Serum Samples Research Articles

Frequency of lyme disease in patients with tick contact and identification of ticks from Bolu province of Turkey.

Lyme disease is a multisystemic disease caused by the bacteria in the Borrelia burgdorferi sensu lato complex, which is transmitted by ticks of Ixodes genus. Although there are seroprevalence studies and case reports of Lyme disease from various regions in Turkey, there is no widespread epidemiological research. This study aimed to determine the frequency of Lyme disease in the cases followed-up after tick contact and to examine the isolated ticks to reveal tick population from Bolu province. The cases who applied to the emergency department due to tick contact between April and September 2020 were firstly evaluated in our infectious diseases and clinical microbiology outpatient clinic on the third day of exposure and antibodies against Lyme disease were investigated with IFA method to exclude the patients who were previously exposed to B. burgdrorferi. Thereafter, patients were requested to continue outpatient visits at the 1st and 3rd month control. At these controls serum samples were taken to study B. burgdorferi antibodies with ELISA method which was stored at -20°C until the study day. Out of 123 patients who came to first control, 69 patients continued later to at least one of the two controls (either at first or third month). Of these 69 patients, only one (1.4%) was diagnosed with Lyme borreliosis according to clinical and laboratory features. Erythema migrans did not occur in any of the cases. Serum samples were assessed by ELISA method. Asymptomatic infection was detected in 22 cases (30.5%). In addition, we could obtain 46 ticks from our cases and two genera were identified. Forty-two (91.3%) were Ixodes spp, and two (4.3%) were Hyalomma spp. In this study, which was carried out for the first time in the province of Bolu, it was concluded that the cases presenting with a history of tick contact were most frequently exposed to Ixodes spp ticks and the probability of developing Lyme borreliosis was low (1.4%) during the three-month follow-up. Further studies with a greater number of cases and more extended follow-up period are needed.

Hepatitis B virus particles in serum contain minus strand DNA and degraded pregenomic RNA of variable and inverse lengths.

This study utilized digital PCR to quantify HBV RNA and HBV DNA within three regions of the HBV genome. Analysis of 75 serum samples from patients with chronic infection showed that HBV RNA levels were higher in core than in S and X regions (median 7.20 vs. 6.80 and 6.58 log copies/mL; p < .0001), whereas HBV DNA levels showed an inverse gradient (7.71 vs. 7.73 and 7.77 log copies/mL, p < .001). On average 80% of the nucleic acid was DNA by quantification in core. The core DNA/RNA ratio was associated with viral load and genotype. In individual patients, the relations between RNA levels in core, S and X were stable over time (n = 29; p = .006). The results suggest that pregenomic RNA is completely reverse transcribed to minus DNA in ≈75% of the virus particles, whereas the remaining 25% contain both RNA and DNA of lengths that reflect variable progress of the polymerase.

Investigation of the Effects of Labware Contamination on Mass Spectrometry-Based Human Serum Lipidome Analysis.

Polypropylene microcentrifuge tubes (MCTs) are increasingly used in lipidome sample preparation. In the absence of a comprehensive study evaluating ramifications of plasticware utilization in mass spectrometry-based lipidomic analyses, we conducted a systematic analysis to elucidate potential negative effects ascribable to labware contamination in serum lipidomics. During serum lipid extractions, tested glassware introduced 24 labware contaminants. In contrast, Eppendorf polypropylene MCTs contributed 485 contaminant features, many of which could be erroneously putatively identified as lipids via their m/z values. Eppendorf MCTs contamination engendered severe ion-suppression of 40 low abundance serum lipids, while generating mild to modest lipid ion-suppression across a multitude of higher abundance coeluting lipids. Less compatible polypropylene MCTs from an alternative manufacturer introduced a staggering 2,949 contaminant m/z values, severely affecting 75 coeluting serum lipids and causing more frequent and pronounced ion-suppression instances. Furthermore, by performing serum extractions with varied initial volumes, it was ascertained that labware-induced lipid ion-suppression is a dynamic phenomenon, contingent on both lipid and labware contaminant concentrations where low-abundance lipids are disproportionately impacted by coelutes of suppressive contaminants. In addition to lipid ion-suppression, the identification and quantification of 7 fatty acid endogenous serum lipids were compromised by the leaching of structurally identical surfactants from MCTs. MCTs artificially introduced 10 additional primary amides extraneous to serum samples. Utmost caution is imperative in interpreting data concerning primary amides and fatty acids when employing plastic labware. Through this investigation, we aspire to elevate awareness regarding the pernicious impact of labware contamination on lipidome analysis.

Analysis of Inflammatory Markers in Response to Induction of Reprometabolic Syndrome by a Eucaloric High Fat Diet in Normal Weight Women.

Obesity is associated with chronic low-level inflammation and is known to contribute to metabolic dysfunction and hypogonadotropic hypogonadism, which we have previously termed the 'Reprometabolic Syndrome.' To investigate potential factors involved in obesity-related reproductive endocrine dysfunction, we conducted a secondary analysis of inflammatory markers in a sample of normal weight women exposed to a one-month eucaloric high-fat diet (HFD), which, as reported earlier, induced the relative hypogonadotropic hypogonadism characteristic of Reprometabolic Syndrome. Eighteen healthy women with a BMI between 18.0-24.9kg/m2 and regular menstrual cycles participated in the study. Frequent blood sampling was performed during the early follicular phase before and after the one-month eucaloric HFD intervention (48% of calories from fat). Serum samples pooled from each participant were analyzed using immunoassay to measure levels of 30 cytokines, interleukins, and chemokines. Differences between pre- and post-HFD intervention measures were examined by one-sample t-tests. Exposure to the eucaloric HFD did not result in changes in body weight. HFD-induction of Reprometabolic Syndrome in normal weight women was associated with a significant elevation only in the anti- inflammatory cytokine IL-10 (p = 0.04). Eotaxin, IL-6 and MIP-1β also increased in response to the HFD, but not statistically significantly (p = 0.07). Results suggest that the increase in multiple inflammatory markers, typically associated with obesity, are not primary mediators of the relative hypogonadotropic hypogonadism of Reprometabolic Syndrome. Clinical Trials Registration Number: NCT02653092; Date of Registration: January 6, 2016.

Exosome-transported circ_0061407 and circ_0008103 play a tumour-repressive role and show diagnostic value in non-small-cell lung cancer

BackgroundCircular RNAs (circRNAs), one of the major contents of exosomes, have been shown to participate in the occurrence and progression of cancers. The role and the diagnostic potential of exosome-transported circRNAs in non-small-cell lung cancer (NSCLC) remain largely unknown.MethodsThe NSCLC-associated exosomal circ_0061407 and circ_0008103 were screened by circRNA microarray. The role of circ_0061407 and circ_0008103 in NSCLC was examined in vitro and in vivo. The encapsulation of the two circRNAs into exosomes and the transport to recipient cells were observed by confocal microscopy. The effects of exosome-transported circ_0061407 and circ_0008103 on recipient cells were investigated using a co-culture device. Bioinformatics analyses were performed to predict the mechanisms by which circ_0061407 and circ_0008103 affected NSCLC. The quantitative polymerase chain reaction was used to quantify the exosome-containing circ_0061407 and circ_0008103 in the serum samples of healthy, pneumonia, benign lung tumours, and NSCLC. The diagnostic efficacy was evaluated using receiver operating characteristic curves.ResultsThe levels of circ_0061407 and circ_0008103 within exosomes were down-regulated in the serum of patients with NSCLC. The up-regulation of circ_0061407 and circ_0008103 inhibited the proliferation, migration/invasion, cloning formation of NSCLC cells in vitro and inhibited lung tumour growth in vivo. Circ_0061407 and circ_0008103 were observed to be packaged in exosomes and transported to recipient cells, where they inhibited the proliferation, migration/invasion, and cloning formation abilities of the recipient cells. Moreover, circ_0061407 and circ_0008103 might be involved in the progression of NSCLC by interacting with microRNAs and proteins. Additionally, lower serum exosomal circ_0061407 and circ_0008103 levels were associated with advanced pathological staging and distant metastasis.ConclusionsThis study identified two novel exosome-transported circRNAs (circ_0061407 and circ_0008103) associated with NSCLC. These findings may provide additional insights into the development of NSCLC and potential diagnostic biomarkers for NSCLC.

Evaluation of humoral and cellular immune responses against Vibrio cholerae using oral immunization by multi-epitope-phage-based vaccine

IntroductionCholera is a severe gastrointestinal disease that manifests with rapid onset of diarrhea, vomiting, and high mortality rates. Due to its widespread occurrence in impoverished communities with poor water sanitation, there is an urgent demand for a cost-effective and highly efficient vaccine. Multi-epitope vaccines containing dominant immunological epitopes and adjuvant compounds have demonstrated potential in boosting the immune response. Material and methodsB and T epitopes of OMPU, OMPW, TCPA, CTXA, and CTXB proteins were predicted using bioinformatics methods. Subsequently, highly antigenic multi-epitopes that are non-allergenic and non-toxic were synthesized. These multi-epitopes were then cloned into the pCOMB phagemid. A plasmid M13KO7ΔpIII containing all helper phage proteins except pIII was created to produce the recombinant phage. Female Balb/c mice were divided into three groups and immunized accordingly. The mice received the helper phage, recombinant phage or PBS via gavage feeding thrice within two weeks. Serum samples were collected before and after immunization for the ELISA test as well as evaluating immune system induction through ELISpot testing of spleen lymphocytes. ResultsThe titer of the recombinant phage was determined to be 1011 PFU/ml. The presence of the recombinant phage was confirmed through differences in optical density between sample and control groups in the ELISA phage technique, as well as by observing transduction activity, which demonstrated successful production of a recombinant phage displaying the Vibrio multi-epitope on M13 phage pIII. ELISA results revealed significant differences in phage antibodies before and after inoculation, particularly notable in the negative control mice. Mice treated with multi-epitope phages exhibited antibodies against Vibrio cholerae lysate. Additionally, ELISpot results indicated activation of cellular immunity in mice receiving both Vibrio and helper phage. ConclusionThis study emphasizes the potential of multi-epitope on phage to enhance both cellular and humoral immunity in mice, demonstrating how phages can be used as adjuvants to stimulate mucosal immunity and act as promising candidates for oral vaccination.

Pharmacokinetics of oral clonazepam in growing commercial pigs (Sus scrofa domestica).

Clonazepam causes sedation and psychomotor impairment in people. Due to similarities between people and swine in response to benzodiazepines, clonazepam may represent a viable option to produce mild-to-moderate tranquillization in pigs. The objective of this study was to determine the pharmacokinetic profile of a single oral dose (0.5 mg/kg) of clonazepam in eight healthy, growing commercial cross pigs. Serial plasma samples were collected at baseline and up to 96 h after administration. Plasma concentrations were quantified using reverse-phase high-performance liquid chromatography, and compartment models were fit to time-concentration data. A one-compartment first-order model best fits the data. Maximum plasma concentration was 99.5 ng/mL, and time to maximum concentration was 3.4 h. Elimination half-life was 7.3 h, mean residence time 7.4 h, and apparent volume of distribution 5.7 L/kg. Achieved plasma concentrations exceeded those associated with psychomotor impairment in people although pharmacodynamic effects have not been investigated in pigs. A simulated oral regimen consisting of 0.35 mg/kg administered every 8 h to pigs would achieve plasma concentrations above 32 ng/mL which are shown to produce psychomotor impairment in people. Further studies to test the clinical efficacy of these dosages in commercial and miniature pigs are warranted.

Downregulation of miR-29a as a possible diagnostic biomarker for schizophrenia

BackgroundMicroRNAs (miRNAs) are epigenetic factors regulating many genes involved in brain development. Dysregulation of miRNA could result in dysregulation of genes which may contribute to diseases affecting the brain and behavior (e.g., schizophrenia). miR-29 family is a miRNA family contributing to brain maturation. miR-29 knockout in animal studies is reported to correlate with psychiatric disorders very similar to those seen in schizophrenia. In this study, we aimed to evaluate the miR-29a level in patients with schizophrenia and its potential value in the diagnosis of schizophrenia.Materials and methodsThe serum sample of 42 patients with schizophrenia and 40 healthy subjects were obtained from the Azeri Recent onset/Acute phase psychosis Survey (ARAS) Cohort study. After preparations, the expression level of miR-29a was investigated by real-time PCR. The SPSS and GraphPad prism software were used to analyze the relation between miR-29a level and clinical parameters and its potential as a biomarker for the diagnosis of schizophrenia.ResultsOur study showed a significantly lower miR-29a level in patients compared to healthy controls (p = 0.0012). Furthermore, miR-29a level was significantly lower in some types of schizophrenia (p = 0.024). miR-29a level was not related to sex, age, or heredity (p > 0.05). miR-29a also showed 80% specificity and 71.43% sensitivity in the diagnosis of schizophrenia.ConclusionDownregulation of miR-29a in schizophrenia is significantly related to the development of this illness. It might have the potential as a biomarker for schizophrenia.

73 Effects of blend of organic acids and monoglycerides on immune responses of weaned pigs experimentally infected with enterotoxigenic Escherichia coli F18

Abstract Our previous research implies that dietary supplementation of organic acids blend, monoglycerides blend, and the combination of both can mitigate diarrhea of weaned pigs infected with Escherichia coli (E. coli) F18. The objective of this study was to investigate the effects of dietary supplementation with a blend of organic acids, a blend of monoglycerides, or the combination of both on immune responses of weaned pigs experimentally infected with E. coli F18. Forty pigs [average body weight (BW) = 7.81 ± 0.84 kg] were individually housed and randomly assigned to one of four treatment groups with 10 replicate pigs per treatment. The four dietary treatments were: () Control: nursery basal diet, 2) Acids: basal diet supplemented with a blend of organic acids (formic acid, lactic acid, and sodium formate) at 0.3%, 3) Monoglycerides: basal diet supplemented with a blend of monoglycerides of fatty acids at 0.3%, or 4) Combination of acids and monoglycerides: basal diet added with the acids blend at 0.2% and the monoglycerides blend at 0.2%. The experiment lasted 28 d, with 7 d before and 21 d after the first inoculation (d 0). All pigs were orally inoculated with E. coli F18 (1010 CFU/3 mL) for 3 consecutive days from d 0. Serum samples were collected on d 0, 2, 5, and 14 post-inoculation (PI) to measure the concentration of C-reactive protein (CRP) and haptoglobin using ELISA. Porcine Cytokine/Chemokine13-Plex Discovery Assay (Eve Technology, Canada) was also conducted with serum samples from d 0, 5, and 14 PI. Data were analyzed by ANOVA using PROC MIXED of SAS with a randomized complete block design. The concentrations of serum CRP and haptoglobin were increased (P &amp;lt; 0.0001) on d 2 PI and then gradually decreased on d 5 and 14 PI. However, no differences were observed in serum CRP and haptoglobin concentrations among all treatment groups throughout the experiment. Pigs fed with monoglycerides tended (P &amp;lt; 0.10) to have decreased serum granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin 1-alpha (IL-1α), and tumor necrosis factor-alpha (TNF-α) concentrations on d 0 PI compared with other treatments. Pigs supplemented with monoglycerides also tended (P &amp;lt; 0.10) to have decreased GM-CSF on d 5 PI and less IL-1α on d 14 PI among all treatments. Supplementation of the combination tended (P &amp;lt; 0.10) to have greater GM-CSF concentration on d 5 PI compared with other treatments. In conclusion, dietary supplementation of monoglycerides blend or the combination of organic acids and monoglycerides alleviates systemic inflammation of weaned pigs during E. coli F18 infection by modulating related cytokines. However, additional research is needed to understand the modulation mechanisms.

220 Litter performance, serum collagen, and hormone concentrations in prolapsed and non-prolapsed sows

Abstract Pelvic organ prolapse is one of the top leading causes of sow mortality today and widely considered a multifactorial issue. Previous reports observed prolapsed sows having decreased serum concentrations of various trace minerals and increased tumor necrosis factor-alpha compared with non-prolapsed sows. Additionally, collagen, hormones, and previous litter performance are other factors proposed to be associated with POP. A study was completed to determine differences in serum collagen type 1 (COL1), matrix metalloproteinase 1 (MMP-1), tissue inhibitor of metalloproteinase 1 (TIMP-1), as well as relaxin and estradiol differences in the serum concentration between prolapsed and non-prolapsed sows. Furthermore, previous litter performance, lactation days, pigs born, stillborns, mummified, and pigs weaned on prolapse rate were evaluated. The study utilized 44 prolapsed sows and 44 non-prolapsed sows of similar parity, location, management, and stage of production. Blood was collected from the prolapsed and non-prolapsed sows upon discovery. Serum samples were analyzed using radioimmunoassay for estradiol content. Additionally, COL1, MMP-1, TIMP-1, and relaxin concentrations were determined from the serum using ELISA testing. Data were analyzed using the PROC GLIMMIX procedure in SAS. Prolapsed sows had decreased (P &amp;lt; 0.05) COL1 and TIMP-1 concentrations in the serum compared with the non-prolapsed sows. Prolapsed sows had marginally greater (P &amp;lt; 0.10) MMP-1 concentration compared with non-prolapsed sows. There were no differences (P &amp;gt; 0.10) between prolapsed and non-prolapsed serum concentraions of sows for estradiol and relaxin. Prolapsed sows had longer lactation days (P &amp;lt; 0.05) in the litter before prolapsing compared with non-prolapsed sows. Additionally, stillborns in the previous litter were marginally greater (P = 0.06) in prolapsed sows when compared with the non-prolapsed sows. There were no differences (P &amp;gt; 0.10) in pigs born, mummified, or pigs weaned between prolapsed and non-prolapsed sows. This study suggests that there is a reduced concentration of serum COL1 and TIMP-1 found in prolapsed sows compared with non-prolapsed sows and a marginally increased MMP-1 concentration in prolapsed sows. This could influence the structural integrity of the connective tissue that supports the pelvic organs. We found no differences in serum estradiol or relaxin levels. Additionally, lactation days and stillborn pigs in the litter prior to prolapsing could be contributing factors to prolapse. This work further supports the idea that prolapse is a multifactorial issue.

Preparation of ruminal fluid and serum samples from beef cattle for nuclear magnetic resonance based–metabolomics

ABSTRACT We evaluated different sample preparation methods for one-dimensional proton nuclear magnetic resonance metabolomics analysis of beef cattle ruminal fluid and serum. After collecting and pooling samples from five bulls (all of which received the same diet and were subjected to the same management conditions), we prepared the samples with the following methods: metabolite extraction using methanol and chloroform (M + C); metabolite extraction using methanol, chloroform and acetone (M + C + A); filtering with a 3-kDa filter; and filtering with a 2-μm filter. We evaluated the methods based on the spectral quality, reproducibility, ease of processing, speed and relative cost. The use of 2-μm filters did not produce good-spectra quality for serum samples, but it was a promising method for ruminal fluid. The yields of metabolites were similar for the the solvent extractions (22 metabolites for ruminal fluid and 29 for serum samples), but lower compared with the filter methods (31 metabolites for ruminal fluid and 47 for serum samples). The filters resulted in a higher concentration of metabolites and better reproducibility. In conclusion, the 2-μm and 3-kDa filter methods represent a suitable way to prepare biological samples from beef cattle for nuclear magnetic resonance – based metabolomics.

41 Is there anything we can do to lessen the severity of PRRSv?

Abstract Veterinarians and the swine industry continue to battle PRRS virus (v), as demonstrated by both the MSHMP (Morrison Swine Health Monitoring Project) and the SDRS (Swine Disease Reporting System). Both systems depict a continued increase in PRRSv prevalence and percentage of positive cases. The PRRSv variants that are circulating today seem to be more severe, especially in the last 2 to 3 yr, with more significant health and production losses in the nursery and finishing phases for prolonged periods of time. The best thing that can be done is to avoid having the virus with a good bioexclusion plan and if sites become infected, a strong biocontainment plan to avoid contaminating more sites should be considered. Realizing that bioexclusion is not always possible and sites will become infected, there are methods to help to reduce the impact of the virus. With the concerns of recombination pushing the virus evolution along faster, more producers are working toward eliminating field viruses from their systems. The use of filtration both on sow farms and nurseries has become more common as well as an additional method of bioexclusion. In the face of an outbreak, obtaining an accurate diagnosis is still one of the most important things that we do. In farrow to wean farms, serum samples are generally used when clinical signs (e.g., abortions, off feed) are observed to target testing to specific animals that are more likely to allow for early detection. In nursery and finishing sites, the use of oral fluid has worked very well as a screening tool when there are clinical signs or problems within a group or flow. Fortunately, there are differential tests that can be used to identify field infection and vaccines strains (since most wean to finish pigs are vaccinated in pig dense areas). With severe strains, depopulation and repopulation strategies have become more popular as a control method, but the majority of herd would use a herd closure. However, the amount of time required for these closures is extending, which lengthens the impacts on sow farms. Among wean to finish systems, the use of depopulating and cleaning up the site is used most, with only routine down times. Anti-inflammatories can also be administered in the acute phase of the outbreak in addition to the use of antibiotics to control secondary bacterial infections, as well as providing a good environment to allow the pigs to better deal with the disease. It’s best to keep the virus out of the herd with a good bioexclusion program. However, when we do have outbreaks, anti-inflammatory and antibiotics to help the pigs through it are important to utilize as we work to eliminate the field virus from the herd.

68 Serological comparisons of IBR-BVD-PI3-BRSV- Mannheimia haemolytica toxoid vaccines

Abstract The objective of this study was to compare serological responses in low-risk calves vaccinated with various IBR-BVD-PI3-BRSV-Mannheimia haemolytica toxoid vaccines. Bovine herpes virus-1 (BHV-1) and M. haemolytica (MH) leukotoxin titers were analyzed to investigate any potential BHV-1 antigen interference to MH leukotoxin levels when administering multi-valent vaccines. Serology responses of BVD, BRSV, and PI3 were not within the scope of this study. Before enrolling in the study, weaned calves born to dams at two University of Arkansas beef cattle research stations that were not previously vaccinated with MH toxoid or BHV-1 were bled and sera harvested. Crossbred steer and heifer calves [n = 364; steers = 168, heifers = 196; body weight (BW) = 187.9 ± 2.8 kg] were used for this study at the two locations. Calves were stratified by MH titers, BHV-1 titers, and sex, and allotted to 1 of 5 treatments: 1) “BS GOLD” = IBR-BVD-PI3-BRSV-MH (Bovi-Shield GOLD One Shot, 2 mL/SQ; Zoetis); 2) “P5 + PRE” = IBR-BVD-PI3-BRSV-MH (Pyramid 5 + Presponse SQ, 2 mL/SQ; Boehringer Ingelheim Animal Health); 3) “BV Once” = IBR-BVD-PI3-BRSV-MH-PM (Bovilis Vista Once SQ, 2 mL/SQ; Merck Animal Health); 4) “IF3 + One Shot” = IBR-BVD-PI3-BRSV (Inforce 3, 2 mL/intranasal; Zoetis) and BVD-MH (One Shot BVD, 2 mL/SQ; Zoetis); and 5) “BN + BV” = IBR-BVD-PI3-BRSV (Bovilis Nasalgen 3-PMH, 2 mL/intranasal; Merck Animal Health) and BVD (Bovilis Vista BVD CFP, 2 mL/SQ; Merck Animal Health). Cattle were weighed on d 0, 7, 14, and 28 and blood samples were taken for MH leukotoxin titers. Serum samples were collected on d 0 and 28 for BHV-1 titers. For statistical analyses, data were analyzed using the PROC MIXED procedure of SAS. Statistical significance was determined at P ≤ 0.05, with tendencies at 0.05 &amp;lt; P ≤ 0.1. There was an overall treatment effect for MH serum antibody titers with BV Once and BN + BV vaccinated calves exhibiting decreased MH serum titers compared with calves vaccinated with BS GOLD, P5 + PRE, and IF3 + One Shot (P = 0.0457). There was a treatment by day interaction for BHV-1 titers (P = 0.0336). On d 28, BHV-1 serum titers were greatest in BS GOLD and BN + BV vaccinated calves, least in calves that received IF3 + One Shot vaccines, and intermediate for BV Once vaccinated calves. Morbidity and average daily gain did not differ between treatments (P &amp;gt; 0.27) throughout the 28-d study period.

187 Polyphenols as a partial replacement for vitamin E in nursery pig diets

Abstract A total of 300 pigs [241×600; DNA, Columbus, NE; initial body weight (BW) = 6.0 ± 0.01 kg] were used in a 42-d trial to determine the effects of vitamin E levels and partially replacing vitamin E with a polyphenol (Cabanin CSD, R2 Argo, Denmark) on growth performance, complete blood count (CBC), serum thiobarbituric acid reactive substances (TBARS), serum superoxide dismutase (SOD), and serum cytokines. Sixty pens of pigs were weighed and allotted to 1 of 5 dietary treatments in a completely randomized design with 12 pens/treatment with 5 pigs/pen. A control treatment was formulated to provide 15 IU/kg of vitamin E. This control treatment was then used as a base for 3 replacement strategies to determine the effects of replacing an additional 60 IU/kg of vitamin E above baseline with polyphenol in diets containing a basal level of vitamin E (15 IU/kg). First, an additional 60 IU/kg of vitamin E was added for a total of 75 IU/kg of vitamin E. Second, 50% of the additional vitamin E (30 IU/kg) was replaced with the vitamin E equivalency of polyphenol. Third, all 60 IU/kg of the additional vitamin E was replaced with the equivalency of polyphenol. To evaluate whether there are negative effects of feeding nursery pigs a high level of polyphenol, a fifth treatment was formulated to provide 575 IU/kg of vitamin E equivalence with 75 IU/kg from vitamin E and 500 IU/kg from polyphenol. Whole blood and serum samples were collected on d 10 and 42, and pig BW and feed disappearance measured on d 10, 21, 31, 38, and 42. For growth performance, there was no evidence of differences (P &amp;gt; 0.10) in average daily gain (ADG) and average daily feed intake (ADFI) as vitamin E equivalence increased or between replacement strategies throughout the entire 42-d experimental period (Table). From d 10 to 21, increasing vitamin E equivalence from 15 to 75 IU/kg increased (quadratic, P = 0.086) gain to feed ratio (G:F) with no further increase at 575 IU/kg. From d 21 to 42 and overall, there was a tendency for increased (linear, P &amp;lt; 0.10) G:F as the vitamin E equivalence increased. There was a vitamin E equivalence × day interaction (P = 0.050) for serum SOD activity where increasing vitamin E equivalence increased (linear, P &amp;lt; 0.05) serum SOD activity on d 42, but not on d 10 (P &amp;gt; 0.10). For serum TBARS, there was no evidence of differences (P &amp;gt; 0.10) between treatments, vitamin E equivalence levels, or day, and negligible differences in complete blood count parameters and serum cytokines. In summary, increasing vitamin E equivalence tended to increase G:F, which may be related to the improved SOD activity. These data indicate that when diets contain requirement levels of vitamin E, additional vitamin E equivalency can be provided by polyphenol or vitamin E to achieve similar levels of performance.

307 Awardee Talk: Effects of dietary valerate glycerides on weanling piglet diarrhea and immunity

Abstract The use of various feed additives for their non-nutrient, immunomodulatory qualities is under investigation for application in swine production, especially during the post-weaning period. Butyrate (C4) is a known modulator of intestinal physiology, produced endogenously by commensal microbes and found in other organic sources. However, the understanding of the role of valerate (C5) as a feed additive for pigs has not been well-evaluated. Two experiments were conducted utilizing glycerides of butyrate or valerate as feed additives for weanling piglets under enterotoxigenic Escherichia coli (ETEC) infection conditions. In both experiments, daily diarrhea scores were recorded to monitor the frequency of diarrhea. The first pilot experiment utilized 20 weanling piglets (21 to 24 d of age) housed individually and assigned to either control diet, or control diet supplemented with 0.10% butyrate glycerides or 0.10% valerate glycerides. This experiment also aimed to establish a coinfection disease model, administering two strains of ETEC at a dose of 0.5 × 109 CFU/1.5 mL for each strain after a 7-d adaptation period (F4 and F18; two highly prevalent strains in swine production). Overall, piglets had significantly (P &amp;lt; 0.05) decreased frequency of diarrhea in either treatment group compared with control, suggesting similar modulatory effects by dietary valerate glycerides and butyrate glycerides. In addition, pigs fed valerate glycerides tended (P = 0.061) to have less neutrophils and had significantly (P &amp;lt; 0.05) less serum TNF-α on d 4 post-inoculation (PI). Moving forward, a full-scale experiment was performed using 60 weanling piglets (21 to 24 d of age) to further investigate the effect of dietary valerate glycerides on piglet health under ETEC F18 infection conditions. Treatments included nursery basal diet (control), 0.075% or 0.10% monovalerin, or 0.1% trivalerin added to control. Pigs fed 0.1% trivalerin exhibited significantly (P &amp;lt; 0.05) reduced frequency of diarrhea and fecal shedding of ETEC F18. Serum samples were collected throughout the experimental period and six replicate pigs per treatment group were sacrificed on d 7 PI to collect intestinal samples for morphological analysis and intestinal mucosa RT-qPCR analysis. Pigs fed trivalerin had reduced (P &amp;lt; 0.05) serum TNF-α on d 0 before inoculation and d 6 PI compared with pigs in control. The mRNA expression of TNFA in ileal mucosa from pigs fed 0.1% monovalerin tended (P &amp;lt; 0.10) to be lower compared with control. In addition, ZO1 mRNA expression tended (P &amp;lt; 0.10) to be increased in jejunal mucosa from pigs fed 0.1% trivalerin. Overall, the second experiment indicated that dietary supplementation of valerate glycerides reduced diarrhea and fecal shedding of ETEC F18 in piglets infected with ETEC. In conclusion, both experiments provide evidence in support of supplementing 0.075 or 0.10% valerate glycerides could enhance disease resistance of weanling piglets experiencing ETEC-associated diarrhea.

PSIII-19 Impact of EnduraPig and EnduraSow supplementation on growth performance of wean to finish pigs naturally infected with porcine reproductive and respiratory syndrome virus

Abstract Porcine reproductive and respiratory syndrome (PRRS) continues to be an economically devastating disease to the pork industry around the world. Apart from vaccination and biosecurity measures, nutritional strategies are needed to mitigate the negative effects of PRRS virus (PRRSV) infection on pork production. The aim of the present study was to determine the effect of EnduraPig and EnduraSow (PMI Additives) on growth performance, viremia, and livability of wean to finish pigs naturally infected with PRRSV. A total of 660 newly-weaned pigs [DNA L600 x 241 genetics; average body weight (BW) = 7.3 ± 0.14 kg) were divided into 33 pens (11 pens per treatment, 20 pigs per pen). Each pen within a location block was randomly assigned to one of three dietary treatments consisting of either a non-supplemented basal diet (NC), basal diet with EnduraPig (EP), or basal diet with EnduraSow (ES). Apart from growth [average daily gain (ADG), average daily feed intake (ADFI), feed to gain ratio (F:G)] and livability metrics, serum sample was also collected from 1 sentinel pig per pen at 0, 21, 42, and 63 d postweaning to monitor recovery from PRRSV infection, based on the change in percentage of viremic pigs. Analysis of variance for growth and mortality was performed using Mixed Procedure in SAS (SAS 9.4, SAS Institute Inc., Cary, NC) and a priori contrasts were used to detect treatment differences at P &amp;lt; 0.05. Association between treatments and % viremic pigs at each time point was assessed using Fisher’s exact test and declared significant at P &amp;lt; 0.05. Mortality and removals were greatest during the first 4 wk postweaning, but no treatment differences were detected in overall wean to finish mortality (23.6, 21.8, and 20.4% for NC, EP, and ES, respectively). At d 21, 89.9% of the pigs sampled were viremic compared with 56.8% on d 0. The average % viremic pigs went down to 57.5 and 18.5%, for d 42 and 63, respectively, with no treatment differences observed in both periods. Feeding EP, however, improved all growth performance metrics compared with NC (+11.8 kg in final body weight, P &amp;lt; 0.01; +13% in ADG, P &amp;lt; 0.01; +7% in ADFI, P &amp;lt; 0.05; and -5.5% in F:G, P &amp;lt; 0.05). Feeding ES also improved performance of wean to finish pigs over NC (+16.4 kg in final body weight, P &amp;lt; 0.01; +18% in ADG, P &amp;lt; 0.01; +12% in ADFI, P &amp;lt; 0.01; and -5.2% in F:G, P &amp;lt; 0.05). In summary, the results of this study indicate that dietary supplementation of EnduraPig or EnduraSow improved growth performance of wean to finish pigs naturally infected with PRRSV, indicating that these nutritional technologies help boost the immune response against PRRSV.

241 Immunophysiology and performance of broilers supplemented with macroalgal sulfated polysaccharides during a subclinical necrotic enteritis challenge

Abstract This study evaluated the effects of a sulfate polysaccharide extracted from marine algae product on performance, intestinal lesion scores, and mRNA abundance of various tight junction proteins and immune response genes. A total of 600 one-day old male broilers (Ross 708) were allocated to one of four treatment groups (6 replicate floor pens, 25 birds/pen): negative control (NC) group fed a corn-soybean meal diet; positive control (PC) fed NC + 15 ppm Avilamycin and 125 ppm Amprolium, an additive group (AGS) fed NC + Algoguard added at 0.1% of the diet, and (AGH) fed NC + Algoguard added at 0.2% of the diet. To predispose the birds to NE, birds were orally gavaged with 2,000 Eimeria maxima sporulated oocysts on d 14 followed by a dose of approximately 1×108 CFU/mL of C. perfringens on d 19. On d 21, four birds/pen were necropsied for NE lesions and on d 14, 21, and 42 jejunal samples were collected to assess mRNA abundance. Birds and feed were weighed on d 14, 21, 28, and 42 on a per pen basis. Average daily gain (ADG), average daily feed intake (ADFI), and feed conversion ratio (FCR) were calculated during each feeding phase and adjusted for daily mortality. The data were analyzed using ANOVA (JMP, Pro 16) and significance (P ≤ 0.05) was determined by the LSD test. During d 14-21, 0-21, 0-28, and 0-42, mortality rate and ADFI were significantly less in AGS, AGH, and PC compared with NC. ADG was greater while FCR was significantly less in PC, AGS, and AGH during d 21-28, 0-28, and 0-42 compared with NC (P ≤ 0.05). Lesion severity was also significantly less in AGS, AGH, and PC compared with NC. On d 14, 21, and 42 there were no toxins present in the tested serum samples. Calprotectin concentration was greater in PC and NC on d 21 and 42 compared with the supplemented groups. C. perfringens toxin was not detected in serum samples of any of the four treatment groups. mRNA abundance of CLDN1, IL1β, IL12B, and IL10 was greater in AGS compared with NC on d 14 and 42. ZO1 was significantly greater on d 21 in PC treatment and was less than both AGS and AGH on d 42 (P ≤ 0.05). Compared with all other treatments, mRNA abundance of OCLDN was greater and those of IFN-γ and ZO2 were less in PC, while that of CLDN3 showing no significant difference among all 4 treatments. To conclude, dietary inclusion of this algal product in broiler diets could be considered as a beneficial alternative to antibiotic growth promoters in the context of this challenge model.

Long-term effects of an egg-protein hydrolysate on cognitive performance and brain vascular function: a double-blind randomized controlled trial in adults with elevated subjective cognitive failures.

Short-term intake of the egg-protein hydrolysate Newtricious (NWT)-03 improved executive function, but underlying mechanisms and long-term effects, including other cognitive domains, are unknown. A 36-week randomized controlled trial involving 44 overweight/obese individuals experiencing elevated Subjective Cognitive Failures (SCF; aged 60-75years) assessed the impact of daily consumption of 5.7g of NWT-03 or placebo powders on cognitive performance (psychomotor speed, executive function, memory) and Cerebral Blood Flow (CBF), a marker of brain vascular function. Cognitive performance was evaluated using a neurophysiological test battery (CANTAB) and CBF was measured using magnetic resonance imaging perfusion method Arterial Spin Labeling (ASL). Serum samples were collected to determine brain-derived neurotrophic factor (BDNF) concentrations. Anthropometrics, and energy and nutrient intakes remained stable throughout the trial. NWT-03 was well tolerated, and compliance was excellent (median: 99%; range: 87-103%). No overall intervention effects were observed on cognitive performance or CBF, but post-hoc analyses revealed significant improvements on executive function in women, but not men. Specifically, a reduction of 74ms in reaction latency on the multitasking task (95% CI: -134 to -15; p = 0.02), a reduction of 9 between errors (95%CI: -14 to -3; p < 0.001), and a reduction of 9 total errors (95%CI: -15 to -3; p < 0.001) on the spatial working memory task were found in women. No intervention effects were observed on serum BDNF concentrations (p = 0.31). Long-term consumption of NWT-03 improved multitasking abilities and working memory in women with elevated SCF. Brain vascular function remained unaffected. Sex differences in executive function require additional clarification.

Effects of short-term moderate intensity exercise on the serum metabolome in older adults: a pilot randomized controlled trial

BackgroundWe previously reported changes in the serum metabolome associated with impaired myocardial relaxation in an asymptomatic older community cohort. In this prospective parallel-group randomized control pilot trial, we subjected community adults without cardiovascular disease to exercise intervention and evaluated the effects on serum metabolomics.MethodsBetween February 2019 to November 2019, thirty (83% females) middle-aged adults (53 ± 4 years) were randomized with sex stratification to either twelve weeks of moderate-intensity exercise training (Intervention) (n = 15) or Control (n = 15). The Intervention group underwent once-weekly aerobic and strength training sessions for 60 min each in a dedicated cardiac exercise laboratory for twelve weeks (ClinicalTrials.gov: NCT03617653). Serial measurements were taken pre- and post-intervention, including serum sampling for metabolomic analyses.ResultsTwenty-nine adults completed the study (Intervention n = 14; Control n = 15). Long-chain acylcarnitine C20:2-OH/C18:2-DC was reduced in the Intervention group by a magnitude of 0.714 but increased in the Control group by a magnitude of 1.742 (mean difference −1.028 age-adjusted p = 0.004). Among Controls, alanine correlated with left ventricular mass index (r = 0.529, age-adjusted p = 0.018) while aspartate correlated with Lateral e’ (r = −764, age-adjusted p = 0.016). C20:3 correlated with E/e’ ratio fold-change in the Intervention group (r = −0.653, age-adjusted p = 0.004). Among Controls, C20:2/C18:2 (r = 0.795, age-adjusted p = 0.005) and C20:2-OH/C18:2-DC fold-change (r = 0.742, age-adjusted p = 0.030) correlated with change in E/A ratio.ConclusionsCorresponding relationships between serum metabolites and cardiac function in response to exercise intervention provided pilot observations. Future investigations into cellular fuel oxidation or central carbon metabolism pathways that jointly impact the heart and related metabolic systems may be critical in preventive trials.

Changes in thyroid hormone concentrations over time in dogs with autoimmune thyroiditis.

The objective of this study was to follow long-term changes in the concentration of thyroid hormones in dogs with subclinical thyroiditis. Samples were obtained from 125 dogs with subclinical thyroiditis. The study population included 70 female and 55 male dogs. The mean testing interval was 3.9 years from initial testing (SD, 2.3 years; range, 1 to 9 years). Dogs with subclinical thyroiditis were identified retrospectively using results from the Orthopedic Foundation for Animals Canine Thyroid Profile performed by the Endocrinology Section of the Michigan State University Veterinary Diagnostic Lab. Owners were invited to submit follow-up serum samples with their veterinarian along with a medical history form, including subsequent treatments. At the time of retesting, 30% of the dogs had progressed to hypothyroidism and/or were treated with thyroxine. Fifty percent maintained positive or equivocal thyroglobulin autoantibody (TgAA) results while remaining euthyroid. Fourteen percent of the dogs became TgAA negative and remained euthyroid. In 6% of the cases tested, proper medical histories were not available, and a final classification could not be determined. These results indicate that most dogs with elevated thyroglobulin autoantibodies either exhibit persistent autoimmune thyroiditis with continued risk of hypothyroidism or progress to hypothyroidism when monitored for more than 1 year. Thyroid function in dogs with subclinical thyroiditis should be monitored every 12 months or if there is change in the clinical presentation.