs - 31 sows, 146 male piglets, and 87 female piglets from commercial indoor farrow-to-finish pig herd were included in this study. 264 serum, 31 colostrum, and 146 processing fluid samples were collected. Serum was collected from all animals, colostrum was collected from sows, and processing fluid was collected from male piglets only. Using commercial ELISA tests, the concentration of various immunoglobulins, cytokines, and acute phase proteins was assessed in each matrix. Statistical analyses were employed to determine differences in the concentration of measured indices between piglets' serum and processing fluid and correlations in the concentration of tested indices between particular sets of matrices. RESULTS: Statistical analyses did not reveal significant differences in the IgG, IgA, IL-1ß, IL-4, IL-6, and IFN-γ concentration between piglets' serum and processing fluid (p > 0.05). A positive correlation (p < 0.05) regarding the concentration of some indices between processing fluid and samples collected from sows was also observed. CONCLUSIONS: Processing fluid can be considered a promising alternative to blood for assessing some immunological indices in piglets, such as IgG, IgA, IL-1ß, IL-4, IL-6, and IFN-γ, and, possibly, in the indirect assessment of some indices in lactating sows, including IgA, IL-1ß, IL-4, IL-6, IL-8, IFN-γ, or Pig-MAP.
Asunto(s)
Calostro , Citocinas , Inmunoglobulinas , Animales , Calostro/química , Calostro/inmunología , Femenino , Masculino , Porcinos/sangre , Citocinas/sangre , Citocinas/análisis , Inmunoglobulinas/sangre , Inmunoglobulinas/análisis , Animales Recién Nacidos/inmunología , Animales Recién Nacidos/sangre , Animales Lactantes/inmunología , Animales Lactantes/sangre , Proteínas de Fase Aguda/análisis , Proteínas de Fase Aguda/metabolismoRESUMEN
BACKGROUND: With the development of intensive farming, long-term exposure of pigs to poor light conditions is not conducive to the production of vitamin D3 , and vitamin D3 deficiency could affect absorption and metabolism of calcium (Ca) and phosphorus (P). 25-Hydroxyvitamin D3 (25OHD3 ) has higher bioactivity than regular vitamin D3 . This study investigated the effects of 25OHD3 on performance, serum parameters, fecal microbiota, and metabolites in weaned piglets fed with low Ca-P diet. RESULTS: It was found that a low Ca-P diet supplemented with 50 µg/kg 25OHD3 (NC + 25-D) improved (P < 0.05) average daily gain (ADG) in phase 2 and in the overall period of the experiment, and increased (P < 0.05) the immunoglobulin G (IgG), immunoglobulin A (IgA), catalase (CAT), bone-specific alkaline phosphatase (BALP), and osteocalcin (OC) serum content on day 28 compared with a low Ca-P diet (NC), but no differences were observed between a normal Ca-P diet (PC) and the NC + 25-D diet. Compared with NC, the abundance of Firmicutes was higher (P < 0.05) in PC and NC + 25-D. NC + 25-D decreased (P < 0.05) the abundance of Streptococcaceae compared with PC and NC, and increased (P < 0.05) the abundance of Lachnospiraceae compared with NC. Serum 25OHD3 was negatively correlated with the abundance of fecal Streptococcaceae (P < 0.05), and positively correlated with the abundance of fecal Lachnospiraceae (P < 0.05). CONCLUSION: Supplementation of 25OHD3 in a low Ca-P diet improved serum immunity, bone biochemical parameters, and fecal microbiota such as decreased Streptococcaceae abundance and increased Lachnospiraceae abundance, which could subsequently promote growth of piglets. The effects were similar to that of a normal Ca-P diet. © 2021 Society of Chemical Industry.
Asunto(s)
Calcifediol/metabolismo , Heces/microbiología , Microbioma Gastrointestinal , Porcinos/crecimiento & desarrollo , Alimentación Animal/análisis , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Bacterias/clasificación , Bacterias/genética , Bacterias/aislamiento & purificación , Desarrollo Óseo , Huesos/metabolismo , Calcio/análisis , Calcio/metabolismo , Suplementos Dietéticos/análisis , Femenino , Masculino , Fósforo/análisis , Fósforo/metabolismo , Porcinos/sangre , Porcinos/metabolismo , Porcinos/microbiología , DesteteRESUMEN
Tools are required for quick and easy preliminary evaluation of functional feeds efficiency on fisheries. The analysis of skin mucus biomarkers is a recent alternative approach providing a faster feed-back from the laboratory which is characterized by being less invasive, more rapid and with reduced costs. The effect of replacing fishmeal and fish protein hydrolysates by means of two porcine by-products, the porcine spray-dried plasma (SDPP) and pig protein hydrolysate (PPH), in compound diets (50.4% crude protein, 16.2% crude protein, 22.1 MJ/kg feed) was evaluated in juvenile meagre (Argyrosomus regius) during a two-months period. To determine the impact of these dietary replacements, growth and food performance were measured together with digestive enzymes activities and filet proximal composition. Additionally, skin mucus was collected and characterized by determining main mucus biomarkers (protein, glucose, lactate, cortisol, and antioxidant capacity) and its antibacterial properties, measured by the quick in vitro co-culture challenges. In comparison to the control group, the inclusion of PPH and SDPP, in meagre diets reduced growth (7.4-8.8% in body weight), increased feed conversion ratios (9.0-10.0%), results that were attributed to a reduction in feed intake values (24.2-33.0%) (P < 0.05). Porcine blood by-products did not modify the activity of gastric and pancreatic digestive enzymes as well as those involved in nutrient absorption (alkaline phosphatase) nor liver oxidative stress condition (P > 0.05). In contrast, a reduction in fillet lipid content associated to an increase in fillet protein levels were found in fish fed SDPP and PPH diets (P < 0.05). As compared to the control diet, the dietary replacement did not alter the levels of the skin mucus biomarkers related to stress (cortisol and antioxidant capacity) or nutritional status (soluble protein, glucose and lactate) (P > 0.05). Interestingly, regardless of the worst performance in somatic growth, meagre fed diets containing both tested porcine by-products showed a significantly improved antibacterial capacity of their skin mucus. This enhancement was more prominent for fish fed with the PPH diet, which may be attributed to a higher content of immunomodulatory bioactive compounds in PPH. Further research will be necessary to provide insights on how the inclusion of SDPP and PPH, at the expense of dietary fishmeal and fish protein hydrolysates, affects feed intake and growth performance in meagre. However, the use of skin mucus biomarkers has been demonstrated to be an excellent methodology for a preliminary characterization of the functional feeds, in particular for their prophylactic properties by the study of mucus antibacterial activity.
Asunto(s)
Alimentación Animal , Antibacterianos , Dieta , Moco , Perciformes , Piel , Porcinos , Animales , Antioxidantes , Biomarcadores , Dieta/veterinaria , Glucosa , Hidrocortisona , Lactatos , Moco/inmunología , Moco/microbiología , Perciformes/microbiología , Perciformes/fisiología , Hidrolisados de Proteína , Piel/inmunología , Piel/microbiología , Porcinos/sangreRESUMEN
BACKGROUND: The widespread popularity of porcine circovirus type 2(PCV2) has seriously affected the healthy development of the pig industry and caused huge economic losses worldwide. A rapid and reliable method is required for epidemiological investigation and evaluating the effect of immunization. However, the current methods for PCV2 antibody detection are time-consuming or very expensive and rarely meet the requirements for clinical application. we have constructed the platform for expressing the nanobody(Nb)horseradish peroxidase(HRP) fusion protein as an ultrasensitive probe to detect antibodies against the Newcastle disease virus(NDV), previously. In the present work, an Nb-HRP fusion protein-based competitive ELISA(cELISA) for rapid and simple detection antibodies against PCV2 was developed using this platform to detect anti-PCV2 antibodies in clinical porcine serum. RESULTS: Using phage display technology, 19 anti-PCV2-Cap protein nanobodies were screened from a PCV2-Cap protein immunized Bactrian camel. With the platform, the PCV2-Nb15HRP fusion protein was then produced and used as a sensitive reagent for developing a cELISA to detect antiPCV2 antibodies. The cutoff value of the cELISA is 20.72 %. Three hundreds and sixty porcine serum samples were tested by both newly developed cELISA and commercial kits. The sensitivity and specificity were 99.68 % and 95.92 %, respectively. The coincidence rate of the two methods was 99.17 %. When detecting 620 clinical porcine serum samples, a good consistent (kappa value = 0.954) was found between the results of the cELISA and those of commercial kits. CONCLUSIONS: In brief, the newly developed cELISA based PCV2-Nb15HRP fusion protein is a rapid, low-cost, reliable and useful nanobody-based tool for the serological evaluation of current PCV2 vaccine efficacy and the indirect diagnosis of PCV2 infection.
Asunto(s)
Anticuerpos Antivirales/inmunología , Infecciones por Circoviridae/veterinaria , Circovirus/inmunología , Ensayo de Inmunoadsorción Enzimática/métodos , Enfermedades de los Porcinos/inmunología , Animales , Anticuerpos Antivirales/sangre , Camelus/inmunología , Infecciones por Circoviridae/sangre , Infecciones por Circoviridae/diagnóstico , Infecciones por Circoviridae/inmunología , Circovirus/aislamiento & purificación , Ensayo de Inmunoadsorción Enzimática/economía , Peroxidasa de Rábano Silvestre/inmunología , Inmunización , Masculino , Proteínas Recombinantes de Fusión/inmunología , Sensibilidad y Especificidad , Anticuerpos de Dominio Único/inmunología , Porcinos/sangre , Porcinos/inmunología , Porcinos/virología , Enfermedades de los Porcinos/sangre , Enfermedades de los Porcinos/diagnóstico , Enfermedades de los Porcinos/virología , Factores de TiempoRESUMEN
Thrombogenic complications are a main issue in mechanical circulatory support (MCS). There is no validated in vitro method available to quantitatively assess the thrombogenic performance of pulsatile MCS devices under realistic hemodynamic conditions. The aim of this study is to propose a method to evaluate the thrombogenic potential of new designs without the use of complex in-vivo trials. This study presents a novel in vitro method for reproducible thrombogenicity testing of pulsatile MCS systems using low molecular weight heparinized porcine blood. Blood parameters are continuously measured with full blood thromboelastometry (ROTEM; EXTEM, FIBTEM and a custom-made analysis HEPNATEM). Thrombus formation is optically observed after four hours of testing. The results of three experiments are presented each with two parallel loops. The area of thrombus formation inside the MCS device was reproducible. The implantation of a filter inside the loop catches embolizing thrombi without a measurable increase of platelet activation, allowing conclusions of the place of origin of thrombi inside the device. EXTEM and FIBTEM parameters such as clotting velocity (α) and maximum clot firmness (MCF) show a total decrease by around 6% with a characteristic kink after 180 minutes. HEPNATEM α and MCF rise within the first 180 minutes indicate a continuously increasing activation level of coagulation. After 180 minutes, the consumption of clotting factors prevails, resulting in a decrease of α and MCF. With the designed mock loop and the presented protocol we are able to identify thrombogenic hot spots inside a pulsatile pump and characterize their thrombogenic potential.
Asunto(s)
Corazón Artificial/efectos adversos , Tromboelastografía/instrumentación , Trombosis/etiología , Animales , Diseño de Equipo , Porcinos/sangre , Tromboelastografía/métodosRESUMEN
Heat stress is major welfare concern during transport of pigs in tropical climates, which can also lead to direct production costs. This study evaluated the dynamics of heat zones through the load and their relationship with heat stress of weaner pigs during road transport in a tropical climate. Both environmental (e.g. temperature and relative humidity) and physiological (e.g. respiratory frequency and lactate) measures were recorded from four vehicle journeys (70 km distance, 216 weaner pigs within each trailer load) within Ceará, northeastern Brazil. Geostatistics and fluid dynamics simulation techniques were applied to understand the dynamics of heat zones and ventilation patterns the truckload. Statistics based on canonical discriminant analysis and ANOVA were performed to verify the relationship between heat zones and heat stress in pigs. The results showed that, during transport, the generation of heat zones occurred with different magnitudes along the load (P < 0.05), which was harmonized by the ventilation dynamics. There was a heat core with high energy content, in the front region of the lower deck (LD) of the trailer. In this zone, weaners pigs had higher rectal temperature (+1.8 °C temperature difference), respiratory frequency (LD = 94 ± 1.3 breaths/min; UD = 86 ± 1.3 breaths/min), and blood cortisol concentration (LD = 32.9 ± 0.8 ng/mL; UD = 30.18 ± 0.6 ng/mL) (all P < 0.05). Weaners pigs transported in the upper deck (UD) compartments had the highest skin temperature (LD = 38.13 ± 0.3 °C; UD = 38.9 ± 0.22 °C) and the highest mean values of blood lactate (LD = 65.5 ± 1.11 m/M; UD = 71.60 ± 1.19 m/M) and Creatine kinase (LD = 3891.23 ± 69U/L; UD = 4107.43 ± 62U/L) (P < 0.05). Weaners transported in compartments of the LD of trailer were more susceptible to heat stress, while weaners in the UD compartments were more susceptible to physical stress and muscle exhaustion. These results provide additional evidence of heat zones within trailer compartments and highlight the requirement for the planning of pig transport operations in tropical climates to mitigate risks of heat stress.
Asunto(s)
Respuesta al Choque Térmico/fisiología , Microclima , Porcinos/fisiología , Transportes , Crianza de Animales Domésticos , Animales , Temperatura Corporal , Brasil , Creatina Quinasa/sangre , Femenino , Trastornos de Estrés por Calor/sangre , Trastornos de Estrés por Calor/fisiopatología , Trastornos de Estrés por Calor/veterinaria , Calor , Hidrocortisona/sangre , Ácido Láctico/sangre , Respiración , Porcinos/sangre , Enfermedades de los Porcinos/sangre , Enfermedades de los Porcinos/fisiopatología , Clima TropicalRESUMEN
The pharmacokinetic behaviours of amoxicillin (AMX) and clavulanic acid (CA) in swine were studied after either an intravenous or oral administration of AMX (10 mg/kg) and CA (2.5 mg/kg). The concentrations of these two medicines in swine plasma were determined using high-performance liquid chromatographic-tandem mass spectrometry, and the data were analysed using a noncompartmental model with the WinNonlin software. After intravenous administration, both substances were absorbed rapidly and reached their effective therapeutic concentration quickly. CA was eliminated more slowly compared with AMX. Moreover, the distribution volume of AMX was larger than that of CA, suggesting that AMX could penetrate tissues better. After oral administration of the granular formulation, no significant difference was observed in the mean elimination half-life value between AMX and CA. The mean maximal plasma concentrations of AMX and CA, reached after 1.14 and 1.32 hr, were 2.58 and 1.91 µg/m, respectively. The mean oral bioavailability of AMX and CA was 23.6% and 26.4%, respectively. After oral administration, the T>MIC50 for three common respiratory pathogens was over 6.12 hr. Therefore, oral administration could be more effective in the clinical therapy of pigs, especially when administered twice daily.
Asunto(s)
Combinación Amoxicilina-Clavulanato de Potasio/farmacocinética , Antibacterianos/farmacocinética , Porcinos/sangre , Administración Oral , Combinación Amoxicilina-Clavulanato de Potasio/administración & dosificación , Combinación Amoxicilina-Clavulanato de Potasio/sangre , Animales , Antibacterianos/administración & dosificación , Antibacterianos/sangre , Área Bajo la Curva , Disponibilidad Biológica , Femenino , Semivida , Inyecciones Intravenosas/veterinaria , MasculinoRESUMEN
BACKGROUND: 25-Hydroxycholecalciferol (25OHD3 ) is a new feed additive, which is a potential alternative to vitamin D3 in swine nutrition. The objective of this study was to determine the effects of different doses of 25OHD3 supplementation on performance, immunity, antioxidant capacity, intestinal morphology and bone quality in piglets. RESULTS: As dietary 25OHD3 supplementation increased, the average daily gain (ADG) improved (P < 0.05) quadratically during days 1-14, and tended to increase (P = 0.06) quadratically during the overall period of the experiment. Increasing 25OHD3 supplementation increased (linear effect, P < 0.05) the serum 25OHD3 level and serum glutathione peroxidase (GSH-Px) activity. On day 14, serum immunoglobulin A (IgA) was increased (linear and quadratic effects, P < 0.05) as dietary 25OHD3 supplementation increased. On day 28, serum IgA level was higher (P < 0.05) linearly and the complement 3 (C3) level was reduced (P < 0.05) linearly as dietary supplementation of 25OHD3 increased. The mucosal GSH-Px activity of the small intestine was higher (quadratic effect, P < 0.05) with increasing 25OHD3 supplementation. Jejunal villus height (P = 0.06) and villus height to crypt depth ratio (P = 0.07) tended to increase quadratically, and the villus height to crypt-depth ratio of the ileum increased (P < 0.05) linearly and quadratically with increasing 25OHD3 supplementation. Dietary supplementation with an increasing level of 25OHD3 increased breaking strength of tibias and femurs (quadratic effect, P < 0.05). CONCLUSION: Increasing dietary 25OHD3 supplementation partly improved performance, immunity, antioxidant status, intestinal morphology, and bone properties of weaned piglets. © 2020 Society of Chemical Industry.
Asunto(s)
Antioxidantes/metabolismo , Huesos/efectos de los fármacos , Calcifediol/administración & dosificación , Suplementos Dietéticos/análisis , Intestino Delgado/crecimiento & desarrollo , Porcinos/inmunología , Animales , Huesos/química , Huesos/metabolismo , Femenino , Glutatión Peroxidasa/sangre , Inmunoglobulina A/sangre , Mucosa Intestinal/crecimiento & desarrollo , Mucosa Intestinal/metabolismo , Intestino Delgado/anatomía & histología , Intestino Delgado/efectos de los fármacos , Intestino Delgado/metabolismo , Masculino , Porcinos/sangre , Porcinos/crecimiento & desarrollo , DesteteRESUMEN
RATIONALE: There are no approved animal drugs for management of inflammation in swine due to lack of validated animal models. To assess efficacy, it was essential to develop proteomics approaches to identify suitable biomarkers of inflammation as presented in this study. METHODS: Serum samples were collected from a group of four pigs prior to (baseline) and 24 and 48 h following lipopolysaccharide (LPS) stimulation to reveal proteomic changes during inflammation. Two other pigs served as untreated controls. Proteins were separated by either one-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) or two-dimensional (2D) gel electrophoresis (2DE) prior to analysis by nano-flow liquid chromatography (nLC) coupled to tandem mass spectrometry (MS/MS). RESULTS: We identified 165 proteins using SDS-PAGE, of which 47 proteins were also detected by 2DE prior to nLC/MS/MS. More than half (72%) of all characterized proteins were modulated as a result of LPS stimulation, many of which are known to be involved with innate and adaptive immunity. Pig serum samples obtained 24 h after LPS initiation of inflammation showed protein modulations of serum albumin, serotransferrin, light and heavy immunoglobulin chains (IGs), and major acute phase proteins including haptoglobin (HPT), serum amyloid A2 (SAA2), C-reactive protein (CRP), ß-2-glycoprotein 1 (B-2GP1), alpha-2-HS-glycoprotein (A2HS), α-1-antitrypsin (A1AT), and α-1-acid glycoprotein (A1AG). SAA2 was distinguished from the other SAA isoforms by the unique peptide sequence of SAA2. CONCLUSIONS: The results provided proteomics analysis of swine serum due to LPS stimulation and indicated the importance of SAA2, which appears to be unique and may be regarded as a potential clinical diagnostic biomarker of inflammation.
Asunto(s)
Proteínas Sanguíneas/análisis , Inflamación/sangre , Lipopolisacáridos/inmunología , Porcinos/sangre , Inmunidad Adaptativa , Animales , Biomarcadores/sangre , Proteínas Sanguíneas/inmunología , Cromatografía Liquida , Electroforesis en Gel Bidimensional , Electroforesis en Gel de Poliacrilamida , Inmunidad Innata , Inflamación/inmunología , Lipopolisacáridos/sangre , Proteómica , Porcinos/inmunología , Espectrometría de Masas en TándemRESUMEN
BACKGROUND: Measurement of adenosine deaminase (ADA) can provide information about cell-mediated immunity. This report's objective was to study the enzymatic activity of total ADA (tADA) and its isoenzymes ADA1 and ADA2 in canine, equine, porcine, and bovine serum and saliva and their changes in different inflammatory situations in each species. Besides, an automated method for ADA2 measurement was developed and validated. RESULTS: tADA was present in serum and saliva of healthy animals of the four species. Erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA) concentration of 0.47 mM was needed for ADA1 inhibition in canine and porcine samples (serum and saliva) and bovine saliva, whereas for equine saliva 0.94 mM was needed. ADA2 activity was not detected in bovine serum and was very low or absent in equine serum and bovine saliva. An automated procedure to measure ADA2 consisting of adding EHNA to a commercial reagent for tADA measurement provided repetitive (coefficients of variation < 8.8% in serum and < 10% in saliva) and accurate (linearity of serial sample dilutions with R2 > 0.90) results, being equivalent to a manual incubation of the sample with EHNA at a similar concentration. Salivary tADA, as well as ADA1 and ADA2, were higher in dogs with leishmaniosis, horses with acute abdominal disease and pigs with lameness than in healthy animals. tADA and isoenzymes in saliva showed a positive significant correlation with serum ferritin in dogs (r = 0.602, P < 0.01; r = 0.555, P < 0.05; and r = 0.632, P < 0.01; respectively for tADA, ADA1 and ADA2) and serum C-reactive protein in pigs (r = 0.700, P < 0.01, for both tADA and ADA1; r = 0.770, P < 0.001, for ADA2), whereas salivary ADA2 significantly correlated with serum amyloid A in horses (r = 0.649, P < 0.01). In cows, salivary tADA and ADA1 significantly increased after calving, correlating with total white blood cell count (r = 0.487, P < 0.05, for both tADA and ADA1). CONCLUSIONS: The activity of total ADA and its different isoenzymes, can be measured in serum and saliva of dogs, horses, pigs and cows by a simple and fast procedure described in this report. When measured in saliva, these analytes correlated with other biomarkers of inflammation and it could potentially be used as a biomarkers of inflammation and immune activation in the species of this study.
Asunto(s)
Adenosina Desaminasa/metabolismo , Bovinos/metabolismo , Perros/metabolismo , Caballos/metabolismo , Inflamación/veterinaria , Saliva/metabolismo , Porcinos/metabolismo , Adenina/análogos & derivados , Adenosina Desaminasa/sangre , Inhibidores de la Adenosina Desaminasa , Animales , Automatización , Biomarcadores/sangre , Biomarcadores/metabolismo , Bovinos/sangre , Pruebas Enzimáticas Clínicas/métodos , Pruebas Enzimáticas Clínicas/veterinaria , Perros/sangre , Caballos/sangre , Inflamación/sangre , Inflamación/enzimología , Isoenzimas/sangre , Isoenzimas/metabolismo , Porcinos/sangreRESUMEN
The first case of African swine fever (ASF) outbreak in China was reported in a suburban pig farm in Shenyang in 2018. Since then, the rapid spread and extension of ASF has become the most serious threat for the swine industry. Therefore, rapid and accurate detection of African swine fever virus (ASFV) is essential to provide effective strategies to control the disease. In this study, we developed a rapid and accurate ASFV-detection method based on the DNA endonuclease-targeted CRISPR trans reporter (DETECTR) assay. By combining recombinase polymerase amplification with CRISPR-Cas12a proteins, the DETECTR assay demonstrated a minimum detection limit of eight copies with no cross reactivity with other swine viruses. Clinical blood samples were detected by DETECTR assay and showed 100% (30/30) agreement with real-time polymerase chain reaction assay. The rapid and accurate detection of ASFV may facilitate timely eradication measures and strict sanitary procedures to control and prevent the spread of ASF.
Asunto(s)
Virus de la Fiebre Porcina Africana/genética , Fiebre Porcina Africana/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificación de Ácido Nucleico/métodos , Porcinos/sangre , Fiebre Porcina Africana/sangre , Fiebre Porcina Africana/virología , Animales , Proteínas Bacterianas/biosíntesis , Proteínas Bacterianas/aislamiento & purificación , Proteínas Asociadas a CRISPR/biosíntesis , Proteínas Asociadas a CRISPR/aislamiento & purificación , Sistemas CRISPR-Cas , China , Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Espaciadas/genética , ADN Viral/genética , Desoxirribonucleasa I/genética , Endodesoxirribonucleasas/biosíntesis , Endodesoxirribonucleasas/aislamiento & purificación , Fluorescencia , Límite de Detección , Reacción en Cadena en Tiempo Real de la Polimerasa , Recombinasas/metabolismo , Sensibilidad y EspecificidadRESUMEN
The medicinal plant Artemisia annua L. is well known for its antimalarial compound artemisinin and the antioxidant capacity of its active ingredients. However, low bioavailability of Artemisia annua L. limits its therapeutic potential, fermentation of Artemisia annua L. can improve its bioavailability. This study was aimed to investigate the effects of dietary supplementation of enzymatically-treated Artemisia annua L. (EA) on reproductive performance, antioxidant status, milk composition of heat-stressed sows and intestinal barrier integrity of their preweaning offspring. 135 multiparous sows of average parity 4.65 (Landrace × large white) at day 85 of pregnancy were randomly distributed into 3 treatments. Sows in the control group were housed at control rooms (temperature: 27.12 ± 0.18 °C, temperature-humidity index (THI): 70.90 ± 0.80) and fed the basal diet. Sows in the HS, HS + EA groups were fed the basal diet supplemented with 0 or 1.0 g/kg EA respectively, and reared at heat stress rooms (temperature: 30.11 ± 0.16 °C, THI: 72.70 ± 0.60). Heat stress increased the malondialdehyde (MDA) content, reduced the activities of total antioxidant capacity (T-AOC) and total superoxide dismutase (T-SOD) of sows and piglets, and seriously compromised the antioxidant capacity of the sows and the intestinal integrity of their offspring. However, dietary supplementation of 1.0 g/kg EA reduced the MDA content, increased the activities of T-SOD and T-AOC in serum, colostrum, and milk of heat-stressed sows, and increased colostrum yield and 14-d milk fat content. EA supplementation also increased piglet weaning weight and the activities of T-SOD and T-AOC in serum. In addition, the abundances of intestinal tight junction proteins claudin-1 and occludin were up-regulated in piglets in EA-supplemented group. In conclusion, dietary EA supplementation at 1.0 g/kg can alleviate the oxidative stress in heat-stressed sows, improve the antioxidant capacity in both sows and their offspring, and promote the intestinal barrier integrity in their offspring. EA may be a potent dietary supplement that ameliorates oxidative stress in livestock production by improving the antioxidant capacity.
Asunto(s)
Artemisia annua , Suplementos Dietéticos , Calor/efectos adversos , Estrés Oxidativo , Reproducción , Alimentación Animal , Animales , Artemisia annua/química , Celulasa/química , Dieta/veterinaria , Femenino , Glutatión/sangre , Trastornos de Estrés por Calor/sangre , Trastornos de Estrés por Calor/genética , Trastornos de Estrés por Calor/veterinaria , Leche/química , Oxidorreductasas/sangre , Poligalacturonasa/química , Embarazo , Porcinos/sangre , Porcinos/genética , Enfermedades de los Porcinos/sangre , Enfermedades de los Porcinos/genética , Proteínas de Uniones Estrechas/genéticaRESUMEN
The pharmacokinetics of carbetocin, which is used to control postpartum hemorrhage after giving birth, was studied in cows and gilts after a single intravenous (IV) or intramuscular (IM) injection. Blood samples from animals were assessed by oxytocin radioimmunoassay, and then the pharmacokinetic parameters were calculated using a noncompartmental model. For gilts, there was no significant difference between half-life (T1/2λZ ), mean residue time (MRT), and maximum concentration (Cmax ) between IM and IV administration. Conversely, the time to reach the Cmax (Tmax ) and MRT were higher following administration of 350 µg/animal in cows via the IM administration compared with IV. The longest T1/2λZ was 0.85 hr, indicating carbetocin was absorbed and eliminated rapidly in both animal species after administration. The Tmax was similar between cows and gilts following IM administration. Moreover, the Cmax after IM injection was about half that of IV administration in both animals. The bioavailability was more than 80% in cows, suggesting administration via the IM route is efficient. This is in agreement with the longer T1/2λZ in cows after IM administration. However, the IV route is recommended for gilts due to a lower bioavailability (35%) and shorter T1/2λZ after IM administration compared with IV.
Asunto(s)
Bovinos/sangre , Oxitócicos/farmacocinética , Oxitocina/análogos & derivados , Porcinos/sangre , Animales , Área Bajo la Curva , Disponibilidad Biológica , Semivida , Inyecciones Intramusculares , Inyecciones Intravenosas , Oxitócicos/administración & dosificación , Oxitocina/administración & dosificación , Oxitocina/farmacocinética , Especificidad de la EspecieRESUMEN
This study aimed to develop one novel meloxicam (MEL) oil suspension for sustained-release and compare the pharmacokinetic characteristics of it with MEL conventional formulation in pigs after a single intramuscular administration. Six healthy pigs were used for the study by a crossover design in two periods with a withdrawal interval of 14 days. Plasma concentrations of MEL were measured by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Pharmacokinetic parameters were calculated by noncompartmental methods. The difference was statistically significant (p < .05) between MEL oil suspension and MEL conventional formulation in pharmacokinetic parameters of mean residence time (6.16 ± 4.04) hr versus (2.66 ± 0.55) hr, peak plasma concentration (Cmax ) (0.82 ± 0.12) µg/ml versus (1.12 ± 0.22) µg/ml, time needed to reach Cmax (Tmax ) (2.33 ± 0.82) hr versus (0.59 ± 0.18) hr, and terminal elimination half-life (t1/2λz ) (3.74 ± 2.66) hr versus (1.55 ± 0.37) hr. The mean area under the concentration-time curve (AUC0-â ) of MEL oil suspension and MEL conventional formulation was 5.35 and 3.43 hr µg/ml, respectively, with a relative bioavailability of 155.98%. Results of the present study demonstrated that the MEL oil suspension could prolong the effective time of drugs in blood, thereby reducing the frequency of administration on a course of treatment. Therefore, the novel MEL oil suspension seems to be of great value in veterinary clinical application.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacocinética , Meloxicam/farmacocinética , Porcinos/sangre , Animales , Antiinflamatorios no Esteroideos/administración & dosificación , Área Bajo la Curva , Disponibilidad Biológica , Semivida , Inyecciones Intramusculares , Meloxicam/administración & dosificación , SuspensionesRESUMEN
In this study, we examined the effects of acute intravenous administration of l-arginine on circulating levels of metabolites in the portal-drained viscera (PDV) of 12 barrows surgically fitted with chronic catheters in the portal vein. At day 14 post-surgery, the pigs were fasted for 12 hr and then randomly allocated to one of three groups to receive administration of normal saline, l-alanine [103 mg/kg body weight (BW), isonitrogenous control] or l-arginine-HCl (61 mg/kg BW), via the portal vein. Blood samples were obtained from the carotid artery before and at 30-min intervals for 5 hr after the administration of saline or amino acid in order to determine metabolic profiles. The results showed that, compared with the saline treatment, arginine infusion increased plasma concentrations of insulin-like growth factor-I, arginine and cystine in the portal vein plasma, whereas plasma concentrations of threonine, serine, leucine and methionine were reduced. These findings indicate that increasing arginine concentrations in the portal vein alters the metabolic profile in swine, an established animal model for studying human nutrition and metabolism.
Asunto(s)
Arginina/farmacología , Porcinos/sangre , Animales , Arginina/administración & dosificación , Esquema de Medicación , Inyecciones Intravenosas , Masculino , Vena Porta , Porcinos/metabolismoRESUMEN
This study was conducted to detect the potential relationship between changed plasma metabolites, intestinal microbiota and the weaning-to-oestrous interval in multiparous sows after weaning. Multiparous sows were allocated to two groups after weaning: the oestrous group (n = 15) with a weaning-to-oestrous interval ≤7 days or the anoestrous group (n = 15) with a weaning-to-oestrous interval >14 days. The levels of plasma reproductive hormones: oestradiol, follicle-stimulating hormone and luteinizing hormone, plasma total protein; blood urea nitrogen; cholesterol; high-density lipoprotein; and ammonia (NH3 ) were significantly lower in the anoestrous sows compared with the oestrous sows (p < .05). The plasma metabolomics analysis identified 14 metabolites (lactose, l-cysteine, cytosine, hydantoin, palmitoleic acid, arachidic acid, linoleic acid methyl ester, α-ketoglutaric acid, N(ε)-trimethyllysine, threo-ß-hydroxyaspartate, 3-(3-hydroxyphenyl) propionic acid and others) with lower concentrations and 12 metabolites (noradrenaline, 5-dihydrocortisone, p-cresol, 1,4-cyclohexanedione, 2,3-dimethylsuccinic acid and others) with higher concentrations in the anoestrous group compared with the oestrous group (p < .05). The 16S rRNA pyrosequencing analysis showed the relative increase in abundance of the Prevotella and the Bacteroides at the genus level in the anoestrous group (p < .05). At the phylum level, lower proportions of Firmicutes and Lentisphaerae were observed in the anoestrous group (p < .05). This study provided a comprehensive assessment of metabolic differences in the blood and differences in the gut microbiome composition between anoestrous and oestrous sows. And suggesting that this profiling approach may offer new insights into explaining the alteration of the gut microbiota and blood metabolomics are correlated with sex hormone secretion and the weaning-to-oestrous interval of sows after weaning.
Asunto(s)
Estro/fisiología , Microbioma Gastrointestinal , Metabolómica , Paridad , Porcinos/sangre , Animales , Estradiol/sangre , Estro/sangre , Femenino , Hormona Folículo Estimulante/sangre , Hormona Luteinizante/sangre , Porcinos/microbiologíaRESUMEN
This study was conducted to determine the effects of oral administration with glutamate on metabolism of suckling piglets based on 1 H-Nuclear magnetic resonance (1 H NMR) spectroscopy through the level of metabolism. Forty-eight healthy [(Yorkshire × Landrace) × Duroc] piglets born on the same day with a similar birth bodyweight (1.55 ± 0.20 kg) were obtained from six sows (8 piglets per sow). The piglets from each sow were randomly assigned into four treatments (2 piglets per treatment). The piglets were given 0.09 g/kg body weight (BW) of sodium chloride (CN group), 0.03 g/kg BW monosodium glutamate (LMG group), 0.25 g/kg BW monosodium glutamate (MMG group) and 0.50 g/kg BW monosodium glutamate (HMG group) twice a day respectively. An 1 H NMR-based metabolomics' study found that the addition of monosodium glutamate (MSG) significantly reduced serum citrate content in 7-day-old piglets, while HMG significantly increased serum trimethylamine content and significantly reduced unsaturated fat content in 7-day-old piglets (p < .05). The content of glutamine, trimethylamine, albumin, choline and urea nitrogen was significantly increased and the creatinine content decreased significantly in the 21-day-old HMG (p < .05). Analysis of serum hormones revealed that glucagon-like peptide-1 (GLP-1) content in the 21-day-old HMG was highest (p < .05). The cholecystokinin (CCK) content in the HMG of 7-day-old piglets was lower than that in the LMG (p < .05), and the CCK content in the serum of the 21-day-old MMG was highest (p < .05). The serum leptin levels in the 21-day-old HMG were the lowest (p < .05). The serum insulin content in the 7-day-old MMG was highest (p < .05). This study suggests that MSG plays an important role in the metabolism of sugar, fat and protein (amino acids). These results provide a theoretical basis for designing piglet feed formulations.
Asunto(s)
Animales Lactantes , Metaboloma/efectos de los fármacos , Metabolómica , Glutamato de Sodio/farmacología , Porcinos/fisiología , Administración Oral , Alimentación Animal , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Dieta/veterinaria , Suplementos Dietéticos , Porcinos/sangreRESUMEN
Moringa oleifera has been considered as a potential functional feed or food, since it contains multiple components beneficial to animal and human. However, little is known about the effects of Moringa oleifera supplementation on productive performances in sows. In the current study, the results showed that dietary Moringa oleifera significantly decreased the farrowing length and the number of stillborn (p < .05), while had an increasing trend in the number of live-born (0.05 < p < .10). Furthermore, 8% Moringa oleifera supplementation significantly elevated protein levels in the colostrum (p < .05); 4% Moringa oleifera lowed serum urea nitrogen of sows after 90 days of gestation (p < .05) and significantly decreased serum glucose on 10 days of lactation (p < .05). Both groups showed significant elevation in serum T-AOC activity (p < .05). The serum malondialdehyde (MDA) of sows declined significantly in 4% Moringa oleifera addition group (p < .05). 8% Moringa oleifera meal significantly elevated serum CAT activity after 60 days of gestation (p < .05), while decreased the serum MDA level and increased the serum GSH-Px activity of sows at 10 days of lactation (p < .05). Of piglets, both two dosages of Moringa oleifera supplementation essentially reduced the serum urea nitrogen (p < .05), and 4% Moringa oleifera meal increased serum total protein (p < .05). In addition, piglets that received 8% Moringa oleifera had the highest serum CAT and SOD activities among all groups (p < .05). The present study indicated that Moringa oleifera supplementation could enhance the reproduction performances, elevate protein levels in the colostrum and improve the serum antioxidant indices in both sows and piglets.