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Use of recombinant porcine ?-defensin 2 as a medicated feed additive for weaned piglets.

ABSTRACT: Post-weaning diarrhoea (PWD) in piglets is associated with colonization of the intestine with bacterial pathogens. In this study, we evaluated the use of recombinant porcine ?-defensin 2 (rpBD2) as a medicated feed additive for weaned piglets. The crude extract from the culture supernatant of rpBD2-expressing Pichia pastoris was used as a medicated feed additive for weaned piglets. Dietary treatments included a positive control (basal diet + antibiotics, designated PC) and three different rpBD2 treatments without antibiotics (basal diet supplemented with 1, 5, or 15 g of crude rpBD2/kg basal diet, designated 1PD, 5PD, and 15PD, respectively). Of all the treatments, 5PD had the greatest impact on the weaned piglets. It increased their body weight, average daily weight gain, average daily feed intake, and intestinal villus height in the duodenum and jejunum, and reduced the incidence of PWD. The diversity of the cecal digesta and mucosa microflora was compared between the weaned piglets in the PC and 5PD groups. Piglets treated with 5PD had lower diversity indices and fewer bacterial pathogens in their cecal digesta and mucosa than the PC group. Our results demonstrate that crude rpBD2 could provide an alternative to the traditional antibiotic feed additives given to weaned piglets.

SUBMITTER: Peng Z

PROVIDER: S-EPMC4880912 | biostudies-literature | 2016 May

REPOSITORIES: biostudies-literature

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Use of recombinant porcine β-defensin 2 as a medicated feed additive for weaned piglets.

Peng Zixin Z Wang Anru A Xie Linqi L Song Weiping W Wang Jie J Yin Zhe Z Zhou Dongsheng D Li Fengqin F

Scientific reports 20160526

Post-weaning diarrhoea (PWD) in piglets is associated with colonization of the intestine with bacterial pathogens. In this study, we evaluated the use of recombinant porcine β-defensin 2 (rpBD2) as a medicated feed additive for weaned piglets. The crude extract from the culture supernatant of rpBD2-expressing Pichia pastoris was used as a medicated feed additive for weaned piglets. Dietary treatments included a positive control (basal diet + antibiotics, designated PC) and three different rpBD2 ...[more]

PMID: 27225034

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Project description:BackgroundEpidermal growth factor (EGF) is an important growth factor in regulation of cell proliferation, differentiation, survival and apoptosis. Studies showed that food-grade Lactococcus lactis (L. lactis) and NICE expression system have superior performance in exogenous protein expression. This study aimed to construct and express porcine EGF (pEGF), and use L. lactis as vehicle for producing and delivering pEGF. Furthermore, investigating biological activity of pEGF and exploring applications feasibility of combination effects of L. lactis and pEGF on early weaned piglets' production.ResultsA recombinant Lactococcus lactis which produced and secreted pEGF at 1000 ng/ml in culture supernatant was generated. Secreted pEGF was a fully biologically active protein, as demonstrated by its capacity to stimulate L929 mouse fibroblast cell line proliferation in vitro. For in vivo study, forty piglets were randomly allocated to control, antibiotic control, empty vector-expressing L. lactis (LL-EV) and pEGF-secreting L. lactis (LL-pEGF). After 14 d of rearing, final body weight and average daily gain in LL-pEGF were greater (P < 0.05, 8.95 vs. 8.37 kg, 206.1 vs. 157.7 g/day, respectively) than those in control, but no significant differences between LL-pEGF, LL-EV and antibiotic control. Overall period average daily feed intake was higher in LL-pEGF, LL-EV and antibiotic control than in control (P < 0.05, 252.9, 255.6, 250.0, 207.3 g/day, respectively). No significant difference was observed on ADFI/ADG. LL-pEGF increased villous height in the duodenum, jejunum and ileum than in control and LL-EV (P < 0.05). Sucrase in the 3 intestinal segments, aminopeptidase A in the duodenum and Jejunum, aminopeptidase N and dipeptidase IV in the duodenum in LL-pEGF were higher than those in control (P < 0.05). Furthermore, Escherichia coli and Enterococcus counts decreased in the ileum and Lactobacillus increased in the ileum and cecum digesta in LL-pEGF compare with the control (P < 0.05). Lactobacillus increased in the cecum in LL-EV compared with control and antibiotic control (P < 0.05).ConclusionWe have generated a recombinant Lactococcus lactis which produced and secreted fully biologically active porcine EGF. Oral administration of pEGF-secreting L. lactis had beneficial effects on intestinal health and performance of early-weaned piglets.

Project description:Zearalenone (ZEA) is widely present in feedstuffs and raw materials, causing reproductive disorders in animals. In this study, Bacillus licheniformis CK1 was used to detoxify ZEA in feed for alleviating its effect in Tibetan piglets. A total of 18 weaned female Tibetan piglets were randomly divided into 3 groups: control group (Control, ZEA-free basal diet); treatment group 1 (T1, ZEA-contaminated diet); and treatment group 2 (T2, ZEA-contaminated but pre-fermented by CK1 diet). There were no significant differences of average daily feed intake (ADFI), average daily gain (ADG), and feed efficiency (FE) among the 3 groups (P > 0.05). The T1 treatment significantly increased the vulva size and relative weight of the reproductive organ (P < 0.05), compared with the Control. However, the T2 treatment caused a significant reduction (P < 0.05) in vulva size and relative weight of the reproductive organ compared with the T1 group. The levels of luteinizing hormone (LH), follicle-stimulating hormone (FSH), progesterone (P), and estradiol (E2) in the T1 group were significantly lower (P < 0.05) than those in the Control, while the levels of LH, P, and E2 in the T2 group were significantly greater (P < 0.05) than those in the T1 group. Zearalenone significantly increased (P < 0.05) the expression of estrogen receptor ? in uterus and ovary and estrogen receptor ? in vagina, while these indicators were not significant different (P > 0.05) between the T2 group and the Control group. In comparison with the Control group, ZEA significantly increased (P < 0.05) expression of several ATP-binding cassette (ABC) transporters: ABCB1 and ABCb4 in the vagina, ABCA1 and ABCb4 in the uterus, and ABCB1, ABCb4, ABCD3, and ABCG2 in the ovary, while these transporters in the T2 group were significantly decreased (P < 0.05) compared with the T1 group. In conclusion, the present study demonstrates that B. licheniformis CK1 could alleviate the harmful effect of ZEA in Tibetan piglets.

Project description:Development of immune competence in pigs is important for resilience, disease resistance, and performance later in life. Dietary interventions in early life can contribute to immune system development. Use of model dietary interventions, such as diets with a high concentration of zinc, can contribute to the understanding of the interactions between the diet, the intestinal microbiota and intestinal tissues. The aim of the present study was to evaluate the effects of provision of a diet with a high concentration of zinc (Zn, 2690 mg/kg), as model intervention, as compared to provision of a control diet with a regular Zn concentration (100 mg/kg) during a period of nine days during the post weaning period (d 14-23 post weaning (pw)), on the composition of the intestinal microbiota and gene expression in the intestinal mucosa on d 23 and 35 pw (12 d after the termination of the Zn intervention). Whole genome gene expression analysis of the jejunal and ileal mucosa revealed a small overall time-treatment interaction effect at d 23 pw in both intestinal tissues. In both jejunal and ileal mucosa various genes/probes were differentially up or down regulated between treatments. In intestinal tissue samples obtained on d 35 no differences in gene expression were observed. Probes of d 23 pw that were differently expressed and had an annotation were subsequently used as input for further functional analysis using DAVID and Gene Decks databases. The analysis did not result in the identification of gene sets that were differentially expressed between dietary treatments. However, it was shown that a number of upregulated genes in the jejunal mucosa on d 23 pw by the high zinc intervention are involved in pathways related to mineral absorption, immune signalling and cell energy metabolism (glycolysis and gluconeogenesis). A few down regulated genes by the Zn intervention are also involved in immune signalling pathways. It was concluded from the present study that provision of a diet with a high concentration of zinc as zinc oxide during a short period of time (9 days) to piglets in the post weaning period (d 14-23 pw) induces differences on the intestinal expression of genes in part related to the functioning of the local innate immune system. The high dietary zinc intervention can therefore be considered as a suitable model for studying relationships between dietary interventions and development of immune competence in post weaning piglets.

Project description:Backgroundβ-defensins have attracted considerable research interest because of their roles in protecting hosts from various pathogens. This study was conducted to investigate the expression profiles of the porcine β-defensin 114 (PBD114) in different breeds and in response to infections. Moreover, the function of PBD114 protein was partially investigated.MethodsSix Tibetan pigs (TP) and six DLY (Duroc×Landrace×Yorkshire) pigs were slaughtered to explore the expression profiles of PBD114 in different breeds and tissues. For infection models, sixteen DLY pigs were divided into two groups and challenged either with sterile saline or E. coli K88. The recombinant protein PBD114 (rPBD114) was obtained by using a heterologous expression system in E. coli.ResultsPBD114 gene was highly expressed in tissues such as the intestine, liver, spleen, and thymus. Interestingly, the expression level of PBD114 gene was higher in the TP pigs than in the DLY pigs (P < 0.05), and was significantly elevated upon E. coli K88 challenge (P < 0.05). The nucleotide sequences of PBD114 from Tibetan and DLY pigs was identical, and both showed a 210-bp open reading frame encoding a 69-amino acid mature peptide. To explaore the function of PBD114 protein, PBD114 gene was successfully expressed in E. coli Origami B (DE3) and the molecular weight of the rPBD114 was estimated by SDS-PAGE to be 25 kDa. The rPBD114 was purified and mass spectrometry verified the protein as PBD114. Importantly, rPBD114 showed antimicrobial activities against E. coli DH5α and E. coli K88, and the minimal inhibitory concentrations (MICs) were 64 and 128 μg/mL, respectively. Hemolytic and cytotoxicity assays showed that rPBD114 did not affect cell viability under physiological concentrations.ConclusionsPBD114 is an infection response gene that is differentially-expressed between different porcine breeds and tissues. The antimicrobial activity of PBD114 protein, against pathogens such as the E. coli K88, suggested that it may serve as a candidate for the substitution of conventionally used antibiotics.

Project description:BackgroundPiglets at weaning suffer many stressors such as sudden change of feed, change in group composition and the end of lactogenic immunity. These stressors may cause poor growth performance. There is a need for alternatives to support piglets during the weaning period. Organic acids are known to have a positive effect on performance through reducing the pH and their antimicrobial action.Context & purposeThe purpose was to study the effect of the inclusion of a free and buffered organic acid blend in drinking water on performance of weaned pigs. Four-hundred and twenty pigs in a conventional herd were allocated after weaning to one of three treatments and monitored during 4 weeks: group (1) Full medication, group (2) organic acid blend + full medication, group (3) organic acid blend + reduced medication. Average daily gain, feed intake and water consumption was recorded at group level.ResultsDuring the overall study period live weight and average daily gain of the piglets was significantly higher (P <0.001) for treatment (3) compared to (1) and (2) (Table 1). Live weight was significantly higher for treatment (3) compared to (1) from week 2 of the study (Fig. 1). No significant differences were found for average daily feed intake. FCR for treatment (3) improved by 1.0 compared to treatment (1) in week 1 (P <0.05), while in week 2 and 3 no significant differences were found (Table 2). Overall, FCR was with 0.3 difference significantly lower (P = 0.001) for treatment (3) than for (1) and (2) (Table 1). Pigs receiving organic acids in drinking water had significantly (P <0.05) higher water consumption than group (1) in weeks 3 and 4 (Table 2).Conclusion & potential implicationsThe use of a blend of free and buffered organic acids together with a reduced medication program improves growth performance during the first month after weaning compared to a control with full medication and a combination between organic acids and full medication. This implies that organic acids could be used as a valid alternative for antibiotic reduction in post-weaning pigs. The treatment also increased the drinking water intake.

Dataset Information

Use of recombinant porcine ?-defensin 2 as a medicated feed additive for weaned piglets.

Publications

Use of recombinant porcine β-defensin 2 as a medicated feed additive for weaned piglets.

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