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1.
Gene ; 731: 144356, 2020 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-31935504

RESUMO

The genetic basis of differential host immune response vis-à-vis transcriptome profile was explored in PBMCs of indigenous (Ghurrah) and crossbred pigs after classical swine fever vaccination and in monocyte derived macrophages (MDMs) challenged with virulent classical swine fever (CSF) virus. The humoral immune response (E2 antibody) was higher (74.87%) in crossbred than indigenous pigs (58.20%) at 21st days post vaccination (21dpv). The rate of reduction of ratio of CD4+/CD8+ was higher in crossbred pigs than indigenous pigs at 7th days post vaccination (7dpv). The immune genes IFIT1, IFIT5, RELA, NFKB2, TNF and LAT2 were up regulated at 7dpv in RNA seq data set and was in concordance during qRT-PCR validation. The Laminin Subunit Beta 1 (LAMB1) was significantly (p ≤ 0.05) down-regulated in MDMs of indigenous pigs and consequently a significantly (p ≤ 0.01) higher copy number of virulent CSF virus was evidenced in macrophages of crossbred pigs than indigenous pigs. Activation of LXR:RXR pathway at 60 h post infection (60hpi) in MDMs of indigenous versus crossbred pigs inhibited nuclear translocation of NF-κB, resulted into transrepression of proinflammatory genes. But it helped in maintenance of HDL level by lowering down cholesterol/LDL level in MDMs of indigenous pigs. The key immune genes (TLR2, TLR4, IL10, IL8, CD86, CD54, CASP1) of TREM1 signaling pathway were upregulated at 7dpv in PBMCs but those genes were downregulated at 60hpi in MDMs indigenous pigs. Using qRT-PCR, the validation of differentially expressed, immunologically important genes (LAMB1, OAS1, TLR 4, TLR8 and CD86) in MDMs revealed that expression of these genes were in concordance with RNA-seq data.


Assuntos
Peste Suína Clássica/genética , Leucócitos Mononucleares/metabolismo , Macrófagos/metabolismo , Suínos , Transcriptoma , Animais , Células Cultivadas , Peste Suína Clássica/sangue , Peste Suína Clássica/imunologia , Peste Suína Clássica/prevenção & controle , Vírus da Febre Suína Clássica/imunologia , Vírus da Febre Suína Clássica/fisiologia , Estudo de Associação Genômica Ampla/veterinária , Interações Hospedeiro-Patógeno/genética , Interações Hospedeiro-Patógeno/imunologia , Hibridização Genética/fisiologia , Imunidade Celular/genética , Imunidade Humoral/genética , Leucócitos Mononucleares/patologia , Leucócitos Mononucleares/virologia , Macrófagos/patologia , Macrófagos/virologia , Suínos/genética , Suínos/imunologia , Suínos/metabolismo , Suínos/virologia
2.
J Anim Sci ; 97(11): 4548-4556, 2019 Nov 04.
Artigo em Inglês | MEDLINE | ID: mdl-31603198

RESUMO

The objectives of the current study were to explore the effects of mannan oligosaccharide (MOS) supplementation in the diets of sow and (or) their offspring on intestinal bacteria, intestinal and systemic inflammation in the piglet. A total of 60 multiparous sows (4 ± 1 parity; Landrace × Yorkshire) were fed either control diet (sCON, n = 30) or a diet containing 400 mg kg-1 MOS (sMOS, n = 30) from day 86 of gestation until weaning (day 20 of postpartum). On day 7 of age, offspring (Duroc × Landrace Yorkshire) were assigned within sow treatments and fed control diet (pCON) or diet containing 800 mg kg-1 MOS (pMOS) for 28 d (end at 35 d of age), resulting in four piglet diet groups (n = 15 litters per diet group): sCON-pCON, sCON-pMOS, sMOS-pCON, and sMOS-pMOS. Results found that piglet diet MOS increased or tend to increase Lactobacillus amount in the ileum digesta (P < 0.01) and jejunum digesta (P = 0.07), respectively; while tend to decrease Escherichia coli amount in jejunum digesta (P =0.06) and cecum digesta (P = 0.08). Both sow and piglet diets add MOS (sMOS-pMOS) increased Lactobacillus amount but decreased E. coli amount in jejunum digesta (P < 0.05) compared with the sCON-pCON diet group. In addition, sow diet MOS (rather than piglet diet MOS) increased sIgA content in piglet jejunum mucosa compared with control (P = 0.04). Sow diet MOS decreased toll-like receptor 2 (TLR2), toll-like receptor 4 (TLR4), and interleukin 8 (IL-8) mRNA levels (P < 0.05) and tended to decrease nuclear factor-κB p65 (NF-κB p65) mRNA level (P = 0.07) in piglet intestinal lymphatic. The interaction effects between sow and piglet diets were found on the mRNA levels of NF- κB p65 (P = 0.03) and IL-8 (P = 0.02) in piglet jejunum. Finally, the sow diet MOS decreased proinflammatory cytokines IL-2 (P < 0.01) and IL-4 (P < 0.01) concentrations in piglet serum. Piglets diet MOS decreased the contents of IL-2 (P = 0.03), IL-4 (P = 0.01) and interferon (IFN)-γ (P < 0.01) while increased anti-inflammatory cytokine IL-10 (P < 0.01) content in serum. The interaction effects between sows and piglet diets on IL-4 (P = 0.02), IL-10 (P < 0.01), and IFN-γ (P = 0.08) were observed. In conclusion, sow and/or piglet diet MOS could improve intestinal microbiota, enhance intestinal mucosal immune competence, and suppress intestinal and systemic inflammation in the piglet.


Assuntos
Suplementos Nutricionais/análise , Microbioma Gastrointestinal/efeitos dos fármacos , Mananas/administração & dosagem , Oligossacarídeos/administração & dosagem , Suínos/fisiologia , Ração Animal/análise , Animais , Animais Recém-Nascidos , Dieta/veterinária , Feminino , Inflamação/veterinária , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/imunologia , Masculino , Paridade , Gravidez , Suínos/imunologia
3.
J Anim Sci ; 97(11): 4557-4566, 2019 Nov 04.
Artigo em Inglês | MEDLINE | ID: mdl-31504564

RESUMO

The present study was conducted to evaluate the effects of dietary garcinol supplementation during late gestation (from the 90th day of pregnancy; day 90) and lactation on the acid-base balance of the umbilical cord blood and performance of sows and piglets. Sixty sows (Duroc × Yorkshire × Landrace; second- or third-parity; n = 20) were randomly divided into 3 gestation (day 90 of pregnancy) or lactation treatments, control diet (CON; basal diet), basal diet with 200 mg garcinol, and basal diet with 600 mg garcinol per kg of feed. The body weight (BW); backfat thickness and litter size of the sows; and birth weight, weaning weight, and mortality of piglets were recorded. Sows' blood and piglets' umbilical cord blood were collected for the measurements of hematological parameters and antioxidative and immune indexes, and acid-base balance parameters, respectively. The colostrum and milk and fecal samples of the sows were also collected for analysis of milk composition and apparent total tract nutrient digestibility. Garcinol had no effect on the BW and backfat thickness of the sows but significantly increased the birth weight and weaning weight of piglets (P < 0.05) and decreased the mortality (P < 0.05). Moreover, the white blood cell counts and neutrophil count, mean cell hemoglobin, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) activity in the plasma of the sows were increased more significantly (P < 0.05) in the garcinol groups than that in the CON group, whereas the malondialdehyde (MDA) content was decreased (P < 0.05). The garcinol treatment significantly increased the pH, HCO3- and base excess values (P < 0.05), whereas it decreased the pCO2 and lactate content (P < 0.05) in the umbilical blood. Dry matter (DM), ash, and ether extract in the colostrum were similar between groups (P > 0.05), whereas the garcinol significantly increased the crude protein (CP) in the milk. In addition, the content of immunoglobulin A (IgA) and immunoglobulin G (IgG) in the plasma of piglets and in colostrum and milk of sows were increased more significantly (P < 0.05) in the garcinol groups than that in the CON group. The apparent total tract nutrient digestibility was similar between treatments. Collectively, this study indicates that sows fed with garcinol in late gestation and lactation showed improved maternal health and antioxidative status, milk protein content, acid-base balance in the umbilical cord blood, and growth performance in piglets, showing promise in natural plant extract nutrition for sows.


Assuntos
Suplementos Nutricionais/análise , Leite/química , Suínos/fisiologia , Terpenos/administração & dosagem , Equilíbrio Ácido-Base/efeitos dos fármacos , Ração Animal/análise , Animais , Colostro/química , Dieta/veterinária , Feminino , Sangue Fetal/efeitos dos fármacos , Imunoglobulina A/sangue , Lactação/efeitos dos fármacos , Tamanho da Ninhada de Vivíparos/efeitos dos fármacos , Paridade , Gravidez , Distribuição Aleatória , Suínos/sangue , Suínos/imunologia , Desmame
4.
J Anim Sci ; 97(10): 4256-4267, 2019 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-31504586

RESUMO

This study was conducted to compare the effects of adding sodium butyrate (SB), medium-chain fatty acids (MCFAs), or n-3 polyunsaturated fatty acids (n-3 PUFAs) to the diet of sows during late gestation and lactation on the reproductive performance of sows and the growth performance and intestinal health of suckling piglets. Twenty-four sows (Landrace × Large-White hybrid; third parity; 200 ± 15 kg) were randomly assigned to receive 1 of 4 diets: basal diet (control group), basal diet + 1 g SB/kg (SB group), basal diet + 7.75 g MCFA/kg (MCFA group), or basal diet + 68.2 g n-3 PUFA/kg (n-3 PUFA group). The experiment began on day 85 of gestation and ended day 22 of lactation. Colostrum samples were collected from each sow. After the experiment, blood and tissue samples were collected from 1 randomly selected piglet. The results showed that the weaning-to-estrus interval of sows in the SB, MCFA, and n-3 PUFA groups was shorter than that of sows in the control group (P < 0.05). The incidence of diarrhea in suckling piglets in the SB, MCFA, and n-3 PUFA groups was lower than that of piglets in the control group (P < 0.05). The fat, protein, IgA, IgG, and IgM concentration in colostrum from sows increased following dietary supplementation with SB, MCFA, or n-3 PUFA (P < 0.05). Comparison with the control group, the mRNA expression of claudin-1, zona occludens 1, and interleukin-10 increased in the jejunum mucosa of suckling piglets in the SB, MCFA, and n-3 PUFA groups, while that of TLR4 decreased (P < 0.05). Compared with the control group, the Chao1 and ACE indexes of microbial flora in the colon contents of piglets in the SB, MCFA, and MCFA groups increased (P < 0.05), while the relative abundance of Firmicutes, Actinobacteria, and Synergistetes decreased at the phylum level (P < 0.05). In conclusion, during late pregnancy and lactation, dietary SB supplementation had a greater effect on intestinal health and caused a greater decrease in preweaning mortality of suckling piglets than did dietary MCFA or n-3 PUFA supplementation; dietary MCFA supplementation shortened the weaning-to-estrus interval of sows to a greater extent than did dietary SB or n-3 PUFA supplementation; and dietary n-3 PUFA supplementation increased the fat and protein content in the colostrum to the greatest extent.


Assuntos
Ácido Butírico/farmacologia , Suplementos Nutricionais/análise , Ácidos Graxos Ômega-3/farmacologia , Ácidos Graxos/farmacologia , Reprodução/efeitos dos fármacos , Suínos/fisiologia , Ração Animal/análise , Animais , Animais Lactentes , Colostro/química , Colostro/imunologia , Dieta/veterinária , Feminino , Lactação/efeitos dos fármacos , Paridade , Gravidez , Distribuição Aleatória , Suínos/crescimento & desenvolvimento , Suínos/imunologia , Desmame
5.
Reprod Domest Anim ; 54 Suppl 3: 12-21, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31512316

RESUMO

The use of hyperprolific sow lines has increased litter size considerably in the last three decades. Nowadays, in some countries litters can reach up to 18-20 piglets being a major challenge for the sow's physiology during pregnancy, parturition and lactation. The increased number of piglets born per litter prolongs sensibly the duration of farrowing, decreases the piglets' average weight at birth and their vitality, increases the competition for colostrum intake and can affect negatively piglets' survival. This review aims to describe how large litters can affect the immune system of the sow and the piglets and proposes measures to improve this condition.


Assuntos
Tamanho da Ninhada de Vivíparos/imunologia , Gravidez/imunologia , Suínos/imunologia , Animais , Animais Recém-Nascidos/imunologia , Peso ao Nascer , Colostro , Feminino , Lactação/imunologia , Parto/fisiologia , Suínos/crescimento & desenvolvimento
6.
Animal ; 13(11): 2736-2744, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31475667

RESUMO

Gaining a deeper understanding into the underlying mechanisms associated with intestinal function and immunity during the weaning transition is critical to help shed new light into applied nutrition approaches to improve piglet performance and health during this critical life-stage transition. The transient anorexia triggered at weaning leads to compromised intestinal barrier function and a localized inflammatory response. Considering barrier function, specific nutrient fractions appear to have a significant impact on the development and function of the immune and microbial systems around weaning. Understanding the specific impact of nutrients in the small intestine and hindgut is important for helping to bring more focus and consistency to nutritional approaches to support health and immunity during the weaning transition period. The challenge continues to be how to translate these modes of action into practical and scalable approaches for swine nutrition. We will focus specifically on practical nutritional approaches to influence intestinal immunity through lipid, protein and antioxidant nutrition.


Assuntos
Estado Nutricional/imunologia , Suínos/imunologia , Animais , Mucosa Intestinal/imunologia , Mucosa Intestinal/fisiologia , Intestinos/imunologia , Intestinos/fisiologia , Suínos/fisiologia , Desmame
7.
Vet Immunol Immunopathol ; 215: 109913, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31420069

RESUMO

The development of a rapid and efficient system to generate porcine monoclonal antibodies (mAbs) is an important step toward the discovery of critical neutralizing targets for designing rational vaccines against porcine viruses. In this study, we established a platform for producing porcine mAbs based on single cell technologies. First, we singled out an optimal donor from 507 pigs based on serum antibody neutralizing activity against porcine reproductive and respiratory syndrome virus (PRRSV). After identifying the contribution of IgG to the neutralizing activity, single CD45R+IgG+Ag+ B cells were sorted from peripheral blood mononuclear cells (PBMCs). Single B cell RT-PCR was performed using primers designed to cover the germline repertoire of the porcine VH/VL gene segments. Paired VH/VLs were cloned into a eukaryotic expression vector and transfected into 293T cells. We demonstrate that full-length porcine mAbs were produced, and antigen-specific mAbs were obtained after further validation. The approach reported in this study can be applied to generate porcine mAbs against any given antigen and may help with the screening of neutralizing antibodies against porcine pathogens.


Assuntos
Anticorpos Monoclonais/biossíntese , Anticorpos Antivirais/biossíntese , Vírus da Síndrome Respiratória e Reprodutiva Suína/imunologia , Suínos/imunologia , Animais , Anticorpos Monoclonais/genética , Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais/isolamento & purificação , Anticorpos Neutralizantes/genética , Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/genética , Anticorpos Antivirais/imunologia , Anticorpos Antivirais/isolamento & purificação , Especificidade de Anticorpos , Linfócitos B/imunologia , Células HEK293 , Humanos , Região Variável de Imunoglobulina/genética , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Transfecção , Recombinação V(D)J
8.
J Anim Sci ; 97(9): 3636-3647, 2019 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-31260524

RESUMO

It was shown previously that high dietary fiber (DF) and immune system stimulation (ISS) with systemic Escherichia coli lipopolysaccharide independently increased the threonine (Thr) requirement to maximize growth performance and protein deposition (PD). However, no additive effects on the Thr requirement were observed when both DF and ISS were present. The objective of the present study was to investigate whether supplementing Thr to meet previously estimated requirements for high DF and systemic immune challenge would maintain performance of pigs exposed to an enteric immune challenge when fed high DF. A total of 128 pigs (22.6 ± SD = 1.6 kg initial BW) were assigned to 1 of 4 dietary treatments in a 2 × 2 factorial arrangement in a randomized complete block design (n = 8 pens/treatment and 4 pigs/pen) for 28 d. Treatments were a low-fiber (LF; 13% total DF) or high-fiber (HF; 20% total DF) diet with either a standard (STD; 0.65% SID) or supplemental (SUP; 0.78% SID) Thr level. After a 7-d adaptation, pigs were orally inoculated with 2 mL (2.3 × 109 CFU/mL) of Salmonella typhimurium (ST). Blood samples and rectal swabs were obtained and rectal temperature recorded to determine clinical responses and ST shedding. On day 7 postinoculation, 1 pig/pen was euthanized and mesenteric lymph nodes, spleen, and digesta (ileum, cecum, and colon) were sampled to assess ST colonization and translocation. Body weight and feed intake were recorded on day 0, 7, and 21 postinoculation to calculate ADG, ADFI, and G:F. Rectal temperature increased (P < 0.05) 24 h postinoculation and remained elevated at day 6. Serum albumin concentration decreased (P < 0.05), whereas haptoglobin concentration increased (P < 0.05) postinoculation. There was no fiber or Thr effect (P > 0.05) on ST counts in the ileum and cecum, but a fiber × Thr interaction (P < 0.05) was observed in the colon. Supplemental Thr improved (P < 0.05) growth performance in LF- and HF-fed challenged pigs. However, performance of supplemented HF challenged pigs was less than (P < 0.05) supplemented LF challenged pigs. These results suggest that Thr supplemented to meet requirements for high DF and systemic immune challenge was not sufficient to maintain growth performance of pigs fed HF diets and challenged with an enteric pathogen.


Assuntos
Fibras na Dieta/metabolismo , Suplementos Nutricionais , Salmonella typhimurium/fisiologia , Suínos/crescimento & desenvolvimento , Treonina/farmacologia , Animais , Derrame de Bactérias/efeitos dos fármacos , Dieta/veterinária , Trato Gastrointestinal/metabolismo , Salmonella typhimurium/imunologia , Suínos/imunologia , Suínos/microbiologia
9.
J Anim Sci ; 97(10): 4140-4151, 2019 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-31310662

RESUMO

This study was conducted to investigate the effects of Clostridium butyricum and Enterococcus faecalis on growth performance, immune function, inflammation-related pathways, and microflora community in weaned piglets challenged with lipopolysaccharide (LPS). One hundred and eighty 28-d-old weaned piglets were randomly divided into 3 treatments groups: piglets fed with a basal diet (Con), piglets fed with a basal diet containing 6 × 109 CFU C. butyricum·kg-1 (CB), and piglets fed with a basal diet containing 2 × 1010 CFU E. faecali·kg-1 (EF). At the end of trial, 1 pig was randomly selected from for each pen (6 pigs per treatment group) and these 18 piglets were orally challenged with LPS 25 µg·kg-1 body weight. The result showed that piglets fed C. butyricum and E. faecalis had greater final BW compared with the control piglets (P < 0.05). The C. butyricum and E. faecalis fed piglets had lower levels of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), IL-1ß, tumor inflammatory factor-α (TNF-α), and had greater level of serum interferon-γ (IFN-γ) than control piglets at 1.5 and 3 h after injection with LPS (P < 0.05). Furthermore, piglets in the C. butyricum or E. faecalis treatment groups had a greater ratio of jejunal villus height to crypt depth (V/C) compared with control piglets after challenge with LPS for 3 h (P < 0.05). Compared with the control treatment, the CB and EF treatments significantly decreased the expression of inflammation-related pathway factors (TLR4, MyD88, and NF-κB) after challenge with LPS for 3 h (P < 0.05). High-throughput sequencing revealed that C. butyricum and E. faecalis modulated bacterial diversity in the colon. The species richness and alpha diversity (Shannon) of bacterial samples in CB or EF piglets challenged with LPS were higher than those in LPS-challenged control piglets. Furthermore, the relative abundance of Bacteroidales-Rikenellanceae in the CB group was higher than that in the control group (P < 0.05), whereas EF piglets had a higher relative abundance of Lactobacillus amylovorus and Lactobacillus gasseri (P < 0.05). In conclusion, dietary supplementation with C. butyricum or E. faecalis promoted growth performance, improved immunity, relieved intestinal villus damage and inflammation, and optimized the intestinal flora in LPS-challenged weaned piglets.


Assuntos
Ração Animal/análise , Clostridium butyricum/fisiologia , Enterococcus faecalis/fisiologia , Microbioma Gastrointestinal , Probióticos/análise , Suínos/fisiologia , Animais , Dieta/veterinária , Inflamação/veterinária , Mucosa Intestinal/microbiologia , Intestinos/microbiologia , Lactobacillus/crescimento & desenvolvimento , Lipopolissacarídeos/administração & dosagem , Distribuição Aleatória , Suínos/crescimento & desenvolvimento , Suínos/imunologia , Suínos/microbiologia
10.
J Anim Sci ; 97(8): 3426-3439, 2019 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-31233597

RESUMO

This study was conducted to investigate the effects of Clostridium butyricum addition to diets in late gestation and lactation on the reproductive performance and gut microbiota for sows. A total of 180 healthy Landrace × Yorkshire sows at 90 d of gestation were randomly assigned to one of four groups, with 45 replicates per group, receiving a basal commercial diet (Control, 0% C. butyricum) or diet added with 0.1% C. butyricum (1 × 108 CFU/kg of feed), 0.2% C. butyricum (2 × 108 CFU/kg of feed), 0.4% C. butyricum (4 × 108 CFU/kg of feed), respectively. The experiment was conducted from 90 d of gestation to weaning at 21 d of lactation. The results showed that the interval between piglet born was linearly (P < 0.05) decreased, and the duration of farrowing was significantly (quadratic, P < 0.05) shortened as C. butyricum addition increased. There was a linear (P < 0.05) increase in litter weight at weaning and litter weight gain. The concentrations of IgG and IgM in colostrum, and IgM in milk were linearly increased (P < 0.05) as C. butyricum addition. Serum MDA concentrations of sows at parturition and 14 d in lactation, and piglets at 14 and 21 d of age were linearly (P < 0.05) decreased, respectively. The serum total antioxidant capacity concentrations of sows at parturition and 14 and 21 d in lactation, and piglets at 14 and 21 d of age were linearly (P < 0.05) increased as C. butyricum addition, respectively. There was a linear decrease in the serum endotoxin concentration of sows on 21 d in lactation (P < 0.05). The serum cortisol concentrations of piglets at 14 and 21 d of age were both significantly (quadratic, P < 0.05) decreased. The 0.2% C. butyricum increased the relative abundance of Bacteroidetes (P = 0.016) at phylum level, Prevotellaceae_NK3B31_group, Prevotella_1, Prevotellaceae_UCG-003, Prevotella_9, Alloprevotella (P < 0.05) at genus level, and decreased the relative abundance of Proteobacteria, Gemmatimonadetes, Actinobacteria (P < 0.001) at phylum level, and Clostridium_sensu_stricto_1, Streptococcus, Escheruchia-Shigella, Sphingomonas, Succinivibrio (P < 0.05) at genus level and Firmicutes/Bacteroidetes ratio (P = 0.020). In conclusion, the present research indicated that dietary addition with C. butyricum could shorten the duration of farrowing and enhance the growth performance of suckling piglets. Moreover, 0.2% C. butyricum administration to sows changed the composition of intestinal microbiota, especially increased the relative abundance of Prevotella.


Assuntos
Ração Animal/análise , Clostridium butyricum/fisiologia , Microbioma Gastrointestinal , Reprodução , Suínos/microbiologia , Animais , Antioxidantes/análise , Colostro/imunologia , Dieta/veterinária , Feminino , Imunoglobulina G/imunologia , Imunoglobulina M/imunologia , Intestinos/microbiologia , Lactação , Leite/imunologia , Parto , Gravidez , Suínos/imunologia , Desmame , Ganho de Peso/efeitos dos fármacos
11.
J Anim Sci ; 97(9): 3795-3808, 2019 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-31231776

RESUMO

Postnatal growth retardation (PGR) is common in piglets. Abnormal development in small intestine was casually implicated in impaired growth, but the exact mechanism is still implausible. The present study unveiled transcriptome profile of jejunal mucosa, the major site of nutrient absorption, in PGR and healthy piglets using RNA-sequencing (RNA-seq). The middle segments of jejunum and ileum, and jejunal mucosa were obtained from healthy and PGR piglets at 42 d of age. Total RNA samples extracted from jejunal mucosa of healthy and PGR piglets were submitted for RNA-seq. Lower villus height was observed in both jejunum and ileum from PGR piglets suggesting structural impairment in small intestine (P < 0.05). RNA-seq libraries were constructed and sequenced, and produced average 4.8 × 107 clean reads. Analysis revealed a total of 499 differently expressed genes (DEGs), of which 320 DEGs were downregulated in PGR piglets as compared to healthy piglets. The functional annotation based on Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) highlighted that most DEGs were involved in nutrient metabolism and immune responses. Our results further indicated decreased gene expression associated with glucose, lipid, protein, mineral, and vitamin metabolic process, detoxication ability, oxidoreductase activity, and mucosal barrier function; as well as the increased insulin resistance and inflammatory response in the jejunal mucosa of PGR piglets. These results characterized the transcriptomic profile of the jejunal mucosa in PGR piglets, and could provide valuable information with respect to better understanding the nutrition metabolism and immune responses in the small intestine of piglets.


Assuntos
Regulação da Expressão Gênica/genética , Transtornos do Crescimento/veterinária , Suínos/genética , Transcriptoma , Animais , Perfilação da Expressão Gênica/veterinária , Ontologia Genética , Mucosa Intestinal/metabolismo , Intestino Delgado/metabolismo , Masculino , Estado Nutricional , Análise de Sequência de RNA/veterinária , Suínos/imunologia , Suínos/fisiologia
12.
J Anim Sci ; 97(9): 3871-3881, 2019 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-31251357

RESUMO

Glutathione (GSH) is the major intracellular thiol that plays a role in numerous detoxification, bio-reduction, and conjugation reactions. The availability of Cys is thought to be the rate-limiting factor for the synthesis of GSH. The effects of immune system stimulation (ISS) on GSH levels and the GSH synthesis rate in various tissues, as well as the plasma flux of Cys, were measured in starter pigs fed a sulfur AA (SAA; Met + Cys) limiting diet. Ten feed-restricted gilts with initial body weight (BW) of 7.0 ± 0.12 kg were injected i.m. twice at 48-h intervals with either sterile saline (n = 4; ISS-) or increasing amounts of Escherichia coli lipopolysaccharide (n = 6; ISS+). The day after the second injection, pigs received a primed constant infusion of 35S-Cys (9,300 kBq/pig/h) for 5 h via a jugular catheter. Blood and tissue free Cys and reduced GSH were isolated and quantified as the monobromobimane derivatives by HPLC. The rate of GSH synthesis was determined by measurement of the specific radioactivity of GSH and tissue free Cys at the end of the infusion period. Plasma Cys and total SAA levels were reduced (16% and 21%, respectively), but plasma Cys flux was increased (26%) by ISS (P < 0.05). Immune system stimulation increased GSH levels in the plasma (48%; P < 0.05), but had no effect on GSH levels in the liver, small and large intestines, heart, muscle, spleen, kidney, lung, and erythrocytes. The fractional synthesis rate (FSR) of GSH was higher (P < 0.05) in the liver (34%), small intestine (78%), large intestine (72%), heart (129%), muscle (37%), and erythrocytes (47%) of ISS+ pigs compared to ISS- pigs. The FSR of GSH tended (P = 0.08) to be higher in the lungs (45%) of ISS+ pigs than in ISS- pigs. The absolute rate of GSH synthesis was increased by ISS (mmol/kg wet tissue/d ± SE, ISS- vs. ISS+; P < 0.05) in the liver (5.22 ± 0.22 vs. 7.20 ± 0.59), small intestine (2.54 ± 0.25 vs. 4.52 ± 0.56), large intestine (0.61 ± 0.06 vs. 1.06 ± 0.16), heart (0.21 ± 0.03 vs. 0.48 ± 0.08), lungs (1.50 ± 0.10 vs. 2.90 ± 0.21), and muscle (0.21 ± 0.03 vs. 0.34 ± 0.04), but it remained unchanged in erythrocytes, the kidney, and the spleen (P > 0.80). The current findings suggest that GSH synthesis is increased during ISS, contributing to enhanced maintenance sulfur amino acid requirements in starter pigs during ISS.


Assuntos
Ração Animal/análise , Cisteína/sangue , Glutationa/biossíntese , Sistema Imunitário , Suínos/fisiologia , Animais , Dieta/veterinária , Escherichia coli/química , Feminino , Glutationa/análise , Lipopolissacarídeos/imunologia , Suínos/imunologia
13.
J Appl Microbiol ; 127(3): 856-866, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31161702

RESUMO

AIMS: We developed a strategy for localized delivery of the LFCA (lactoferricinlactoferrampin), which is actively synthesized in situ by Lactococcus lactis (pAMJ399-LFCA/LLMG1363), and explored the possibility of using pAMJ399-LFCA/LLMG1363 as an alternative additive diet to antibiotics. METHODS AND RESULTS: The antimicrobial activities of the LFCA derived from pAMJ399-LFCA/LLMG1363 were tested in vitro. The results showed that LFCA had an inhibitory effect on Staphylococcus aureus, Escherichia coli and Salmonella enteritidis. Then, the pAMJ399-LFCA/LLMG1363 was used as an additive diet for piglets. Our data demonstrated that oral administration of pAMJ399-LFCA/LLMG1363 significantly improved the average daily gain, feed-to-gain ratio, intestinal mucosal integrity and decreased the serum endotoxin and d-lactic acid levels. The mRNA expression levels of intestinal tight junction proteins (including occludin, Claudin-1 and ZO-1) were significantly upregulated by pAMJ399-LFCA/LLMG1363 administration. The serum immunoglobulin G (IgG) levels, intestinal secretory immunoglobulin A (sIgA) levels, IL-2, IL-10 and TGF-ß levels were significantly increased by pAMJ399-LFCA/LLMG1363. Furthermore, our data revealed that oral administration of pAMJ399-LFCA/LLMG1363 significantly increased the number of general Lactobacillus, and decreased the total viable E. coli counts in the ileum and cecum contents. CONCLUSIONS: We developed a novel pAMJ399-LFCA/LLMG1363 secreting LFCA, which had probiotic effects on the growth performance, intestinal morphology, intestinal barrier function and immunological indices of weaned piglets. SIGNIFICANCE AND IMPACT OF THE STUDY: pAMJ399-LFCA/LLMG1363, with probiotic effects on the health of weaned piglets, may be a promising feed additive for weaned piglets.


Assuntos
Antibacterianos/farmacologia , Lactococcus lactis/metabolismo , Lactoferrina/farmacologia , Fragmentos de Peptídeos/farmacologia , Suínos/imunologia , Animais , Bovinos , Escherichia coli/efeitos dos fármacos , Interleucina-10/metabolismo , Interleucina-2/metabolismo , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/metabolismo , Intestinos/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Probióticos/farmacologia , Salmonella enteritidis/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos , Fator de Crescimento Transformador beta/metabolismo
14.
J Sci Food Agric ; 99(13): 6076-6083, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31233219

RESUMO

BACKGROUND: This study was conducted to evaluate the health benefits to weaning pigs, raised under low sanitary conditions, of dietary supplementation with a multi-strain yeast fraction product (Cyberlindnera jadinii and Saccharomyces cerevisiae). In total, 160 weaning pigs (7.21 ± 1.05 kg) were randomly allotted to two dietary treatments in a 6-week feeding trial. The dietary treatments included a corn-soybean meal-based basal diet (CON) and CON + 2 g kg-1 multi-strain yeast fraction product (MsYF) during weeks 1-2 and 0.4 g kg-1 MsYF during weeks 3-6. RESULTS: The MsYF supplementation increased (P < 0.05) body weight (BW) at day 42 and average daily gain (ADG) during days 1-14 and days 1-42 (P < 0.05) compared to CON. The total tract digestibility of dry matter (DM), fecal Lactobacillus counts, and serum immunoglobulin G (IgG) concentration at day 42 were higher (P < 0.05) in pigs fed a MsYF supplemented diet. The concentration of serum haptoglobin in pigs receiving a MsYF-supplemented diet was higher (P < 0.05) at days 7, 14, and 42 than those receiving CON. The mRNA expression for INF-γ and TNF-α genes were lower (P < 0.05) at days 14 and 7 respectively and the expression of IL-6 and TLR-2 genes was lower (P < 0.01) at days 7 and 14 in pigs fed an MsFY supplemented diet than those fed CON. CONCLUSION: Supplementation with a multi-strain yeast fraction product had a positive effect on ADG during the early post-weaning period and led to better health in weaning pigs. © 2019 Society of Chemical Industry.


Assuntos
Saccharomyces cerevisiae/química , Suínos/crescimento & desenvolvimento , Leveduras/química , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Animais , Suplementos Nutricionais/análise , Fezes/microbiologia , Microbioma Gastrointestinal , Trato Gastrointestinal/metabolismo , Trato Gastrointestinal/microbiologia , Higiene , Interferon-alfa/genética , Interferon-alfa/imunologia , Interleucina-6/genética , Interleucina-6/imunologia , Suínos/genética , Suínos/imunologia , Suínos/microbiologia , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologia , Desmame
15.
J Anim Sci ; 97(7): 2940-2951, 2019 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-31081510

RESUMO

The present study used Escherichia coli lipopolysaccharide (LPS) to investigate whether maternal immune challenge during late gestation altered programming of the offspring hypothalamus and hypothalamic-pituitary-adrenal axis (HPAA). In addition, interactions of maternal diet, supplementation with fish oil (FO) or microalgae (AL), and complex vs. simple weaning diets were investigated. Briefly, Landrace × Yorkshire sows (N = 48) were randomly assigned to diets supplemented with FO, AL, or a standard gestation control diet (CON) from day 75 of gestation (gd 75) until parturition. On gd 112, half the sows from each dietary treatment were immune challenged with LPS (10 µg/kg BW) or saline as a control. At 21 d postpartum, the offspring were weaned, and half the animals from each maternal treatment were allocated to either a complex or simple weaning diet. At 28 d postpartum, the offspring's hourly fever and 2-h cortisol responses to LPS immune challenge (40 µg/kg BW) were measured to assess hypothalamus and HPAA function. Results indicated that the maternal temperature of sows on the FO diet returned to baseline levels faster than sows on the AL and CON diets after LPS immune challenge (P < 0.05). In contrast, there was no difference in the maternal cortisol response across the dietary treatments (P > 0.10). Regardless of the dietary treatments, the maternal LPS immune challenge induced a greater cortisol response in male offspring (P = 0.05) and a greater fever response in female offspring (P = 0.03) when they were LPS immune challenged post-weaning. Male offspring from LPS-immune-challenged sows fed the FO and AL diets had a greater fever response than male offspring from the maternal CON diet group (P ≤ 0.05). Last, no effect of the complex or simple weaning diets was observed for the nursery pig cortisol or fever responses to LPS immune challenge. In conclusion, LPS immune challenge during late pregnancy altered responsiveness of the offspring hypothalamus and HPAA to this same microbial stressor, and a sex-specific response was influenced by maternal dietary supplementation with FO and AL.


Assuntos
Suplementos Nutricionais , Óleos de Peixe/farmacologia , Microalgas , Suínos/fisiologia , Animais , Óleo de Milho/farmacologia , Dieta/veterinária , Escherichia coli/química , Feminino , Hidrocortisona/sangue , Sistema Hipotálamo-Hipofisário/efeitos dos fármacos , Sistema Hipotálamo-Hipofisário/metabolismo , Lipopolissacarídeos/administração & dosagem , Masculino , Projetos Piloto , Sistema Hipófise-Suprarrenal/efeitos dos fármacos , Sistema Hipófise-Suprarrenal/metabolismo , Gravidez , Distribuição Aleatória , Fatores Sexuais , Suínos/imunologia , Desmame
16.
Dev Comp Immunol ; 99: 103398, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31121186

RESUMO

Emerging evidence suggests that CD83, a dendritic cells (DCs) maturation marker in humans and mice, may prossess immunomodulatory capacities. Although porcine CD83 shares ∼75% sequence homology with its human counterpart, whether it functions as an immunoregulatory molecule remains unknown. To investigate porcine CD83 function, we deleted it in porcine DCs by RNA intereference. Results show that membrane-bound CD83 (mCD83) promotes DC-mediated T cell proliferation and cytokine production, thus confirming its immunoregulatory capacity. Intriguingly, porcine soluble CD83 (sCD83) treatment instead led to inhibition of DC-mediated T cell activation. Moreover, porcine sCD83 also inhibited differentiation of prepheral blood mononuclear cells (PBMCs) into DCs. These results collectively indicate that in addition to being a DC maturation maker, both membrane bound and souble porcine CD83 serve as immunoregulatory molecules with opposite effects on DC-mediated T cell activation and DC differentiation.


Assuntos
Antígenos CD/imunologia , Células Dendríticas/imunologia , Glicoproteínas/imunologia , Imunoglobulinas/imunologia , Glicoproteínas de Membrana/imunologia , Suínos/imunologia , Linfócitos T/imunologia , Animais , Antígenos CD/genética , Diferenciação Celular/imunologia , Células Cultivadas , Citocinas/metabolismo , Glicoproteínas/antagonistas & inibidores , Glicoproteínas/genética , Células HEK293 , Humanos , Imunoglobulinas/genética , Leucócitos Mononucleares/imunologia , Ativação Linfocitária , Glicoproteínas de Membrana/antagonistas & inibidores , Glicoproteínas de Membrana/genética , RNA Interferente Pequeno
17.
Vet Immunol Immunopathol ; 211: 19-24, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31084889

RESUMO

Porcine reproductive and respiratory syndrome (PRRS) is one of the most common diseases in the global swine industry. PRRSV infection is highly restricted to cells of the monocyte-macrophage lineage. However, the lack of antibodies to swine monocyte-macrophage lineage markers significantly hampers PRRSV research. In this study, we have developed a monoclonal antibody against the swine leukocyte antigen (SLA)-DRα chain and confirmed its reactivity with endogenous expressed SLA-DR in a variety of cell lines and primary swine antigen-presenting cells (PAMs, PBMC and BM-DCs). Moreover, the level of SLA-DR expression after PRRSV infection were evaluated by our homemade Mab and a commercial anti-SLA-DR antibody. Based on our result, the protein level of SLA-DRα expression is increased after PRRSV infection in DC, while the mRNA of both SLA-DRα and SLA-DRß were significantly inhibited by PRRSV replication. In conclusion, we successfully developed a MAb reactive with endogenous SLA-DR in western blotting, and this MAb could be a useful tool for further research and analysis. Moreover, the inconsistency of SLA-DR expression between protein and mRNA levels may suggest a novel role of DC played during the immune response after PRRSV infection.


Assuntos
Anticorpos Monoclonais/imunologia , Células Dendríticas/metabolismo , Cadeias alfa de HLA-DR/imunologia , Síndrome Respiratória e Reprodutiva Suína/imunologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/imunologia , Animais , Western Blotting , Medula Óssea/imunologia , Medula Óssea/metabolismo , Linhagem Celular , Células Dendríticas/imunologia , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , Imunofluorescência , Células HEK293 , Cadeias alfa de HLA-DR/metabolismo , Humanos , Imunoprecipitação , Camundongos , Camundongos Endogâmicos BALB C/imunologia , Proteínas Recombinantes , Suínos/imunologia
18.
Vet Immunol Immunopathol ; 211: 35-37, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31084891

RESUMO

In mammals, the immune system is undergoing significant changes during development, which has many impacts on the individual's capacity to cope with infectious diseases or other pathologic conditions, where the immune system is involved. Especially in livestock, it is important to know in detail about these changes, including shifts in the composition of systemic leukocyte populations, as this knowledge may help to focus on relevant cell populations when developing novel vaccines for use in juvenile versus adult animals. In this mini-review, a synoptic comparison of published PBMC populations, which were analysed in healthy weaned piglets as well as multiparous non-gestating sows, shows remarkable shifts within leukocyte populations. γδ T cells increase by factor 1.5, plasmacytoid dendritic cells and T helper cells more than double, and cytotoxic T cells as well as regulatory T cells increase more than four fold, whereas NK cells as well as B cells in adult sows comprise only 40% and monocytes 70% of the relative population sizes in weaned piglets. In summary, these insights into age-dependent shifts of porcine leukocyte populations indicate a principal increase of acquired immunity-associated leukocyte populations, whereas primarily innate immunity-associated cell types (NK cells, monocytes) are diminished.


Assuntos
Envelhecimento/imunologia , Leucócitos Mononucleares/imunologia , Suínos/imunologia , Animais , Leucócitos Mononucleares/fisiologia , Suínos/sangue , Suínos/fisiologia
19.
Vet Immunol Immunopathol ; 211: 6-9, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31084896

RESUMO

Helminth parasites are highly prevalent in swine production, causing chronic infections and considerable morbidity due to growth retardation. Moreover, helminths actively modulate host immune responses to other pathogens and/or vaccines. Here, we investigated the modulatory effects of Ascaris suum adult body fluid (ABF) and Trichuris suis Soluble Products (TsSP) on the cytokine response in porcine peripheral blood mononuclear cells (PBMCs) and the intestinal epithelial cell line IPEC-J2. In PBMCs, TsSP induced the secretion of IL-6, IL-10 and IL-1ß, but not TNF-α. Moreover, TsSP significantly enhanced the production of bacterial lipopolysaccharide (LPS)-induced IL-6 and IL-10 but suppressed the production of LPS-induced TNF-α. ABF did not induce cytokine secretion from PBMC, but suppressed LPS-induced secretion of TNF-α and IL-6. ABF did not have any effect on cytokine production in IPEC-J2 cells. In contrast, TsSP selectively induced the secretion of IL-6, and enhanced the IL-6 response induced by LPS. The IL-6 response appeared to be a conserved response to T. suis products, as significant secretion was also observed in alveolar macrophages. Thus, T. suis products have diverse modulatory effects on cytokine secretion in vitro, with IL-6 production a consistent feature of the innate host response.


Assuntos
Antígenos de Helmintos/imunologia , Ascaris suum/imunologia , Citocinas/metabolismo , Células Epiteliais/imunologia , Leucócitos Mononucleares/imunologia , Doenças dos Suínos/parasitologia , Trichuris/imunologia , Animais , Ascaríase/imunologia , Ascaríase/parasitologia , Ascaríase/veterinária , Citocinas/imunologia , Células Epiteliais/metabolismo , Células Epiteliais/parasitologia , Feminino , Interleucina-10/metabolismo , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Leucócitos Mononucleares/metabolismo , Leucócitos Mononucleares/parasitologia , Masculino , Suínos/imunologia , Suínos/parasitologia , Doenças dos Suínos/imunologia , Tricuríase/imunologia , Tricuríase/parasitologia , Tricuríase/veterinária
20.
Dev Comp Immunol ; 99: 103396, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31125574

RESUMO

Porcine thymus contains three independent populations of cells that have rearranged immunoglobulin heavy chain VDJH genes. The first population can be found exclusively in medulla and it consists of existing mature B cells and plasma cells. The second consists of developing B cells characterized by the presence of selected VDJH rearrangement, similar to B cell lymphogenesis in the bone marrow. The third population is entirely unaffected by selection mechanism for productive VDJH rearrangement and represents T lineage cells that rearrange immunoglobulin genes. Transcription of unselected VDJH repertoire is not allowed in T cells. Sequence analysis of unselected VDJH repertoire from T cells also revealed important consequences for B cell lymphogenesis and selection of B cell repertoire. As far as we know, this is the first evidence that some species completely rearrange VDJH genes in T cells. Our results also support the finding that B cells actively develop in the thymus.


Assuntos
Genes de Cadeia Pesada de Imunoglobulina/genética , Subpopulações de Linfócitos/imunologia , Suínos/imunologia , Linfócitos T/imunologia , Animais , Linfócitos B/imunologia , Feto/imunologia , Humanos , Especificidade da Espécie , Suínos/genética , Suínos/crescimento & desenvolvimento , Timo/crescimento & desenvolvimento , Timo/imunologia , Recombinação V(D)J/genética
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