326FOLLICULAR SIZE, BUT NOT STAGE OF REPRODUCTION OR SEASON, INFLUENCES 
MEIOTIC MATURATION OF DOMESTIC DOG OOCYTES

The current in vitro maturation system (IVM) for dog oocytes is inefficient. On the average, only 15% of ovarian oocytes complete nuclear maturation in vitro. For unknown reasons, the ability of oocytes to develop to the metaphase II stage (MII) varies markedly among bitches (Songsasen et al., 2002, Mol. Reprod. Dev. 62, 407–415). The objective of this study was to identify the cause(s) underlying these significant variations in nuclear maturation. Initially, we retrospectively analyzed data obtained during the past 3 years;; 1661 oocytes were obtained from 74 bitches where stage of reproduction for the donor was known based on ovarian morphology. Oocytes were cultured in TCM 199+0.1% polyvinyl alcohol at 38.5°C in 5% CO2 in humidified air under various experimental conditions. Analysis of variance (ANOVA) was performed to compare differences in meiotic competence of oocytes obtained at various reproductive stages and during different seasons. Stage of reproduction did not influence meiotic abilities of oocytes. Percentages of oocytes obtained during proestrus/estrus (n=468 oocytes), diestrus/metestrus (n=333), anestrus (n=331) or prepuberty (6–8 months of age, n=479) and developing to MII were 17.9±2.9%, (mean±SEM), 24.0±6.0%, 20.8±4.7%, and 17.8±5.2%, respectively (P>0.05). A similar analysis across seasons (spring, summer, fall, winter) also indicated no influence of time of year on nuclear maturation (P>0.05). Because there is a known strong link between follicular growth and meiotic competence of goat oocytes (De Smedt et al., 1994 J. Exp. Zool. 269, 128–139), we also examined the impact of follicular size on nuclear maturation. The cortex of ovaries from 15 bitches was horizontally dissected (5mm thickness) so follicles could be observed and divided into three classes: (1) <0.5mm diameter (n=60); (2) ≥0.5 to <1mm (n=110); and (3) 1–2mm (n=72). Follicles were separated according to these size classes;; oocytes were recovered and cultured in TCM 199+0.25mM pyruvate, 2mM glutamine, 25mM β-mercaptoethanol, 10ng/mL epidermal growth factor (Basal TCM) supplemented with 0.5IU/mL equine chorionic gonadotropin for 1h. Oocytes then were cultured in Basal TCM for 48h before staining with 1% orcein to assess nuclear status. Follicular size influenced meiotic competence of the oocytes (ANOVA, P<0.05). Mean percentages of MII oocytes were 14.2±7.2, 15.6±4.5, and 30.9±8.2, for oocytes recovered from <0.5-mm, ≥0.5 to <1-mm and 1–2-mm diameter follicles, respectively. This study revealed that stage of reproduction and season have no impact on in vitro nuclear maturation of the dog oocyte. However, the findings demonstrate that dog oocytes acquire meiotic competency during follicular development. Because the source of most dog oocytes for IVM are small follicles, results suggest that oocytes may be incapable of completing nuclear maturation under in vitro conditions that are designed for fully-grown oocytes.

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The potential of microdialysis to estimate rifampicin concentrations in the lung of guinea pigs

PLoS ONE ◽

10.1371/journal.pone.0245922 ◽

2021 ◽

Vol 16 (1) ◽

pp. e0245922

Author(s):

Faye Lanni ◽

Neil Burton ◽

Debbie Harris ◽

Susan Fotheringham ◽

Simon Clark ◽

...

Keyword(s):

Drug Development ◽

Guinea Pigs ◽

Sampling Technique ◽

Tissue Fluid ◽

Experimental Conditions ◽

Continuous Sampling ◽

Drug Concentrations ◽

The Impact

Optimised pre-clinical models are required for TB drug development to better predict the pharmacokinetics of anti-tuberculosis (anti-TB) drugs to shorten the time taken for novel drugs and combinations to be approved for clinical trial. Microdialysis can be used to measure unbound drug concentrations in awake freely moving animals in order to describe the pharmacokinetics of drugs in the organs as a continuous sampling technique. The aim of this work was to develop and optimise the microdialysis methodology in guinea pigs to better understand the pharmacokinetics of rifampicin in the lung. In vitro experiments were performed before progressing into in vivo studies because the recovery (concentration of the drug in the tissue fluid related to that in the collected dialysate) of rifampicin was dependent on a variety of experimental conditions. Mass spectrometry of the dialysate was used to determine the impact of flow rate, perfusion fluid and the molecular weight cut-off and membrane length of probes on the recovery of rifampicin at physiologically relevant concentrations. Following determination of probe efficiency and identification of a correlation between rifampicin concentrations in the lung and skeletal muscle, experiments were conducted to measure rifampicin in the sacrospinalis of guinea pigs using microdialysis. Lung concentrations of rifampicin were estimated from the rifampicin concentrations measured in the sacrospinalis. These studies suggest the potential usefulness of the microdialysis methodology to determine drug concentrations of selected anti-TB drugs to support new TB drug development.

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206 EFFECT OF ADDITION OF FOLLICULAR FLUID OR GROWTH DIFFERENTIATION FACTOR-9 ON IN VITRO MATURATION OF PORCINE OOCYTES DENUDED 20h AFTER THE START OF IN VITRO MATURATION

Reproduction Fertility and Development ◽

10.1071/rdv28n2ab206 ◽

2016 ◽

Vol 28 (2) ◽

pp. 234

Author(s):

P. Ferré ◽

T. T. M. Bui ◽

M. T. Tran ◽

T. Wakai ◽

H. Funahashi

Keyword(s):

Follicular Fluid ◽

In Vitro Maturation ◽

Cumulus Cells ◽

Developmental Competence ◽

Dapi Staining ◽

Experimental Conditions ◽

Nuclear Maturation ◽

Differentiation Factor ◽

Growth Differentiation Factor 9

The interruption of communication between oocyte and cumulus cells (CC) can trigger meiotic resumption and exogenous additives, such as follicular fluid (FF) and growth differentiation factor-9 (GDF9), can improve oocyte quality and the developmental competence. This study was undertaken to examine if the absence and presence of FF from medium follicles (MF; 3–6 mm in diameter) or recombinant human GDF9 (Biovision, Milpitas, CA, USA) during the first or/and second half of in vitro maturation (IVM) had any effects on IVM of oocytes from small follicles (SF; 0.5–2 mm in diameter) or MF when the oocytes were denuded at 20 h after the start of IVM. Cumulus-oocyte complexes (COC) were aspirated from SF or MF of slaughtered prepubertal gilt ovaries. Groups of ~30 COC were cultured in a 300-μL drop of porcine oocyte medium containing 50 µM β-mercaptoethanol (mPOM) with or without 10% (v/v) FF and/or 100 ng mL–1 GDF9 at 39°C and 5% CO2 in air. During the first 20 h after the start of IVM, the medium was supplemented with 1 mM dibutyryl c-AMP, 10 IU mL–1 eCG and 10 IU mL–1 hCG. After the first period of IVM, the CC surrounding the oocytes were removed and the denuded oocytes continued culture for IVM with or without FF or/and GDF9 in the absence of dibutyryl c-AMP and gonadotropins in the same medium for another 24 h. At the end of IVM, meiotic progression of the oocytes was examined by DAPI staining. Statistical analyses from at least 4 replicates data were performed by a 2-way ANOVA and a Tukey’s multiple comparisons test. Removal of CC 20 h after the start of IVM significantly improved the incidence of mature oocytes derived from SF (59.2–64.1% v. 41.6–43.1% in controls, P < 0.05) but not from MF (73.1–78.5% v. 70.6–71.8% in controls), whereas regardless of supplementation with FF or GDF9, the maturation rates were always significantly higher in the denuded oocytes from MF (72.4–83.6%) than SF (57.8–66.2%; P < 0.05). Despite of the origin of COC (SF or MF), maturation rates of oocytes denuded 20 h after the start of IVM were not affected by supplementation with FF or GDF9 during the first and/or second half of IVM (P > 0.05). In summary, CC removal from COC 20 h after the start of IVM promotes nuclear maturation of oocytes from SF. Exogenous additives such as GDF9 and follicular fluid from MF do not seem to affect the promotion of nuclear maturation in our experimental conditions.

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The Novel Quantitative Assay for Measuring the Antibiofilm Activity of Volatile Compounds (AntiBioVol)

Applied Sciences ◽

10.3390/app10207343 ◽

2020 ◽

Vol 10 (20) ◽

pp. 7343

Author(s):

Malwina Brożyna ◽

Anna Żywicka ◽

Karol Fijałkowski ◽

Damian Gorczyca ◽

Monika Oleksy-Wawrzyniak ◽

...

Keyword(s):

Volatile Compounds ◽

Cost Effective ◽

Antibiofilm Activity ◽

Experimental Conditions ◽

Laboratory Equipment ◽

Tea Tree ◽

Technical Parameters ◽

The Impact ◽

Standard Disc

Herein, we present a new test, dubbed AntiBioVol, to be used for the quantitative evaluation of antibiofilm activity of volatile compounds in vitro. AntiBioVol is performed in two 24-well plates using a basic microbiological laboratory equipment. To demonstrate AntiBioVol usability, we have scrutinized the activity of volatilized eucalyptus, tea tree, thyme essential oils, and ethanol (used for method suitability testing) against biofilms of Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans. We have also compared AntiBioVol with the standard disc volatilization method, placing a special stress on evaluating the impact of various technical parameters on the outcomes of the latter method. The obtained results indicate that AntiBioVol allows analyzing the antibiofilm activity of volatile compounds in a high number of repeats and provides semi-quantitative or quantitative results of high repeatability. In comparison to disc volatilization, AntiBioVol is a more space- and cost-effective method that allows analyzing various types of microbial aggregates. Moreover, we have indicated that the possible reasons for the discrepancies in the results obtained by means of the standard disc volatilization method may be related to various parameters of the testing dishes used (height, volume, diameter) and to various volumes of the agar medium applied. In turn, the application of a 24-well plate and a strictly defined AntiBioVol protocol provide a higher control of experimental conditions. Therefore, the application of AntiBioVol may enable an optimization of and introduction of volatile compounds to the fight against infective biofilms.

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Dietary compounds as potential modulators of microRNA expression in psoriasis

Therapeutic Advances in Chronic Disease ◽

10.1177/2040622319864805 ◽

2019 ◽

Vol 10 ◽

pp. 204062231986480 ◽

Cited By ~ 13

Author(s):

Hristina Kocic ◽

Giovanni Damiani ◽

Bojana Stamenkovic ◽

Michael Tirant ◽

Andrija Jovic ◽

...

Keyword(s):

Vitamin D ◽

Omega 3 ◽

Experimental Conditions ◽

Keratinocyte Proliferation ◽

Methyl Donors ◽

Beneficial Effects ◽

Small Noncoding Rnas ◽

The Impact

Nutrigenomic DNA reprogramming in different chronic diseases and cancer has been assessed through the stimulation of gene expression and mRNA synthesis versus DNA silencing by CpG DNA modification (methylation); histone modification (acetylation, methylation) and expression of small noncoding RNAs, known as microRNAs (miRNAs). With regard to the specific nutrigenomic effects in psoriasis, the influence of specific diets on inflammatory cell signaling transcriptional factors such as nuclear factor (NF)-κB and Wnt signaling pathways, on disease-related specific cytokine expression, pro/antioxidant balance, keratinocyte proliferation/apoptosis and on proliferation/differentiation ratio have been documented; however, the influence of dietary compounds on the balance between ‘good and bad’ miRNA expression has not been considered. This review aims to summarize knowledge about aberrant microRNAs expression in psoriasis and to emphasize the potential impact of some dietary compounds on endogenous miRNA synthesis in experimental conditions in vivo and in vitro. Among the aberrantly expressed miRNAs in psoriasis, one of the most prominently upregulated seems to be miR-21. The beneficial effects of phenolic compounds (curcumin and resveratrol), vitamin D, methyl donors, and omega-3 fatty acids (eicosapentaenoic acid and docosahexaenoic acid) are discussed. Highly expressed miR-155 has been downregulated by flavonoids (through a quercetin-rich diet) and by vitamin D. Quercetin has been effective in modulating miR-146a. On the other hand, downregulated miR-125b expression was restored by vitamin D, Coenzyme Q10 and by microelement selenium. In conclusion, the miRNA profile, together with other ‘omics’, may constitute a multifaceted approach to explore the impact of diet on psoriasis prevention and treatment.

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Follicular size affects the meiotic competence of in vitro matured prepubertal and adult oocytes in sheep

Reproduction Nutrition Development ◽

10.1051/rnd:19990409 ◽

1999 ◽

Vol 39 (4) ◽

pp. 503-508 ◽

Cited By ~ 28

Author(s):

Sergio Ledda ◽

Luisa Bogliolo ◽

Giovanni Leoni ◽

Salvatore Naitana

Keyword(s):

Follicular Size ◽

Meiotic Competence

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Time-dependent and force-dependent vasoreactivity of isolated human umbilical arteries

SCRIPTA MEDICA ◽

10.5937/scriptamed51-28131 ◽

2020 ◽

Vol 51 (3) ◽

pp. 134-140

Author(s):

Đorđe Đukanović ◽

Milica Gajić ◽

Ranko Škrbić

Keyword(s):

Incubation Period ◽

Cold Storage ◽

Dose Response ◽

In Vitro Studies ◽

Time Dependent ◽

Response Curves ◽

Experimental Conditions ◽

Umbilical Arteries ◽

The Impact

Background/Aim: There have been different experimental conditions for in vitro studies on human umbilical arteries (HUA) in tissue bath system. This diversity was mainly reflected in variables such as stretching tension, incubation period and initial constriction challenging with potassium (KCl). The aim of the study was to establish optimal experimental conditions which will provide better responsiveness of HUA preparations, as well as to examine the impact of 24 h cold storage on viability and responsiveness of HUA to KCl and serotonin. Methods: The KCl-induced constrictions at different stretching tensions (0.5 g, 1.0 g, 2.0 g, 4.0 g), incubation times (30 min, 60 min, 120 min), and after multiple initial constriction challenging were compared. Dose response curves for serotonin were obtained under different conditions (1.0 g and 60 min vs. 2.0 g and 120 min). The influence of 24 h cold storage on KCland serotonininduced vasoconstriction of HUA preparations was examined as well. Results: The strongest constrictions induced by serotonin or KCl were obtained when preparations were adjusted at 2.0 g and incubated for 120 min. The KCl-induced constrictions observed after 120 min were statistically higher (p < 0.05) when preparations were challenged three times (30 min, 60 min, 120 min), compared to those challenged only once. The preparations that were stored at 4 ⁰C for 24 h showed significantly stronger serotonin-induced constrictions (p < 0.01). The cold storage had no influence on KCl-induced constriction. Conclusion: For performing in vitro studies on HUA preparations in tissue bath, we propose stretching tension of 2.0 g, incubation period of 120 min and multiple initial constriction challenging with KCl as optimal experimental condition. We also showed that HUA preparations retained functional viability even after 24 h of cold storage.

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43 WARMING TEMPERATURE AFFECTS THE VIABILITY AND MEIOTIC COMPETENCE OF IMMATURE PORCINE OOCYTES VITRIFIED IN A CHEMICALLY DEFINED SOLUTION

Reproduction Fertility and Development ◽

10.1071/rdv26n1ab43 ◽

2014 ◽

Vol 26 (1) ◽

pp. 135

Author(s):

D. Takahashi ◽

H. Funahashi

Keyword(s):

In Vitro Maturation ◽

Fluorescent Microscopy ◽

Dibutyryl Cyclic Amp ◽

Nuclear Maturation ◽

Porcine Oocyte ◽

Different Temperatures ◽

Maturation Rate ◽

Post Hoc ◽

Meiotic Competence

The aim of this study was to examine the viability and meiotic competence of porcine oocytes when immature porcine cumulus-oocyte complexes (COC) were pretreated for vitrification at different temperatures (25 and 39°C), vitrified in a chemically defined solution, and warmed at different temperatures (39 and 60°C). Cumulus-oocyte complexes were aspirated from middle-size follicles (3–6 mm in diameter) of abattoir-derived porcine ovaries. After collection, the COC were pretreated with cryoprotectants at different temperatures (25 and 39°C) and vitrified in a serum-free chemically defined solution containing 0.6 mg mL–1 of hydroxypropyl cellulose, basically according to a commercial protocol (Cryotop, Kitazato BioPharma Co. Ltd., Fuji, Japan). The vitrified COC were warmed in 1 M trehalose solution at 39 for 60 s or at 60°C for 30 s. The COC were cultured for in vitro maturation (IVM) in modified porcine oocyte medium (POM) supplemented with 50 μM β-mercaptoethanol, 10 IU mL–1 of eCG, 10 IU mL–1 of hCG, and 1 mM dibutyryl cyclic AMP (dbcAMP) for 20 h and then in the fresh medium without hormonal supplements and dbcAMP for another 24 h. Viability of COC was evaluated under fluorescent microscopy after stain with fluorescein diacetate and propidium iodide. Nuclear maturation of the oocytes was evaluated after 44 h of IVM. Statistical analyses of results from 5 replicated trials were performed by ANOVA with a Bonferroni/Dunn post-hoc test (significance, P < 0.05). Although viabilities of vitrified oocytes after 44 h of IVM [6.0% (9/149) to 37.8% (59/155)] were significantly lower than fresh controls [98.8% (158/160)], the viabilities of vitrified oocytes warmed at 60°C [32.0% (49/160) to 37.8% (59/155)] were significantly higher than those warmed at 39°C [6.0% (9/149) to 10.0% (16/160)]. Maturation rates in vitrified oocytes [2.7% (4/149) to 19.8% (31/155)] were also significantly lower than fresh controls [74.8% (120/160)]. Regardless of temperature during pretreatment for vitrification (25 and 39°C), maturation rate of the oocytes warmed at 60°C after vitrification [16.4% (25/154) to 19.8% (31/155)] was significantly higher than that warmed at 39°C [3.1% (5/160) to 2.7% (4/149)]. In conclusion, these results demonstrate that warming at 60°C for 30 s maintains the viability and meiotic competence of immature porcine COC.

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231 EFFECT OF DIFFERENT EQUINE CHORIONIC GONADOTROPIN CONCENTRATIONS ON IN VITRO-PRODUCED BOVINE EMBRYOS

Reproduction Fertility and Development ◽

10.1071/rdv25n1ab231 ◽

2013 ◽

Vol 25 (1) ◽

pp. 263

Author(s):

C. Decanine ◽

E. M. Pioltine ◽

I. P. Emanuelli ◽

R. Z. Puelker ◽

M. F. G. Nogueira

Keyword(s):

Calf Serum ◽

Bovine Embryos ◽

In Vitro Production ◽

Experimental Conditions ◽

Essential Components ◽

Chi Squared ◽

Equine Chorionic Gonadotropin ◽

And Control ◽

Hatching Rates

In vitro maturation (IVM) is one of the most challenging steps in the in vitro production of bovine embryos. The IVM medium must provide the necessary conditions for the occurrence of nuclear and cytoplasmic maturation, close to the physiological conditions. The pituitary gonadotropins are essential components for generating competent oocytes; however, whether these hormones (pituitary, placental, or both) are essential and which concentrations should be used are still controversial. Our work aimed to compare the effect of different concentrations of the placental gonadotropin (eCG) in the IVM medium on the in vitro-produced bovine blastocysts. Cumulus–oocyte complexes (n = 1341, grades I and II), obtained from ovaries from an abattoir were selected and distributed into six groups: (1) eCG (4 IU mL–1; n = 192); (2) eCG (1.5 IU mL–1; n = 204); (3) eCG (1.3 IU mL–1; n = 203); (4) eCG (0.15 IU mL–1; n = 202); (5) eCG (0.015 IU mL–1; n = 199); (6) control: FSH (0.1 mg mL–1), LH (50 µg mL–1), and 10% of fetal calf serum (FCS; n = 341). Medium from groups 1 to 5 also contained LH (50 µg mL–1) and BSA (6 mg mL–1). The cumulus–oocyte complexes were matured in TCM-199 for 24 h and were IVF (Day 0) in TALP-IVF for 22 to 24 h. Viable spermatozoa were selected by Percoll gradient, and they were evaluated (motility and spermatozoa concentration) to provide the insemination concentration (106 spermatozoa mL–1). Presumptive zygotes were cultured in SOF medium supplemented with FCS (2.5%) and BSA (5 mg mL–1) in an incubator (38.3°C, 5% CO2, and maximum humidity). Embryo development was evaluated in terms of cleavage (Day 3), blastocyst (Day 7), and hatching rates (Day 10). Mean rates were analysed by chi-squared test and ANOVA, and significance was considered when P < 0.05. The results obtained from the different groups are shown in Table 1. Cleavage, blastocyst, and hatching rates were not different among groups. We conclude that, under our experimental conditions, even minimal concentrations of eCG (0.015 IU) may be a viable and effective alternative in the replacement of FSH for the IVM of bovine oocytes. Table 1.Cleavage, blastocyst, and hatching rates of the experimental groups (1, 2, 3, 4, and 5) and control group1 Fellowships and support by CAPES and FAPESP.

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Effect of administering a crude equine gonadotrophin preparation to mares on follicular development, oocyte recovery rate and oocyte maturation in vivo

Reproduction ◽

10.1530/reprod/118.2.351 ◽

2000 ◽

pp. 351-360 ◽

Cited By ~ 15

Author(s):

I Bruck ◽

J Bezard ◽

M Baltsen ◽

B Synnestvedt ◽

I Couty ◽

...

Keyword(s):

Oocyte Maturation ◽

Recovery Rate ◽

In Vitro Maturation ◽

Follicular Development ◽

Vitro Fertilization ◽

Nuclear Maturation ◽

Oocyte Yield ◽

Oocyte Recovery

In mares, the shortage of oocytes and the variability in nuclear maturation at a certain time of the oestrous cycle hinders the optimization of methods for in vitro maturation and in vitro fertilization. Increasing the number of small-to-medium-sized follicles available for aspiration in vivo may increase the overall oocyte yield. The aims of the present study were to investigate whether administration of crude equine gonadotrophins affects follicular development, oocyte recovery rate, in vivo oocyte maturation and follicular concentrations of meiosis-activating sterols. During oestrus, all follicles >/= 4 mm were aspirated from 19 pony mares (first aspiration: A1). Over the next 8 days, the mares were treated daily with either 25 mg crude equine gonadotrophins (n = 10) or physiological saline (n = 9). Between day 1 and day 8, follicular growth was monitored by ultrasonography. On day 8, all follicles >/= 4 mm were evacuated (second aspiration: A2) and nuclear maturation of the recovered oocytes was assessed after orcein staining. Follicular growth between A1 and A2, as well as the number and size of follicles at A2 were similar for control mares and mares treated with crude equine gonadotrophins. The oocyte recovery rates at A1 and A2 were similar. At A2, the oocyte recovery rate and oocyte maturation in vivo were not affected by treatment with crude equine gonadotrophins. The number of expanded cumulus oophorus complexes recovered from follicles </= 29 mm was significantly higher at A1 than at A2. The number of oocytes at the germinal vesicle stage was significantly higher at A2 (41.5%) than at A1 (17.8%). Meiosis-activating sterols (FF-MAS and T-MAS) were identified in follicular fluid recovered at A2. Follicular concentrations of FF-MAS and T-MAS were unaffected by treatment with crude equine gonadotrophins. The present study demonstrates that follicular aspiration during oestrus allowed a new follicular population to develop and resulted in a higher degree of synchronization of oocyte development with respect to cumulus expansion and nuclear maturation. The availability of a more homogeneous population of oocytes might facilitate a better optimization of in vitro maturation and in vitro fertilization techniques in mares. Administration of crude equine gonadotrophins during early dioestrus did not affect the growth of small follicles, the oocyte yield after aspiration or oocyte maturation in vivo.

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Intestinal fermentation in vitro models to study food-induced gut microbiota shift: an updated review

FEMS Microbiology Letters ◽

10.1093/femsle/fnaa097 ◽

2020 ◽

Vol 367 (12) ◽

Author(s):

Lorenzo Nissen ◽

Flavia Casciano ◽

Andrea Gianotti

Keyword(s):

Gut Microbiota ◽

In Vitro Models ◽

Experimental Conditions ◽

Applied Microbiology ◽

Microbial Groups ◽

Technical Features ◽

State Of The Science ◽

The Impact ◽

Small Working

ABSTRACT In vitro gut fermentation models were firstly introduced in nutrition and applied microbiology research back in the 1990s. These models have improved greatly during time, mainly over the resemblance to the complexity of digestion stages, the replication of experimental conditions, the multitude of ecological parameters to assay. The state of the science is that the most competitive models shall include a complex gut microbiota, small working volumes, distinct interconnected compartments and rigorous bio-chemical and ecological settings, controlled by a computer, as well as a free-hands accessibility, not to contaminate the mock microbiota. These models are a useful tool to study the impact of a given diet compound, e.g. prebiotics, on the human gut microbiota. The principal application is to focus on the shift of the core microbial groups and selected species together with their metabolites, assaying their diversity, richness and abundance in the community over time. Besides, it is possible to study how a compound is digested, which metabolic pathways are triggered, and the type and quantity of microbial metabolites produced. Further prospective should focus on challenges with pathogens as well as on ecology of gut syndromes. In this minireview an updated presentation of the most used intestinal models is presented, basing on their concept, technical features, as well as on research applications.

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