Fresh Feces Research Articles

Integration of gut microbiome and serum metabolome revealed the effect of Qing-Wei-Zhi-Tong Micro-pills on gastric ulcer in rats

Ethnopharmacological relevanceQing-Wei-Zhi-Tong Micro-pills (QWZT) is herbal compound used in the treatment of GU, whose functions include clearing the stomach and fire, softening the liver and relieving pain. However, its mechanistic profile on host intestinal microbiota and metabolism has not been determined. Aim of the studyThe present study aimed to observe the healing effect of QWZT on acetic acid-induced gastric ulcer in a rat model and to preliminarily elucidate its possible therapeutic mechanism from the perspective of host intestinal microbiota and metabolism. Materials and methodsThe Wistar male rats (7 weeks old; weight 180–200 g) were randomly divided into normal control group (NC), acetic acid-induced gastric ulcer group (GU), and QWZT treatment group (High dose: 1250 mg/kg/day, Middle dose: 625 mg/kg/day, Low dose: 312.5 mg/kg/day) of 6 rats each. An acetic acid-induced gastric ulcer rat model was constructed based on anatomical surgery. QWZT (High dose, Middle dose, and Low dose) was used to treat gastric ulcer rats for 7 days by gavage. At the end of treatment, the body weight, macroscopic condition of gastric tissue ulcers, pathological changes (HE staining), inflammatory factors, oxidative stress factors, and endocrine factors were assessed in each group of rats. Fresh feces and serum from each group of rats were collected for microbiome and metabolome analysis on the machine, respectively. Drug-disease common targets and functional pathways were captured based on network pharmacology. The complex network of Herbs-Targets-Pathways-Metabolites-Microbiota interactions was constructed. Ultimately, Fecal Microbiota Transplantation (FMT) evaluated the contribution of gut microbiota in disease. ResultsQWZT increased the abundance of beneficial bacteria (Bacteroides, Alloprevotella, Rikenellaceae_RC9_gut_group, Lactobacillus, Lachnospiraceae_NK4A136_group, Parabacteroides, etc.), reduced the abundance of harmful bacteria (Micromonospora, Geobacter, Nocardioides, and Arenimonas, etc.), reduced the levels of inflammatory mediators (12,13-EpOME, 9,10-Epoxyoctadecenoic acid, SM(d18:1/16:0) and Leukotriene A4, etc.), restored host metabolic disorders (Linoleic acid metabolism, Glycerophospholipid metabolism, and Arachidonic acid metabolism), and regulated the level of cytokines (IL-6, TNF-a, SOD, MDA, PEG-2 and NO), ultimately exerting an anti-ulcer effect. Apart from that, FMT improved acetic acid-induced gastric ulcers in rats. ConclusionQWZT improved acetic acid-induced gastric ulcers in rats by remodeling intestinal microbiota and regulating host metabolism. This work may promote the process of developing and utilizing clinical applications of QWZT.

Health impacts of poor water quality on an endangered shorebird breeding programme in Aotearoa New Zealand

ABSTRACT Case history Two clusters of mortality among endangered tūturuatu/tchūriwat’/shore plover (Thinornis novaeseelandiae) have occurred at captive breeding facilities around New Zealand in recent years. In the first, four chicks died at Pūkaha National Wildlife Centre (Mount Bruce, NZ) in February 2016, and in the second five adult birds at the Cape Sanctuary (Cape Kidnappers, NZ) died in 2022. Clinical findings In 2016, four chicks were noted to become weak, have increased vocalisations and closed eyes prior to death. The remaining chicks were treated for 5 days with amoxycillin/clavulanate orally twice daily. Water containers and brooders were cleaned and disinfected with chlorhexidine. No further mortality was seen. In the 2022 cluster, three adult breeding birds died acutely and five others showed inappetence, weight loss and diarrhoea approximately 10 days after heavy rains flooded the local river. The five birds were treated with amoxycillin/clavulanate orally twice daily and oral fluids for 5 days. Two birds died and three survived. No breeding occurred in the aviaries in the following season. Pathological findings In 2016, the chicks showed pulmonary changes ranging from congestion and oedema to heterophilic inflammation consistent with septicaemia. In 2022, the adult birds showed proliferation of bacteria in the distal small intestine associated with mucosal ulceration and heterophilic infiltration. Acid-fast staining of the caecal contents in one bird showed organisms consistent with Cryptosporidium spp. Laboratory findings Aerobic bacterial cultures of the lung and liver of two affected chicks carried out in 2016 showed heavy growth of Plesiomonas shigelloides. The same organism was cultured from water trays and holding tanks containing water boatmen (Sigara arguta) on which the chicks were fed. In 2022, cultures from the livers of three dead birds each showed a mixed bacterial growth with differing dominant organisms (Aeromonas sobria, Hafnia alvei, Citrobacter freundii and an Enterococcus sp.). PCR and sequencing confirmed Cryptosporidium parvum in the caecum of one bird. Fresh faeces from 24 breeding birds from the captive breeding facilities were negative by PCR for Cryptosporidium spp. The captive breeding facilities obtain water for the aviaries and aquatic invertebrates to feed to the chicks from local freshwater sources. Water quality testing at the Cape Sanctuary revealed concentrations of faecal indicator bacteria in excess of safe drinking water guidelines, with peaks following heavy rainfall. Clinical relevance Fluctuations in water quality associated with mammalian faecal bacteria can adversely affect bird health and impact on captive rearing of endangered wildlife.

Damage and ethology of Cryptarcha sp. in the cactus Opuntia ficus-indica(L.) Mill. Ayacucho, Perú

In prickly pear (Opuntiaficus-indica(L.) Mill.) of the semiarid forest of Ayacucho, Chryptarchasp. infests and damages the tender organs of the cactus. The objective of the study was to determine the damage of the insect during a year and its behavior. Fortnightly during the year, in 60 prickly pear plants, the number of young leaves, flower buds, flowers and green fruits, healthy and damaged by the insect and its behavior were monitored. Using the PAST 4.12 software, the annual cycle of damage occurrence and the annual cycle of the vegetative period of the prickly pear were diagrammed, to relate them to the climatic seasons; likewise, the principal component analysis (PCA) to establish the most affected tender organs and the period of infestation to the cactus. It turned out that during theyear the flower bud and the green fruit were significantly damaged. It infests the tuna mainly from November to March of the following year, which corresponds to the hot and humid season; in which it managed to damage 35.9% of flowers, 14.7% of tender leaves, 9.7% of green fruits and 0.7% of flowers. It has phytophagous and saprophagous behavior, being attracted by the juice of ripe fruit damaged by other agents, by the ferments of the stems of the prickly pear and other cacti in the process of putrefaction, and by the fresh feces of the pig.

Alterations in gut microbiome and metabolite profile of patients with Schistosoma japonicum infection

BackgroundSchistosoma infection is a significant public health issue, affecting over 200 million individuals and threatening 700 million people worldwide. The species prevalent in China is Schistosoma japonicum. Recent studies showed that both gut microbiota and metabolome are closely related to schistosomiasis caused by S. japonicum, but clinical study is limited and the underlying mechanism is largely unclear. This study aimed to explore alterations as well as function of gut microbiota and metabolite profile in the patients with S. japonicum infection.MethodsThis study included 20 patients diagnosed with chronic schistosomiasis caused by S. japonicum, eight patients with advanced schistosomiasis caused by S. japonicum and 13 healthy volunteers. The fresh feces of these participators, clinical examination results and basic information were collected. 16S ribosomal RNA gene sequencing was used to investigate gut microbiota, while ultraperformance liquid chromatography-mass spectrometry (UHPLC-MS) was applied to explore the metabolome of patients in different stages of schistosomiasis.ResultsThe study found that gut microbiota and metabolites were altered in patients with different stages of S. japonicum infection. Compared with healthy control group, the gut microbial diversity in patients with chronic S. japonicum infection was decreased significantly. However, the diversity of gut microbiota in patients with chronic schistosomiasis was similar to that in patients with advanced schistosomiasis. Compared with uninfected people, patients with schistosomiasis showed decreased Firmicutes and increased Proteobacteria. As disease progressed, Firmicutes was further reduced in patients with advanced S. japonicum infection, while Proteobacteria was further increased. In addition, the most altered metabolites in patients with S. japonicum infection were lipids and lipid-like molecules as well as organo-heterocyclic compounds, correlated with the clinical manifestations and disease progress of schistosomiasis caused by S. japonicum.ConclusionsThis study suggested that the gut microbiota and metabolome altered in patients in different stages of schistosomiasis, which was correlated with progression of schistosomiasis caused by S. japonicum. This inter-omics analysis may shed light on a better understanding of the mechanisms of the progression of S. japonicum infection and contribute to identifying new potential targets for the diagnosis and prognosis of S. japonicum infection. However, a large sample size of validation in clinic is needed, and further study is required to investigate the underlying mechanism.

Experiences in Peru for surveillance in the fight against highly pathogenic H5N1 avian influenza. Proceeding of The First International Avian Influenza Summit, University of Arkansas- October 16-17, 2023"

The emergence of highly pathogenic H5N1 avian influenza poses a significant threat to human and animal health worldwide. This summary provides an overview of the surveillance efforts implemented in Peru to combat the spread of H5N1 avian influenza. Joint efforts between the authority, the productive sector, and academia date back to the first influenza outbreak in Chile in 2002. Since then, the government has been developing intense clinical and serological surveillance in the domestic birds and poultry industry. In parallel, the university has been carrying out important research to identify and typify the viruses that circulate in the resident and migratory wild bird populations of the Peruvian coast, having detected and identified during these years around 50 strains of influenza viruses of 10 different hemagglutinin subtypes and 8 neuraminidases, a single strain of the low pathogenicity H7N3 subtype. Until then, an H5-type virus had never been detected in wild birds. With the arrival of the virus in North America, a comprehensive response was developed in which the key components of the surveillance program included active and passive clinical surveillance in wild bird populations, commercial poultry, and backyard and game bird populations. Regarding surveillance of wild birds, the authority has received 238 notifications, of which 64 were confirmed positive, from 15 regions. In the Avian Pathology Laboratory of the UNMSM, actions were carried out to support the active surveillance of wild birds of the Peruvian coast in the impact zones for poultry farming, analyzing 405 samples of fresh feces from 41 species of asymptomatic coastal aquatic wild birds, being positive 12 (2.96\%). Additionally, 45 samples of wild birds with suspicious symptoms were analyzed and sent by different government institutions (SENASA, SERFOR, SERNAMP), with 19 (42\%) being confirmed positive. Regarding domestic bird populations, 1,676 notifications have been made to SENASA, of which 194 were positive for the H5N1 virus in 13 country regions. On the other hand, the Avian Pathology Lab received 16 confirmed positive cases by PCR and viral isolation in SPF embryonated eggs from ducks, quails, layers, native birds, and one case of turkeys. The complete genome analysis of 09 strains of wild birds, 04 of domestic birds, and one of sea lions has been carried out. Analysis of new samples is in process. NGS analysis has identified the IA H5N1 clade 2.3.4.4b virus; however, it has been found that the Peruvian strains are lining up in a group together with the Chilean strains. All these activities are possible to carry out thanks to the financial support of national and international institutions: NAMRU, Concytec-World Bank, Peruvian Poultry Association, and ALA-US Poultry. The high vulnerability of poultry populations in the country is due to its location along the coast, where tens of thousands of dead wild birds have fallen, in addition to being surrounded by a very high population of ducks, fighting, and backyard birds. The government determined emergency vaccination, as well as the continuation of clinical and virological surveillance in vaccinated and unvaccinated poultry populations. SENASA has been evaluating the protection of approved vaccines, both by the serological response with hemagglutination inhibition tests using local antigens and by challenge. The industry has responded by significantly increasing the levels of structural and operational biosafety, as well as directly supporting control actions at the regional level. Since March, no outbreaks have been reported in industrially farmed birds.

Fecal microbiota transplantation from HUC-MSC-treated mice alleviates acute lung injury in mice through anti-inflammation and gut microbiota modulation.

Acute lung injury (ALI) is a severe respiratory tract disorder facilitated by dysregulated inflammation, oxidative stress and intestinal ecosystem. Fecal microbiota transplantation (FMT) is a rapid method for gut microbiota (GM) reconstruction. Furthermore, our previous studies have confirmed that human umbilical cord mesenchymal stromal cells (HUC-MSCs) can alleviate ALI by improving GM composition. Therefore, we aimed to explore the efficacy and mechanism of FMT from HUC-MSCs-treated mice on ALI. In brief, fresh feces from HUC-MSCs-treated mice were collected for FMT, and the mice were randomly assigned into NC, FMT, LPS, ABX-LPS, and ABX-LPS-FMT groups (n = 12/group). Subsequently, the mice were administrated with antibiotic mixtures to deplete GM, and given lipopolysaccharide and FMT to induce ALI and rebuild GM. Next, the therapeutic effect was evaluated by bronchoalveolar lavage fluid (BALF) and histopathology. Immune cells in peripheral blood and apoptosis in lung tissues were measured. Furthermore, oxidative stress- and inflammation-related parameter levels were tested in BALF, serum, lung and ileal tissues. The expressions of apoptosis-associated, TLR4/NF-κB pathway-associated, Nrf2/HO-1 pathway related and tightly linked proteins in the lung and ileal tissues were assessed. Moreover, 16S rRNA was conducted to assess GM composition and distribution. Our results revealed that FMT obviously improved the pathological damage of lung and ileum, recovered the immune system of peripheral blood, decreased the cell apoptosis of lung, and inhibited inflammation and oxidative stress in BALF, serum, lung and ileum tissues. Moreover, FMT also elevated ZO-1, claudin-1, and occludin protein expressions, activating the Nrf2/HO-1 pathway but hindering the TLR4/NF-κB pathway. Of note, the relative abundances of Bacteroides, Christensenella, Coprococcus, and Roseburia were decreased, while the relative abundances of Xenorhabdus, Sutterella, and Acinetobacter were increased in the ABX-LPS-FMT group. FMT from HUC-MSCs-treated mice may alleviate ALI by inhibiting inflammation and reconstructing GM, additionally, we also found that the TLR4/NF-κB and Nrf2/HO-1 pathways may involve in the improvement of FMT on ALI, which offers novel insights for the functions and mechanisms of FMT from HUC-MSCs-treated mice on ALI.

Dietary Inclusion of Yellow Mealworms (T. molitor) and Lesser Mealworms (A. diaperinus) Modifies Intestinal Microbiota Populations of Diet-Induced Obesity Mice

BackgroundInsect-based proteins are high-quality alternatives to support the shift toward more sustainable and healthy diets. Additionally, insects contain chitin and have unique fatty acid profiles. Studies have shown that mealworms may beneficially affect metabolism, but limited information is known regarding their effects on gut microbiota. ObjectivesWe determined the effects of defatted yellow mealworm (Tenebrio molitor) and whole lesser mealworm (Alphitobius diaperinus) meals on the intestinal microbiota of diet-induced obesity mice. MethodsMale C57BL/6J mice were fed a high-fat diet (HFD; 46% kcal) to induce obesity. Obese mice were then randomly assigned to treatments (n = 10/group) and fed for 8 wk: HFD, HFD with casein protein; B50, HFD with 50% protein from whole lesser mealworm; B100, HFD with 100% protein from whole lesser mealworm; Y50, HFD with 50% protein from defatted yellow mealworm; Y100, HFD with 100% protein from defatted yellow mealworm. Lean mice (n = 10) fed a low-fat-diet (10% kcal) were included. Fresh feces were collected at baseline and every 2 wk, with cecal digesta collected at kill. Fecal and cecal DNA was analyzed for microbiota using 16S rRNA MiSeq Illumina sequencing. ResultsIn feces and cecal digesta, mice fed mealworms had greater (P < 0.05) bacterial alpha diversity, with changes occurring in a time-dependent manner (P < 0.05). Beta diversity analyses of cecal samples showed a clear separation of treatments, with a time-based separation shown in fecal samples. Widespread microbial differences were observed, with over 45 genera altered (P < 0.05) by diet in cecal digesta. In feces, over 50 genera and 40 genera were altered (P < 0.05) by diet and time, respectively. ConclusionMealworm consumption changes the intestinal microbiota of obese mice, increasing alpha diversity measures and shifting bacterial taxa. More investigation is required to determine what mealworm components are responsible and how they may be linked with the metabolic benefits observed in mealworm-fed mice.

The Effects of Dietary Chromium Supplementation along with Discontinuing a High-Fat Diet on the Microbial Enzymatic Activity and the Production of SCFAs in the Faeces of Rats.

The present study assessed the changes in faecal microbial activity in obese Wistar rats fed high-fat or low-fat diets supplemented with various forms of chromium (picolinate or nanoparticles). The 18-week study was divided into two phases: an introductory period (9 weeks; obesity status induction via a high-fat diet) and an experimental period (9 weeks; maintained on a high-fat diet or switched to a low-fat diet and Cr supplementation). During the experimental period (10-18 weeks of feeding), samples of fresh faeces were collected on chosen days. The bacterial enzymatic activity and short-chain fatty acids (SCFAs) concentration were assessed to characterise the dynamism of the changes in faecal microbial metabolic activity under the applied dietary treatments. The results indicated that faecal microbial metabolic activity displayed several adaptation mechanisms in response to modifications in dietary conditions, and a beneficial outcome resulted from a pro-healthy dietary habit change, that is, switching from a high-fat to a low-fat diet. Dietary supplementation with chromium nanoparticles further modulated the aforementioned microbial activity, i.e., diminished the extracellular and total enzymatic activities, while the effect of chromium picolinate addition was negligible. Both the high-fat diet and the addition of chromium nanoparticles reduced SCFA concentrations and increased the faecal pH values.

Evaluation of Giardiasis in Companion Birds in Mashhad, Iran

Introduction: Giardia spp. are parasitic protozoans that infect the digestive system of ornamental birds, especially in Psittaciformes, with different prevalence in various species. Symptoms of Giardia infection vary from no noticeable signs in adult birds to poor growth, weakness, anorexia, ruffled feathers, and diarrhea in chicks. This study aimed to evaluate the prevalence of giardiasis in some parrots referred to a private veterinary clinic in Mashhad, Iran, during 2022. Case report: Feces of 215 birds suspected of being infected with Giardia spp. were collected. The birds were visited at one of the clinics in Mashhad, Iran, in 2022. Wet smear was prepared from their fresh feces and then sent to the laboratory department of the clinic for diagnosis. The parrots included adult cockatiels, cockatiel chicks, lovebirds, African grey parrots, and Green-cheeked parakeets. After the final diagnosis, metronidazole was administered orally at a dosage of 50 milligrams per kilogram of body weight every 12 hours for 5 consecutive days. Sick birds were tested again after the treatment to confirm they were parasite-free. The obtained results of the laboratory indicated 60 positive Cockatiels (47.6%), 10 positive cockatiel chicks (43.4%), 4 positive lovebirds (14.2%), 2 Green-cheeked parakeets (8.6%), and only 1 positive African grey parrot (6.6%). Among the positive cases, the severity of infection varied, with some birds showing severe, moderate, or mild infection levels. Upon a second examination, birds with mild and moderate contamination were found to be disease-free, while those with severe contamination required further treatment. Conclusion: The results suggested that treatment with metronidazole for 5 consecutive days was effective against mild to moderate infection in sick parrots. However, birds with severe conditions need to continue the treatment procedure.

Detection of Escherichia coli and other Enterobacteriales members in seabirds sampled along the Brazilian coast.

Escherichia coli, an Enterobacterales member, is a normal representative of the microbiota of homeothermic animals. Most strains are commensal, but several pathotypes can cause disease, and numerous antimicrobial resistance factors have been identified. These bacteria have spread rapidly in recent years, highlighting the importance of screening the environment and non-human reservoirs for virulent strains and/or those presenting resistance factors, in addition to other microorganisms of public health importance. In this context, this study aimed to survey Enterobacteriales present in seabirds sampled along the Brazilian coast, comparing findings between migratory and resident birds, as well as between wrecked and non-wrecked animals. Escherichia coli pathotypes were also characterized through rapid seroagglutination and polymerase chain reaction techniques and antimicrobial resistance profiles were investigated through the disc agar diffusion method. Cloacal, ocular, oral, tracheal, and skin lesion swabs, as well as fresh feces, were collected from 122 seabirds. The findings indicate these animals as important hosts for opportunistic human pathogens. Escherichia coli strains were identified in 70 % of the analyzed seabirds, 62 % of which displaying resistant or intermediate profiles to at least one antimicrobial, while 7% were multiresistant. Resistance to tetracycline (22 %), nalidixic acid (15 %), trimethoprim-sulfamethozaxol (14 %) and ampicillin (12 %) were the most prevalent. Resistance to cefoxitin, a critically important antimicrobial for human medicine, was also detected. Virulence genes for one of the EAEC, ETEC or EPEC pathotypes were detected in 30 % of the identified strains, the first two described in seabirds for the first time. The EAEC gene was detected in 25 % of the sampled seabirds, all resident, 8 % of which exhibited a multidrug-resistant profile. Thus, seabirds comprise important reservoirs for this pathotype. Escherichia coli was proven an ubiquitous and well-distributed bacterium, present in all evaluated bird species and sampling sites (except Marajó Island). According to the chi-square test, no significant differences between E. coli prevalences or antimicrobial resistance profiles between migratory and resident and between wrecked and non-wrecked seabirds were observed. Thus, migratory birds do not seem to contribute significantly to E. coli frequencies, pathotypes or antimicrobial resistance rates on the Brazilian coast.

Non-invasive specimen collections for Mycobacterium tuberculosis detection in free-ranging long-tailed macaques (Macaca fascicularis).

Surveillance of infectious diseases in free-ranging or wild animals has been widely conducted in many habitat-range countries after the COVID-19 episode. Thailand is located in the center of the distribution range of long-tailed macaques (Macaca fascicularis; Mf) where the animals have both frequent human contact and a high prevalence of human tuberculosis. For the large-scale detection of Mycobacterium tuberculosis complex (MTBC) using IS6110-nested PCR in free-ranging Mf, non-invasive sampling was developed using oral (via rope bait) and fecal (direct swabs of fresh feces) specimen collection. Firstly, the MTBC-IS6110-nested PCR was validated in non-invasively collected specimens, in terms of its specificity and sensitivity, and then compared with those of the invasively collected oral and rectal swabs in 24 captive MTBC-suspected Mf. After validation, these methods were applied to survey for the prevalence of shed MTBC (MTBCS) in four previously reported MTBC-infected populations. A total of 173 baited rope specimens and 204 freshly defecated excretions were collected. The limit of detection of the IS6110-nested PCR technique was 10 fg/μL and the 181-bp PCR amplicon showed 100% sequence similarity with the MTB H37Rv genome sequence. Comparing the MTBCS detection between the invasive and non-invasive collected specimens in captive suspected Mf revealed a significant correlation between the two types of oral specimens (oral swabs and baited ropes; n = 24, r2 = 1, p-value < 0.001), but fresh fecal swabs showed higher MTBCS frequencies than the rectal swabs. Moreover, the proportion of MTBCS-positive free-ranging Mf were significantly higher in the fresh fecal swabs (8.82%; 95% CI; 4.9-12.7%) than in the baited ropes (5.20%; 95% CI; 1.9-8.5%). This result indicates that oral sampling via baited ropes and fecal sampling via defecated excretion swabs can serve as ancillary specimens for MTBCS detection in free-ranging non-human primates.

Fecal nitrogen excretion in dairy cows supplemented with feed additive containing active dry yeast

The aim of the study was to determine the fecal excretion of N (TN and NH4 +-N) in dairy cows receiving the feed supplement Biolife® (Eurovix S.p.A., Italy) containing active dry yeast Saccharomyces cerevisiae. Two groups of 14 Holstein cows (B – experimental; C – control) at tie-stalled breeding participated in the experiment. Group B received the feed supplement through the total mixed ration (TMR). Fresh feces samples from both groups were collected on d 7, 14, 21, 28, 35 from the start of supplementation. Mean values of fecal pH and excreted DM did not differ between groups. The concentration of NH4 +-N in the fecal samples of group B was reduced by 28.05%, average for the entire studied period (P&lt;0.05). In the period d 21-35, this reduction was 35% (P=0.095). Excreted TN, averaged over the entire period, was reduced by 16.18% in group B, but the difference was not statistically significant. The reduction of TN content in feces from group B was more pronounced in the period d 21-35, when the reduction was 31.99% compared to group C (P=0.084). A period of at least 14 days is required for adaptation to the supplement and for an occur effect on N excretion.

Bioactive Compounds and In Vitro Antioxidant and Anticoccidial Activities of Opuntia ficus-indica Flower Extracts.

The objective of the present study is to identify the biochemical compounds extracted from OFI flowers using ultra-high-performance liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry and to evaluate their in vitro antioxidant activities and anticoccidial effects on the destruction of Eimeria oocysts isolated from naturally infected chickens. A domestic microwave was used with a refrigerant to condense the vapors generated during the extraction. The flavonoid and phenolic compound contents of the OFI flowers were determined according to standard methods. DPPH radical and H2O2 scavenging capacities were used to assess the antioxidant activity. Regarding the anticoccidial activity, the Eimeria spp. oocysts used were isolated from the fresh feces of infected broilers and were determined in triplicate by incubation at an ambient temperature for 24 h. The results highlighted the considerable influence of the optimized acetone concentration, ratio, irradiation time, and microwave power parameters on the phenolic content and antioxidant activities. Our results revealed significant matches between the predicted and experimental values of the models. Molecular analysis revealed the presence of several biophenol classes such as quercetin, isorhamnetin 3-O-rutinoside, and quercetin-3-O-rutinoside. OFI flower extracts inhibited sporulation and damaged the morphology of Eimeria oocysts compared with normal sporulated Eimeria oocysts containing sporocysts. In conclusion, the optimized conditions were validated and found to fit very well with the experimental values. These findings suggest that the flowers of OFI should be considered sources of antioxidants. The results of the present study revealed that OFI flower extracts have anticoccidial activities against Eimeria-spp.-induced infection in broiler chickens.

Identification of Antimicrobial Resistance in Faecal Microbes from Wild Dugongs (Dugong dugon)

Estuarine and coastal waters are areas of potential concern for antimicrobial resistance because of the discharge of wastewater from sewage treatment plants and the run-off from urban and agricultural lands. Herein, we evaluate the antimicrobial resistance profiles in bacteria from dugongs (Dugong dugon), mammals that inhabit and feed in shallow coastal regions and, thus, are vulnerable to encountering water and sediment contaminated by human activities. Bacterial isolates were cultured from fresh faeces of four wild dugongs, as well as from one sediment sample from a dugong foraging ground in Queensland, Australia. Ten bacterial isolates underwent phenotypic antimicrobial susceptibility testing using disc diffusion and minimum inhibitory concentration testing, and genotypic resistance and virulence gene identification through whole genome sequencing. Four Staphylococcus warneri isolates and one Bacillus cereus isolate from dugong faeces were resistant to penicillin, with two S. warneri isolates also displaying resistance to trimethoprim. Four Escherichia coli isolates were all resistant to ampicillin. Resistance genes, including fosB, BcII, dfrC, blaZ, and mdfA, were identified in the isolates cultured from dugong faeces with two virulence genes (gad and lpfA) identified in all E. coli isolates. Lysinibacillus sphaericus cultured from the marine sediment and B. cereus from dugong faeces displayed phenotypic multidrug resistance (across categories of non-extended spectrum cephalosporins, penicillins and beta-lactamase inhibitors, and clindamycin; and for L. sphaericus, phosphonic acids). These results demonstrate the role that dugongs can play as a sentinel species for antimicrobial resistance in the coastal waters across their range, which includes both disturbed urban and rural regions.

Exploring the potential links between gut microbiota composition and natural populations management in wild boar (Sus scrofa)

We surveyed wild boar (Sus scrofa) populations using 16S rRNA gene analysis of the gut microbiota in fresh faeces taken from 88 animals hunted in 16 hunting estates. The wild boar is a very convenient model system to explore how environmental factors including game management, food availability, disease prevalence, and behaviour may affect different biological components of wild individuals with potential implications in management and conservation. We tested the hypotheses that diet (according to stable carbon isotopes analyses), gender (i.e., animal behaviour studying males and females), and both health (analyses of serum samples to detect exposure to several diseases) and form statutes (i.e., thoracic circumference in adults) are reflected in changes in the intestinal microbiota. We focused on a gut functional biomarker index combining Oscillospiraceae and Ruminococcaceae vs. Enterobacteriaceae. We found that gender and the estate (population) were explanatory variables (c.a. 28% of the variance), albeit a high degree of overlapping among individuals was observed. The individuals with higher abundance of Enterobacteriaceae showed a gut microbiota with low diversity, mostly in males. Significant statistical differences for thoracic circumference were not found between males and females. Interestingly, the thoracic circumference was significantly and inversely related to the relative abundance of Enterobacteriaceae in males. Overall, we found that diet, gender, and form status were major factors that could be related to the composition and diversity of the gut microbiota. A high variability was observed in the biomarker index for populations with natural diet (rich in C3 plants). Although, we noticed a marginally significant negative trend between the index (higher abundance of Enterobacteriaceae) and the continuous feeding of C4 plants (i.e., supplementary maize) in the diet of males. This result suggests that continuous artificial feeding in hunting estates could be one of the factors negatively influencing the gut microbiota and the form status of wild boars that deserves further investigations.

Screening of skatole-degrading bacteria and control of human fecal odor by compound bacteria

PurposeThe biodegradation of skatole was used as a starting point in this study, and existing strains of degrading NH3 and H2S in the laboratory were combined to create a composite deodorant. The deodorization effect of the composite deodorant on human feces was investigated in order to provide a foundation for fecal odor gas treatment.MethodsSkatole-degrading bacteria were identified, degradation conditions were optimized, and skatole metabolites were identified using liquid chromatography-mass spectrometry (LC-MS). The skatole-degrading bacteria were combined with the existing strains of degrading NH3 and H2S in the laboratory to make a compound deodorizer, and the deodorizing effect of the compound deodorizer on fresh human feces and old human feces was compared. Liquid chromatography, specific sensors, and gas chromatography were used to determine the contents of skatole, NH3, H2S, and volatile organic compounds, and the microbial diversity was examined.ResultsMicrobial agents H and Y capable of utilizing skatole were screened out under aerobic and facultative anaerobic conditions, respectively. Within 48 h, the degradation rate of skatole by microbial agent Y was 88.52%. Following condition optimization, the optimum temperature for skatole degradation by microbial agent Y was 33 °C, and the optimum pH was 7. The main functional bacteria were Acinetobacter xiamenensis. The metabolites of skatole were determined by liquid chromatography-mass spectrometry (LC–MS), and six possible metabolites were found, including 3-aldehyde indole, 3-carboxyl indole, 1H-indole-2,3-dione, and 3-methyl indole pyruvate. The skatole-degrading bacteria Lactobacillus rhamnosus and Saccharomyces cerevisiae were then combined in proportion to form a composite deodorant. The 5-day degradation rate of skatole in the old manure group was 88.11%, while the 5-day degradation rate in the fresh manure group was 47.56%.ConclusionsThe compound microbial agent developed in this study has a solid deodorizing effect, particularly in the control of the odor of obsolete feces. The use of composite microbial agents can efficiently degrade skatole in human feces, providing a theoretical foundation for the use of microbial remediation in the actual world.Graphical

#3454 IMPACT OF SYMBIOTICS ON UREMIC TOXINS PRODUCTION, RENAL FUNCTION AND PEW SYNDROME IN SHIME AND CKD MICE MODELS

Abstract Background and Aims Alterations in gut microbiota in CKD patients have been linked to CKD progression, cachexia, and mortality. Gut dysbiosis is associated by modification of gut-microbiota-derived metabolites, including a decrease of short-chain fatty acids (SCFAs), an increase in gut-derived uremic toxins. Therefore, modulation of gut microbiota seems to be an attractive therapeutic approach. In silico studies led to the selection of bacterial species able to increase SCFAs production, improve gut function without having the enzymatic machinery for producing uremic toxins. The propose of this study is to indentify the impact of two different symbiotics containing the selected bacteria: 1) on both uremic toxins and SCFAs production using a Simulator of the Human Intestinal Microbial Ecosystem (SHIME) colonized with fresh feces from CKD or healthy controls (HC) and subsequently 2) on renal function, metabolic parameters, intestinal microbiota and gut-microbiota-derived metabolites in a mouse model of CKD (5/6 nephrectomy). Method 1) The SHIME® system manufactured by ProDigest (Ghent, Belgium) consists of systems comprising five double-wall biovessels simulating the stomach (ST), small intestine (SI), ascending colon (AC), transverse colon (TC), and descending colon (DC). The supernatant of the homogenized fecal slurry was inoculated into the AC (25 mL), TC (40 mL), and DC (30 mL), respectively. After fecal inoculation, SHIME system were fed in ST twice day with two different symbiotics (P1 or P2) for 10 days. After 7 days, we applied a diet amino acid (AA) challenge to all the groups. SCFAs and uremic toxins were quantified every day. 2) CKD mice received either P1 or P2 admix for 6 weeks. We measured food intake, body composition and renal function including renal fibrosis quantification. Plasma metabolomic and feces metagenomic analyses were performed. Results In vitro, the concentration of precursor of uremic toxins (indols and p-cresol) tended to be higher in DC colonized by feces from CKD patients and AA challenge increased uremic toxins production more significantly in CKD compared to HC. CKD microbiome showed a lower butyrate concentration at baseline and higher propionate and acetate production. Both P1 and P2 significantly limited uremic toxins production during AA challenge and increase SCFA production. In vivo, both symbiotics increased body weight (+14%) and food intake (+31%) compared to CKD mice on chow diet. Treatment did not impact lipid or glucose metabolism but fat accretion in epididymal was restored with P1. Both diets, but particularly P1, significantly reduced plasmatic urea, proteinuria, and kidney fibrosis. Symbiotics improved intestinal barrier with a greater expression of Occludin in the ileum. Metagenomic analyses of the gut microbiota indicated that the alfa-diversity was not different across groups while the beta-diversity was similar to the sham group if CKD mice were treated with symbiotics. Both symbiotics reduced significantly plasma levels of deleterious uremic toxins such as p-cresyl sulfate and indoxyl sulfate and symbiotics reduce fecal module of trypotophan degradation and other modules related to AA metabolism. Both diets increased fecal bacteria modules related to SCFAs production. Conclusion We have shown both in vitro and in vivo that a symbiotic association can reduce uremic toxins production and increase SCFA production by inducing a significant change in gut microbiota composition and function. These metabolic changes are associated with improved appetite, decreased uremic cachexia, and preserved renal function. This data needs be confirmed in a human clinical trial.

Camel potentiality in survival and germination of wild pastoral species: The case of Fabaceae in Sahara rangelands of Algeria

Previous studies conducted in the Sahara desert have reported the consumption of wild native pastoral plants by camels. To test the hypothesis that camels may effectively break seed dormancy, we collected fresh faeces from different Saharan rangelands frequented by camels.Seeds were extracted from camel faeces and were quantified. Germination of extracted seeds was compared with that of freshly collected seeds under controlled conditions. Sixteen native pastoral species in 8 families and two unidentified species were quantified, with a high proportion of Fabaceae seeds recovered especially Astragalus ghysensis. The Digestive system of camels, effectively (1) broke seed physical dormancy exceptionally for Argyrolobium uniflorum, Astragalus cruciatus and Astragalus ghysensis there by promoting seed germination; (2) enhanced the percentage of germination of non-dormant seeds in Bassia muricata, Erodium glaucophyllum, Fagonia glutinosa and Plantago ciliata and (3) maintained viability but did not contribute to a prompt germination of Cornulaca monacantha seeds. Therefore, camels act as effective dispersers of these Fabaceae species by scattering seeds and promoting germination. We conclude that it is necessary to better understand the role of camels in the maintenance and conservation of the studied species by endozoochory.

Shaoyao-Gancao-Tang regulates the T-helper-type 1/T-helper-type 2 ratio in the lung and gut and alters gut microbiota in rats with ovalbumin-induced asthma.

Shaoyao-Gancao Tang (SGT) is a traditional Chinese medicine formulation. It has been used to treat kinds of pain and to alleviate asthma in clinic. However, the mechanism of action is not known. To investigate the anti-asthma effect of SGT involving modulation of the T-helper type 1 (Th1) Th1/Th2 ratio in the gut-lung axis and alteration of the gut microbiota (GM) in rats with ovalbumin (OVA)-induced asthma. The main constituents of SGT were analyzed by high-performance liquid chromatography (HPLC). A model of asthma was established in rats by OVA-induced allergen challenge. Rats suffering from asthma (RSAs) were treated with SGT (2.5, 5.0 and 10.0 g/kg), dexamethasone (1 mg/kg) or physiologic saline for 4 weeks. The level of immunoglobulin (Ig)E in bronchoalveolar lavage fluid (BALF) and serum was determined by enzyme-linked immunosorbent assay. Histology of lung and colon tissues was investigated using staining (hematoxylin and eosin and periodic acid-Schiff). The Th1/Th2 ratio and levels of cytokines (interferon (IFN)-γ and interleukin (IL)-4) in the lung and colon were detected by immunohistochemistry. The GM in fresh feces was analyzed by 16 S rRNA gene sequencing. Twelve main constituents (gallic acid, albiflorin, paeoniflorin, liquiritin apioside, liquiritin, benzoic acid, isoliquiritin apioside, isoliquiritin, liquiritigenin, glycyrrhizic acid, isoliquiritigenin and glycyrrhetinic acid) of SGT were simultaneously determined by HPLC. SGT treatment (5.0 and 10.0 g/kg) was found to reduce the IgE level (a vital marker of hyper-responsiveness) in BALF and serum, improve typical morphological changes (inflammatory-cell infiltration and goblet cell metaplasia) in the lung and colon, alleviate airway remodeling (including bronchiostenosis and basement membrane-thickening) in the lung, significantly decrease the IL-4 level and increase the IFN-γ level in the lung and colon, which led to restoration of the IFN-γ/IL-4 ratio. The dysbiosis and dysfunction of GM in RSAs were modulated by SGT. The abundance of bacteria of the genera Ethanoligenens and Harryflintia was increased in RSAs and was decreased upon SGT treatment. The abundance of Family_XIII_AD3011_group was decreased in RSAs and increased upon SGT treatment. Moreover, SGT therapy increased the abundance of bacteria of the genera Ruminococcaceae_UCG-005 and Candidatus_Sacchrimonas, and decreased that of Ruminococcus_2 and Alistipes. SGT ameliorated rats with OVA-induced asthma via regulation of the Th1/Th2 ratio in the lung and gut, and modulated the GM.

Increased abundance of bacteria of the family Muribaculaceae achieved by fecal microbiome transplantation correlates with the inhibition of kidney calcium oxalate stone deposition in experimental rats.

The incidence of nephrolithiasis is increasing rapidly worldwide. Calcium oxalate is the most common constituent, contributing to approximately 80% of all kidney stones. The gut microbiome, through its oxalate-degrading ability, may play a role in decreasing morbidity due to urinary calculus. Fecal microbiome transplantation (FMT) has been reported to be effective in restoring the gastrointestinal microbial community in different conditions. The transplantation of whole communities that have oxalate-degrading function may be a more effective strategy than the transplantation of isolated strains. FMT was carried out in male guinea pigs and male Sprague-Dawley laboratory rats (SDRs). Fresh feces were collected from guinea pigs housed in metabolic cages. SDRs were divided into four groups: two groups received standard rat chow (SC) (groups SC and SC + FMT), and two groups were fed a 5% potassium oxalate diet (OD) (groups OD + phosphate-buffered saline (PBS) and OD + FMT). On day 14, groups OD + PBS, OD + FMT, and SC + FMT received either PBS or guinea pig feces by esophageal gavage. The composition of the microbiota of guinea pigs and SDRs was analyzed using a 16S rRNA gene sequencing approach. Biochemical analysis of urine samples from SDRs revealed the presence of calcium oxalate (CaOx) crystals, which were presumed to originate from kidney stones. Renal function was examined using real-time PCR analysis and immunohistochemical staining for renin, angiotensin-converting enzyme, and osteopontin (OPN) expression. FMT resulted in a gut microbiota that was a mixture of guinea pig and SDR bacteria. A microbial network involving Muribaculaceae, Lactobacillus, and Bifidobacterium was activated by FMT in group OD + FMT. As a result, urinary oxalate, calcium, uric acid, creatinine and urea in urine samples were reduced significantly. Similarly, significant reduction of uric acid and blood urea nitrogen to creatinine ratio in serum samples was observed (p < 0.05). Microscopic observations revealed a high CaOx crystal score (4+) in the kidneys of rats in group OD + PBS, whereas a lower score (2+) was observed in the rats in group OD + FMT. Up-regulation of OPN and down-regulation of renin were also associated with FMT. A microbial network involving Muribaculaceae and other oxalate-degrading bacteria achieved by FMT was capable of reducing urinary oxalate excretion and CaOx crystal deposition in the kidney through increasing intestinal oxalate degradation. FMT may exert a renoprotective function in oxalate-related kidney stones.