Evaluation of ruminal methane and ammonia formation and microbiota composition as affected by supplements based on mixtures of tannins and essential oils using Rusitec

Background Dietary supplements based on tannin extracts or essential oil compounds(EOC)have been repeatedly reported as a promising feeding strategy to reduce the environmental impact of ruminant husbandry.A previous batch culture screening of various supplements identified selected mixtures with an enhanced potential to mitigate ruminal methane and ammonia formation.Among these,Q-2(named after quebracho extract and EOC blend 2,composed of carvacrol,thymol,and eugenol)and C-10(chestnut extract and EOC blend 10,consisting of oregano and thyme essential oils and limonene)have been investigated in detail in the present study with the semi-continuous rumen simulation technique(Rusitec)in three independent runs.For this purpose,Q-2 and C-10,dosed according to the previous study,were compared with a non-supplemented diet(negative control,NC)and with one supplemented with the commercial EOC-based Agolin^(R) Ruminant(positive control,PC).Results From d 5 to 10 of fermentation incubation liquid was collected and analysed for pH,ammonia,protozoa count,and gas composition.Feed residues were collected for the determination of ruminal degradability.On d 10,samples of incubation liquid were also characterised for bacterial,archaeal and fungal communities by high-throughput sequencing of 16S rRNA and 26S ribosomal large subunit gene amplicons.Regardless of the duration of the fermentation period,Q-2 and C-10 were similarly efficient as PC in mitigating either ammonia(-37%by Q-2,-34%by PC)or methane formation(-12%by C-10,-12%by PC).The PC was also responsible for lower feed degradability and bacterial and fungal richness,whereas Q-2 and C-10 effects,particularly on microbiome diversities,were limited compared to NC.Conclusions All additives showed the potential to mitigate methane or ammonia formation,or both,in vitro over a period of 10 d.However,several differences occurred between PC and Q-2/C-10,indicating different mechanisms of action.The pronounced defaunation caused by PC and its suggested consequences apparently determined at least part of the mitigant effects.Although the depressive effect on NDF degradability caused by Q-2 and C-10 might partially explain their mitigation properties,their mechanisms of action remain mostly to be elucidated.

STABILITY ANALYSIS OF EQUILIBRIUM FOR MICROORGANISMS IN CONTINUOUS CULTURE

The problem of equilibrium of non-linear dynamic system for microorganism in continuous culture is considered in this paper. The existence of equilibrium point is proved. The conclusion that the equilibrium is the continuous function of the dilution rate and substrate concentration in medium in a certain range is concluded. And the stability criterion of equilibrium is obtained. In theory, it explains the multiplicity shown in laboratory.

Environmental control of growth and BChl a expression in an aerobic anoxygenic phototrophic bacterium, Erythrobacter longus (DSMZ6997)

anoxygenic phototrophic bacteria （AAPB）, which form a unique functional group of heterotrophic bacteria, have the abilit,to utilize light energy. The impact of carbon source and light intensity on the growth and bacteriochlorophyn a （ BChl a） expression of a typical strain of AAPB, Erythrobacter longus strain DSMZ6997 was examined during batch culture and continuous culture. The results showed that the expression of BChl a in DSMZ6997 was regulated by both carbon-source and light conditions, and was stimulated by low availability of carbon but inhibited by light to a certain extent at 300 lx and completely at 1 500 lx. In contrast, cell abundance, and even cell size of this strain, was substantially enhanced under light/dark cycle cultivation conditions over dark conditions, indicating the promotion of growth by light. These results led to the conclusion that utilization of light through BChl a helps AAPB to survive under carbon stress, while light at high intensity is harmful to the synthesis of BChl a in AAPB.

Ultra-thin temperature controllable microwell array chip for continuous real-time high-resolution imaging of living single cells

Single-cell imaging,a powerful analytical method to study single-cell behavior,such as gene expression and protein profiling,provides an essential basis for modern medical diagnosis.The coding and localization function of microfluidic chips has been developed and applied in living single-cell imaging in recent years.Simultaneously,chip-based living single-cell imaging is also limited by complicated trapping steps,low cell utilization,and difficult high-resolution imaging.To solve these problems,an ultra-thin temperature-controllable microwell array chip(UTCMA chip)was designed to develop a living single-cell workstation in this study for continuous on-chip culture and real-time high-resolution imaging of living single cells.The chip-based on ultra-thin ITO glass is highly matched with an inverted microscope(or confocal microscope)with a high magnification objective(100×oil lens),and the temperature of the chip can be controlled by combining it with a home-made temperature control device.High-throughput single-cell patterning is realized in one step when the microwell array on the chip uses hydrophilic glass as the substrate and hydrophobic SU-8 photoresist as the wall.The cell utilization rate,single-cell capture rate,and microwell occupancy rate are all close to 100%in the microwell array.This method will be useful in rare single-cell research,extending its application in the biological and medical-related fields,such as early diagnosis of disease,personalized therapy,and research-based on single-cell analysis.