There is demand from women for alternatives to giving birth in a standard hospital setting however access to these services is limited. This systematic review examines the literature relating to the economic evaluations of birth setting for women at low risk of complications.Searches of the literature to identify economic evaluations of different birth settings of the following electronic databases: MEDLINE, CINAHL, EconLit, Business Source Complete and Maternity and Infant care. Relevant English language publications were chosen using keywords and MeSH terms between 1995 and 2015. Inclusion criteria included studies focussing on the comparison of birth setting. Data were extracted with respect to study design, perspective, PICO principles, and resource use and cost data.Eleven studies were included from Australia, Canada, the Netherlands, Norway, the USA, and the UK. Four studies compared costs between homebirth and the hospital setting and the remaining seven focussed on the cost of birth centre care and the hospital setting. Six studies used a cost-effectiveness analysis and the remaining five studies used cost analysis and cost comparison methods. Eight of the 11 studies found a cost saving in the alternative settings. Two found no difference in the cost of the alternative settings and one found an increase in birth centre care.There are few studies that compare the cost of birth setting. The variation in the results may be attributable to the cost data collection processes, difference in health systems and differences in which costs were included. A better understanding of the cost of birth setting is needed to inform policy makers and service providers. Maternity services are the third most common specialist service in Australia and single spontaneous delivery is the most common principle diagnosis among acute overnight admissions to hospital. There arIn 2009, the Department for Health and Ageing released the National Review of Maternity Services which contained feedback from stakeholders outlining issues relating to limited access to different models of care.  As a resThe majority of Australian women do not have access to alternative birth settings. Access to birth centres remains at around 5% of women and homebirth less than 1%. In a recThere is evidence that alternative models of care and settings for birth are a safe, highly acceptable option for childbearing women. Within the last decade, Australian research has shown significantly lower perinatal mortality for women cared for in birth centres compared to hospital births.  SignificAccess to alternative birth settings internationally varies both within and between countries and this is closely linked to status and role of the midwife in that country , 11 whicIn Norway, midwives provide care to all childbearing women in labour in and out of hospital with referral to medical specialists for complicated cases. In-hospital birth accounts for 98.4% of women, the remaining 1.6% of women giving birth out of hospital including at home or in “freestanding birth homes”.  The propThe total number of out-of-hospital births in the Unites States (US) in 2012 was 1.36%, two-thirds of which occurred at home and the remaining third in birth centres.  While thWhilst evidence of the safety of alternative birth settings in Australia and internationally is growing, information about the comparative costs and cost-effectiveness is less easy to find. The most comprehensive study to date has been the Birthplace in England Study , in whicThe aims of this review are firstly to identify economic evaluations or cost analyses comparing places of birth which include home, birth centres, both freestanding and alongside, and standard hospital labour wards. Secondly, this review aimed to explore the methodological approaches employed by the selected studies. The purpose of this aim was to assess the factors that inform the cost and cost effectiveness of these settings and to determine the most appropriate approach to inform future cost analyses on place of birth.Searches of the literature to identify economic evaluations of different birth settings were made in February 2015 of the following electronic databases: MEDLINE, CINAHL, EconLit, Business Source Complete and Maternity and Infant care. The PICO principles  were used to inform keyword search terms. , 27 RefeOriginal, English language studies between the years 1995 and 2015 were included. Studies that reported on costs and/or incorporated an economic analysis of models of care and place of birth for low risk women at term were reviewed. To achieve this, the search included alternative birth settings, models of care, and economic analyses. After testing a number of combinations, the search terms associated with the concepts of place of birth and model of care were paired by group with economic analyses to achieve a reasonable number of papers to review see . RetrievThe Critical Appraisal Skills Programme (CASP) Economic Evaluations Checklist was used to appraise the methodologies of the included papers. This checklist has twelve questions adapted from Drummond et al (2005). The tool consists of questions with some guidance comments where applicable and “Yes, Can’t tell, No” responses see . The queData were extracted by one reviewer using a template agreed on by all authors that included the following areas of interest: Study design and perspective, population, intervention, comparison and outcome (PICO), source of resource use data, details of cost included in the study, results of the cost analyses, and overall interpretation the results .Of the 11 articles reviewed, three were from the United States of America (USA), two from Canada, two from the United Kingdom (UK), two from Europe, one from Ireland and one from Australia. Six of the studies used a cost-effectiveness model, with the remaining five studies reporting a combination of cost analyses (four) and a cost comparison. Eight of the 11 studies found a cost saving in the alternative settings, namely at home or in midwifery-led units . Two found no difference in the cost of the alternative settings and one found an increase in cost in providing care to women in a birth centre. The intervention compared in this review was the alternative setting for birth and this included home (four studies), birth centres or midwifery-led units  which were situated either alongside a standard hospital facility (six) or as free-standing facilities separate from a hospital campus (three).While papers were selected on the basis of including women at low risk of complications, the assessment of low risk status varied among the studies. Women were largely chosen using the criteria for selection to ‘birth centre care’ or ‘midwife-led care’ or ‘eligibility for a homebirth’ and this criteria was used to match the comparison cohort attending the standard hospital labour ward or ‘consultant led unit’. These criteria were not routinely described in the papers therefore it is difficult to attest to the comparability of the cohorts. Schroeder et al (2012) employed the National Institute for Health and Care Excellence (NICE) definition of a woman at low risk of complications.The perspectives of the studies included societal , 28, MedMany of the studies included in this review have reported clinical outcomes, either from Randomised Controlled Trials (RCTs) or associated studies, however this review has focussed on the cost outcomes of each study. These will be further divided by comparisons of birthplace and cost analysis methodology.Four studies described cost analysis or economic analysis of homebirth, birth centres and labour wards for low-risk women. , 21, 29 In another large study of 11,592 women who gave birth at home and 11,788 in hospital In the United States, Anderson and Anderson 1999) compared mortality and caesarean section rates and costs of homebirth and hospital birth. The authors concluded that homebirth was less expensive at US$1711 compared to birth in hospital estimated at US$5382. Janssen et al (2015) in British Columbia linked data from six administrative databases from 2001 to 2004 of 9864 women who planned to give birth at home under the care of a “regulated” midwife, or in a hospital under the care of a midwife or a physician. All women met the criteria for home birth eligibility. This study demonstrated significant cost saving between birth settings, home birth being the least costly at CAN$2275, followed by planned hospital birth with a midwife CAN$4613 and hospital birth with a physician CAN$4816 99 comparThe remaining studies examined cost in birth centres and standard labour wards. , 30, 33 Toohill (2012) in a small Australian cohort study of 102 women compared the costs and outcomes of women receiving Midwifery Group Practice (MGP) care, who gave birth in a birth centre, with women receiving ‘standard care’ by midwives or GPs, who gave birth in a standard labour ward. When the costs were compared overall, there was a significant reduction in the cost of giving birth in the birth centre under MGP care compared to standard care at AU$4722 and AU$5614 respectively (p<0.001). Further differences in costs between the settings were found in studies by Bernitz (2012) in Norway, and Howell (2014) in the USA. Bernitz studied 1100 women in a 4-year RCT, calculated costs per woman (using a top-down and bottom-up approach), and found women who had care in an alongside midwifery unit (AMU) were less costly to care for than those in the standard obstetric unit (SCU) (€1672 vs €1950 p<0.001) in the same hospital complex. An RCT in Ireland also found the cost of care of women in alongside Midwifery-Led Units (MLU) was €182 lower than in the hospital setting led by consultant doctors. The cost-analysis by Howell (2014) involved 43,859 participants, 872 of whom gave birth in one birth centre and 42,987 who gave birth in a ‘usual care’ setting who were matched on observable variables on birth certificate data by propensity score modelling. This study found a 16% reduction in costs for every pregnancy conducted within a birth centre; a saving of US$1163 per birth. In Canada, Reinharz (2000) in a control-matched cohort study measured the cost effectiveness of 2000 women in seven pilot projects and found differing results across all settings. These projects examined the processes and outcomes of women who chose to give birth in midwife-led birth centres compared to women giving birth under the care of midwives or physicians in a hospital setting. The findings varied in cost and effectiveness, with overall conclusions suggesting that midwives are less costly in three of the seven birth centres and in the remaining four, cost overlaps suggested no real cost difference. Sensitivity analyses revealed that the upper limit of the midwifery groups’ costs resulted in a marginally lower cost than the physician costs. The Scottish RCT by Hundley et al (1995) with 2844 participants examined intrapartum costs of introducing a midwifery-led birth centre compared to the standard consultant-led labour ward. Nine sensitivity analyses of differing factors were tested and the result indicated an additional cost of 10.5% for the introduction of the midwifery-led service . The birth centre was newly established and the results of the trial showed that the setup costs of the birth centre and the higher level of staff employed to staff the new unit factored significantly in the overall cost thus making it a slightly more costly option. This was the only study that showed the costs associated with an alternative birth setting were higher than the standard hospital setting.Using a decision analytic model, Stone and Walker (1995) measured the cost-effectiveness of birth centre birth compared to hospital birth for women with matched low risk profiles . The analysis concluded that it was less costly to give birth in a birth centre (US$3385) compared to a hospital labour ward (US$4673). Effectiveness results favoured the birth centre for this low risk cohort, calculating that only when the transfer rate from the birth centre to the hospital exceeded 62%, would the cost-effectiveness reverse.and consequences are examined simultaneously, the result is a ‘cost-effectiveness’ study.[Economic analysis is broadly characterised by two features: addressing the inputs and outputs (costs and consequences) of a given activity and identifying the critical criteria that inform the decisions around how resources are distributed or spent . The bass’ study.Six of the eleven appraised studies performed a cost-effectiveness analysis. , 31, 33 Three of the cost effectiveness analyses in this review were published since 2012, two of which examined the cost of providing alternative birth settings at one health service or campus , 31 alonSchroeder et al. (2012) performed a cost-effectiveness study from the perspective of the National Health Service (NHS) and calculated the incremental cost and incremental effects of births planned to be in the different birth settings and were expressed as mean cost per woman in each birth setting and mean net benefit. Unit cost estimation combined top-down/ bottom-up costing methods and resources costed were listed and augmented by an expert advisory group involving focus groups of midwives. This data was obtained through the questionnaires and datasheets associated with the larger Birthplace in England Study. Bootstrapping and sensitivity analyses were used to address uncertainty around ratios and variation in costs.The three remaining cost-effectiveness studies were published between the years 1995 and 2000. Stone and Walker (1995) and Anderson and Anderson (1999) examined the “National Birth Centre Study”. The National Birth Centre Study (1989) studied 11, 814 low-risk women admitted for labour and delivery at 84 free-standing birth centres in the United States. They were followed though their labour, birth, transfers and for at least four weeks after birth A decisiAnderson and Anderson (1999) took the perspective of the cost to Medicaid and reported charges by CNMs from 29 US states through a survey mailed out to every CNM known to the American College of Nurse-Midwives (ACNM). The Medicaid Program in the USA provides free or low-cost medical benefits to low-income people who have no medical insurance or have inadequate medical insurance. These charges were compared to obstetrician charges from published work  and the In Canada, an observational study by Reinhartz et al (2000) used data collected from perinatal medical records, client postnatal questionnaires and physician billing information from the Regie de l’assurance du Quebec (RAMQ), the Quebec health insurance board. The costs were considered from four perspectives in total—the Health Ministry, the regional board, the client and her family. Other costs were obtained from non-physician professional associations to value visits made by women, for example a chiropractor or dietician and unit costs for drugs were calculated on generic prices where available.Hundley et al (1995) and Kenny et al (2105) also performed an economic analysis alongside an RCT. Kenny et al (2015) estimated costs associated with medical and midwifery staff for antenatal, intrapartum and postnatal care as well as investigations and interventions and hospitalisation costs. The financial data were collected by an experienced hospital accountant and costs relating to resource use estimated and verified with financial and services managers. Midwifery managers provided advice on the various pathways women in the service may take throughout their pregnancy. Hundley (1995) performed the cost analysis from the perspective of the hospital by costing the settings separately and classifying the costs into four main groups: staff costs; consumables; capital costs; and overheads. The main method of data collection was by questionnaire to the midwife managing the birth and the women on discharge from each facility. Medical record review also elicited information regarding consumables and interventions such as monitoring, pharmaceuticals, operative delivery and vaginal examination.In the Netherlands Hendrix et al (2009) used the Dutch manual for costing in healthcare, which is a methodological reference for performing cost studies. This six-step reference included regarding the scope of the research, the choice of cost categories , resources to be included, the volume of resources, the valuation of the resources, and the calculation of the unit costs. Hendrix et al took the societal perspective and collected data identifying health care costs such as contact with health care professionals, medications, medical interventions, analgesia and hospitalisation. Non health care costs were regarded as costs to the woman and family and were collected using cost diaries, questionnaires given to the women and birth registration forms. The second study in this review from the perspective of Medicaid is by Howell et al (2014). This study estimated costs based on a previous study that used propensity score reweighting and instrumental variable analysis. Major components of the cost of care were estimated for the antenatal, intrapartum and postnatal care and these included physician costs (from Medicaid fees), midwife costs  and Maternal and infant hospital costs were calculated from the National Inpatient Sample and the National Perinatal Information Centre for costs incurred in the Neonatal Intensive Care Unit (NICU). Howell et al. (2014) went on to report that the most costly component of maternity care was the maternity facility cost regardless of the model of care.The results varied among the evaluations largely due to the differing costing methods, the data collection, the measurement and valuation of the resources identified. The authors concluded in eight of the eleven evaluations that it was less costly to give birth in a setting alternate to a standard labour ward. , 31–33 TThis review included international and Australian studies spanning two decades. Due to the variations in timing, method of economic evaluation, currency and local economies, it was not possible to meaningfully compare the outcomes of the evaluations between the jurisdictions and the Australian setting.Labour and birth consume significant resources in every country and in Australia it is the primary reason for acute hospital admission. Transferring the cost of place of birth is critical for policy, practice and management. In light of this we aimed to locate and assess the quality of economic evaluations of place of birth and the different associated costs.Systematic reviews of economic analyses can be difficult due to the varying study designs employed.  There haIt is important to note that all costs in this review were reported in local currency and at varying times, thus the value of reporting the monetary values rests in the comparisons between the birth settings rather than the generalisability of the actual costs reported. In fact, with increasing health care costs and greater competition for finite resources, it would seem negligent to avoid performing economic analyses on service delivery sectors; the question is which economic analysis is going to provide the best evidence? Is economic data a driver of reform? There is demand from women to expand birthing options  however This review has presented a variety of economic analyses that have been conducted on place of birth between 1995 and 2015; however the methods tend to vary in their clarity and scope of the measurement of resource use and costs included in the calculations see . OvercomThere is high quality evidence that midwifery-led care in pregnancy, labour and birth is safe, with comparative or improved outcomes for women at low risk of complications. –44 The cHomebirth costs included midwifery time and some consumables whereas the charges in the hospital group included accommodation for the women as well as the fixed costs of facility use and variable costs including staff time and consumables. Broadly speaking, most studies included in this review reported lower costs associated with giving birth at home or in birth centres. Given the ethical implications of performing an RCT to measure the differences in cost and outcomes of place of birth, the cohort study by Schroeder et al (2012) provided the most methodologically robust results. Prospective data collection in real time allowed the data collectors to document detailed use of resources, staff time and consumables, a method that could provide high quality data in future detailed cost analyses.The difference in costs for women attending birth centres were associated with fewer interventions and procedures , 31 shorCost savings can be found by moving low-risk childbearing women away from hospital settings where there are established fixed costs to consider, as well as the availability of the convenient use of technology and interventions that are motivated by increased throughput and medical practices. Logically, home births require fewer resources and do not incur accommodation costs.  The inteUndeniably, standard labour wards carry the highest fixed costs and staff costs. Standard labour wards are necessary but the level of care is not required for all women. Many low risk women give birth spontaneously, in Australia around 50 percent , and proOf the eleven studies reviewed here, eight reported a cost saving associated with giving birth in an alternative birth setting, namely at home or in a birth centre. Two of the studies found no significant difference in the cost of providing care to women who chose to give birth in an alternate setting, however the benefits for the women and the staff were notable with higher satisfaction reported by both the women and the staff working in these settings and no increase in adverse outcomes. One study reported a cost increase in providing care in a birth centre. The differences reported in these studies highlight the differences in location, health system, methods of analysis and resources included in the costing. More research needs to be conducted on cost of alternative birth settings to support the growing demand for these services for women however the difficulty in accurately measuring these costs requires careful planning of methodology if the results are to be useful to service planners.S1 PRISMA Checklist(DOC)Click here for additional data file.


Thus, the pumping effect increased as the electrode diameter decreased. Interestingly in CWEs, the initiating voltage for liquid pumping did not change with increasing diameter, up to 20 μm. We classified into three pumping zones, according to the initiating voltage and faradaic reaction. Liquid pumping using the CWEs could provide a new method for biological studies with adoptable flow rates and a larger ‘Recommended pumping zone’.Liquid pumping can occur along the outer surface of an electrode under a DC electric field. For biological applications, a better understanding of the ionic solution pumping mechanism is required. Here, we fabricated CNT wire electrodes (CWEs) and tungsten wire electrodes (TWEs) of various diameters to assess an ionic solution pumping. A DC electric field created by a bias of several volts pumped the ionic solution in the direction of the negatively biased electrode. The resulting electro-osmotic flow was attributed to the movement of an electric double layer near the electrode, and the flow rates along the CWEs were on the order of picoliters per minute. According to electric field analysis, the  The manipulation of droplets or liquids is important in various applications such as printing or patterning13579101213Transported droplets or liquids can be controlled using a variety of approaches, including architectural tapering with conical fibers18192122et al. in 1839, under tens of kilovolts biasingLiquid pumping of dielectric liquids along the outer surfaces of electrodes was first introduced by Faraday In this study, we fabricated CNT wire electrodes (CWEs) and sharpened tungsten wire electrodes (TWEs) having various diameters to study the liquid pumping of an ionic solution. The electric field between two electrodes was analyzed by COMSOL and the flow rate of the transported liquid was assessed by changing the applied voltage and distance between the two electrodes. The ionic solution pumping mechanism along the electrode was described, based on our experimental results and analysis.Sharpened TWEs were fabricated using a dynamic electrochemical etching. A 300 μm diameter TWE and a Pt electrode were immersed in a NaOH electrolytic solution. An electric potential of 7 V was applied between the TWE and Pt electrode. The TWE was moved up and down slowly to establish smooth morphology.CWEs were fabricated on sharpened TWEs using dielectrophoresis (DEP) and surface tension. A TWE was submerged in a solution, containing CNTs. An AC electric field was applied between the tungsten tube and the TWE . The CNT4 · 4H2O and 500 mM HBO3 was introduced by means of a tungsten tube. The fabricated CWE was submerged in the gold solution. A DC electric bias of 1 V was applied between the two electrodes for 20 sec. Generally in this method, the CNTs of the CWE were combined by van der Waals force. When a DC electric field was applied to the immersed CWE, the gathered CNTs could be re-dispersed. To strengthen the bonding force between the CNTs, gold nanoparticles were coated on to CWEs by electrodeposition. A gold solution of 5 mM HAuCl+ and Cl– ions have similar electric mobility. The CWE was manipulated by a three-axis stage and placed into an ionic solution droplet. A KCl ionic solution was used in this study because KThe fact that dielectric liquids can be pumped by a DC electric potential has been known for at least 100 yearsR1 + 1/R2 = C = Δp/ϒ, the meniscus profile can be expressed as [z(x) = b·cosh(x/b)] when gravity is neglected. R1 and R2 represent the radii of curvature of the surfaces, Δp is the pressure difference, b is the electrode diameter, and ϒ is the surface tension. According to the above equation, the smaller the electrode diameter, the smaller and sharper the meniscus surrounding the electrode. In this study, we numerically analyzed the electric field as a function of the radius of curvature (ROC) of the meniscus, using COMSOL software. To simplify our model, the presence of an electric double layer (EDL) around the electrode was neglected, and the electrode was assumed to be a perfect conductor. An electric potential of 5 V was applied to the entire surface of the electrode, and the electrical ground condition was applied to the bottom of the droplet. z-directional electric field in the droplet. The z-directional electric field exhibited considerable strength around the meniscus and at the electrode tip. For a ROC of 1 μm  and cathode (CWEs) proceed according to the following equations:The current response between a CWE or a TWE and the gold electrode was measured using a modulab system . A scan rate of 100 mVsA faradaic reaction of the CWEs and TWEs was generated above 2.7 V, and the current of the TWEs increased sharply compared with that of the CWEs . To assez-directional electric field around the meniscus of small electrode was more concentrated than that of the larger electrode. As described in the electric field analysis, the The faradaic reactions of CWEs and TWEs were generated over 2.7 V, and hydrogen bubbles were evident in optical microscopy imaging at ~3.9 and 3 V for the CWEs and TWEs, respectively. During the faradaic reaction, the liquid pumping was continuous. The CWEs and TWEs having various diameters were classified into three zones: a ‘No pumping zone’, ‘Recommended (R.) pumping zone’, and ‘Not recommended (N.R.) pumping zone’, according to the initial voltage and faradaic reaction . In the In this study, flow rates associated with 800 nm diameter CWEs were evaluated over a voltage bias range of 1.5 to 3.5 V. When a droplet is generated at a certain position on the CWE, the droplet increased in size at this position or crept along the CWE. Note that when the droplet size increased, we assumed that the transported liquid was used to increase the droplet volume. The flow rates of the transported liquids were analyzed for two conditions: the first condition corresponded to a change in the bias voltage while the distance between the electrodes remained fixed, and the second condition involved a change in the separation distance of the electrodes for a fixed bias.εξE/η) describes the EOF, in which ε is the dielectric constant, ξ is the zeta potential, E is the applied electric field, and η is the solution viscosity. According to this equation, the flow rate of transported liquid is linearly proportional to the applied voltage, and inversely proportional to the distance between the two electrodes. In our results, the flow rates of the pumped liquid along the CWE increased with increasing applied voltage, with the exception of the 3.5 V bias. At 3.5 V, the liquid was pumped along the CWE, and small beads were expelled from the mother droplet. This phenomenon is caused by the repulsive force in the ionic solution close to the electrodeR2 = 0.97).For the first condition, the applied voltage was varied from 1.5 V to 3.5 V, while the other conditions remained fixed . The HelR2 = 0.97). These two results demonstrate that the EOF played a significant role in liquid pumping along the CWE by a DC electric field. The flow rate using the CWEs was on the order of picoliters per minute; this range is applicable to various biological studies, including single cells .In the second part of the experiment, the distance between the two electrodes was changed, while the other conditions remained fixed . The disz-directional electric field around the meniscus of the small electrode was found to be more concentrated than that of the larger electrode. Therefore, liquid pumping with a smaller electrode can be initiated at lower voltages. Interestingly in CWEs, the initiating voltage for liquid pumping did not change, up to 20 μm diameter, because the liquid crept along the individual CNT surfaces. Regarding the initiating voltage and faradaic reaction, the pumping zone could be classified into three zones: a ‘No pumping zone’, ‘R. pumping zone’, and ‘N.R pumping zone’. The liquid pumping of an ionic solution for biological studies should be applied within the ‘R. pumping zone’ to prevent faradaic reactions. Similar to the Helmholtz-Smoluchowski slip velocity, the flow rate of the transported liquid was linearly proportional to the applied voltage, and inversely proportional to the distance between the two electrodes. The flow rates using the CWEs were on the order of picoliters per minute. We anticipate that the CWEs could provide new devices for biological studies to manipulate liquid with adaptable flow rates.In summary, CWEs and sharpened TWEs having various diameters were fabricated to study liquid pumping of an ionic solution. When a DC electric field was applied between two electrodes dipped into an ionic solution, liquid was pumped along the electrode having a negative bias. During liquid pumping, the liquid gathered and formed beads at several positions along the electrode. The liquid pumping of an ionic solution is caused by the movement of mobile ions (EDL) gathered around the electrode, and the resulting flow is the EOF. According to Laplace’s theorem, the smaller the electrode diameter, the smaller and sharper the meniscus around the electrode. Upon analysis of the electric field by COMSOL, the How to cite this article: Jung Hwal Shin et al. Ionic liquid flow along the carbon nanotube with DC electric field. Sci. Rep.5, 11799; doi: 10.1038/srep11799 (2015).


Objective. DHA supplementation was compared to nutrition education to increase DHA consumption from fish and DHA fortified foods. Design. This two-part intervention included a randomized double-blind placebo controlled DHA supplementation arm and a nutrition education arm designed to increase intake of DHA from dietary sources by 300 mg per day. Setting. Denver Health Hospitals and Clinics, Denver, Colorado, USA. Population. 871 pregnant women aged 18–40 were recruited between16 and 20 weeks of gestation of whom 564 completed the study and complete delivery data was available in 505 women and infants. Methods. Subjects received either 300 or 600 mg DHA or olive oil placebo or nutrition education. Main Outcome Variable. Gestational length. Results. Gestational length was significantly increased by 4.0–4.5 days in women supplemented with 600 mg DHA per day or provided with nutrition education. Each 1% increase in RBC DHA at delivery was associated with a 1.6-day increase in gestational length. No significant effects on birth weight, birth length, or head circumference were demonstrated. The rate of early preterm birth (1.7%) in those supplemented with DHA (combined 300 and 600 mg/day) was significantly lower than in controls. Conclusion. Nutrition education or supplementation with DHA can be effective in increasing gestational length. Evidence from both human and animal studies suggests that essential fatty acids of the n-6 and n-3 series play important and modifiable roles in maintaining gestation. In early epidemiological studies , 2 erythThe study was a two-part intervention which included a randomized double-blind placebo controlled supplementation arm in which subjects received either 300 or 600 mg of algal derived DHA or olive oil placebo and a nutrition education arm designed to increase intake of DHA by 300 mg per day from fish and other dietary sources. Approval was obtained from the Human Research Committee of Colorado State University and Colorado Multiple Institutional Review Board.p < 0.05 level. 871 subjects were recruited from Denver Health Hospitals  antenatal clinics. Four of the largest clinics were selected to participate in the study. Three clinics were randomized to receive supplements and one to receive nutrition education. For the supplemental arm, 662 subjects were recruited and of these 28 were ineligible, 634 subjects were randomly allocated to treatment, 289 withdrew, and complete delivery data was available on 345 subjects in the supplemental arm. 209 subjects were recruited and consented to participate in nutrition education. Birth data was available on 191 of these and 18 were lost to follow-up. In total, complete delivery data was available for 536 of the 563 subjects  using a stratified block randomization schedule, generated using a randomization table by staff at Martek Biosciences, to insure equal group assignment from each of three clinics participating in the supplement trial. DHA was provided in the form of 300 Kcal supplement bars containing DHASCO-S oil. Gel capsules containing the test oil or olive oil were available for those who refused the bars. Bars and gel capsules were provided by Martek Biosciences, Columbia, MD. Supplementation was initiated at week 20 of gestation and continued until delivery. Both subjects and all study personnel were blinded as to treatment. DHA content of the bars and capsules was verified by gas chromatography. Prepackaged supplements were stored in cool secured locations, accessible only to the investigative team. A four- to six-week supply of supplements was dispensed at each prenatal visit and subjects were instructed to return any unused supplements. Records of the supplements dispensed and returned were used to calculate adherence with treatment. In addition, each subject kept a supplement diary and research assistants reviewed supplement diaries at each prenatal visit and collected data on side effects, dietary intake of DHA-rich foods, and adverse events including vaginal infections, vaginal bleeding, and preterm labor occurring during pregnancy. Following delivery, maternal and infant medical records were reviewed for adverse antenatal and intrapartum events in the supplemental arm. The maternal record provided date of delivery, estimated delivery date, maternal height and weight at first prenatal visit, mode of delivery, type of rupture of membranes (ROM), type of labor onset, estimated blood loss, and complications of labor and delivery. The newborn record provided information on infant gender, birth weight, birth length, and head circumference. Gestational length was determined using the difference between the estimated delivery date and the actual delivery date. Gestational length was determined using the difference between the delivery date established using a combination of the LMP method and ultrasound, where available, and the actual delivery date.Blood samples were obtained at enrollment between 16 and 20 weeks of gestation and at delivery. Seven mL blood samples were drawn in EDTA-containing vacutainer tubes. Samples were separated by centrifugation  within four hours of delivery and flushed with nitrogen gas, frozen in liquid nitrogen, and stored at −80°C for later batched analysis of phospholipid fatty acids. Plasma lipids were extracted in chloroform/methanol (2 : 1 v : v). Fatty acid methyl esters (FAME) will be prepared by transmethylation with 14% boron trifluoride in methanol . FAME were analyzed by temperature-programmed gas liquid chromatography using a 30 m × 0.25 mmID microcapillary column and identified using reference standards .r = 0.39, p < 0.005). The inventory contained a finite list of foods which are the rich sources of long chain n-3 fatty acids (≥1.0 mg/100 g edible portion of DHA +/or EPA) and DHA-enriched functional foods and eggs. The food inventory was quantitated using the USDA Nutrient Data Base (release 15).Dietary intake of long chain n-3 fatty acids was estimated at enrollment (16–20 weeks), using a pictorial food frequency inventory developed for this study. The food frequency was validated against RBC DHA in 340 subjects shown in Nutrition Education Intervention: subjects were recruited from a fourth Denver Health, Women Infants and Children's (WIC) clinic, which was not participating in the clinical trial to avoid contamination of the supplemental groups by nutrition information. Convenience sampling was used to recruit subjects at WIC clinic visits between 18 and 20 weeks of gestation to participate in the nutrition education component of the study. Subjects received educational materials which were developed using focus group data collected in this population and based upon the Health Belief Model. This process and resulting materials are described elsewhere . The eduThe primary outcome variables tested were gestational length (in days). Secondary outcomes, infant birth weight, birth length and head circumference, percent preterm birth, and percent postterm birth were evaluated. Continuous variables were analyzed using analysis of covariance (ANCOVA) and regression analysis using the SAS statistical package version 14. An intent to treat analysis was conducted using SAS phreg and lifetest procedures. Covariates used in the ANCOVA, maternal prepregnancy BMI, and ethnicity were those shown to have significant effects on gestational days in the univariate analysis. Dichotomous variables were analyzed using the IBM SPSS Statistics 20, Pearson Chi-Square analysis.p = 0.773). The intent to treat analysis showed that there were no significant differences in ethnicity or other characteristics of those who completed the study compared to those who withdrew (p = 0.025 for the model) 4-day increase in gestational length with 600 mg (p = 0.025 DHA supplement) and 4.5 days with nutrition education (p = 0.003) compared to controls (p = 0.065). The differences between 600 mg DHA supplement and nutrition education groups were not significant.To examine the difference in effectiveness of DHA supplements in the form of gel capsules compared to the food bars, analysis of differences in gestational length by supplement type was conducted. Fifty-one subjects consumed capsules as a substitute for bars. There was no difference in response within each group attributable to supplement type (withdrew . Ethnicicontrols . There wp = 0.3). In order to test the effect of any amount of DHA in the controlled trial, 300 and 600 mg DHA treatment groups were combined. When supplement groups were combined, the preterm birth rate in those receiving any DHA (300 or 600 mg/day) was not significantly different from placebo. The early preterm birth rate of 1.7% in those supplemented with any DHA (300 or 600 mg/day) was significantly lower than the early preterm birth rate of 5.7% in the placebo group .Four infants were born postterm (>294 days), one in each of the supplement groups and two in the nutrition education group. Labor was induced at 41 weeks in thirteen subjects, four supplemented subjects  compared to six in the placebo and three in the education group . There wp < 0.01  r = 0.15) and birth weight (p = 0.05  r = 0.11). Each 1% increase in RBC DHA was associated with a corresponding 1.6-day increase in gestational length. Maternal RBC % total arachidonic acid at delivery was not correlated with gestational length or birth weight.Mean birth weight, birth length, and head circumference with supplementation or nutrition education did not differ from the control or among groups. Maternal RBC DHA significantly increased in supplemented groups and decreased in controls. Maternal RBC % total DHA at delivery positively correlated with gestational length  were not significantly different at entry . Mean DHGiven the broad inclusion criteria and the potential for pregnancy complications from altering maternal fatty acid consumption, we recorded and analyzed a large variety of adverse effects. The most serious of these included late miscarriage, stillbirth, and early neonatal demise and each of these is due to extreme immaturity at delivery. In each of these cases, delivery occurred after enrollment but before consumption of supplements. In each instance women removed themselves from the study and were thus included only in the “intent to treat” analysis. Other obstetrical outcomes included cervical insufficiency, primary intra-intrauterine infection, placenta previa necessitating expedited surgical delivery, and an undiagnosed uterine anomaly and were deemed not to be study related. There were only 3 cases of severe preeclampsia (1 in each group of the supplement arm) and a very low incidence of postpartum bleeding and no difference in blood loss among supplementation groups. Events at delivery  includinThe data are in agreement with several studies of fish oil supplementation at higher doses in which average 4- to 4.5-day increases in gestational length were observed . It is pp < 0.05) 68 g increase in birth weight corresponding to a one-day increase in gestational length with the DHA supplement [To date, two systematic analyses did not show a statistically significant effect of n-3 supplements using combined power of several studies on gestational length, reduction of PT labor, birth weight, or birth length. However, a meta-analysis which included four studies, three of which used long chain fatty acids, showed a significant decrease in early preterm birth (<34 weeks) and no decrease in late preterm birth . An earlpplement . This inAlthough some differences in DHA intake among Hispanics and non-Hispanic Whites and African American subjects were seen in this study population, intakes were similar to those previously reported. The 2007-2008 National Health and Nutrition Examinations Survey (NHANES) and What We Eat In America (WWEIA) show that women aged 20–29 years consume 0.05 grams (50 mg) DHA/day from food sources and women aged 30–39 years consume on average 0.06 grams (60 mg) DHA/day . Judge uOverall, small to moderate fish consumption has a greater impact on preterm delivery than fish oil supplements. Studies of FO supplementation had greatest effects on women who eat little or no fish , 8 and tThe physiological significance of a 4-day increase in gestational length is unknown. However, a four-day increase may have a great impact on long term outcomes since recent data suggests birth between 37 and 38 completed weeks of gestation is associated with a 10% greater risk for attention-deficit hyperactivity disorder (ADHD) in school age children which increases to 30% in those born between 35 and 36 weeks of gestation . LimitatAnother limitation was that baseline blood DHA levels and obstetric risk factors were not obtained in the nutrition education arm and these may have affected the outcome. However, there were no differences in initial RBC DHA levels or significant differences in demographics or obstetrical variables among any of the other three clinics serving the same population nor were there initial differences among treatment groups. Comparison of early preterm birth rates was only significant when the treatment groups were pooled. This result should be regarded with caution since the number of early preterm births was small and these were not preplanned comparisons. The strength of the study is that the effects of n-3 DHA from supplements and from food sources on gestational length were comparable in the same population group. Although the population studied was low income, the subjects were largely Hispanic, a group known to be at lower risk for preterm birth compared to other low income groups, so the results may not be generalizable to other populations.Nutrition education or supplementation with DHA was shown to be effective in increasing gestational length in a low income population at relatively low risk for preterm birth. Improving DHA status through education or supplements is vital since maternal DHA levels are correlated with fetal DHA status and also represent enhanced DHA stores to support lactation and early infant growth and cognitive development. The nutrition intervention in this study would be expensive for clinics to implement but WIC programs routinely provide nutrition education at 3-month intervals to low risk pregnant participants and more frequently to high risk participants. Many WIC programs currently offer omega-3 DHA counseling and some even offer DHA-enriched foods. Alternative education methods may be explored, including existing programs such as Text4Baby which delivers gestational age appropriate health messages to pregnant and postnatal subscribers.


There are few, limited, and to some extent contradictory, reports on the cellular and subcellular morphology of arachnoid cysts. In the literature cyst membranes are described as similar to, or as vastly different from, normal arachnoid membranes.This paper reports electron microscopic analyses of symptomatic cysts from 24 patients , that underwent fenestration surgery. Fourteen cysts were located in the middle cranial fossa , one in the interpeduncular cistern, five in the posterior fossa, and four were overlying the frontal cortex.Microscopic findings confirmed the diverse nature of this clinical condition. Twelve cyst walls resembled normal arachnoid, four had a conspicuous core of dense fibrous tissue with a simple epithelial lining, and the remaining aberrant cysts exhibited non-arachnoid luminal epithelia with plentiful microvilli and/or cilia, and also nervous tissue components in the wall. The possible identity and origin of various cyst types are discussed. We hypothesize that cysts are formed mostly at an early stage of embryonic development, as a teratological event.Cysts with various epithelial linings and extracellular components most likely have different barrier properties and fluid turnover characteristics. Further studies are needed to elucidate relations between cyst morphology, fluid composition, pathogenesis, and clinical behaviour including growth rate and relapse tendency. Arachnoid cysts (AC) are fluid filled cyst-like structures that are anatomically connected with the arachnoid mater and can be found throughout the cranial-spinal axis with a preponderance in the middle cranial fossa . Severalet al. [The ultrastructure of the arachnoid cyst membrane has been sparsely studied previously. Rengachary and Watanabe described the ultrastructure of three arachnoid cysts and found the cyst wall to be very similar to normal arachnoid mater . A cyst et al. ,19. Howeet al. -23. We dThe study was performed as a prospective explorative study. It was approved by the Regional Ethical Committee (380–09) and all patients gave their written consent to participate in the study. In the pediatric cases written consent was given by the patients’ parents.Twenty-four consecutive patients  with symptomatic ACs, who after evaluation were offered and accepted surgical treatment for their cysts, were included. Patients were both pediatric and adult cases, ages ranging from 10 to 79 years. Fourteen cysts were located in the middle cranial fossa , one in the interpeduncular cistern, five in the posterior fossa, and four were overlying the frontal cortex , and lumbar puncture sampling of 10 mL CSF before surgery. The MRI protocol included flair-, diffusion- and T1-weighted sequences for closer differential diagnosis of the cysts. The T1 scan was to determine cyst volume  fitted with diamond knives. Semithin sections 1 μm thick were examined by light microscopy after treatment with Richardson’s stain (0.5% Azur II and 0.5% Methylene blue). Ultrathin sections were contrasted with uranyl acetate and lead citrate before examination in a digitized LEO 912AB Omega electron microscope . Digital image files were acquired with a MegaView III or Veleta CCD camera . For samples prepared for SEM the aldehyde fixation was followed by repeated osmification according to the OTOTO protocol [Patients were operated under general anaesthesia with a craniotomy and microsurgical fenestration of the cyst wall. The procedure commenced with puncture and cyst fluid collection followed by excision of part of the cyst wall. This membrane was subjected to microscopical analyses. All patients were re-examined 3 months after surgery with the same test battery and MRI. Reference (control) material was obtained from normal-looking arachnoid mater from the cisterna magna of 5 consecutive patients with Chiari type I malformation undergoing first surgery . The surgical samples were divided into two parts. One small piece, for routine histopathological diagnosis, was fixed in 4% formaldehyde; no material was reserved for immunohistological analyses. The major part was immersed for 24 to 72 h in a mixture of 2% formaldehyde + 2.5% glutaraldehyde + 0.02% sodium azide in 0.05 M Na cacodylate buffer, pH 7.2. This sample was prepared for transmission and scanning electron microscopy (TEM and SEM). Pieces intended for TEM were postfixed in 1% osmium tetroxide + 1% potassium hexacyanoferrate in 0.1 M cacodylate followed by Patient history revealed no instances of complications during birth or early childhood. Only four patients reported head trauma. All head traumas were minor and there was no correlation between head trauma and cyst location or type Table . All patSamples from 20 patients underwent routine pathological-anatomical diagnosis . The structure of these samples was considered to agree with arachnoid cyst morphology.A macroscopically-evident variation in cyst wall character was confirmed by subsequent light microscopy of 1 μm sections of plastic-embedded specimens. Cyst wall thickness ranged from <10 μm to ≈ 800 μm and there were considerable differences in cellularity, connective tissue elements and vascularisation Figure . This dii) Cysts composed of arachnoid-like tissue (12 cases).en face SEM examination as a rule revealed a smooth and structure-less continuous surface (not shown). The core of the cyst wall consisted of a trabecular connective tissue with widely spaced cells and scattered microvessels. The cyst lumen  Cysts composed of fibrous connective tissue (4 cases).These cyst walls were generally thicker than in the other groups and had a dominating core of dense connective tissue with scattered cellular elements. The epithelium on both aspects of the cyst was single-layered Figure b except iii) Cysts with aberrant structure (8 cases).These samples deserve a more detailed description since they presented individually unique features. Most cysts in this heterogeneous category had a luminal surface that was richly equipped with microvilli of uniform length Figure . One of One cyst located in posterior fossa, had sub-epithelial tissue of neuropil appearance, i.e. a complex packing of unmyelinated nerve fibres Figure . This tiThe luminal cyst epithelium constitutes a primary barrier for cyst fluid exchange. Regardless of type  the epithelial lining was ordered and continuous in the examined surfaces of all samples except one. The latter refers to one specimen where a denuded area exposing the underlying collagenous core was encountered in the SEM Figure c. HoweveSamples from this patient group do not constitute an ideal control for normal arachnoid tissue. Nevertheless, these membranes agreed well with the expected morphology with a thickness up to 60–65 μm and organisation into three layers . MeningoThis is, to the best of our knowledge, the first prospective study on the detailed morphology of the walls of intracranial cysts, diagnosed as arachnoid cysts (ACs) from clinical criteria. The study material is also the largest hitherto reported with 24 patients. The most striking overall results are that (i) only half of the samples had a cyst wall arrangement in good agreement with arachnoid tissue; (ii) four cysts made up a morphologically distinct group with a dominant core of cell-poor dense connective tissue lined by a thin squamous epithelium; and (iii) the architecture and cellular composition of the remaining group was variable with an obvious admixture of various elements of an expected intracerebral origin or location, e.g. ciliated epithelia, glial and neuronal components.per se that these cysts are spontaneous and formed at an early stage, probably as a teratological event [None of these findings is unique, a number of case reports exist based on few patients that demonstrate the variability of the arachnoid cyst entity -23,25. Tal event -29. It ial event -37 it isThe fibrous cysts in the present report in one respect share morphology with normal dura through the dominance of densely organised cell-poor connective tissue. It might thus be assumed that they reflect erroneous positioning of mesenchymal islands destined for development into dura mater. Experimental evidence based on transgenic techniques indicates that the barrier-forming arachnoid and adjacent dural layers differentiate from a common cell lineage expressing prostaglandin D2 synthase (PGD2S) of neural crest origin at forebrain level and mesodermal origin from midbrain level and posteriorly . In addiAn alternative interpretation of the fibrous cysts is that instead they reflect an equivalent of mature scar or capsule tissue, for example after an early and limited hemorrhage. A recent study presented a “2 hit” process for development of arachnoid cysts with both a congenital histological defect in the development of the arachnoid membrane and a later event of head trauma or hemorrhage ,40. ThisCyst-lining epithelial cells equipped with cilia or numerous microvilli indicate a possible origin from ependymal and choroid lineages, respectively. Similarly, the presence of nervous tissue in the group of aberrant extracerebral cysts, anatomically connected with the arachnoid, reasonably represents true malpositioning of neural tube progenitor cells. We lack knowledge of relevant reference literature in the neuro-developmental field that would give clues on possible pathogenetic mechanisms behind such failures in the control of normal development. In a few case reports, authors discuss the possibility that CNS cysts lined by strongly-ciliated epithelia, due to the similarity to respiratory epithelium, are even derived from endodermal progenitors ,42. Fromet al. [Regionally the ciliated epithelia also presented a probable secretory morphology that differed from normal ependyma. Whether or not such a differentiation would result in effects on cyst fluid composition or turnover is beyond the scope of the present study. Nevertheless, this finding points at the possibility that cyst wall epithelium can actively modify cyst fluid. This hypothesis is also supported in a recent study by Berle et al. . We planet al. [The examined tissue samples invariably had a tight and continuous epithelial lining with seemingly ordered junctions between cells. This would implicate that the luminal epithelium, the primary exchange barrier, limits free exchange of cyst fluid by restricting the possibility for paracellular flow. The degree of resistance should, on the other hand, vary considerably between the attenuated blood capillary endothelium-like junctions of the lowest single layered epithelium, and the elaborate intercellular pathways with multiple specific junctional elements of more complex epithelia. In addition, the variations of stromal thickness and composition including vascularization, and of the arrangement of cells at the dural aspect of cysts, must influence the net fluid exchange properties of cyst walls. Recent studies have focused interest on active mechanisms for cyst fluid accumulation. For example, Berle et al.  reportedet al.  whereas All patients in this study were operated with microsurgical decompression through craniotomy. It is well known that intracranial cysts can re-occlude years after surgery requiring secondary surgery ,48. To oThis study on 24 patients established that arachnoid cysts have variable organization of epithelial linings and extracellular components suggesting different barrier properties and fluid turnover characteristics. Further studies are needed to elucidate relations between cyst morphology, cyst pathogenesis, and clinical behavior, including growth rate and relapse tendency. Cyst morphology might be a relevant factor since some cysts become symptomatic -51 whileCarsten Wikkelsø receives honorarium for lecturing and consulting by Codman, Johnson&Johnson, and Likvor AB.KR carried out the compiling of patients’ data, participated in collection and preparation of microscopic specimens, performed light and electron microscopy and interpretation thereof, participated in manuscript preparation and took main responsibility in literature search; MT performed all neurosurgical procedures including initial specimen harvesting and drafted the manuscript; CW initiated the study and was the senior medical officer in charge for the clinical presurgical exploration of patients, drafted the manuscript; BRJ supervised and performed light and electron microscopy, assisted KR in preparation of manuscript, performed graphical work of micrographs. All authors have read and approved the final version of the manuscript.


The effect of maternal dietary treatment on the piglet gene expression profile of inflammatory cytokines in the colon following a lipopolysaccharide (LPS) challenge was also investigated. Dietary SDP reduced sow faecal Enterobacteriaceae gene numbers at parturition. Small-intestinal morphology, nutrient transporter and cytokine gene expression in newborn piglets did not differ between maternal dietary treatments (P > 0·10). At 48 h after birth, sodium–glucose-linked transporter 1 gene expression was down-regulated in the ileum of piglets suckling the SDP-supplemented sows compared with those suckling the basal sows (P = 0·050). There was a SDP × LPS challenge interaction on IL-1 and IL-6 gene expression in the colon of piglets (P < 0·05). The gene expression of IL-1 and IL-6 was down-regulated in the LPS-challenged colon of piglets suckling the SDP sows compared with those suckling the basal sows (P < 0·05). However, there was no difference in IL-1 and IL-6 gene expression in the unchallenged colon between treatment groups. At weaning, piglets suckling the SDP-supplemented sows had increased villus height in the jejunum and ileum compared with those suckling the basal-fed sows (P < 0·05). In conclusion, maternal dietary SDP supplementation enhanced the immune response of suckling piglets and improved gut morphology, making them more immune competent to deal with post-weaning adversities.The experiment investigated the effect of maternal dietary supplementation of seaweed-derived polysaccharides (SDP) (–SDP  The gastrointestinal tract (GIT) of the piglet encounters numerous challenges at birth and weaning. Immediately after birth, the neonatal GIT is rapidly colonised by bacteria originating from the mother or the environment. It has become clear that this primary colonisation is important for the right development and programming of the animal's local and systemic immune system. Furthermore, the GIT takes complete responsibility for the provision of nutrients, and shifts from processing amniotic fluid swallowed during and after birth to a nutrient-enriched colostrum ingested after birth. At weaning, pigs are exposed to a large number of stressors, such as separation from the dam, transition from milk to a diet based on plant polysaccharides, and withdrawal of maternal IgA from milk that acts locally in the intestine of suckling piglets. These factors combined can disturb the intestinal immune system and microbiota equilibrium, which contribute toward a decrease in daily gain immediately post-weaning. In order to adapt to the changes encountered at birth and weaning, the GIT undergoes accelerated tissue growth (birth) and functional maturation (birth and weaning), characterised by increases in intestinal weight and length, villus height (VH) and crypt depth (CD), cell migration rate, RNA and DNA contents, and the adaptation of intestinal enzymic activity. The intestinal epithelium is not only a physical barrier and a nutritional site; it also acts as a crucial regulator of intestinal immune homeostasis. In response to bacteria colonisation at birth and weaning, the enterocytes can act as a component of the immune system by secreting cytokines.Piglets are born in an agammaglobulinaemic state(–). However there is no information about the effect of maternal dietary treatment of SDP on intestinal dysfunction immediately after birth and after ingestion of colostrum. Furthermore, Leonard et al. found no effect of maternal SDP supplementation on piglet body weight (BW) at birth. The starting date of the supplementation (day 107 of gestation) may have been too late to promote an effect on piglet birth weight. Studies have been shown that the nutrient needs for fetal growth increase from day 69 of gestation, and the transfer of nutrients to the fetus increases between days 90 and 100 of gestation.Recent research has indicated that supplementing sows with seaweed-derived polysaccharides (SDP), containing laminarin and fucoidan, bioactives containing antimicrobial, prebiotic and immunomodulatory properties, from day 107 of gestation influenced piglet immune status at weaning, and had beneficial effects on weaning-associated intestinal dysfunction and growth depression immediately after weaningThus, the objective of the present study was to investigate the effect of maternal dietary supplementation of SDP from day 83 of gestation until weaning (day 28) on piglet BW at birth, and small-intestinal morphology, nutrient transporter and inflammatory cytokine gene expression, and colonic microbiota population of piglets at birth (0 h), 48 h after birth and weaning. The effect of maternal dietary treatment on the piglets’ gene expression profile of inflammatory cytokines in the colon following a lipopolysaccharide (LPS) challenge was also investigated. The hypothesis of this study is that maternal SDP supplementation from day 83 of gestation would improve piglet BW at birth and modulate selected intestinal microbial populations, the inflammatory response and aspects of intestinal health of piglets at birth and during the suckling period, making them more immune competent to deal with post-weaning adversities.All procedures described in the present experiment were conducted under experimental license from the Irish Department of Health in accordance with the Cruelty to Animals Act 1876 and the European Communities  Regulations.et al.. The SDP supplement (10·0 g/d) contained laminarin (1·0 g), fucoidan (0·8 g) and ash (8·2 g), and was extracted from a Laminaria spp. according to the procedure described by Lynch et al.. The gestation diet contained 140 g/kg of crude protein (CP), 13·5 MJ/kg of digestible energy (DE) and 4·4 g/kg of standardised ileal digestible (SID) lysine. The lactation diet contained 190 g/kg of CP, 14·5 MJ/kg of DE and 8·5 g/kg of SID lysine. All amino acid requirements were met relative to lysine. The ingredient composition and chemical analysis of the diets are given in A total of twenty crossbred pregnant gilts  were randomly assigned to one of the two dietary treatments (ten gilts/treatment): (T1) basal gestation/lactation diet (control) and (T2) basal gestation/lactation diet supplemented with 10·0 g SDP/d from day 83 of gestation until weaning (day 28). The quantity of SDP (Bioatlantis Ltd) used was based on previous work by Leonard Prior to day 83 of gestation, the gilts were housed in groups of ten. From day 83 until day 106 of gestation, they were housed individually in crates (2·0 × 0·6 m) in the gestation house. In the farrowing house, the gilts were housed individually in farrowing pens (2·2 × 2·4 m). The gestation and farrowing houses were maintained at 20°C throughout the experiment.ad libitum consumption of the diet, which was adjusted for each sow depending on daily intake. The sows were fed in three equal meals provided at 09·00, 13·00 and 17·00 hours. They had ad libitum access to drinking water throughout the experimental period.The experimental supplement (SDP) was top-dressed on the gestation diet and added to the trough prior to feeding the lactation diet each morning (09·00 hours) to ensure consumption. The dams received specific amounts of feed in the following quantities: 2·5 kg/d of gestation diet from day 83 until day 106 of gestation. They were fed 2·0 kg/d of lactation diet from day 107 of gestation until the day of farrowing and then the feed supply was increased by 1·0 kg/d until day 3 post-farrowing and by 0·5 kg/d until day 6 post-farrowing. Afterwards, the sows were allowed semi-sd 1·0) g) were collected from the ground into sterile containers (Sarstedt) and stored at −20°C for quantification  of Lactobacillus spp. and Enterobacteriaceae.On the expected farrowing date, fresh sow faecal samples (approximately 10 (n 12 piglets/sow), and this was maintained throughout the suckling period. Cross-fostering performed on average at 20 h after birth has no adverse effects on growth performance and IgG serum concentration of both adopted and biological piglets. The individual piglet BW was recorded at birth and weaning and the average daily gain calculated from these data. The piglets received an intramuscular injection of Fe-dextran  on day 7 after birth. No creep feed was offered to the piglets throughout the lactation period, and piglets did not have access to the sows’ feed.All farrowings were supervised. At parturition, piglets (Meat line boars × Large White × Landrace genetic lines gilts) were individually weighed and ear-tagged. Between 6 and 12 h after the birth of the last piglet, litter size was adjusted by cross-fostering piglets within sow dietary treatments to ensure that sows nursed a similar number of piglets  into the marginal ear vein of the sow. The colostrum samples were immediately frozen at −20°C before analysis for IgG concentration.. Diarrhoea score was performed by one trained person with no prior knowledge of the dietary treatment of the sows.A diarrhoea score was recorded daily on litter basis from birth to weaning using a scale from 0 to 3: (0) no diarrhoea; (1) slight; (2) middle; (3) acuten 8 piglets/treatment group, one piglet/sow) were selected at birth, with an average BW of 1·3 (sd 0·3) kg. The selection was based on the parturition of the third piglet. One trained operator was positioned at the back of the sow from the initiation of parturition, in order to make sure that the third piglet would not touch anything other than the sow vaginal tract and the gloved hand of the operator.A total of sixteen piglets , with an average BW of 1·8 (sd 0·1) kg.A total of sixteen piglets were selected at 48 h after birth , with an average BW of 7·4 (sd 0·9) kg. The selection of piglets at 48 h after birth and weaning was based on the litter average BW.A total of sixteen piglets were selected at weaning (The piglets were humanely killed on each day (immediately after birth (0 h), at 48 h after birth and at weaning) by lethal injection with pentobarbital sodium  at a rate of 0·71 ml/kg BW to the cranial vena cava, and the entire intestinal tract was immediately removed.et al.. The feed and dried faeces samples were milled through a hammer mill provided with a 1-mm screen (Christy and Norris). The DM of dried faeces and feed was determined after drying overnight at 103°C. Ash was determined after ignition of a known weight of concentrates or faeces in a muffle furnace (Nabertherm) at 500°C. The N content of both feed and faeces was determined using the LECO FP 528 instrument . Neutral-detergent fibre was determined using a Fibertec extraction unit (Tecator). The gross energy of the feed and faeces was determined using a Parr 1201 oxygen bomb calorimeter (Parr). The total laminarin content of the SDP supplement was determined using a Megazyme kit . Fucoidan levels were determined using the method of Usov et al..The feed samples were milled through a hammer mill provided with a 1-mm screen . The gestation and lactation feed samples were analysed for N, DM, ash and gross energy as described by Heim 2, which had been stripped of the overlying smooth muscle, were cut from each tissue. Then, one section from each tissue was placed in 1 ml of Dulbecco's modified Eagle's medium  in the presence or absence of bacterial LPS  at a concentration of 10 mg/ml. Both LPS-challenged and unchallenged tissues were incubated at 37°C for 120 min before being removed, blotted dry and weighed. Approximately 1–2 g of the ileum and colon tissues were cut into small pieces and stored in 15 ml RNAlater® solution (Applied Biosystems) overnight at 4°C. RNAlater® was then removed before storing the samples at −80°C.Immediately after slaughter, the entire intestinal tract was removed by blunt dissection and sections of the duodenum (10 cm from the stomach), jejunum (60 cm from stomach) and ileum (15 cm from caecum) were excised and fixed in 10 % phosphate-buffered formalin for VH and CD measurements. Ileum and colon tissues  were excised, emptied by dissecting them along the mesentery and rinsed using sterile PBS (Oxoid). Two tissue sections of 1 cmLactobacillus spp. and Enterobacteriaceae.Colon digesta samples (approximately 2 g) were collected into sterile containers (Sarstedt) and stored at −20°C for quantification  of g; Rotanta 460 R; Hettich Lab Technology) and 10 µl of the defatted fraction containing the IgG were added into 990 µl of 1× Dilution Buffer B to give a 1:100 dilution. The serial dilutions were repeated until the dilution recommended for colostrum: 1:1 000 000. The IgG was quantified according to the manufacturer's instructions.An assay for colostrum concentration of IgG was performed using a specific pig-ELISA IgG quantification kit (Bethyl Laboratories Inc.). For colostrum sample preparation, 10 ml of colostrum were centrifuged  using Lactobacillus spp. and Enterobacteriaceae primers. The primer sequences were as follows: Lactobacillus spp. – forward 5′–AGCAGTAGGGAATCTTCCA–3′ and reverse 5′–CACCGCTACACATGGAG–3′, 58°C; 341 bp and Enterobacteriaceae – forward 5′–CATTGACGTTACCCGCAGAAGAAGC–3′ and reverse 5′–CTCTACGAGACTCAAGCTTGC–3′, 58°C; 190 bp. All primers were designed using Primer Express™ Software (Applied Biosystems) and synthesised by MWG Biotech. Serial dilutions of these amplicons served to generate standard curves using RT-qPCR  permitting estimations of absolute quantification based on gene copy number (GCN). The RT-qPCR were performed in a final reaction volume of 20 µl containing 2 µl template DNA, 1 µl of forward (100 pm) and 1 µl of reverse primers (100 pm), 10 µl SYBR Green PCR Master Mix (Applied Biosystems) and 6 µl nuclease-free water. The thermal cycling conditions involved an initial denaturation step at 95°C for 10 min followed by forty cycles of 95°C for 15 s and 65°C for 1 min. Dissociation analyses of the RT-qPCR product were performed to confirm the specificity of the resulting RT-qPCR products. All samples were prepared in duplicate. The mean cycle threshold (Ct) values of duplicates of each sample were used for calculations.Microbial genomic DNA was extracted from sow faeces and piglet colon digesta samples using a QIAamp DNA stool kit (Qiagen) in accordance with the manufacturer's instructions. Quantity and quality of DNA were assessed using a NanoDrop Spectrophotometer . Standard curves were prepared as described by O'Shea . Measurements of fifteen well-orientated and intact villi and crypts were taken for each segment. The VH and the CD were measured as described by Heim et al.. The results are expressed as mean VH or CD in μm. The VH:CD ratio was calculated.The preserved segments  were prepared using standard paraffin-embedding techniques. The samples were sectioned at 5 µm thickness and stained with haemotoxylin and eosin® . The cDNA synthesis was performed using 1 µg of total RNA and oligo(dT)20 primers in a final reaction volume of 20 µl using a Superscript™ III First-Strand synthesis kit  following the manufacturer's instructions. The final reaction volume of 20 µl was then adjusted to 250 µl using nuclease-free water. All primers for the selected nutrient transporter (peptide transporter 1 (PEPT1/SLC15A1), sodium–glucose-linked transporter 1 (SGLT1/SLC5A1), glucose transporter 1 (GLUT1/SLC2A1), GLUT2/SLC2A and fatty acid binding proteins 2 (FABP2/I-FABP)) and inflammatory cytokine , IL-17R, TNF-α, interferon γ (IFN-γ), transforming growth factor β1 (TGF-β1) and forkhead box P3 (FOXP3)) genes were designed using Primer Express™ (Applied Biosystems) and synthesised by MWG Biotech. Primer sequences are presented in et al.. The efficiency of all primer sets was established using a semi-log curve of quantity v. control of 2-fold serial dilutions of cDNA as reported previously by Smith et al.. The following porcine reference genes were used as described previously by Ryan et al.: hydroxymethyl-bilane synthase (HMBS) and peptidylprolylisomerase A (PPIA) used for 0 h (immediately after birth) and weaning samples, and HMBS and tyrosine 3-mono-oxygenase/tryptophan 5-mono-oxygenase activation protein, zeta polypeptide (YWHAZ) used for 48 h after birth samples. The RT-qPCR was then carried out on cDNA using the ABI PRISM 7500 Fast sequence detection system for ninety-six-well plates (Applied Biosystems). All samples were prepared in duplicate using the SYBR Green Fast PCR Master Mix (Applied Biosystems), cDNA as the template and specific primers for the genes selected. For each reaction, 5 µl cDNA, 1·2 µl forward and reverse primer mix (300 nm), 3·8 µl nuclease-free water and 10 µl Fast SYBR Green PCR Master Mix were added and made up to a final volume of 20 µl. The two-step PCR programme was as follows: 95°C for 10 min for one cycle, followed by 95°C for 15 s and 60°C for 1 min for forty cycles.RNA was extracted from approximately 50 mg of ileum and colon tissue samples using the GenElute Mammalian Total RNA Miniprep Kit (Sigma Aldrich Corporation) according to the manufacturer's instructions. The total RNA was quantified using 1·5 µl of total RNA on a NanoDrop Spectrophotometer  and samples with a 260:280 ratio ≥2·0 were considered suitable for complementary DNA (cDNA) synthesis. Total RNA integrity  was assessed by analysing 1 µl of total RNA using the Agilent 2100 Bioanalyser version A.02.12  using RNA Nano LabChipsCt values for the reference genes were converted to relative quantities using the formula Q = EΔCt where E is the PCR efficiency of the assay and ΔCt is the value calculated for the difference between the lowest Ct value and the Ct value of the sample in question for each gene. The relative quantities of the endogenous controls were then analysed for stability in geNorm. The stability ‘M’ value generated by the geNorm application for the selected endogenous controls (β-actin (ACTB), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and PPIA) which was less than 1·5 indicated their suitability as endogenous controls for these intestinal samples. The geometric mean of the relative quantities for HMBS, YWHAZ and PPIA  was then calculated using geNorm. The relative quantities were divided by the normalisation factor (obtained in geNorm) for that sample to give the final normalised relative expression for each target gene.The raw . All data were initially checked for normality using the Univariate procedure in SAS. Inflammatory cytokine and nutrient transporter gene expression in ileum tissue, small-intestinal morphology and Lactobacillus spp. and Enterobacteriaceae quantification data were analysed as a complete randomised design, with the piglet (sow pen) as the experimental unit. GCN of Lactobacillus spp. and Enterobacteriaceae were log-transformed before statistical analysis. Inflammatory cytokine gene expression in colon tissue data was analysed as a 2 × 2 factorial arrangement, with the piglet (sow pen) as the experimental unit. The statistical model used included the effects of maternal dietary treatment and challenge (LPS or PBS), and their associated two-way interactions. The sow lactation data were analysed as a complete randomised design, with the sow pen as the experimental unit. For piglet diarrhoea score, data were analysed by repeated-measures analysis using the PROC MIXED procedure of SAS. The model included the effects of maternal dietary treatment and time (days after birth) and the associated two-way interactions. The pdiff function of SAS was used to separate means. Simple regression models were used to determine the relationships between BW and small-intestinal morphology, between small-intestinal morphology and nutrient transporter gene expression, and between inflammatory cytokine and nutrient transporter gene expression. Animal (piglet) was modelled as a random effect. All data presented in the tables are expressed as least-square means with their standard errors. The probability level that denotes significance is P < 0·05, a numerical trend is P > 0·05 and P < 0·10, and not significant P > 0·10.All data were analysed by using the General Linear Model procedure of SASv. 113·5 (se 0·27) d; P < 0·05). Colostrum IgG concentration did not differ between maternal dietary treatments; however, it did tend towards significance (57·0 v. 92·4 (se 16·9) mg/ml, for basal-fed and SDP-supplemented sows, respectively; P = 0·07). Litter size (14·4 (se 0·72) piglets) and the number of live born piglets (12·9 (se 0·62) piglets) were not influenced by maternal dietary treatments (P > 0·10). Piglet BW at birth (1·76 (se 0·66) kg) and weaning (8·33 (se 0·22) kg), and average daily gain from birth to weaning (0·24 (se 0·07) kg/d) (P > 0·10) did not differ between sow dietary treatments.Gilts offered diet supplemented with SDP had a longer gestation period compared with the basal-fed gilts , 48 h after birth and weaning did not differ between maternal dietary treatments . Piglet small-intestinal morphology at birth and 48 h after birth did not differ between maternal dietary treatments  and jejunum . There was also a positive linear effect of piglet BW at 48 h after birth on the VH in the jejunum .The VH and the CD in the ileum showed a numerical increase from birth to weaning (VH average: 143 P < 0·05). Piglets suckling the SDP-supplemented sows had reduced CD in the ileum compared with those suckling the basal-fed sows (P < 0·05). Piglets suckling the SDP-supplemented sows had an increased VH:CD ratio in the ileum compared with those suckling the basal-fed sows (P < 0·05). There was a positive linear effect of piglet BW at weaning on the VH in the duodenum .At weaning, piglets suckling the SDP-supplemented sows had increased VH in the jejunum and ileum compared with those suckling the basal-fed sows , followed by a decline to the end of lactation . The gene expression of FAPB2 and SGLT1 was lowly expressed at birth , followed by an increase to the end of lactation .P > 0·10; SGLT1 was down-regulated in the ileum of piglets suckling the SDP-supplemented sows compared with those suckling the basal-fed sows .At weaning, the gene expression of P > 0·10; Inflammatory cytokine gene expression in ileum tissue of piglets at birth and 48 h after birth did not differ between maternal dietary treatments . There was a negative linear effect of TNF-α gene expression on GLUT1 gene expression in the ileum of piglets at birth (0 h) . There was a negative linear effect of piglet BW at 48 h after birth on TNF-α  and IL-10  gene expression in the ileum.There was a positive linear effect of piglet birth weight on IL-1, IL-12A (p35), IFN-γ, TGF-β1 and TNF-α was up-regulated and the gene expression of IL-10, IL-17R, IL-6 and IL-8 was down-regulated in the ileum of piglets suckling the SDP-supplemented sows compared with those suckling the basal-fed sows , GLUT2  and PEPT1  gene expression in the ileum of piglets.At weaning, the gene expression of ex vivo LPS challenge at birth did not differ between maternal dietary treatments (P > 0·10) , IL-17R, IFN-γ and TGF-β1 in the colon tissue of piglets at birth (P < 0·05) (Inflammatory cytokine gene expression in piglet colon tissue following an  > 0·10) . The LPS < 0·05) .Table 7IL-1 and IL-6 (P < 0·05) . However, there was no difference in the gene expression of IL-1 and IL-6 in the unchallenged colon tissue between treatment groups (P > 0·10). The LPS challenge up-regulated the gene expression of IL-6 in the colonic tissue of piglets at weaning (P < 0·05)  . The gen < 0·05) .P > 0·10)    . The gen < 0·05) . The LPS < 0·05) .. Pig litters are unique: large litters at birth and the piglets are born relatively well developed. Much of this development takes place during the last few days of gestation. It seems likely that prolonged gestation would improve piglet maturation at birth, as the nutrient needs for fetal growth increase from day 69 of gestation, and the transference of nutrients to the fetus increases between days 90 and 100 of gestation. SDP contain sugars normally not present in plant dietary fibres. It was expected that these sugars, particularly laminarin, would provide nutrients to the fetus and consequently increase piglet birth weight. However, in the present study the increase in gestation length did not influence piglet BW at birth. There was also no effect of maternal dietary treatment on piglet BW at weaning. In agreement, Leonard et al. reported that sow dietary supplementation of a similar SDP from day 109 of gestation did not favour the growth performance of piglets during the lactation period.The hypothesis of this study is that maternal SDP supplementation from day 83 of gestation would improve piglet BW at birth and modulate selected intestinal microbial populations, the inflammatory response and aspects of intestinal health of piglets at birth and during the suckling period, making them more immune competent to deal with post-weaning adversities. The positive response observed in piglets suckling the SDP-supplemented sows, such as down-regulation of pro-inflammatory cytokine gene expression, and improved intestinal morphology in the ileum, partially supports this hypothesis. Dietary supplementation of SDP increased the gestation length. Gestation length of supplemented animals was prolonged by 1 d compared with the basal-fed gilts. The sow's gestation length is an average of 114 d, with 85 % of farrowing concentrating between 114 and 116 d of gestationP = 0·07). This is probably due to immunomodulatory property of laminarin. Laminarin is absorbed from the ileum, as recent work by Heim et al. showed that there was an increase in glucose transporters in the ileum following the ingestion of laminarin in weaned pigs. According to Bourne & Curtis, all colostral IgG is derived from the serum of the sow. The IgG uptake from serum is then mediated by specific Fc-dependent receptors on mammary epithelial cells. Previously, Leonard et al. reported that maternal supplementation of a similar SDP from day 109 of gestation increased colostrum IgG concentration.Maternal dietary SDP supplementation numerically increased colostrum IgG concentration . In the small intestine, the nutrients undergo a series of degradative steps carried out by digestive enzymes that can then be efficiently absorbed via nutrient transporters expressed along the crypt–villus axis in the enterocytes. Under conventional practice, abrupt weaning is often associated with undesirable morphological changes in the small-intestinal architecture, which include villous atrophy and crypt hyperplasia. There was no effect of maternal dietary treatment on nutrient transporter gene expression in the ileum of newborn piglets (0 h). There was also no effect of maternal dietary treatment on small-intestinal morphology. There was a positive linear effect of piglet birth weight on VH in the ileum and jejunum. Maternal dietary supplementation of SDP resulted in a down-regulation of the gene expression of SGLT1 in the ileum of piglets at 48 h after birth, and PEPT1, GLUT1 and GLUT2 at weaning. It is still not clear why maternal dietary supplementation of SDP down-regulated protein and glucose transporter gene expression in the ileum of suckling piglets. First, it may be related to an increase in VH in the ileum of these piglets. According to the regression analysis, there was a negative relationship between VH and the gene expression of GLUT1 at weaning. However, in this study, only the mRNA abundance of these nutrient transporters was measured, and unfortunately the nutrient transporter proteins were not measured. The expression of SGLT1 is tightly linked to the villus architecture. Regulation of glucose transport by diet may involve increased transcription of SGLT1 mainly in crypt cells. As cells migrate to the villus, the mRNA is degraded, and transporter proteins are then inserted into the membrane, leading to increases in glucose transport about 1 d after an increase in carbohydrate levels. Endocrine factors regulate the development of intestinal brush-border enzymes during weaning, but do not regulate the development of intestinal glucose transporters. The absence of regulation of glucose transport in suckling animals that nurse on milk is due to animals consuming a diet that does not vary in carbohydrate composition. It is possible that SGLT1 in cells present in the small intestine since birth cannot be regulated, and dietary regulation begins only when these cells are eventually replaced. The difference in SGLT1 expression may have been due to difference in milk composition. Unfortunately, milk composition was not measured in this experiment. However, Leonard et al. demonstrated that sow dietary supplementation of a similar SDP altered milk composition during lactation.The intestine is a major site of digestion, nutrient absorption, harbouring a complex microbiota and a highly evolved mucosal immune systemPEPT1 gene expression in the ileum of piglets suckling the SDP-supplemented sows at weaning may be attributed to the up-regulation of pro-inflammatory cytokine gene expression. This is supported by a negative relationship between TNF-α gene expression and PEPT1 gene expression in the ileum at weaning. According to Shu et al., bacterial infections alter PEPT1 expression level in the intestine. Furthermore, in the current study, a panel of pro-inflammatory cytokines was up-regulated in the ileum of piglets suckling SDP-supplemented sows at weaning. Third, the down-regulation of nutrient transporter gene expression may be attributed to the polysaccharide fucoidan. Heim et al. reported that when gilts were supplemented with fucoidan, from day 107 of gestation to weaning (day 24), the gene expression of SGLT1 was down-regulated in the ileum of piglets at weaning.Second, the down-regulation of GLUT1, GLUT2 and PEPT1 are highly expressed at birth followed by a decline to the end of lactation. On the other hand, the gene expression of FAPB2 and SGLT1 was lowly expressed at birth, followed by an increase to the end of lactation. VH and CD in the ileum showed a numerical increase from birth to weaning. Interestingly, there was a positive linear effect of piglet BW at birth on VH in the ileum. However, no maternal dietary treatment effect was observed on piglet BW at birth and VH in the ileum. In addition, maternal dietary SDP supplementation positively changed piglet small-intestinal architecture at weaning (increased VH and VH:CD ratio). The VH:CD ratio is a useful criterion for assessing intestinal health and function. This increase in VH and the VH:CD ratio may be attributed to the reduced diarrhoea score during the lactation period. Reduction in VH and the VH:CD ratio have been associated with increased incidences of scouring in weaned pigs challenged with enterotoxigenic E. coli.The present data also show that  and this primary colonisation is important for the right development and programming of the animal's local and systemic immune system. Dietary provision of SDP reduced dam Enterobacteriaceae GCN/g faeces on the expected parturition day, indicating an antimicrobial property of SDP. Previous research indicated that supplementation of a similar SDP to sows decreased their faecal Enterobacteriaceae numbers at parturition. The reduction in sow Enterobacteriaceae GCN/g faeces may be attributed to the potential agglutination properties of the SDP. β-Glucans (laminarin) have the capacity to agglutinate certain bacterial species, thus inhibiting subsequent attachment and colonisation of epithelial mucosal surfaces. Furthermore, fucoidan possesses numerous biological functions, including antimicrobial properties. Demecková et al. reported that sow faecal microbiota composition at farrowing influences subsequent bacterial colonisation of the neonatal intestinal tract. In the current study, lactobacilli were selected as bacterial indicators of beneficial bacteria, while Enterobacteriaceae were chosen based on their potential association with gastrointestinal disequilibrium. Piglets suckling the SDP-supplemented sows had reduced diarrhoea score during the lactation period. However, the decline in sow Enterobacteriaceae GCN/g faeces did not affect piglet Enterobacteriaceae GCN/g colonic digesta at birth, 48 h after birth and weaning. This reduction in diarrhoea score may be attributed to the immunomodulatory effects of SDP in enhancing the cellular and humoral immune function, as well as suppressing the E. coli population. Leonard et al. demonstrated that sow SDP supplementation reduced piglet colonic E. coli numbers at weaning. A suppressed colonic E. coli number may ultimately alleviate the incidence and severity of diarrhoea. Unfortunately, E. coli GCN was not measured in the current study.Immediately after birth the neonatal intestine is rapidly invaded by bacteria, originating from the mother and the environment. To mimic an immunological stress and the response of pigs exposed to a microbial challenge, colon tissue of piglets was LPS challenged ex vivo at birth, 48 h after birth and weaning. The biological effects mediated by LPS are attributed to inflammatory cytokine synthesis and release by stimulated macrophages. The LPS challenge up-regulated the gene expression of a panel of pro-inflammatory cytokines at birth , 48 h after birth (IL-6) and weaning (IL-8 and TNF-α), proving the LPS challenge worked.LPS is a component of the outer membrane of Gram-negative bacteria and a commonly used immunostimulantIL-1 and IL-6 gene expression down-regulated at 48 h after birth. There are a number of ways these findings could be interpreted. Porcine epithelial cells respond to various pathogens with the production of TNF-α and IFN-γ which block pathogen replication and cytokines are also involved in recruitment and activation of immune cells. The fact that piglets suckling the basal fed sows displayed increased expression of IL-1 and IL-6 in response to LPS suggests that these animals retained a capacity to respond more adequately to infection relative to the piglets suckling SDP-supplemented sows. An alternative hypothesis is that piglets suckling the SDP-supplemented sows possess a number of inherent energy-saving mechanisms, one of which is exhibited as a reduced inflammatory response following infection, as a result of maternal feeding of SDP. While both pig populations had no differences in their pathogenic bacteria levels at 48 h, experimental infection would be necessary to establish if this reduction in cytokine expression results in a measurable decrease in their resistance to pathogenic bacteria.The colonic tissue of piglets suckling the SDP-supplemented sows, following LPS challenge had TGF-β1 was up-regulated and the gene expression of the pro-inflammatory cytokines IL-6 and IL-8 was down-regulated in the ileum and colon tissues of piglets suckling the SDP-supplemented sows. IL-6 is produced by a number of cell types including macrophages, endothelial cells, B cells and mast cells, and is up-regulated in most, if not all inflammatory states. IL-8 is known to be regulated by inflammatory signals. IL-6 and IL-8 are cytokines classically involved in E. coli-caused diarrhoea. Down-regulation of the gene expression of these cytokines may be related to the reduced faecal scores observed in piglets suckling the SDP-supplemented sows. If the maternal SDP supplementation suppresses the secretion of pro-inflammatory cytokines, this could lower the activity of the immune system resulting in an energy-saving mechanism and increased growth performance during the suckling and post-weaning periods, as suggested by Li et al..At weaning, the gene expression of the anti-inflammatory cytokine IL-10 gene expression in the ileum. IL-10 is considered a potent anti-inflammatory cytokine that strongly inhibits the production of pro-inflammatory cytokines. There was negative linear relationship between piglet BW at 48 h after birth and gene expression of the pro-inflammatory TNF-α in the ileum, enforcing the concept of maximising piglets’ ingestion of colostrum and milk in the first 48 h after birth.There was a positive linear relationship between piglet BW at birth and the anti-inflammatory . β-Glucans can stimulate macrophages, neutrophils and natural killer cells, and could promote T cell-specific responses by induction of pro-inflammatory cytokines like IFN-γ, IL-8 and IL-12A (p35) from those cells(–). This pro-inflammatory response could be of particular importance because piglets often encounter presenting pathogens immediately post-weaning. Leonard et al. reported that maternal SDP supplementation enhanced the expression of TNF-α in the ileum after an ex vivo LPS challenge at weaning.A panel of pro-inflammatory cytokines was also up-regulated in the ileum and colon tissues of piglets suckling the SDP-supplemented sows at weaning. However, this pro-inflammatory response did not affect piglet growth performance during the lactation period. These cytokines play an important role in acute inflammation and are responsible for neutrophil recruitment and activation to the initial site of infectionE. coli-caused diarrhoea in piglets at 48 h after birth and weaning, indicating an important immunomodulatory effect of SDP. Maternal SDP supplementation also had positive effect in the small-intestinal architecture of suckling piglets, represented by the increased VH in the ileum of piglets at weaning. Even though little positive effect was observed in nutrient transporter gene expression and the microbiota population of suckling piglets, the results provide new insights into the protective activity of maternal SDP supplementation, making the suckling piglets more immune competent to deal with pathogens commonly encountered during the immediate post-weaning period.Maternal dietary SDP supplementation down-regulated the gene expression of a panel of pro-inflammatory cytokines, classically involved in