Lactobacillus crispatus thrives in pregnancy hormonal milieu in a Nigerian patient cohort

AbstractSteroid hormones are one of the presumed modulators of Lactobacillus abundance in the vaginal epithelium. We set out to characterize the vaginal microbiome (VMB) and also provide an in-depth understanding of the relative contribution of estradiol (E2) and progesterone (P1) in shaping the vaginal microbiome of Nigerian women (n = 38) who experienced both uncomplicated term delivery and preterm delivery using samples longitudinally collected during pregnancy (17–21, 27–31, 36–41 weeks gestation) and 6 weeks postpartum. Vaginal swabs and blood samples were aseptically collected. Vaginal swabs were used for microbiome assessment using 16S ribosomal RNA (rRNA) gene sequencing. Blood samples were used for hormonal measurement using a competitive-based enzyme-linked immunosorbent assay (ELISA). Across several maternal covariates, maternal age, pregnancy status and delivery mode were not significantly associated with the vaginal microbiota whereas maternal E2 level (pE2 = 0.006, Omnibus), and P1 level (pP1 = 0.001, Omnibus) were significantly associated with the vaginal microbiome. E2 and P1 concentrations increased throughout pregnancy commensurately with increasing proportions of L. crispatus (pE2 = 0.036, pP1 = 0.034, Linear Mixed Model). An increasing trend of α-diversity was also observed as pregnancy progressed (pobserved ASV = 0.006, LMM). A compositional microbiome shift from Lactobacillus profile to non-Lactobacillus profile was observed in most postnatal women (pCST IV < 0.001, LMM). Analysis of our data shows a species-specific link between pregnancy steroid hormone concentration and L. crispatus abundance.

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The effects of gestational age on the composition and predicted function of gut microbiota in neonates and early infants

World Journal of Advanced Research and Reviews ◽

10.30574/wjarr.2021.12.2.0648 ◽

2021 ◽

Vol 12 (2) ◽

pp. 567-573

Author(s):

Kaiyu Pan ◽

Lianfang Yu ◽

Chengyue Zhang ◽

Jianhua Zhan ◽

Rongliang Tu

Keyword(s):

Gut Microbiota ◽

Gestational Age ◽

Full Term ◽

Rrna Gene ◽

Delivery Mode ◽

16S Sequencing ◽

Α Diversity ◽

Preterm Group ◽

Stool Samples ◽

Predicted Function

Gut microbiota can influence cell differentiation, metabolism, and immune function and is key for the normal development and future health of early infants. Several factors have been reported to be related to the microbiota composition of neonates, such as gestational age, delivery mode, feeding method, antibiotics consumption, and ethnicity, among others. So we investigated the relationship between gestational age and the composition and predicted function of the gut microbiota of neonates and early infants by sequencing the 16S rRNA gene present in stool samples obtained from 100 prospectively enrolled full-term and preterm newborns. In the 3-day-old neonates samples, the prominent genera in the full-term group were Escherichia-Shigella, Streptococcus, Bifidobacterium, and Bacteroides, while in the preterm group, Staphylococcus, Streptococcus, Escherichia-Shigella and Clostridium were the most abundant genera identified. There were statistical difference between two groups(P<0.05). Moreover, the predominant genera in the full-term group were Bifidobacterium, Lactobacillus, Bacteroides, and Clostridium , whereas the main genera in the preterm group were Escherichia-Shigella, Clostridium, Bifidobacterium and Bacteroides, in stool samples from 30-42-day-old infants. We found the α-diversity in 3-day-old group was significantly lower than in the 30-42-day-old group whether it’s full-term or preterm (P<0.001). Functional inference analysis revealed higher levels of biodegradation and metabolism of carbohydrates, vitamins in the full-term group than in the preterm group, both in neonates and early infants, which may contribute to the stability of the microbiota in the full-term group. There were significant differences in the composition and predicted function of the gut microbiota of early infants due to gestational age. The 16S sequencing technology was an effective and reliable tool in the detection of gut microbiota in early infants.

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The nasopharyngeal, ruminal, and vaginal microbiota and the core taxa shared across these microbiomes in virgin yearling heifers exposed to divergent in utero nutrition during their first trimester of gestation and in pregnant beef heifers in response to mineral supplementation

10.1101/2021.06.03.446997 ◽

2021 ◽

Author(s):

Samat Amat ◽

Devin B Holman ◽

Kaycie Schmidt ◽

Ana Clara B Menezes ◽

Friederike Baumgaertner ◽

...

Keyword(s):

Mixed Model ◽

Linear Mixed Model ◽

First Trimester ◽

Nervous System Development ◽

Vaginal Microbiota ◽

Rrna Gene ◽

Beef Heifers ◽

Mineral Supplement ◽

Alpha And Beta Diversity ◽

Significant Difference

Emerging evidence has indicated that microbial transmission from the bovine dam to her fetus may take place before birth, and that the maternal microbiota during pregnancy modulates programming of fetal metabolic and nervous system development, highlighting the potential and extended role of the maternal microbiome in calf health and development. In the present study, we characterized the nasopharyngeal, ruminal and vaginal microbiota from two cohorts of beef heifers managed at the same location: 1) virgin yearling heifers (9 months old) born from dams received gestational diets which resulted in low (LG, n = 22) or medium (MG, n = 23) weight gain during the first 84 days of gestation; and 2) pregnant replacement heifers that received a vitamin and mineral supplement (VTM, n = 17) or not (Control, n = 15) during the first 6 months of gestation. Nasopharyngeal and vaginal swabs as well as ruminal fluid were collected from both cohorts and the microbiota of each sample was assessed using 16S rRNA gene sequencing. In addition to the comparison between treatment groups within each cohort, the similarity of the microbiota of the three sample types were evaluated, and shared taxa amongst these communities were identified. The bacterial genera present in the rumen and vagina that can influence methanogenic archaeal genera were predicted using a stepwise-selected generalized linear mixed model. No significant difference was observed in the alpha and beta diversity in any of the nasopharyngeal, ruminal and vaginal microbiota between LG and MG offspring virgin heifers, or between the control and VTM pregnant heifers (p > 0.05). Subtle compositional changes in the vaginal microbiota in yearling heifers, and in the nasopharyngeal and ruminal microbiota of pregnant heifers were detected in response to treatments. Forty-one archaeal and bacterial OTUs were shared by over 60% of all samples from both virgin and pregnant heifers. Two taxa within the Methanobrevibacter genus were identified as core taxa and this genus was more relatively abundant in pregnant heifers compared to virgin heifers. Among the 25 top genera, Prevotella and Prevotella UCG-003 (negative) and Christensenellaceae R-7 group (positive) were predicted to have a significant effect on ruminal Methanobrevibacter spp. The results of this study indicate that there is little impact of divergent gestational nutrition during the first trimester on the calf microbiome at 9 months postnatal, and that VTM supplementation during pregnancy may not alter the maternal microbiome. This study provides evidence that there are several microbial taxa, including methanogenic archaea, that are shared across the respiratory, gastrointestinal, and reproductive tracts, suggesting the need for a holistic evaluation of the bovine microbiota when considering potential maternal sources for seeding calves with pioneer microbiota.

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Abstract WMP106: Soluble CD163 as a Prognostic Marker for Perihematomal Edema Formation and Functional Outcomes in Intracerebral Hemorrhage

Stroke ◽

10.1161/str.48.suppl_1.wmp106 ◽

2017 ◽

Vol 48 (suppl_1) ◽

Author(s):

Meaghan Roy-O’Reilly ◽

Davis So ◽

Glenda Torres ◽

Liang Zhu ◽

Jaroslaw Aronowski ◽

...

Keyword(s):

Intracerebral Hemorrhage ◽

Functional Outcomes ◽

Mixed Model ◽

Linear Mixed Model ◽

Tissue Injury ◽

Enzyme Linked Immunosorbent Assay ◽

Healthy Controls ◽

Edema Formation ◽

Soluble Cd163 ◽

Perihematomal Edema

Introduction: Macrophages are the predominant cell capable of removing toxic hemoglobin at sites of tissue injury, and CD163 has been recognized as the hemoglobin scavenger receptor present on the macrophage cell surface. In this study, we explored the levels of soluble CD163 (sCD163) in patients with intracerebral hemorrhage (ICH) to ascertain whether sCD163 was associated with clinicoradiologic features and long-term functional outcomes. Methods: Our ICH cohort was comprised of 50 patients with moderate-sized basal ganglia hematomas. We collected serial serum and cerebrospinal fluid (CSF) at pre-specified timepoints (24 hours, 48 hours, 3-5 days, 6-8 days, and greater than 10 days post-ictus). We also obtained samples from 10 healthy controls. Levels of sCD163 were measured by enzyme-linked immunosorbent assay. A linear mixed model was used to compare sCD163 values among various groups, using a Bonferroni correction for multiple test adjustment. The method of generalized estimating equations was used to determine associations with dichotomized outcomes (modified Rankin Scale score 0-3 versus 4-6). Results: Compared to healthy controls, serum sCD163 was higher in the ICH patients (40.6 versus 128.4 ng/mL). Within the ICH cohort, early values (24 hours to 5 days post-ictus) of serum sCD163 were significantly higher in patients who elaborated minimal perihematomal edema (PHE) (200.3 in patients with less than 10 mL PHE versus 71.8; p = 0.046). 6 to greater than 10 days post-ictus, sCD163 levels tailed off for patients with less PHE whereas levels rose in patients with greater PHE. Continued subacute elevation of sCD163, particularly in the CSF, was highly associated with poorer outcomes, both at discharge and at 90 days (p < 0.001). These associations were independent of age, gender, peak hematoma volume, and ICH score; there was a statistically significant association of CSF sCD163 values with degree of intraventricular hemorrhage (p = 0.04). Conclusions: sCD163 may be a dynamic marker in ICH, with acute levels distinguishing edema patterns and subacute levels predicting functional outcome. Further studies are needed to confirm these findings and explore the pathophysiology behind these observations.

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Oral Cholera Vaccination as a Model to Assess Dietary Modulation of Gut Inflammation and Immunity

Current Developments in Nutrition ◽

10.1093/cdn/nzab061_013 ◽

2021 ◽

Vol 5 (Supplement_2) ◽

pp. 1129-1129

Author(s):

Alwine Kardinaal ◽

Min Young Park ◽

Jong Eun Jeon ◽

Hee Jung Choi ◽

Ji Yeon Kim ◽

...

Keyword(s):

Inflammatory Response ◽

Antibody Response ◽

Mixed Model ◽

Linear Mixed Model ◽

Intestinal Inflammation ◽

Serum Levels ◽

Fecal Calprotectin ◽

Food Ingredients ◽

Blood Samples ◽

Cholera Vaccination

Abstract Objectives The aim was to develop a human challenge model in which modulation of immune and inflammatory response by food ingredients can be evaluated. We hypothesized that oral cholera vaccination, in addition to inducing a specific antibody response, induces a significant increase in gut inflammatory response. Methods Twenty healthy men (age 30.6 ± 1.8 y; BMI 24.9 ± 0.6) were enrolled in the study. Fecal and blood samples were collected at baseline. After a 2-week run-in period (D0-D14), subjects were vaccinated with oral cholera vaccine Dukoral (D15). After a period of 2 weeks, a second vaccination was administered (D29). Fecal samples were collected the day before (D14; D28) and the days (D16/D17; D30/D31) after each vaccination, as well as on D42. Blood samples were collected before vaccination (D15; D29), and on D16 and D17/D31, on D43. Primary outcome was fecal calprotectin concentration, secondary outcomes were serum levels of cholera toxin (CTB)-specific IgA and IgG. Other markers of local and systemic inflammatory response included beta-defensins, IP-10, IL-1 ra and hsCRP. Changes over time were tested by means of a linear mixed model. Outliers were identified with the 1.5xIQR rule and excluded from analysis. Results Fecal calprotectin did not increase after the first vaccination. After the second vaccination, a significant increase was observed: from 12.8 ± 2.5 μg/g feces (mean ± SEM) on day 29 to 18.0 ± 2.9 μg/g on day 31 (P = 0.017). Plasma CTB-specific IgA and IgG were strongly increased after the first vaccination, with a further increase after the second vaccination. Plasma CRP slightly decreased on D17, compared to D15 (P = 0.016). IL-1ra significantly decreased 2 days after the first vaccination (P = 0.011), whereas no change was observed after the second vaccination. Beta-defensin was significantly increased at D31 compared to D29 (from 42.7 ± 7.0 to 80.7 ± 16.1 (P = 0.014)). IP-10 did not show any response to vaccination. Conclusions In addition to the expected antibody response, oral cholera vaccination induces an increase in fecal calprotectin and beta defensin, pointing to vaccine induced intestinal inflammation. These readouts may be added to intervention studies with dietary compounds to evaluate the potential for modulating immune responsiveness. Funding Sources Bio&Medical Technol Developm Program of the Natl Res Foundation, Min Science & ICT of Rep Korea.

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Vaginal microbiome and serum metabolite differences in late gestation commercial sows at risk for pelvic organ prolapse

Scientific Reports ◽

10.1038/s41598-021-85367-3 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Zoë E. Kiefer ◽

Lucas R. Koester ◽

Lucas Showman ◽

Jamie M. Studer ◽

Amanda L. Chipman ◽

...

Keyword(s):

Pelvic Organ Prolapse ◽

16S Rrna Gene Sequencing ◽

Pelvic Organ ◽

Late Gestation ◽

Rrna Gene ◽

Vaginal Microbiome ◽

Swine Industry ◽

Vaginal Microflora ◽

Vaginal Swabs ◽

Rrna Gene Sequencing

AbstractSow mortality attributable to pelvic organ prolapse (POP) has increased in the U.S. swine industry and continues to worsen. Two main objectives of this study were, (1) to develop a perineal scoring system that can be correlated with POP risk, and (2) identify POP risk-associated biological factors. To assess POP risk during late gestation, sows (n = 213) were scored using a newly developed perineal scoring (PS) system. Sows scored as PS1 (low), PS2 (moderate), or PS3 (high) based on POP risk. Subsequently, 1.5, 0.8, and 23.1% of sows scored PS1, PS2, or PS3, respectively, experienced POP. To identify biomarkers, serum and vaginal swabs were collected from late gestation sows differing in PS. Using GC–MS, 82 serum metabolite differences between PS1 and PS3 animals (P < 0.05) were identified. Vaginal swabs were utilized for 16S rRNA gene sequencing and differences in vaginal microbiomes between PS1 and PS3 animals were detected on a community level (P < 0.01) along with differences in abundances of 89 operational taxonomic units (P < 0.05). Collectively, these data demonstrate that sows with greater POP risk have differential serum metabolites and vaginal microflora. Additionally, an initial and novel characterization of the sow vaginal microbiome was determined.

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Detection of Genetically Modified Corn (Bt176) in Spiked Cow Blood Samples by Polymerase Chain Reaction and Immunoassay Methods

Journal of AOAC International ◽

10.1093/jaoac/88.2.654 ◽

2005 ◽

Vol 88 (2) ◽

pp. 654-664 ◽

Cited By ~ 4

Author(s):

Laetitia Petit ◽

Fabienne Baraige ◽

Yves Bertheau ◽

Philippe Brunschwig ◽

Annick Diolez ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Whole Blood ◽

Limit Of Detection ◽

Enzyme Linked Immunosorbent Assay ◽

Rrna Gene ◽

Chain Reaction ◽

Cry1ab Protein ◽

Blood Samples ◽

Polymerase Chain ◽

Whole Blood Samples

Abstract The fate of DNA and protein transgenic sequences in products derived from animals fed transgenic crops has recently raised public interest. Sensitive molecular tests targeting the Bt176 genetic construct and the transgenic Cry1Ab protein were developed to determine whether plant sequences, especially transgenic sequences, are present in animal products. A protocol for total DNA extraction and purification from cow whole blood samples was first drawn up and assessed by spiking with known amounts of DNA from Bt176 maize. The limit of detection for transgenic sequences (35S promoter and Bt176-specific junction sequence) was determined by both the polymerase chain reaction–enzyme-linked immunosorbent assay (PCR–ELISA) and the 5′-nuclease PCR assay. Four additional PCR systems were built to substantiate the results. The first detects a mono-copy maize-specific sequence (ADH promoter). Two others target multi-copy sequences from plant nucleus (26S rRNA gene) and chloroplast (psaB gene). The last one, used as a positive control, targets a mono-copy animal sequence (αs1-casein gene). Both methods detected a minimum spiking at 25 copies of Bt176 maize/mL in 10 mL whole blood samples. The sandwich ELISA kit used detected down to 1 ng transgenic Cry1Ab protein/mL spiked whole blood.

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Bean pod mottle virus Spread in Insect-Feeding-Resistant Soybean

Plant Disease ◽

10.1094/pdis-94-2-0265 ◽

2010 ◽

Vol 94 (2) ◽

pp. 265-270 ◽

Cited By ~ 10

Author(s):

Margaret G. Redinbaugh ◽

Julio E. Molineros ◽

Jean Vacha ◽

Sue Ann Berry ◽

Ronald B. Hammond ◽

...

Keyword(s):

Mixed Model ◽

Linear Mixed Model ◽

Seed Quality ◽

Mottle Virus ◽

Enzyme Linked Immunosorbent Assay ◽

Growth Stages ◽

Insect Feeding ◽

Inoculum Level ◽

Bean Pod Mottle Virus ◽

Soybean Genotypes

Bean pod mottle virus (BPMV) infection reduces yield and seed quality in soybean. To test the hypothesis that virus incidence and movement within plots would be reduced in soybean with resistance to feeding by the virus' bean leaf beetle (Cerotoma trifurcata) vector, BPMV spread was evaluated in five soybean genotypes at two inoculum levels over 2 years at two locations in Ohio. Soybean genotypes included two insect-feeding-susceptible genotypes (Williams 82 and Resnik), two insect-feeding-resistant, semidwarf genotypes (HC95-15 and HC95-24), and an insect-feeding-susceptible, semidwarf genotype (Troll). BPMV incidence was assessed in individual plants at growth stages R5/R6 and R7/R8 using enzyme-linked immunosorbent assay. Beetle feeding was visually assessed in 2004. Data for infection of individual plants were analyzed using a generalized linear mixed model, with a binomial distribution and logit-link. Within plots, BPMV incidence was highest in Resnik and Williams 82 and significantly lower in Troll. Incidence in HC95-15 was not significantly different than in Williams 82 and Resnik but incidence in HC95-24 was lower than in Resnik. BPMV incidence was also significantly (P < 0.05) affected by year, location, inoculum level and sampling date, with increasing incidence over time and higher incidence at the higher inoculum level. Beetle feeding damage was affected by the interaction of location–genotype. Significant spatial aggregation of infected plants was found for most plots but aggregation was independent of host genotype and inoculum level. Although the results indicate that BPMV infection varied by genotype, they do not support the hypothesis that insect-feeding resistance is sufficient to reduce the incidence and spread of BPMV.

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The Value of Vaginal Microbiome in Patients with Endometrial Hyperplasia

Journal of Healthcare Engineering ◽

10.1155/2021/4289931 ◽

2021 ◽

Vol 2021 ◽

pp. 1-7

Author(s):

Hu Zhang ◽

Qiao Feng ◽

Zhanpeng Zhu ◽

Haiyan Dai ◽

Hua Hu

Keyword(s):

Endometrial Hyperplasia ◽

Abnormal Uterine Bleeding ◽

Rrna Gene ◽

Vaginal Microbiome ◽

Bacterial Populations ◽

Α Diversity ◽

Significant Difference ◽

Proliferative Endometrium ◽

Microbial Dna ◽

Insight Into

Objective. To investigate the profiles of the vaginal microbiome in patients with endometrial hyperplasia and to explore the potential value of vaginal microbiome in the diagnosis of endometrial hyperplasia. Materials/Methods. 26 patients suffering from abnormal uterine bleeding (AUB) with thickened endometrium revealed by transvaginal ultrasonography were enrolled. Based on pathology, 12 patients with endometrial hyperplasia were classified as the Veh group and 14 patients with proliferative endometrium were classified as the Vne group. The vaginal samples were collected for the presence of microbial DNA by high-throughput next-generation sequencing of the 16S rRNA gene. The α-diversity and ß-diversity of vaginal microbiome were analyzed and compared between bacterial populations. The ROC curve was made to evaluate the feasibility of flora as a biomarker. Results. The diversity of vaginal microbiome in the Veh group was significantly lower than that in the Vne group ( P < 0.05 ). Lactobacillus was the most represented genus in the Veh group. The study’s t-test between the two groups showed that Lactobacillus has the only significant difference in the abundance of the first 15 genera ( P < 0.01 ). ROC analysis of the abundance of Lactobacillus showed that the area of AUC was 0.83, the sensitivity was 93.00%, and the specificity was 75.00%. Conclusion. The study offers insight into the nature of the vaginal microbiome and suggests that surveying the vaginal microbiota might be useful for detection of endometrial hyperplasia.

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Evaluation of the Precision Xtra meter for monitoring blood β-hydroxybutyrate concentrations in late-gestation ewes

Journal of Veterinary Diagnostic Investigation ◽

10.1177/1040638718819688 ◽

2018 ◽

Vol 31 (1) ◽

pp. 17-22 ◽

Cited By ~ 1

Author(s):

Niorn Ratanapob ◽

John VanLeeuwen ◽

Shawn McKenna ◽

Maureen Wichtel ◽

Juan C. Rodriguez-Lecompte ◽

...

Keyword(s):

Mixed Model ◽

Linear Mixed Model ◽

Point Of Care ◽

Standard Test ◽

Late Gestation ◽

Medical Decision ◽

Standard Laboratory ◽

Limits Of Agreement ◽

Serum Samples ◽

Blood Samples

Blood samples were collected from late-gestation ewes to determine the agreement of a point-of-care (POC) Precision Xtra meter and a standard laboratory test for β-hydroxybutyrate (BHBA). Fresh whole blood samples were immediately tested with the POC instrument, and serum samples were analyzed with a standard commercial biochemical analyzer. Ewes were classified as having ketonemia if their BHBA concentrations were ≥800 µmol/L. Scatter plots, paired t-tests, Bland–Altman limits of agreement, and Gwet AC1 tests were used to compare results. The 2 tests had very good agreement. The values between instruments were not statistically different based on paired t-tests ( p = 0.312). The intercept and slope of a linear mixed model, containing the standard test results as an outcome and the POC meter results as a predictor, were 0.02 (95% CI: 0.00, 0.04) and 0.98 (95% CI: 0.96, 1.01), respectively. When the samples were classified into ketonemic classes (non-ketonemic and ketonemic) based on BHBA concentrations obtained from each test, the Gwet AC1 statistic was 0.94 (95% CI: 0.91, 0.97; p < 0.001). The ketosis classification agreed in 95% of samples. Based on the Bland–Altman plot and limits of agreement, the optimal cutoff to diagnose ketonemia with the POC meter was 1,000 µmol/L, which is 200 µmol/L higher than the laboratory BHBA medical decision limit. The Precision Xtra meter provided excellent correlation and substantial agreement with the standard laboratory technique for measuring blood BHBA in late-gestation ewes.

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243 Digestibility, fecal characteristics, and blood parameters of adult dogs fed high fat diets

Journal of Animal Science ◽

10.1093/jas/skz258.127 ◽

2019 ◽

Vol 97 (Supplement_3) ◽

pp. 61-61

Author(s):

Logan R Kilburn

Keyword(s):

Health Status ◽

Fixed Effects ◽

Mixed Model ◽

Linear Mixed Model ◽

Body Condition Score ◽

Energy Requirement ◽

Blood Samples ◽

Linear Discriminant ◽

Commercial Diet ◽

Fat Level

Abstract The objective was to evaluate the effects of feeding adult dogs increasing levels of fat in low carbohydrate diets on apparent digestibility, fecal characteristics, and health status. Eight adult beagles were used in a 4x4 replicated Latin square design. Dogs were randomly assigned to 1 of 4 fat levels within a phase: 32% (T1), 37% (T2), 42% (T3), and 47% (T4). Fat levels were adjusted with inclusion of canola oil to a commercial diet. Each dog was fed to exceed their energy requirement based on NRC recommendation. Dogs were fed their respective diet twice daily for 15 days. Total fecal collections were done d 11–15 and blood samples were taken on d 15. Data were analyzed using PROC MIXED of SAS (diet and room as fixed effects; period and dog were random). Covariates of baseline, initial body weight, and/or initial body condition score were dependent on trait. Digestibility improved linearly with fat level for dry matter (P = 0.021), organic matter (P = 0.019), and fat (P = 0.001). Fecal output decreased as levels of fat increased in the diet (P = 0.002). There was no effect of fat level on stool quality or short chain fatty acid and ammonia concentrations in fecal samples (P > 0.1). Blood samples were analyzed for complete blood counts and chemistry profiles. All parameters remained within normal reference intervals. Blood urea nitrogen levels decreased with increased fat level (P = 0.03). No significant differences were seen in canine pancreatic lipase immunoreactivity. In addition, serum metabolomics were analyzed among T1 and T4. Metabolite data were log2-transformed and changes among diets were detected using a linear mixed model, the metabolomic profile among diets was characterized using linear discriminant analysis. 19 metabolites, primarily phosphatidylcholines, were statistically significant among treatments. Increase of dietary fat improved digestibility, did not disrupt fecal characteristics, and maintained the health status of each dog.

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