Rat Feces Research Articles

Effects of ultra-processed foods on the liver: insights from gut microbiome and metabolomics studies in rats

PurposeHigh consumption of Ultra-processed foods (UPF) have been identified as a potential risk factor for Non-alcoholic fatty liver disease (NAFLD). Nevertheless, there is limited empirical evidence regarding the impact of UPF, which are typical combination of processed foods, on liver health through alterations in gut microbiota and metabolic processes. We aim to examine the potential impact of UPF on liver health and to explore the role of gut microbiota and metabolites.MethodsThis study used Sprague–Dawley rats to mimic modern UPF diets for 90 days. Some serum biochemical indices, inflammatory factors, oxidative stress markers, hematoxylin–eosin (HE) staining of the liver, 16S ribosomal RNA (rRNA) and Liquid chromatography-mass spectrometry (LC–MS) of rat feces were detected.ResultsThe UPF diet-induced simple steatosis of the liver in rats without affecting the levels of IL-6, GSH, MDA, and SOD. Additionally, it modified the gut microbiota, increasing potentially harmful bacteria, such as norank_f__Desulfovibrionaceae and Staphylococcus, while also elevating the relative abundance of potentially beneficial bacteria, including Dubosiella and Allobaculum. Furthermore, the consumption of UPF led to a metabolomic disorder characterized by disruptions in the sphingolipid signaling pathway, sulfur relay system, and arachidonic acid metabolism.ConclusionIn conclusion, the findings of this study indicate that the consumption of UPF influences the development of simple hepatic steatosis, potentially through alterations in gut microbiota and metabolomics.

Metabolic characteristics of saponins from Panax notoginseng leaves biotransformed by gut microbiota in rats.

Saponins are responsible for the clinical effects of Panax notoginseng leaves, which are traditionally produced as the single herb resource of 'Qiye Shenan Pian' in Chinese patent medicine. In this study, the metabolic characteristics of PNLSs were explored in rat feces. PNLSs as well as their metabolites were analyzed by ultra-performance liquid chromatography tandem/quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS/MS). Subsequently, seventy-five metabolites were tentatively identified in the control group mainly due to the deglycosylation and dehydration biopathways, but only twenty low yields were determined in the pseudo-germ-free (GF) group. Ginsenoside compound K was the predominant metabolite in the control group. The data presented that gut microbiota played a pivotal role in the metabolic kinetics of PNLSs.

Suplementação oral de prebiótico, mananoligossacarídeo, mitiga disbiose intestinal em ratos jovens cronicamente expostos ao tabagismo passivo

In humans, passive smoking has been shown to alter the microbiota and increase the risk of infections by pathogens and/or opportunistic bacteria. The objective of this study was to evaluate the effect of MOS supplementation on the concentration of E. coli in the feces of passive smoking rats. Sixty young rats (23 days) were randomly allocated into four groups (n=15): supplemented or not with MOS in the diet; and chronically exposed or not to cigarette smoke. Exposure to cigarette smoke was performed twice a day for 180 days and fecal samples were collected four times (days 0, 60, 120, and 180). After fecal collection, bacterial populations were amplified by real-time PCR. Data means were analyzed using one-way ANOVA followed by Student-Newman-Keuls post-hoc analysis, considering 5% as the level of significance. The results revealed that MOS supplementation significantly (P<0.05) reduced the E. coli population resulting from chronic exposure to cigarette smoke. The mean results of the group of rats supplemented with MOS and exposed to cigarette smoke did not differ (P>0.05) from the groups not exposed to cigarette smoke and supplemented or not with MOS. The results allow us to conclude that MOS supplementation mitigates the chronic effect of cigarette smoke on the concentration of E. coli bacteria in young rats, as a preclinical model.

Studies on the Stability and Microbial Biotransformation of Five Deschloroketamine Derivatives as Prerequisite for Wastewater-Based Epidemiology Screening.

Wastewater (WW)-based epidemiology (WBE) is a powerful tool for screening and surveillance of drugs (of abuse) or new psychoactive substances (NPSs) in larger population. Since the drug market changes frequently, it is crucial for WBE to define screening and surveillance biomarkers considering drug metabolism and (microbial) stability. The aims of the presented work were first to identify metabolites, potentially serving as a WBE biomarker of five deschloroketamine derivatives (DCKDs) in rat feces samples after oral administration in addition to already known urinary metabolites, and second to elucidate the microbial biotransformation and WW stability of five DCKDs and their metabolites detected in urine and feces. Microbial biotransformation and stability of DCKD and their metabolites in WW were assessed by incubating parent compounds at 0.1 mg/L or rat urine or rat feces samples in freshly collected, untreated, influent WW over a period of 24 h. All samples were analyzed using liquid chromatography-high-resolution tandem mass spectrometry. All parent compounds, seven Phase I, and one Phase II metabolite were detected in rat feces samples. After WW incubations, all tested DCKD and their metabolites were still detectable at least in trace amounts, but particularly, peak areas of the Phase II N- and O-glucuronides showed a markable decrease. This is in line with previous findings where Phase II conjugates were identified to be unstable in WW and thus not recommended as a WW biomarker. Hence, incubations demonstrated that the five DCKD and most of their metabolites were sufficiently stable in WW influent and can thus be used as analytical targets in the context of WBE.

Total Alkaloids of Rhizoma Corydalis regulates gut microbiota and restores gut immune barrier to ameliorate cognitive dysfunction in diabetic rats.

Given the widespread dysbiosis of gut microbiota in patients with T2DM, it has been found that the microbiota-gut-brain axis plays an influential regulatory role in diabetic cognitive dysfunction, and improving gut dysbiosis may be a potential strategy for treating diabetic cognitive dysfunction. Total Alkaloids of Rhizoma Corydalis (TAC) is the main active component extracted from Rhizoma Corydalis. Pharmacological studies have demonstrated its significant pharmacological effects on the cardiovascular and cerebrovascular systems, and berberine, the main component of TAC, has a certain regulatory effect on gut microbiota. Rats were randomly divided into Control group, Model group, TAC-low group, TAC-mid group and TAC-high group. Cognitive function of diabetic rats was evaluated through behavioral testing using the Morris water maze experiment. The relative abundance of gut bacteria in rat feces was determined via 16S rRNA analysis. IHC and Western blot techniques were employed to assess IL-22, IL-23, Reg3g, ZO-1, occludin 1 expression in the colon tissue; GPX4, xCT, NLRP3, Caspase-1 p20, GSDMD-N were detected in the hippocampus. The cognitive function of diabetic rats decreased significantly. TAC demonstrated a significant reduction in inflammatory factors in serum, hippocampus, and colon, thus alleviating inflammation. Additionally, it effectively decreased ferroptosis induced by NLRP3 and reduced pathological damage in the hippocampus of diabetic rats. After treatment, the differential microbiota such as Lachnoclotridium and Bacteroides. TAC improved gut barrier permeability and integrity in rats while remodeling gut mucosal homeostasis. Moreover, pyroptosis and ferroptosis caused by the inflammatory cascade in the rat hippocampus were also significantly inhibited. The combination of high lipid and high glucose with STZ can result in gut microbiota disturbance, damage gut immune barrier, decreased gut mucosal permeability and integrity, aggravated gut inflammation, further spread inflammatory factors to brain tissue, cause inflammatory cascade reaction of encephalopathy, and ultimately resulting in neuronal ferroptosis and cognitive dysfunction in diabetes mellitus. Our study suggests that TAC may regulate gut microbiota, restore gut immune homeostasis, improve gut barrier permeability and integrity, inhibit brain tissue inflammatory cascade, reduce neuronal ferroptosis, and thus improve diabetes. This provides new targets for its treatment strategy.

Comparative study on metabolites in rat liver microsomes, urine, feces and bile between Shuganning Injection and Scutellariae Radix extract

This study aims to compare the metabolic differences of baicalin and its analogues between Shuganning Injection and Scutellariae Radix extract. Twelve SD rats were randomly divided into a Shuganning Injection group and a Scutellariae Radix extract group, with 6 rats in each group. Their liver microsomes were incubated with the drugs, and then the samples were collected. Ultra performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry(UPLC-Q-Exactive Orbitrap-MS) was used to analyze the prototype components and metabolites of the drugs in liver microsomes of each group. Another 12 SD rats were also divided into a Shuganning Injection group and a Scutellariae Radix extract group, with 6 rats in each group. The rats were administrated with 4.2 mL·kg~(-1) Shuganning Injection or Scutellariae Radix extract by tail vein injection. After 48 h, the rat urine, feces, and bile were collected, and UPLC-Q-Exactive Orbitrap-MS was used to analyze the prototype components and metabolites in each biological sample. The results showed that 5 prototype components and 8 metabolites of Shuganning Injection and Scutellariae Radix extract were identified in liver microsomes. A total of 5 prototype components were identified in rat urine, feces, and bile separately. Fifteen metabolites were identified in the urine, 9 metabolites in the feces, and 12 metabolites in the bile. The differences of metabolic pathways and number of metabolites of baicalin were compared between Shuganning Injection and Scutellariae Radix extract. For both Shuganning Injection and Scutellariae Radix extract, the metabolites of baicalin or baicalein in rat liver microsomes, urine, bile, and feces were mainly formed glucuronic acid conjugates, and there were a small amount of glucose conjugates and methylation products. Differences were found in the number and types of metabolites of baicalin in urine samples between Shuganning Injection and Scutellariae Radix extract, indicating that differences existed in metabolism between the two. This suggests that the other components in the formula lead to changes of metabolites in vivo.

Mechanism of inflammatory microecological response to TAS2R14/SIgA/TSLP in regulating epithelial cell barrier in cold asthma rats through lung-gut axis by using Shegan Mahuang Decoction and bitter and purging Chinese herbs

This study aimed to investigate the mechanism by which Shegan Mahuang Decoction(SGMH) and its bitter Chinese herbs(BCHs) regulated the lung-gut axis through the bitter taste receptor 14(TAS2R14)/secretory immunoglobulin A(SIgA)/thymic stromal lymphopoietin(TSLP) to intervene in the epithelial cell barrier of cold asthma rats. Fifty SD rats were randomly divided into the following five groups: normal group, model group, dexamethasone group, SGMH group, and BCHs group. A 10% ovalbumin(OVA) solution was used to sensitize the rats via subcutaneous injection on both sides of the abdomen and groin, combined with 2% OVA atomization and cold(2-4 ℃) stimulation to induce a cold asthma model in rats. The SGMH, BCHs, and dexamethasone groups were given corresponding treatments by gavage and nebulization, while the normal and model groups received normal saline by gavage and nebulization. After the final stimulation, pathological changes in the lung and intestine tissues were observed using hematoxylin-eosin(HE) and periodic acid-Schiff(PAS) staining. Lung function was assessed by measuring the ratio of forced expiratory volume in the first second to forced vital capacity(FEV1/FVC), the ratio of the average flow rate at 25%-75% of forced vital capacity to foned vital capacity(FEV25%-75%/FVC), the peak expiratory flow(PEF), and pulmonary resistance(RL). The levels of IL-4, IL-5, IL-13, and TNF-α in serum, and sIgA in serum, intestinal, and bronchial mucosa were detected by enzyme-linked immunosorbent assay(ELISA). The expression of TAS2R14 protein in lung tissue was detected by Western blot(WB). The content of short-chain fatty acids(SCFAs) in rat feces was determined by gas chromatography-mass spectrometry(GC-MS). The effect of TAS2R14/TSLP on lipopolysaccharide(LPS)-induced inflammation in epithelial cells in the BCHs group was observed, and the expression of TAS2R14 and TSLP in cells was detected by WB. Compared with the normal group, the model group showed reduced water intake, diet, and body weight, increased infiltration of inflammatory cells in the lung and intestinal tissues, goblet cell hyperplasia, significantly decreased FEV1/FVC, FEV25%-75%/FVC, and PEF, and significantly increased RL. Moreover, serum levels of IL-4, IL-5, IL-13, and TNF-α were elevated, and sIgA levels in serum, intestine, and bronchial mucosa were significantly decreased. TAS2R14 expression in lung tissues was inhibited, and the content of acetic acid, propionic acid, and butyric acid in feces was significantly reduced. In the LPS group, TSLP expression increased, and TAS2R14 expression decreased. Compared with the model group, the general condition of rats in the SGMH and BCHs groups improved, with reduced infiltration of inflammatory cells and goblet cell hyperplasia in the lung and intestinal tissues. FEV1/FVC, FEV25%-75%/FVC, and PEF significantly increased, and RL significantly decreased. Serum levels of IL-4, IL-5, IL-13, and TNF-α decreased, while sIgA levels in serum, intestine, and bronchial mucosa significantly increased, and TAS2R14 expression was activated in lung and intestinal tissues. The content of acetic acid, propionic acid, and butyric acid in feces significantly increased. Compared with the model group, the BCHs group and the agonist group showed inhibited TSLP expression and increased TAS2R14 expression. The results showed that both SGMH and BCHs could reduce lung and intestinal inflammatory reactions, improve lung function, and regulate the content of intestinal SCFAs in asthmatic rats. There was no significant difference in TAS2R14 protein expression between the SGMH and BCHs groups, indicating that the clinical efficacy of BCHs may be related to the activation of the bitter receptor TAS2R14 and the regulation of immune inflammatory mediators in lung and intestinal epithelial cells.

Dietary Fiber with Functional Properties Counteracts the Thwarting Effects of Copper Nanoparticles on the Microbial Enzymatic Activity and Short-Chain Fatty Acid Production in the Feces of Rats

Dietary Fiber with Functional Properties Counteracts the Thwarting Effects of Copper Nanoparticles on the Microbial Enzymatic Activity and Short-Chain Fatty Acid Production in the Feces of Rats

To explore pharmacodynamic substances and mechanism of Xuanfei Baidu Decoction on LPS-induced ALI / ARDS by pharmacochemistry and metabolomics

Ethnopharmacological relevanceXuanfei Baidu Decoction (XFBD) is made up of five classic prescriptions, which is composed of 13 traditional Chinese medicines, which have the effects of dispersing lung and resolving dampness, clearing heat and evil, purging lung and detoxification. It is used for epidemic diseases caused by damp toxin obstructing lung. In this paper, the endogenous and exogenous metabolites of ALI/ARDS rats after oral administration of XFBD were studied and the material basis and metabolic pathway of XFBD in relieving ALI/ARDS were explained. Materials and methodsIn this study, UPLC-Q-TOF/MS qualitative analysis method was established, and plasma-urine-feces pharmacochemistry was used to identify metabolites in plasma, urine and feces after oral administration of XFBD in rats. RT-PCR and immunohistochemistry were used to study the expression of CYP protein in XFBD and monomeric compounds. In addition, the functional mechanism of XFBD in ALI/ARDS rats was elucidated by non-targeted metabolomics. ResultsA total of 77 prototype components and 389 metabolites in plasma, urine and feces were identified by exogenous components, mainly including oxidation, reduction, hydrolysis, glucuronidation, sulfation and other reactions. The results of RT-PCR and immunohistochemistry showed that the activity of CYP was inhibited under the pathological state of ALI/ARDS. At the same time, XFBD and its monomeric compounds can change the metabolic process of drugs in vivo by inducing or inhibiting the activity of CYPs. The metabolic process of drugs in vivo is the result of the combined action of different CYPs. In addition, a total of 33 differential metabolites were identified in plasma, 45 in urine and 14 in feces, which were mainly related to the synthesis and degradation of ketone bodies, phenylalanine metabolism, tyrosine metabolism, histidine metabolism, butyric acid metabolism and other metabolic pathways. ConclusionsThis study conducted a relatively scientific and systematic analysis of XFBD. A UPLC-Q-TOF/MS qualitative analysis method was established to identify 77 prototype components and 389 metabolites in plasma, urine and feces of rats after oral administration of XFBD.An ARDS rat model was established to analyze the pharmacokinetic differences of XFBD in normal and ARDS model rats and its regulation effect on CYPs.Non-targeted metabolomics was used to identify the pattern recognition of ARDS pathological model, the diagnosis of disease and the intervention effect of XFBD on ARDS. Preliminary discussion was conducted to provide a theoretical basis for clinical rational drug use.

Polypeptides from Sea Cucumber Acaudina molpadioides Ameliorate Glucolipid Metabolism Disorder and Reverse Gut Microbiota Dysbiosis in Type 2 Diabetic Rats

ABSTRACT The therapeutic effects of polypeptides isolated from sea cucumber Acaudina molpadioides (AP) on the intestinal flora and glycolipid metabolic disturbance in high-fat diet/STZ-induced T2DM rats were investigated. The results showed that AP intervention notably ameliorated blood glucose, lipid levels, and pancreas injuries in T2DM rats. The diversity of intestinal microbial communities in T2DM rats was also enhanced by AP. Moreover, microbially derived short-chain fatty acids concentrations markedly improved in the feces of diabetic rats after AP treatment. These results suggest that AP played a protective role in T2DM rats by regulating glucose, lipid metabolism, and altering intestinal microbial.

Study on regulating AQP1, AQP3, AQP4, 5-HT, NOS1 in slow transit constipation rats by Liqi Tongbian mixture.

Liqi Tongbian is a traditional Chinese medicine (TCM) preparation that contains herbs that may treat slow transit constipation (STC). Atractylodes macrocephala, Astragalus membranaceus, Fructus aurantii, radish seed, uncooked Polygonum multiflorum, and Agastache rugosa were included in the formula for their unique qualities. The control of water transfer in the colon is greatly influenced by aquaporin 3 (AQP3). Based on this, the Liqi Tongbian mixture was used to detect the concentrations of aquaporins (AQPs), 5-HT and nitrix oxide synthase 1 (NOS1) in STC rats, and explore its effect, in order to provide a theoretical basis for the remedy of STC with TCM. Zhejiang University of Traditional Chinese Medicine provided 32 three-week-old Sprague Dawley rats of SPF-grade. The pairs licensed under SYXK (Zhejiang) 2021-0012 were kept at 20-25°C and humidity of 50-65%. The compound diphenoxylate caused constipation in the control, model, Liqi laxative (LQTB), and mosapride groups. The Liqi laxative rats were administered a mixture of traditional Chinese herbs after modeling, while mosapride was given to the other group. The levels of 5-HT, NOS1 and AQPs were tested in the feces and intestinal tissues. Comparing the condition of rat feces, it was found that the model group had significantly lower overall bulk, score and particles within 24 h compared to the control group. In comparison to mosapride, LQTB performed better. The model group had higher levels of 5-HT and NOS1 in intestinal tissue, while the LQTB and mosapride groups had decreased levels of these AQPs. LQTB had lower levels of AQP1, AQP3 and AQP4 than mosapride, while the model group had higher levels of these AQPs. Liqi Tongbian mixture works better than mosapride in improving constipation symptoms in rats with STC, and its mechanism is related to regulating the level of intestinal AQPs and neurotransmitters.

Metabolic fate of the natural anticancer agent cucurbitacin B: an LC-MS/MS-enabled profiling of its major phase I and II conjugates in vivo.

Cucurbitacin B (CuB) is a natural triterpenoid with diverse pharmacological effects including potent anticancer activity. However, its oral bioavailability is hampered by limited metabolism in vivo. We characterized CuB's in vivo metabolism in rats to uncover bioactive metabolites retaining therapeutic potential, using a robust UHPLC-Q-TOF-MS/MS workflow. This workflow combined molecular networking, fragmentation filtering, and mass defect filtering to identify CuB metabolites in rat urine, plasma, and feces following oral administration. Thirteen metabolites were identified and seven were confirmed. Major phase I transformations involved hydrolysis, reduction, epoxidation, and amination. Phase II conjugation included cysteine, glutathione, glucuronide, and gluconic acid conjugates. Notably, one of the main metabolites formed was the cysteine conjugate CuB-Cys. CuB-Cys maintained similar in vitro antiproliferative activity to CuB on HepG2, MCF-7, and PANC-1 cancer cell lines. However, it demonstrated lower cytotoxicity towards non-cancerous L02 cells, highlighting improved therapeutic selectivity. Mechanistically, CuB-Cys induced greater apoptotic signaling in HepG2 cells than CuB via enhanced caspase activation and disrupted BAX-Bcl-2 balance. This represents the first systematic characterization of CuB's in vivo metabolic pathway. The identification and confirmation of CuB-Cys provide insight for drug development efforts aiming to maintain therapeutic efficacy while reducing toxicity, via metabolite-based approaches. Our findings shed light on strategies for improving CuB's clinical potential.

Sex differences and testosterone interfere with the structure of the gut microbiota through the bile acid signaling pathway.

The gut microbiome has a significant impact on human wellness, contributing to the emergence and progression of a range of health issues including inflammatory and autoimmune conditions, metabolic disorders, cardiovascular problems, and psychiatric disorders. Notably, clinical observations have revealed that these illnesses can display differences in incidence and presentation between genders. The present study aimed to evaluate whether the composition of gut microbiota is associated with sex-specific differences and to elucidate the mechanism. 16S-rRNA-sequencing technology, hormone analysis, gut microbiota transplantation, gonadectomy, and hormone treatment were employed to investigate the correlation between the gut microbiome and sex or sex hormones. Meanwhile, genes and proteins involved bile acid signaling pathway were analyzed both in the liver and ileum tissues. The composition and diversity of the microbiota from the jejunum and feces and the level of sex hormones in the serum differed between the sexes in young and middle-aged Sprague Dawley (SD) rats. However, no similar phenomenon was found in geriatric rats. Interestingly, whether in young, middle-aged, or old rats, the composition of the microbiota and bacterial diversity differed between the jejunum and feces in rats. Gut microbiota transplantation, gonadectomy, and hormone replacement also suggested that hormones, particularly testosterone (T), influenced the composition of the gut microbiota in rats. Meanwhile, the mRNA and protein level of genes involved bile acid signaling pathway (specifically SHP, FXR, CYP7A1, and ASBT) exhibited gender-specific differences, and T may play a significant role in mediating the expression of this pathway. Sex-specific differences in the structure of the gut microbiota are mediated by T through the bile acid signaling pathway, pointing to potential targets for disease prevention and management techniques by indicating that sex differences and T levels may alter the composition of the gut microbiota via the bile acid signaling pathway.

Rapid Characterization of the Potential Active of Sinomenine in Rats by Ultra-High-Performance Liquid Chromatography-Quadrupole-Exactive Orbitrap Mass Spectrometry and Molecular Docking.

Sinomenium acutum (Thunb.) Rehd. et Wils is widely used in the treatment of rheumatoid arthritis, with its alkaloid compound sinomenine (SIN) being renowned for its significant anti-inflammatory properties. However, despite its widespread application, the in vivo anti-inflammatory mechanisms and metabolic pathways of SIN remain incompletely understood. This study established a rapid and reliable method based on an ultra-high-performance liquid chromatography method coupled with Quadrupole-Exactive Orbitrap mass spectrometry and molecular docking to identify and characterize SIN and 69 metabolites in rat plasma, urine, and feces, revealing primary metabolic pathways of hydroxylation, demethylation, sulfation, and glucuronidation. Molecular docking results revealed that phase I reactions, including dedimethylation, demethylation, dehydrogenation, and dihydroxylation, along with their composite reactions, were pivotal in influencing SIN's in vivo anti-inflammatory activity. M28, M36, and M59 are potentially the most anti-inflammatory active metabolites of SIN in vivo. This comprehensive analysis unveils SIN's metabolic pathways, offering insights into its biological processes and suggesting a novel approach for exploring active drug constituents. These findings pave the way for further understanding SIN's anti-inflammatory mechanisms, contributing significantly to the development of new therapeutic strategies.

Molecular Diagnosis Of Clostridium piliforme, The Causative Agent Of Tyzzer’s Disease In Mice And Rats

Tyzzer’s disease, caused by Clostridium piliforme (C. piliforme), occurs in many species of mammals mainly in rodents, rabbits and foals is an acute, epizootic bacterial disease that can result in mortality. In our study, C. piliforme was investigated in feces samples from mice and rats in a laboratory experimental animals center by PCR technique. C. piliforme was detected in 44 (83.01%) of a total of 53 mouse feces of different ages. Of the mice found positive, 28 (73.6%) were female and 16 (88.8%) were male. C. piliforme was detected in 40 (90.9%) of a total of 44 rats feces of different ages. Of the rats found positive, 25 (96.1%) were female and 15 (83.3%) were male. C. piliforme, the causative agent of Tyzzer's disease, was detected at a high rate in mice and rats examined by the classical PCR method. When evaluated in terms of gender, 28 (73.6%) of the 40 mice positive for Tyzerr's disease were female and 16 (88.8%) were male; While 25 (96.1%) of the rats were female, 15 (83.3%) were male. As a result, Tyzerr's disease caused by C. piliforme was detected at a high rate in mice and rats by classical PCR. When evaluated by age, the disease was detected to be higher in mice at the ages of 4, 5 and 8, and in rats at the ages of 4, 5, 6 and 7. Due to the reliability of the analyses and the high risk of transmission to other animals in centers where laboratory animals are raised, Tyzzer's disease should be checked and necessary precautions should be taken to prevent the disease.

The combination of oxalic acid and uric acid degrading probiotic from traditional Chinese fermented food reduces calcium accumulation and prevents kidney stones formation in rats

Kidney stones, often resulting from a complex interplay of factors including oxalic acid, uric acid, and calcium. Current treatments lack comprehensive consideration of these factors and come with side effects. In this study, Lactobacillus paracasei GR-10 and Pediococcus acidilactici GR-11 were isolated from the fermented food “Jiangshui”. GR-10 demonstrated the ability to degrade 62.70% of oxalic acid (10 mM) within 24 h, while GR-11 degraded 28.60% of uric acid (4 mM). Combined treatment with GR-10 and GR-11 significantly reduced the number of stones in rats with ethylene glycol-induced kidney stones. This reduction was accompanied by a 57.19% decrease in urinary oxalate and a 52.80% decrease in serum uric acid levels. Simultaneously, urinary calcium level decreased by 27.18%. Inflammatory markers and indicators of oxidative stress associated with kidney stones were restored. Furthermore, the combined treatment effectively regulated the microbiota imbalance induced by kidney stones, resulting in reduced purine nucleoside accumulation in rat feces. This study offers promising insights into probiotics as an adjunct therapy for the prevention of kidney stones in the future.

Gastrointestinal Parasites in Non-Human Primates in Zoological Gardens in Northern Italy.

Non-human primates (NHPs) host a variety of helminth and protist parasites that are able to cause infection in humans. Gastrointestinal parasites in NHPs living in two zoological gardens of Northern Italy were studied. An total of 96 faecal pools were collected from 26 groups of NHPs. The mini-Flotac method was applied to fecal samples to detect gastrointestinal helminthiases, while the detection of the protists Cryptosporidium spp., Blastocystis sp. and Giardia duodenalis was performed by targeting SSU rRNA through nested PCR and real-time PCR; they were further studied by sequencing the same gene for Blastocystis and βgiardine and triosephosphate isomerase (TPI) genes for Giardia. Twenty-two out of the 96 examined fecal pools (22.9%) were positive for one or more helminth species, including Hymenolepis diminuta, Trichurid, Capillariid and Strongylid eggs. All samples were negative for Cryptosporidium spp., while 16/26 (61.5%) animals were positive for G. duodenalis in the real-time PCR; the sequences obtained assigned them all to sub-assemblage BIV. Blastocystis sp. was detected in 22/26 of the NHPs (84.6%); molecular analyses attributed the isolates to ST 4, allele 92. Analyses of the feces of sympatric rats revealed the presence of the same allele, as well as of Hymenolepis diminuta eggs, raising concern about their role as parasite reservoirs in the facilities.

Excretion characteristics of main compounds of Yigong San in urine, feces, and bile of rats.

Yigong San (YGS) is a traditional Chinese medicine formula used for pediatric anorexia, chronic atrophic gastritis, and irritable bowel syndrome. In this study, the excretion of eight main compounds, including liquiritin; isoliquiritin; hesperidin;ginsenosides Rb1, Re, and Rg1; and atractylenolides I and II, in rat urine, feces, and bile, was investigated by ultra-high performance liquid chromatography-tandem mass spectrometry. The results showed that the cumulative excretion rates of the compounds in rat urine, feces, and bile were 0.018-1.15%, 0.024-19.89%, and 0.0025-0.72%, respectively. Among the eight compounds detected, liquiritin was the richest in urine, and ginsenosides Re and Rg1 and atractylenolide I were mainly found in feces and bile. In summary, the main components of YGS are excreted via multiple approaches. Liquiritin is mainly through urine, whereas isoliquiritin;hesperidin;ginsenosides Rb1, Re,and Rg1; and atractylenolides I and II are mainly through feces. The excretion of these compounds in bile is usually positively correlated with that in feces. This study lays a foundation for further pharmacological research and application of YGS.

Metabolic profiling of triptophenolide in rat plasma, urine, bile and faeces after intragastric administration

Triptophenolide, a major diterpenoid extracted from Tripterygium wilfordii Hook. f., has been reported to possess significant anti-tumour, anti-androgen and anti-inflammatory activities. However, the metabolic fate of triptophenolide remains unknown. Therefore, this study focused on the metabolic profiling of triptophenolide in rat plasma, urine, bile and faeces following intragastric administration. An ultraperformance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry with combination of extracted ion chromatogram strategy based on 71 typical metabolic reactions was established to comprehensively profile the metabolites of triptophenolide. This strategy allowed for the identification of 17 metabolites from the biosamples. Reduction, oxidation, glucuronide conjugation, and hydroxylation were considered as its main metabolic pathways in vivo. The present study will be greatly helpful for the further pharmacological studies on triptophenolide and would provide valuable information for its clinical application.

Development of an integrated strategy for comprehensive characterization of Sinomenii Caulis extract and metabolites in rats based on UPLC/Q-TOF-MS

Sinomenii Caulis (SC), a commonly used traditional Chinese medicine for its therapeutic effects on rheumatoid arthritis, contains rich chemical components. At present, most studies mainly focus on sinomenine, with little research on other alkaloids. In this study, a comprehensive profile of compounds in SC extract, and biological samples of rats (including bile, urine, feces, and plasma) after oral administration of SC extract was conducted via ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF-MS). The fragmentation patterns and potential biotransformation pathways of six main types of alkaloids in SC were summarized, and the corresponding characteristic product ions, relative ion intensity, and neutral losses were obtained to achieve rapid classification and identification of complex components of SC from in vitro to in vivo. As a result, a total of 114 alkaloid compounds were identified, including 12 benzyl alkaloids, 4 isoquinolone alkaloids, 32 aporphine alkaloids, 28 protoberberine alkaloids, 34 morphinan alkaloids and 4 organic amine alkaloids. After administration of SC extract to rats, a total of 324 prototypes and metabolites were identified from rat plasma, urine, feces and bile, including 81 aporphines, 95 protoberberines, 117 morphinans and 31 benzylisoquinolines. The main types of metabolites were demethylation, hydrogenation, dehydrogenation, aldehydation, oxidation, methylation, sulfate esterification, glucuronidation, glucose conjugation, glycine conjugation, acetylation, and dihydroxylation. In summary, this integrated strategy provides an additional approach for the incomplete identification caused by compound diversity and low abundance, laying the foundation for the discovery of new bioactive compounds of SC against rheumatoid arthritis.