The concept that the gut microbiota serves as a virtual endocrine organ arises from a number of important observations. Evidence for a direct role arises from its metabolic capacity to produce and regulate multiple compounds that reach the circulation and act to influence the function of distal organs and systems. For example, metabolism of carbohydrates results in the production of short-chain fatty acids, such as butyrate and propionate, which provide an important source of nutrients as well as regulatory control of the host digestive system. This influence over host metabolism is also seen in the ability of the prebiotic inulin to influence production of relevant hormones such as glucagon-like peptide-1, peptide YY, ghrelin, and leptin. Moreover, the probiotic Lactobacillus rhamnosus PL60, which produces conjugated linoleic acid, has been shown to reduce body-weight gain and white adipose tissue without effects on food intake. Manipulating the microbial composition of the gastrointestinal tract modulat...

Minireview: Gut Microbiota: The Neglected Endocrine Organ

Background: Several studies suggested an important role of the gut microbiota in the pathophysiology of neurological disorders, implying that alteration of the gut microbiota might serve as a treatment strategy. Fecal microbiota transplantation (FMT) is currently the most effective gut microbiota intervention and an accepted treatment for recurrent Clostridioides difficile infections. To evaluate indications of FMT for patients with neurological disorders, we summarized the available literature on FMT. In addition, we provide suggestions for future directions. Methods: In July 2019, five main databases were searched for studies and case descriptions on FMT in neurological disorders in humans or animal models. In addition, the ClinicalTrials.gov website was consulted for registered planned and ongoing trials. Results: Of 541 identified studies, 34 were included in the analysis. Clinical trials with FMT have been performed in patients with autism spectrum disorder and showed beneficial effects on neurological symptoms. For multiple sclerosis and Parkinson's disease, several animal studies suggested a positive effect of FMT, supported by some human case reports. For epilepsy, Tourette syndrome, and diabetic neuropathy some studies suggested a beneficial effect of FMT, but evidence was restricted to case reports and limited numbers of animal studies. For stroke, Alzheimer's disease and Guillain-Barre syndrome only studies with animal models were identified. These studies suggested a potential beneficial effect of healthy donor FMT. In contrast, one study with an animal model for stroke showed increased mortality after FMT. For Guillain-Barre only one study was identified. Whether positive findings from animal studies can be confirmed in the treatment of human diseases awaits to be seen. Several trials with FMT as treatment for the above mentioned neurological disorders are planned or ongoing, as well as for amyotrophic lateral sclerosis. Conclusions: Preliminary literature suggests that FMT may be a promising treatment option for several neurological disorders. However, available evidence is still scanty and some contrasting results were observed. A limited number of studies in humans have been performed or are ongoing, while for some disorders only animal experiments have been conducted. Large double-blinded randomized controlled trials are needed to further elucidate the effect of FMT in neurological disorders.

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Fecal Microbiota Transplantation in Neurological Disorders.

Small mammalian herbivores have a limitation in their supply system of nutrients to their energy and protein demands because they need much more energy and protein per unit body mass than larger herbivorous animals. Therefore, small herbivores need to have characteristic strategies in their digestive systems to overcome the limitation of their small body mass compared with larger animals. Although small herbivorous mammals commonly have an enlarged cecum, the pattern of flow and mixing of digesta in the large intestine varies among them. Distinct separation of the larger fiber particles from smaller and liquid contents which are retained in the cecum can be recognized in some species. Coprophagy, practiced by many small herbivores, has nutritional significance providing a source of vitamins, amino acids, and other nutrients which are excreted with feces. Among coprophagous mammals, several species produce two types of feces: soft feces, which are eaten; and hard, which are not eaten. Soft feces contain more water than hard feces and dry matter includes more protein and less fiber. Coprophagic behavior must be supported by the colonic separation mechanism, which operates retrograde transport of fluid and fine particle digesta or bacteria trapped in the mucus, resulting in high density bacteria in the cecum contents, which is successively consumed as cecotroph. These mechanisms must be necessary for small herbivores to survive on the feed in their habitat.

Digestive strategies of small hindgut fermenters

In mice, maternal dietary folate, a cofactor in 1-carbon metabolism, modulates neurogenesis and apoptosis in the fetal brain. Similarly, maternal dietary choline, an important methyl-donor, also influences these processes. Choline and folate are metabolically interrelated, and we determined whether choline supplementation could reverse the effects of folate deficiency on brain development. Timed-pregnant mice were fed control (CT), folate-deficient (FD), or folate-deficient, choline-supplemented (FDCS) AIN-76 diets from d 11 to 17 (E11-17) of pregnancy, and on E17, fetal brains were collected for analysis. Compared with the CT group, the FD group had fewer neural progenitor cells undergoing mitosis in the ventricular zones of the developing mouse brain septum (47%; P < 0.01), hippocampus (29%; P < 0.01), striatum (34%; P < 0.01), and anterior and mid-posterior neocortex (33% in both areas; P < 0.01). In addition, compared with CT, the FD diet almost doubled the rate of apoptosis in the fetal septum and hippocampus (P < 0.01). In the FDCS group, the mitosis rates generally were intermediate between those of the CT and FD groups; mitosis rates in the septum and striatum were significantly greater compared with the FD group and were significantly lower than in the CT group only in the septum and neocortex. In the FDCS group, the hippocampal apoptosis rate was significantly lower than in the FD group (P < 0.01) and was the same as in the CT group. In the septum, the apotosis rate in the FDCS group was intermediate between the CT and FD groups' rates. These results suggest that neural progenitor cells in fetal forebrain are sensitive to maternal dietary folate during late gestation and that choline supplementation can modify some, but not all, of these effects.

/pdf/dietary-choline-reverses-some-but-not-all-effects-of-folate-3dxkp1m5d1.pdf

Dietary Choline Reverses Some, but Not All, Effects of Folate Deficiency on Neurogenesis and Apoptosis in Fetal Mouse Brain

The upper gastrointestinal tract plays a prominent role in human physiology as the primary site for enzymatic digestion and nutrient absorption, immune sampling, and drug uptake. Alterations to the small intestine microbiome have been implicated in various human diseases, such as non-alcoholic steatohepatitis and inflammatory bowel conditions. Yet, the physiological and functional roles of the small intestine microbiota in humans remain poorly characterized because of the complexities associated with its sampling. Rodent models are used extensively in microbiome research and enable the spatial, temporal, compositional, and functional interrogation of the gastrointestinal microbiota and its effects on the host physiology and disease phenotype. Classical, culture-based studies have documented that fecal microbial self-reinoculation (via coprophagy) affects the composition and abundance of microbes in the murine proximal gastrointestinal tract. This pervasive self-reinoculation behavior could be a particularly relevant study factor when investigating small intestine microbiota. Modern microbiome studies either do not take self-reinoculation into account, or assume that approaches such as single housing mice or housing on wire mesh floors eliminate it. These assumptions have not been rigorously tested with modern tools. Here, we used quantitative 16S rRNA gene amplicon sequencing, quantitative microbial functional gene content inference, and metabolomic analyses of bile acids to evaluate the effects of self-reinoculation on microbial loads, composition, and function in the murine upper gastrointestinal tract. In coprophagic mice, continuous self-exposure to the fecal flora had substantial quantitative and qualitative effects on the upper gastrointestinal microbiome. These differences in microbial abundance and community composition were associated with an altered profile of the small intestine bile acid pool, and, importantly, could not be inferred from analyzing large intestine or stool samples. Overall, the patterns observed in the small intestine of non-coprophagic mice (reduced total microbial load, low abundance of anaerobic microbiota, and bile acids predominantly in the conjugated form) resemble those typically seen in the human small intestine. Future studies need to take self-reinoculation into account when using mouse models to evaluate gastrointestinal microbial colonization and function in relation to xenobiotic transformation and pharmacokinetics or in the context of physiological states and diseases linked to small intestine microbiome and to small intestine dysbiosis.

https://link.springer.com/content/pdf/10.1186/s40168-020-0785-4.pdf

Self-reinoculation With Fecal Flora Changes Microbiota Density and Composition Leading to an Altered Bile-Acid Profile in the Mouse Small Intestine

Germfree mice died when they were fed a purified diet of AIN-76 formula sterilized by gamma-irradiation. Vitamin K deficiency was suspected and this study was performed to confirm the cause of the death. Germfree mice were fed purified diets of AIN-76 or AIN-93M formula, which were pelleted and sterilized by gamma-irradiation at a dose of 50 kGy. One half of the mice fed the AIN-76 diet died within two weeks and the surviving animals were also in poor health, while 91% of mice fed the AIN-93M diet survived. No hemorrhage was observed grossly in any organs of the surviving animals. Histologically, degeneration with inflammatory cell infiltration was observed as well as hemorrhage and fibrosis in the heart muscles of mice fed the AIN-76 diet. No microscopic lesions were observed in the other organs. Prothrombin time (PT) and activated partial thromboplastin time (APTT) were extremely prolonged when mice were fed the AIN-76 diet. The animals totally recovered when they were intragastrically administered 1 microg/day of vitamin K(3) from the third day of feeding of the AIN-76 diet, except for PT and APTT which were still slightly longer than in mice fed the AIN-93M diet. The concentration of vitamin K(3) supplied in the AIN-76 diet decreased to an undetectable level after gamma-irradiation, while the AIN-93M diet contained 240 microg/kg of vitamin K(1). These results indicate that the deaths of the germfree mice fed the gamma-irradiated AIN-76 diet were caused by vitamin K deficiency. Vitamin K deficiency may cause fatal degeneration of cardiac muscle cells.

Vitamin K deficiency of germfree mice caused by feeding standard purified diet sterilized by gamma-irradiation.

Changes in coprophagy with age were investigated in male ICR mice during their life span Sucklings showed coprophagy at 17 to 18 days old, ie, a few days after they began to excrete feces autonomously The number of fecal pellets ingested peaked at 5 to 6 weeks old (13 pellets/day) and gradually decreased, thereafter (21 pellets at 78 weeks old, 15 pellets at 104 weeks old) The diurnal pattern of coprophagy also changed with age Growing mice showed vigorous coprophagous activity in both light and dark phases, whereas animals over 30 weeks old exhibited less activity in both phases, especially in the dark phase Feces proved to be abundant in vitamin B12 and folic acid throughout the life span These results suggest that the frequency of coprophagy changes in association with the nutritional requirements of mice during the process of growth or aging

Lifelong coprophagy in male mice

Electron beam sterilization for laboratory animal diets was examined as an alternative to 60Co gamma rays. Solid, powder diets for "mice and rats" and solid diets for "rabbits and guinea pigs" which are the main products sterilized by 60Co gamma rays were irradiated with 10-MeV electrons from a linear accelerator at the Research Institute for Advanced Science and Technology, Osaka Prefecture University. At least 20 kGy was required to sterilize the samples irrespective of solid or powder diets, which was in good accordance with the results for 60Co gamma rays. Using a set dose of 30 kGy, a thickness of 45 mm for solid diets and 30 mm for powder diets could be sterilized by "one-sided" irradiation. "Dual-sided" irradiation could sterilize all the solid diets and the powder diets contained in the thicknesses of 90 mm and 75 mm, respectively. Irradiation effects of 10-MeV electrons on the nutrient quality of each diet were almost equivalent to those of 60Co gamma rays. These results suggest that commercially adopted sterilization doses for 60Co gamma rays are applicable to electron sterilization without modification if the depth-dose profile and the minimum dose of irradiated samples are precisely assessed.

Electron-beam sterilization of laboratory animal diets--sterilizing effect of 10-MeV electrons from a linear accelerator

Coprophagy was observed in germfree (GF) ICR mice of both sexes, and the results were compared with those of conventional mice. Frequency of coprophagy per animal per day in GF mice was 5.1 in males and 5.8 in females. In conventional (CV) mice, the frequencies were 6.2 in males and 5.3 in females (data from Zoological Science 2:249-255, 1985), with no significant differences compared with GF mice. Coprophagy in CV mice was frequently observed during 6-8 hr after lighting, whereas such close time relationships tended to weaken in GF animals. In a comparison of levels of constituents per unit weight between feces and diet, fecal crude protein and crude fat exhibited lower values than those in the diet. Levels of fecal crude ash and crude fiber were higher than those in the diet, and nitrogen-free extract was almost equal to that in the diet. No essential difference in these tendencies was found compared with CV mice. Levels of fecal vitamin B1, B2, B12 and folic acid were lower than those in the diet. In CV mice, except for vitamin B1, these vitamins exhibited either almost equal or much higher levels compared with those in the diet (data from Experimental Animals 35: 381-386, 1986). From the fact that coprophagy was observed in GF mice, it is suggested that the behavior is inherent in the mouse.

Coprophagy in the germfree mouse

The effects of prevention of coprophagy on reproductive performance were examined in ICR mice. Females were treated with restrainers in order to prevent them from ingesting their feces from day 1 through day 17 of pregnancy. The restrained animals fed a commercial diet did not show any clear adverse effects. In contrast, restrained dams fed a purified diet deficient in vitamin B12 exhibited stillbirths (14%) and abortions (7%). Restrained dams fed a diet lacking in vitamin B12 and folic acid also experienced frequent abortions (27%). In addition, six out of 14 restrained dams (43%) aborted when fed a vitamin B complex-deficient diet. Sham-restrained animals, fed the vitamin B complex deficient-diet, but able to ingest their feces trapped by smaller-mesh floors, escaped these adverse effects. Sham-restrained animals fed the commercial diet, however, showed only a slight improvement in their reproductive performance. In conclusion, coprophagy has nutritional significance as long as the diet is lacking at least B vitamins, especially vitamin B12 and folic acid, whereas it almost entirely loses its nutritional significance when the mouse has access to a balanced diet such as the one made available to the laboratory mice in the present study.

Yoshibumi Kuwabara

Papers

Vitamin K deficiency of germfree mice caused by feeding standard purified diet sterilized by gamma-irradiation.

Lifelong coprophagy in male mice

Electron-beam sterilization of laboratory animal diets--sterilizing effect of 10-MeV electrons from a linear accelerator

Coprophagy in the germfree mouse

Effects of prevention of coprophagy on pregnant mice--is coprophagy beneficial on a balanced diet?