The Intestinal Eukaryotic Virome in Healthy and Diarrhoeic Neonatal Piglets

Neonatal porcine diarrhoea of uncertain aetiology has been reported from a number of European countries. The aim of the present study was to use viral metagenomics to examine a potential viral involvement in this diarrhoea and to describe the intestinal virome with focus on eukaryotic viruses. Samples from the distal jejunum of 50 diarrhoeic and 19 healthy piglets from 10 affected herds were analysed. The viral fraction of the samples was isolated and nucleic acids (RNA and DNA fractions) were subjected to sequence independent amplification. Samples from diarrhoeic piglets from the same herds were pooled whereas samples from healthy piglets were analysed individually. In total, 29 clinical samples, plus two negative controls and one positive control consisting of a mock metagenome were sequenced using the Ion Torrent platform. The resulting sequence data was subjected to taxonomic classification using Kraken, Diamond and HMMER. In the healthy specimens, eight different mammalian virus families were detected (Adenoviridae, Anelloviridae, Astroviridae, Caliciviridae, Circoviridae, Parvoviridae, Picornaviridae, and Reoviridae) compared to four in the pooled diarrhoeic samples (Anelloviridae, Circoviridae, Picornaviridae, and Reoviridae). It was not possible to associate a particular virus family with the investigated diarrhoea. In conclusion, this study does not support the hypothesis that the investigated diarrhoea was caused by known mammalian viruses. The results do, however, indicate that known mammalian viruses were present in the intestine as early as 24–48 hours after birth, indicating immediate infection post-partum or possibly transplacental infection.     

Probiotic Lactobacillus johnsonii BS15 Promotes Growth Performance,Intestinal Immunity,and Gut Microbiota in Piglets

Probiotics and Antimicrobial Proteins - Numerous studies have investigated the beneficial effects of Lactobacillus johnsonii strain BS15 on mice and broilers. This study aimed to understand the...     

酶解乳蛋白对初生仔猪小肠组织学结构及隐窝细胞增殖的影响(英文)

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Effects of Recombinant Human Lactoferrin on Osteoblast Growth and Bone Status in Piglets

Lactoferrin (LF), an ~80?kDa iron-binding glycoprotein, modulates many biological effects, including antimicrobial and immunomodulatory activities. Recently, it was shown that LF also regulates bone cell activity, suggesting its therapeutic effect on postmenopausal bone loss. However, a minimal amount is known regarding the effects of recombinant human LF (rhLF) supplementation on bone status in young healthy infants. We found osteoblast cell differentiation was significantly promoted in vitro. Furthermore, treatment of human osteoblast cells with rhLF rapidly induced phosphorylation of p44/p42 mitogen-activated protein kinase (p44/p42 MAPK, ERK1/2). In order to investigate the effects of rhLF on bone status in vivo, we used a piglet model, which is a useful model for human infants. Piglets were supplemented with rhLF milk for 30 days. Bone formation markers, Serum calcium concentration, bone mineral density (BMD), bone mineral content (BMC), tibia bone strength, and the overall metabolite profile analysis showed that rhLF was advantageous to the bone growth in piglets. These findings suggest that rhLF supplementation benefits neonate bone health by modulating bone formation.     

Roles of Probiotic Lactobacilli Inclusion in Helping Piglets Establish Healthy Intestinal Inter-environment for Pathogen Defense

The gastrointestinal tract of pigs is densely populated with microorganisms that closely interact with the host and with ingested feed. Gut microbiota benefits the host by providing nutrients from dietary substrates and modulating the development and function of the digestive and immune systems. An optimized gastrointestinal microbiome is crucial for pigs’ health, and establishment of the microbiome in piglets is especially important for growth and disease resistance. However, the microbiome in the gastrointestinal tract of piglets is immature and easily influenced by the environment. Supplementing the microbiome of piglets with probiotic bacteria such as Lactobacillus could help create an optimized microbiome by improving the abundance and number of lactobacilli and other indigenous probiotic bacteria. Dominant indigenous probiotic bacteria could improve piglets’ growth and immunity through certain cascade signal transduction pathways. The piglet body provides a permissive habitat and nutrients for bacterial colonization and growth. In return, probiotic bacteria produce prebiotics such as short-chain fatty acids and bacteriocins that benefit piglets by enhancing their growth and reducing their risk of enteric infection by pathogens. A comprehensive understanding of the interactions between piglets and members of their gut microbiota will help develop new dietary interventions that can enhance piglets’ growth, protect piglets from enteric diseases caused by pathogenic bacteria, and maximize host feed utilization.     

不同来源乳铁蛋白对幼鼠肠道发育的影响

目的 探究哺乳期乳铁蛋白（lactoferrin，LF）的缺失及不同来源LF补充后对幼鼠肠道发育的影响。方法 以LF基因敲除型雌鼠作为哺乳母鼠造成幼鼠哺乳期无LF的摄入，且从幼鼠出生第3~21天每日人工饲喂100 mg/kg 牛血清白蛋白（BSA）、牛源乳铁蛋白（bovine Lactoferrin，bLF）及重组人源乳铁蛋白（recombinant human Lactoferrin，rhLF），于幼鼠21日龄取样，测定各组小鼠小肠发育指标。结果 在本实验周期下，哺乳期rhLF的补充显著性增加小鼠回肠绒毛长度/隐窝深度值（P<0.05），且上调回肠Occludin和ZO-1基因的表达（P<0.05），增加小鼠十二指肠、空肠和回肠麦芽糖酶酶活/乳糖酶酶活比值（P<0.05），表明哺乳期rhLF的补充能够增强小鼠肠道消化吸收能力和肠屏障功能；哺乳期bLF的补充显著增加小鼠十二指肠及回肠麦芽糖酶活性/乳糖酶活性比值（P<0.05）。结论 对于哺乳期无LF摄入的乳鼠来说，哺乳期间LF的补充能够增强乳鼠肠道对营养物质的消化吸收能力、促进肠道的发育成熟、增强肠道屏障功能，并且，本实验中rhLF表现出比bLF更加有效的作用。     

Effects of Copper-Loaded Chitosan Nanoparticles on Intestinal Microflora and Morphology in Weaned Piglets

The objective of the present study was to investigate the effects of dietary supplementation with copper-loaded chitosan nanoparticles (CNP-Cu) on growth performance, intestinal microflora, and morphology in weaned piglets. A number of 90 weaned piglets (Duroc × Landrace × Yorkshire), weaned at 21?days with body weight of 7.2?±?0.81?kg, were randomly divided into three groups by weight and sex, each treatment including three replicates of ten pigs. The piglets were fed the same basal diet supplemented with 0 (the control group), 100?mg/kg CNP-Cu, and 100?mg/kg chlortetracycline (the positive group). The results showed that 100?mg/kg CNP-Cu significantly increased average daily gain and feed intake and decreased feed/gain ratio and diarrhea rate (P?<?0.05). Compared with the control group, the amount of Escherichia coli in duodenum, jejunal, and caecum were significantly decreased by 100?mg/kg CNP-Cu; the number of lactobacillus in jejunal and caecum were increased (P?<?0.05), and the amount of bifidobacterium in duodenum and caecum were also increased (P?<?0.05). Moreover, the villous height of duodenum, jejunum, and ileum mucosa was significantly increased (P?<?0.05), and the crypt depth was significantly decreased (P?<?0.05). The results indicated that CNP-Cu is beneficial to growth and intestinal microflora and morphology and could be a potential substitution of chlortetracycline in diets of weaned piglets.     

Intestinal Microbiota Succession and Immunomodulatory Consequences after Introduction of Lactobacillus reuteri I5007 in Neonatal Piglets

Seventy-two, suckling piglets, obtained from 9 litters standardized to 8 piglets, were assigned to 1 of 3 treatments (n = 24) to compare short-term, early administration with intermittent, longer-term administration of Lactobacillus reuteri I5007. The treatments were a control (given a placebo of 0.1% peptone water from day 1 to 5) or treatments in which 1.7 × 10¹⁰ CFU L. reuteri was administrated either daily for 4 days starting on day 1 or every 4th day from day 1 to 17. Five piglets per treatment were killed at 3 time points (day 7, 14 and 21). Denaturing Gradient Electrophoresis of ileal digesta revealed an increase in the presence of L. reuteri I5007 and Clostridium lentocellum (on day 14 and 21) in the every 4th-day treatment and Actinobacillus porcinus (on day 7 and 14) in both L. reuteri treatments, while reducing the abundance of E. coli on day 21 in the every 4th-day treatment. Real-time qPCR of ileal digesta showed an increase in Bifidobacterium spp. on day 14 for both L. reuteri I5007 treatments. An increase in the concentration of lactic acid and a lower pH was observed in the first 4-day treatment on day 7 and the every 4^th day treatment on day 14. The relative abundance of mRNA for TGF-β was increased while that for IFN-γ was decreased in the mesenteric lymph nodes of piglets treated with L. reuteri every 4^th day. In conclusion, early intervention with L. reuteri increases the presence of beneficial bacteria and decreases the presence of undesirable microbes in the lower gastrointestinal tract. The changes appear to be mediated by altering the intestinal pH through lactic acid production resulting in favorable bacterial species colonization. A prolonged duration of treatment (i.e. every 4^th day) would appear to be superior to treatment only during the first 4 days.     

Diversity of Intestinal Bifidobacteria in Patients with Japanese Cedar Pollinosis and Possible Influence of Probiotic Intervention

This study was conducted to evaluate the potential association between intestinal bifidobacteria and Japanese cedar pollinosis (JCPsis) and possible influences of probiotic intervention. In this study, fecal samples were the collected from 29 JCPsis patients. The qualitative and quantitative analyses of fecal bifidobacteria were conducted by quantitative real-time PCR with 16S rRNA-gene-targeted species-specific primers before cedar pollen spread and after a 10-week intervention with fermented milk prepared with Lactobacillus GG and L. gasseri TMC0356 during pollen spread. Each JCPsis patient had a unique diversity of bifidobacteria, which varied qualitatively and quantitatively in an individual-dependent manner during pollen spread. The serum IgE concentration of JCPsis patients with more than 3 detectable Bifidobacterium species was significantly lower than that of patients with less than 2 detected species. The prevalence of B. adolescentis, B. longum, and B. catenulatum increased after probiotic intervention, although the changes were not statistically significant. These results suggest that lower diversity of intestinal Bifidobacterium species might be a pathological aspect of JCPsis. The diversity of intestinal bifidobacteria could be a prospective target for using probiotics in the management of IgE-mediated allergic disorders including JCPsis.     

Effects of Weaning on Intestinal Upper Villus Epithelial Cells of Piglets

The intestinal upper villus epithelial cells represent the differentiated epithelial cells and play key role in digesting and absorbing lumenal nutrients. Weaning stress commonly results in a decrease in villus height and intestinal dysfunction in piglets. However, no study have been conducted to test the effects of weaning on the physiology and functions of upper villus epithelial cells. A total of 40 piglets from 8 litters were weaned at 14 days of age and one piglet from each litter was killed at 0 d (w0d), 1 d (w1d), 3 d (w3d), 5 d (w5d), and 7 d (w7d) after weaning, respectively. The upper villus epithelial cells in mid-jejunum were isolated using the distended intestinal sac method. The expression of proteins in upper villus epithelial cells was analyzed using the isobaric tags for relative and absolute quantification or Western blotting. The expression of proteins involved in energy metabolism, Golgi vesicle transport, protein amino acid glycosylation, secretion by cell, transmembrane transport, ion transport, nucleotide catabolic process, translational initiation, and epithelial cell differentiation and apoptosis, was mainly reduced during the post-weaning period, and these processes may be regulated by mTOR signaling pathway. These results indicated that weaning inhibited various cellular processes in jejunal upper villus epithelial cells, and provided potential new directions for exploring the effects of weaning on the functions of intestine and improving intestinal functions in weaning piglets.     

菊粉和刺五加提取物对早期断奶仔猪生长性能、血液指标和肠道形态的影响

试验选用120头21日龄5.64kg左右的三元杂交断奶仔猪,分别饲喂基础日粮(对照组)、基础日粮+抗生素(抗生素组)和基础日粮+5%菊粉+0.1%刺五加提取物(实验组),试验期为14d,观察日粮中复合添加菊粉和刺五加(AS)提取物对早期断奶仔猪生长性能、血液生化指标和肠道形态的影响。试验结果表明,实验组与对照组相比,仔猪日增重(ADG)显著提高(P0.05),而料肉比(F/G)则显著降低(P0.05),但平均日采食量(ADFI)差异不显著(P0.05);实验组仔猪血清中血糖(GLU)、低密度脂蛋白(LDL)、乳酸脱氢酶(LDH)和尿素氮(BUN)与对照组差异不显著(P0.05),但试验第14d,碱性磷酸酶(ALP)、超氧化物歧化酶(SOD)和还原型谷胱甘肽(GSH)浓度均显著高于对照组(P0.05);实验组仔猪十二指肠和回肠的隐窝深度显著高于对照组(P0.05),空肠和回肠的绒毛高度显著高于对照组(P0.05)。试验显示,日粮中添加菊粉和刺五加提取物对提高早期断奶仔猪的生长性能、改善小肠消化和吸收功能有重要作用。     

益生菌干预治疗对非小细胞肺癌化疗患者肠道菌群、免疫功能及相关并发症的影响

目的:研究观察益生菌干预治疗对非小细胞肺癌化疗患者肠道菌群、免疫指标及相关并发症的影响。方法:选取自2016年1月-2018年12月赤峰学院附属医院收诊的80例非小细胞肺癌(NSCLC)需要化疗患者作为观察对象,将其随机分为益生菌给药组及安慰剂对照组,每组各40例。测量两组患者化疗前后体格指标,观察患者化疗相关并发症的发生情况,定量检测治疗前后双歧杆菌属、乳酸杆菌属、大肠杆菌和肠球菌属,血清CD3^+、CD4^+、CD8^+T细胞水平,粪便悬浮液中的粪便SIg A含量的变化。结果:化疗后,益生菌给药组患者的BMI、WHR均显著高于安慰剂对照组(P<0.05);益生菌给药组的双歧杆菌、乳酸杆菌含量及粪便SIg A含量与安慰剂对照组相比显著增加,肠球菌、大肠杆菌的含量显著减少(P<0.05);化疗后,益生菌给药组CD3^+、CD4^+T细胞比例显著增加(P<0.05),CD8^+T细胞比例显著下降(P<0.05),安慰剂对照组无明显改善(P>0.05);益生菌给药组的CD4^+/CD8^+比例与安慰剂对照组相比较显著升高(P<0.05)。结论:益生菌干预能调节NSCLC化疗患者的肠道菌群失衡,提高其免疫功能,并减少患者化疗期间相关并发症。     

Studies on Immunoglobulins and Trypsin Inhibitor in Colostrum and Milk from Sows and in Serum of Their Piglets

The concentrations of IgG, IgM, IgA and the specific sow colostrum trypsin inhibitor (SCTI) were measured by radial immunodiffusion in colostrum and milk samples from sows and in serum samples from their offspring during the suckling period. A clear time dependence was found for all the measured variates in both whey and serum. Statistically significant positive correlations were found between, on the one hand, concentrations of IgG and IgA, but not IgM, in sera from 39 suckling piglets 1 and 3 days old, and, on the other hand, concentrations of the same immunoglobulins and of the trypsin inhibitor in maternal colostrum (n = 7). Multiple regression analyses showed that at day 1 and day 3 the levels of both IgG and IgA in serum samples from the suckling piglets were positively influenced by both the SCTI and the IgG or IgA contents in maternal colostrum.     

Intestinal Integrity and Akkermansia muciniphila, a Mucin-Degrading Member of the Intestinal Microbiota Present in Infants, Adults, and the Elderly

Fluorescence in situ hybridization and real-time PCR analysis targeting the 16S rRNA gene of Akkermansia muciniphila were performed to determine its presence in the human intestinal tract. These techniques revealed that an A. muciniphila-like bacterium is a common member of the human intestinal tract and that its colonization starts in early life and develops within a year to a level close to that observed in adults (10⁸ cells/g) but decreases (P < 0.05) in the elderly.     

Maternal and Neonatal Mortality in South-West Ethiopia: Estimates and Socio-Economic Inequality

Introduction

Ethiopia has achieved the fourth Millennium Development Goal by reducing under 5 mortality. Nevertheless, there are challenges in reducing maternal and neonatal mortality. The aim of this study was to estimate maternal and neonatal mortality and the socio-economic inequalities of these mortalities in rural south-west Ethiopia.

Methods

We visited and enumerated all households but collected data from those that reported pregnancy and birth outcomes in the last five years in 15 of the 30 rural kebeles in Bonke woreda, Gamo Gofa, south-west Ethiopia. The primary outcomes were maternal and neonatal mortality and a secondary outcome was the rate of institutional delivery.

Results

We found 11,762 births in 6572 households; 11,536 live and 226 stillbirths. There were 49 maternal deaths; yielding a maternal mortality ratio of 425 per 100,000 live births (95% CI:318–556). The poorest households had greater MMR compared to richest (550 vs 239 per 100,000 live births). However, the socio-economic factors examined did not have statistically significant association with maternal mortality. There were 308 neonatal deaths; resulting in a neonatal mortality ratio of 27 per 1000 live births (95% CI: 24–30). Neonatal mortality was greater in households in the poorest quartile compared to the richest; adjusted OR (AOR): 2.62 (95% CI: 1.65–4.15), headed by illiterates compared to better educated; AOR: 3.54 (95% CI: 1.11–11.30), far from road (≥6 km) compared to within 5 km; AOR: 2.40 (95% CI: 1.56–3.69), that had three or more births in five years compared to two or less; AOR: 3.22 (95% CI: 2.45–4.22). Households with maternal mortality had an increased risk of stillbirths; OR: 11.6 (95% CI: 6.00–22.7), and neonatal deaths; OR: 7.2 (95% CI: 3.6–14.3). Institutional delivery was only 3.7%.

Conclusion

High mortality with socio-economic inequality and low institutional delivery highlight the importance of strengthening obstetric interventions in rural south-west Ethiopia.     

Natural Pig Plasma Immunoglobulins Have Anti-Bacterial Effects: Potential for Use as Feed Supplement for Treatment of Intestinal Infections in Pigs

There is an increasing demand for non-antibiotics solutions to control infectious disease in intensive pig production. Here, one such alternative, namely pig antibodies purified from slaughterhouse blood was investigated in order to elucidate its potential usability to control post-weaning diarrhoea (PWD), which is one of the top indications for antibiotics usage in the pig production. A very cost-efficient and rapid one-step expanded bed adsorption (EBA) chromatography procedure was used to purify pig immunoglobulin G from slaughterhouse pig plasma (more than 100 litres), resulting in >85% pure pig IgG (ppIgG). The ppIgG thus comprised natural pig immunoglobulins and was subsequently shown to contain activity towards four pig-relevant bacterial strains (three different types of Escherichia coli and one type of Salmonella enterica) but not towards a fish pathogen (Yersinia ruckeri), and was demonstrated to inhibit the binding of the four pig relevant bacteria to a pig intestinal cell line (IPEC-J2). Finally it was demonstrated in an in vivo weaning piglet model for intestinal colonization with an E. coli F4+ challenge strain that ppIgG given in the feed significantly reduced shedding of the challenge strain, reduced the proportion of the bacterial family Enterobacteriaceae, increased the proportion of families Enterococcoceae and Streptococcaceae and generally increased ileal microbiota diversity. Conclusively, our data support the idea that natural IgG directly purified from pig plasma and given as a feed supplement can be used in modern swine production as an efficient and cost-effective means for reducing both occurrence of PWD and antibiotics usage and with a potential for the prevention and treatment of other intestinal infectious diseases even if the causative agent might not be known.     

Different Tissue Uptake after Administration of Proline and Hydroxyproline in Neonatal Piglets

Using whole body autoradiography and impulse counting technique, distribution of intramuscularly administered 14C-DL-hydroxyproline and 14C-L-proline in newborn piglets was compared. The main excretion route of hydroxy-proline was via the kidneys. Hydroxyproline or its metabolic products were excreted at quite another rate by the liver than was proline. Skeletal muscles and myocardium showed a lowered ratio of proline to hydroxyproline. A difference in favour of hydroxyproline in passing the blood-brain barrier was shown. Contrary to proline hydroxyproline yielded a low 14C-level in the skeletal tissues. The possibilities of an incomplete hydroxylation of proline as an aetiological factor in the origin of skeletal disorders are discussed.     

Effects of the Dietary Probiotic,Enterococcus faecium NCIMB11181, on the Intestinal Barrier and System Immune Status in Escherichia coli O78-Challenged Broiler Chickens

Probiotics and Antimicrobial Proteins - The effects of Enterococcus faecium on growth, intestinal barrier function, and immune response in Escherichia coli O78-challenged broiler chickens were...