组学视角下养殖动物消化道微生物组结构与功能研究进展北大核心

养殖动物消化道中含有大量的微生物,不仅参与动物对营养物质的消化和吸收,还对宿主生长发育及免疫起重要调节作用。动物消化道微生物组研究是目前国内外的热点领域,取得了一系列重要研究进展。深入了解养殖动物消化道微生物组的结构与功能,将为今后调控和应用消化道微生物、提高动物生产性能、改善动物胃肠道健康和实现绿色健康养殖奠定理论基础。本文以4种代表性养殖动物(牛、羊、猪和鸡)为主体,对组学视角下其消化道微生物群落结构、功能等研究进展进行总结和分析;并对未来研究方向进行展望。     

Evolutionary patterns of Perissodactyla and Artiodactyla (Mammalia) with different types of digestion

Described and discussed are the comparative and functional anatomy of the digestive tract in ungulates, as well as the retention of digesta in the total digestive tract and in its compartments. Additionally, the history of distribution of ungulates under special consideration of the connections between continents and the development of different biomes is discussed. The last 40 million years are considered. During the Caenozoic the forestomach-fermenting Artiodactyla were strongly radiating, which is in contrast to conditions in hindgut-fermenting Artiodactyla (Suidae) and Perissodactyla (Equidae, Rhinocerotidae, Tapiridae). The Pecora have been especially successful forestomach-fer-menters because they are able to reduce particle size of digesta through the process of rumination and thus increase digesta passage. This is advantageous because not only the metabolic products (mainly volatile fatty acids) of microbial activity are made available to the mammal, but also a higher percentage of those nutrients that can be autoenzymatically digested by the ungulate.     

Phylogenetic constraints on digesta separation: Variation in fluid throughput in the digestive tract in mammalian herbivores

The relevance of the mean retention time (MRT) of particles through the gastrointestinal tract (GIT) is well understood and MRT(particle)GIT is an important parameter in digestion models. Solute markers have been used to estimate MRT(solute)GIT (or 'fluid passage') in animals, but the relevance of this measure is less evident and is usually sought in its relation to MRT(particle)GIT. The ratio between the two measures indicates the degree of 'digesta washing', with little washing occurring at ratios of 1, aborad washing at ratios >1 (where the solute marker travels faster than the particle marker), and orad (retrograde) washing at ratios <1 (where the solute marker travels slower than the particle marker). We analysed digesta washing in a dataset of 98 mammalian species including man of different digestion types (caecum, colon and nonruminant foregut fermenters, and ruminants), controlling for phylogeny; a subset of 72 species allowed testing for the influence of food intake level. The results indicate that MRT(solute)GIT and the degree of digesta washing are related to digestion type, whereas variation in MRT(particle)GIT is influenced mainly by effects of body mass and food intake. Thus, fluid throughput and digesta washing emerge as important correlates of digestive anatomy. Most importantly, primates appear constrained to little digesta washing compared to non-primate mammalian herbivores, regardless of their digestion type. These results may help explain the absence of primates from certain herbivore niches and represent a drastic example of a physiologic limitation in a phylogenetic group. More experimental research is required to illuminate relative benefits and costs of digesta washing.     

Animal digestive strategies versus anaerobic digestion bioprocesses for biogas production from lignocellulosic biomass

Herbivorous mammals and wood-eating insects are fairly effective in the digestion of plant polymers, such as lignocellulosics. In order to improve methane production from the lignocellulosic biomass, several kinds of anaerobic digestion processes derived from animal models have been devised. However, the rates of biodegradation occurring in the anaerobic bioreactors still remain lower than in animal guts. The effectiveness of the digestive systems of those animals results from the concerted action of the various enzymes (e.g. cellulases, xylanases, esterases, ligninases) produced in their guts as well as their integration with mechanical and chemical actions. Powerful pretreatment (prefermentation) operations are integrated to and support efficiently the microbial fermentation system, e.g. the rumination (i.e. mechanical) in ruminants and the secretion of endogenous cellulases (i.e. enzymatic) or the alkaline treatment (chemical) at mid-way in xylophagous insects. The oxygen gradients along the gastrointestinal tract may also stimulate the hydrolytic activities of some microbial populations. In addition, the solid retention time, the digesta flow and the removal of the end-products are well ordered to enable animals to thrive on a complex polymer such as lignocellulose. At the same time, technologies were developed to degrade lignocellulosic biomass, such as the rumen derived anaerobic digestion (RUDAD) process and the rumen simulating technique (RUSITEC), more elaborated and using rumen microbial consortia. Overall, they showed that the fermentation taking place in the rumen fermentation and even in the hindgut are biological processes that go beyond the limited environmental conditions generally found in anaerobic digesters. Hence, knowledge on herbivores' digestion mechanisms might be better exploited in the design and operation of anaerobic digesters. This literature review is a cross-analysis of the relevant information about the digestive strategies of herbivorous and wood-eating animals and the bioengineering techniques in lignocelluloses degradation. The aim is to highlight strategies of animals' digestion simulation for more effective anaerobic digestion of lignocellulosic compounds and other solid residues.     

Ruminal fermentation and fill change with season in an arctic grazer: responses to hyperphagia and hypophagia in muskoxen (Ovibos moschatus)

We studied castrated adult muskoxen fed a standard diet of grass hay and supplement throughout the year to determine seasonal changes in digesta passage, fill, and fermentation without the confounding effects of reproductive demands or changes in food quality. Although food intake increased by 74% between spring and autumn, mean retention times of fluid and particulate digesta markers were maintained between seasons in both the rumen (9-13 h) and the intestines (27-37 h). The rumen contained 84.5% of digesta and accounted for 79% of dry matter digestion in the whole digestive tract. Ruminal fluid space and whole-gut digesta fill increased by 31%-34%, while ruminal rates of in situ degradation increased by more than 100% between spring and autumn for cellulose and hemicellulose. Hyperphagia in autumn was accompanied by increased bacterial counts in ruminal fluid (30%), declines in ruminal pH, and increases in the concentration of fermentation acids (16%) when compared with spring hypophagia. Consumption of fresh hay and supplement increased the concentrations of acids most markedly during winter and spring when bacterial counts were low. Low food intakes in winter and spring may limit the microbial population, whereas hyperphagia in autumn may foster a much more active microflora that requires consistent supplies of substrate. Plasticity of fill and fermentation in muskoxen minimizes winter costs and maximizes nutrients and energy gained from coarse forages in small home ranges throughout the year.     

Flow rates of components in digesta of pigs prepared with re-entrant cannulas in the proximal duodenum and terminal ileum, and fed semipurified, hard wheat, and soft wheat diets.

Four pigs prepared with re-entrant cannulas in the proximal duodenum and terminal ileum were used to study flow rates of total digesta, insoluble dry matter, nitrogen, and amino acids entering and leaving the small intestine. The pigs received a semipurified diet, a hard wheat diet, or a soft wheat diet. These were approximately isonitrogenous. A higher rate of passage of digesta through the proximal duodenum and terminal ileum were measured in pigs receiving the hard wheat diet. Peak flow of digesta at the duodenum of all pigs occurred at 1 h post feeding. Peak flow of digesta at the ileum occurred at 9 h post feeding on the soft wheat diet, but somewhat earlier on the hard wheat and semipurified diet. More nitrogen and essential amino acids flowed in the solid fraction of duodenal digesta during the first 2 h post feeding for the wheat diets and 4 h post feeding for the semipurified diet. It was concluded that flow rate of most nutrients from the stomach and through the small intestine of pigs is modified by the composition and texture of the food ingested. It is postulated that efficiency of mixing of digesta with digestive secretions in the stomach is a major factor influencing rate of flow.     

Digesta passage rates in the Florida manatee (Trichechus manatus latirostris)

The Florida manatee, Trichechus manatus latirostris (Sirenia: Trichechidae), is an herbivorous marine mammal found within coastal areas throughout the state of Florida, which feeds on both fresh and salt water sea grasses. Manatees, like other Sirenians, are a tropical species with little tolerance for water temperatures below 20°C, rely on a relatively poor nutritional food source, and have a low metabolic rate. Although manatees are hindgut fermenting herbivores, they are very efficient at extracting nutrients from the plants on which they feed. Slow passage rates of digesta have been suggested to be a factor in this increased efficiency. Two studies monitored the digesta passage times and mixing of particulate digesta within the manatee digestive tract using MicroGrits colored corncob grit as a fecal marker. Fecal samples were collected subsequently from four manatees in Study 1 and 3 manatees in Study 2, grit pieces removed, counted, and measured. The digesta passage times ranged from 6 and 10 days in Study 1, and 4.3 and 8.3 days in Study 2, supporting data presented in previous studies. When two different colored markers were administered on sequential days, minimal to no mixing was seen in recovered feces, suggesting that the digesta from a given day traveled through the tract as a bolus. Less than 1% of the marker fed was recovered and we hypothesize that perpendicular folds of the large intestine may be the major contributing factor, with pieces being retained and eventually digested. Zoo Biol 26:503–515, 2007. © 2007 Wiley‐Liss, Inc.     

Digesta Passage,Digestibility and Behavior in Captive Gorillas Under Two Dietary Regimens

Gorilla adaptation has been debated in recent years given the wide variation among diets of gorillas in different habitats. Gorillas are the largest of living primates, have large colons and should be capable of processing tough foods. Preliminary captive studies have suggested that they may well have long average gut retention times relative to smaller hominoids, which should facilitate digestive efficiency in their wild counterparts. Indeed, wild gorillas consume large amounts of fibrous foods as staples or fall-back foods across their range, in response to habitat-related or seasonal changes in fruit availability. Fluctuations in diet might be matched by changes in digesta passage and digestibility, with possible selective retention of harder to digest items. We further studied digestive processes via chemical cobalt and chromium markers to track liquid and solids, as they passed through the guts of gorillas at the San Francisco Zoo (SFZ). In addition, we examined the effects of variation in captive diets on intake, digesta passage, digestion and behavior. The SFZ gorillas exhibited high digestibility coefficients, and gut passage was long relative to those of smaller-bodied hominoids. The results permit us to understand more fully the relationships of digestive processes to adaptation and dietary flexibility in the wild and to inform the development of dietary recommendations to improve the well-being of captive gorillas.     

Transfer of sulphur to the digestive tract of sheep.

The transfer of sulphate from plasma to digestive tract and from digestive tract to plasma in crossbred sheep was estimated by the use of isotope dilution techniques with Na235SO4. The passage of 35S along the digestive tract was simultaneously measured by reference to two inert radioactive markers infused intraruminally. In the first experiment, three sheep given a roughage-based diet containing 174 +/- 7 mg S/day received an intravenous infusion of Na235SO4 for 7 days before collections were made of plasma and of digesta from the rumen, abomasum and terminal ileum. Similar collections were made in the second experiment in which four sheep received intraruminal infusions of Na235SO4. From estimates of infusion rate of 35S, specific radioactivity of 35S in plasma and digesta and rate of flow of sulphur in the digestive tract the following calculations were made: The transfer of sulphate from the plasma to the rumen was calculated as 29 mg S/day. Of this only 12 mg S/day passed as organic sulphur in digesta from the stomach. As the net gain of sulphur in the stomach in this experiment was 153 mg/day, sulphate transferred from the plasma contributed only a small amount of sulphur derived from endogenous sources in the stomach. In contrast, the substantial passage of 35S into the intestinal lumen during intravenous infusion of 35SO4 suggested that 38 and 41 mg S/day of the 236 and 145 mg organic S/day flowing from the small and large intestine respectively was derived from plasma sulphate, corresponding to about 26% of the dose.     

哺乳动物的消化策略（英文）

理解动物的营养生态位是充分理解其整个生态学的基础,对于害兽控制和物种保护也很重要,食肉动物的小肠很发达,这可能与对食物的高消化能力有关;杂食性动物有更复杂的胃肠器官,其后端有可进行发酵的盲肠,消化物的平均滞留时间（mean retention times,MRTs)更长;最长的平均滞留时间见于食草动物,其消化道内高密度的微生物种群对不同滞留区内的消化物进行发酵,但是,并不是所有的食草动物都能够最大程度地消化植物纤维,只有反刍动物、骆驼和个体较大的后肠发酵动物（hindgut fermenter)能够具有这种能力,对比而言,许多其它的食草动物,如前肠发酵的有袋类和小型的后肠发酵动物如兔子、田鼠和负鼠等,它们具备可以使植物纤维消化效率最大的消化系统,可以在食物中的纤维素含量非常高的情况下仍能处理大量的食物。这些不同的消化策略使哺乳动物具有广幅的营养生态位。     

Differences among captive callitrichids in the digestive responses to dietary gum

Although many members of the Callitrichidae, a monophyletic family of small, New World monkeys, have been observed to feed on plant exudates, available field data support the generalization that pygmy and common marmosets (Cebuella pygmaea and Callithrix jacchus) feed on gums to a greater extent than most other callitrichids. Because microbial fermentation is required for vertebrates to digest gums, gum-feeding primates may react differently to dietary gum from their relatives that eat little gum. To test this hypothesis, digestion trials were conducted on animals from the two marmoset species, two tamarin species (Saguinus fuscicollis and S. oedipus), and a species of lion tamarin (Leontopithecus rosalia). These species span the range of body sizes within the Callitrichidae. All animals were fed two variations of a homogeneous diet, which differed only in that gum arabic was added to one. Transit time of digesta (TFA) and digestive efficiency (as measured by the coefficients of apparent digestibility of dry matter and energy [ADDM and ADE, respectively]) were compared between diets for each individual. As predicted, the digestive responses of marmosets differed from the responses of the other study species. In marmosets, TFA tended to be longer when gum was added to the diet, while TFA did not change in the other three species. Digestive efficiency decreased in tamarins and lion tamarins with the addition of gum to the diet; marmoset digestive efficiency was unaffected by diet. The results of this research are consistent with the hypothesis that marmosets have digestive adaptations that aid in the digestion of gum that other callitrichids lack. © 1996 Wiley-Liss, Inc.     

Physiology of intermittent feeding: integrating responses of vertebrates to nutritional deficit and excess

Food intakes of wild animals may not match their requirements for nutrients and energy but may vary between periods of nutritional excess (hyperphagia) and nutritional deficit (hypophagia) at timescales that vary from days to months. We present a simple model of feeding patterns and requirements of vertebrates. Frequent fasts and high intakes are typical of endothermic predators and migratory birds, whereas slow cycles and long deficits typify feeding patterns of ectothermic predators and ungulates in seasonal environments. We propose that hyperphagia is constrained by the ability to increase processes of digestion, absorption, intermediary metabolism, net deposition in tissue, and excretion to match loads of digesta and metabolites. Hyperphagia on high-quality diets is limited by the clearance of metabolites, whereas digestive tract capacity and flow limit consumption of low-quality diets. Of all digestive strategies, small omnivores with simple digestive systems may be the most tolerant of frequent hyperphagia. Tolerance of hypophagia favors large endogenous stores or low mass-specific rates of metabolism and reproductive output. Large animals may be most able to sustain reproduction during prolonged deficits in seasonal environments. Responses to excessive and deficient intakes of food are constrained by the length of the feeding cycle. Animals adapted to short feeding cycles may be best suited to unpredictable food supplies but at the energetic cost of maintaining spare capacity for digestion and absorption. Predictions of the response to food disruption are best evaluated in the context of body size, nutritional physiology, and life history of the species and the time for internal response.     

Digestive performance and selective digesta retention in the long-nosed bandicoot,Perameles nasuta,a small omnivorous marsupial

Bandicoots are opportunistic omnivores that feed on invertebrates, fungi and both epigeal and hypogeal plant parts. We examined the performance of the digestive tract of the long-nosed bandicoot (Perameles nasuta) in terms of intake and total digestibility, patterns of excretion of inert digesta markers, and likely sites of digesta retention, on two diets designed to mimic part of their natural plant and insect diets. On the insect diet (mealworm larvae), bandicoots virtually maintained body mass at a digestible energy intake of 511 kJ · kg^-0.75 · day^-1 and were in strongly positive nitrogen balance. In contrast, on the plant diet (shredded sweet potato), bandicoots ate only one-third as much digestible energy, lost 7% body mass, and were in negative nitrogen balance. Mean retention times of two particle markers on the plant diet (27.5 and 27.0 h) were more than double those on the insect diet (12.4 and 11.2 h), and on both diets the mean retention time of the fluid digesta marker was greater than those of the particle markers, indicating consistent selective retention of fluid digesta in the gut. It was seen radiographically than in mealwormfed bandicoots major sites of digesta retention were the distal colon and rectum, whereas in the sweet potato-fed animals the caecum and proximal colon were principal sites. It was concluded that retention of plant material in the caecum and proximal colon (the main sites of microbial digestion) and the preferential retention of fluid digesta (together with bacteria and small feed particles) in the caecum were important factors in the ability of bandicoots to switch between insect and plant foods, depending on relative availabilities, and thus to exploit nutritionally unpredictable environments.Abbreviations ADF acid-detergent fibre - bm body mass - Co-ED-TA cobalt-ethylenediaminetetra-acetic acid - CWC cell wall constituents - DE digestable energy - dm dry matter - EUN endogenous urinary nitrogen - ICP inductively-coupled plasma atomic emission spectroscopy - MFN metabolic faecal nitrogen - MRT mean retention time - NDF neutral-detergent fibre - ww wet weight     

Optimal digestive strategies for arboreal herbivorous mammals in contrasting forest types: Why Koalas and Colobines are different

Abstract The eucalypt forests of temperate eastern Australia contrast with many forests in tropical Australia, Asia, Africa and Central America in terms of degrees of sclerophylly, concentrations of nutrients in foliage, types and amounts of chemical challenges posed for herbivores, and the range of alternatives to foliage as food. Comparisons between digestive strategies employed by arboreal herbivorous marsupials and primates inhabiting these forests reveal several trends: (i) foregut-fermenters occur only in tropical and sub-tropical forests, whereas caecum and caecum-colon fermenters occur in both temperate eucalypt and the tropical forests; (ii) the arboreal marsupials and primates (koalas, greater gliders, ringtail possums and sportive lemurs) that are the most folivorous are caecum or caecum-colon fermenters with colonic specializations for selective retention of small panicles of digesta; (iii) no caecum or caecum- colon fermenters lacking colonic separation mechanisms achieve consistently high degrees of folivory; (iv) even the most folivorous foregut-fermenting arboreal marsupials (tree kangaroos) or primates (colobus monkeys) include a substantial proportion of high-quality components (fruits or seeds) in their diet. Caecum or caecum-colon fermentation with colonic separation is postulated to be essential for a high degree of folivory in small mammals because it offsets potential limits on intake of high-fibre food and minimizes losses of faecal nitrogen that otherwise would be prohibitive. This digestive strategy is common among folivores in eucalypt forests because the low abundance of large seeds and fleshy fruits limits folivory-frugivory and folivory-granivory. Foregut fermentation employing a sacciform/ tubiform forestomach (as in tree kangaroos and colobus monkeys) is postulated to be optimal for mixed leaf/fruit and leaf/seed diets that are available in many tropical forests, assuming it allows high-fibre meals to be retained for microbial fermentation while permitting low-fibre meals to pass rapidly to the hindstomach for acid/enzymic digestion. However, foregut fermentation appears innappropriate for either primary frugivory or exclusive folivory due to the inefficiency of microbial digestion of simple sugars and constraints on microbial fermentation as a primary energy source for small mammals.     

Opportunities and challenges with transitioning to non-lethal sampling of wild fish for microbiome research

The microbial communities of fish are considered an integral part of maintaining the overall health and fitness of their host. Research has shown that resident microbes reside on various mucosal surfaces, such as the gills, skin, and gastrointestinal tract, and play a key role in various host functions, including digestion, immunity, and disease resistance. A second, more transient group of microbes reside in the digesta, or feces, and are primarily influenced by environmental factors such as the host diet. The vast majority of fish microbiome research currently uses lethal sampling to analyse any one of these mucosal and/or digesta microbial communities. The present paper discusses the various opportunities that non-lethal microbiome sampling offers, as well as some inherent challenges, with the ultimate goal of creating a sound argument for future researchers to transition to non-lethal sampling of wild fish in microbiome research. Doing so will reduce animal welfare and population impacts on fish while creating novel opportunities to link host microbial communities to an individual's behavior and survival across space and time (e.g., life-stages, seasons). Current lethal sampling efforts constrain our ability to understand the mechanistic ecological consequences of variation in microbiome communities in the wild. Transitioning to non-lethal sampling will open new frontiers in ecological and microbial research.     

Organic acid concentrations and digesta movement in the gastrointestinal tract of the bushbaby (Galago crassicaudatus) and Vervet monkey (Cercopithecidae pygerythrus)

Studies were conducted to compare digestive functions in two species of sub-human primate, the bushbaby (Galago crassicaudatus) and the Vervet monkey (Cercopithecidae pygerythrus). Major differences were observed in the rate at which digesta markers moved through their respective gastrointestinal tracts, and in the concentration of lactic acid present in the gut. In both species, the caecum and colon were the principle sites of microbial activity and organic acid production.     

Trade-off between digestion and respiration in two airbreathing callichthyid catfishes Holposternum littorale (Hancock) and Corydoras aeneus (Gill)

In callichthyid catfishes, the posterior intestine is modified to function as an air breathing organ by being air-filled, thin-walled and highly vascularized. These modifications make it unsuitable for digestive functions and digesta has to be transported quickly through this region to minimize disruption of vital respiratory functions. However, the weak muscles of the wall of the respiratory intestine make this problematic. It is hypothesized that the unidirectional ventilatory air current within the respiratory intestine is responsible for the quick transport of digesta through the respiratory intestine. To verify this, movement of digesta through the alimentary tract was examined in Hoplosternum littorale and Corydoras aeneus that were either allowed to breathe air or prevented from air breathing. When air breathing was prevented, digesta was not transported to the rectum in H. littorale and there was a 94% reduction in the amount of digesta in the rectum of C. aeneus. This study suggests that the anterior digestive intestine facilitates the passage of air although it is filled with digesta. The anterior digestive intestine packages digesta into a string of slightly compressed boluses, creating an air channel in the digestive intestine thus allowing air to pass unimpeded.     

Flow and passage rate studies at the ileal level in the rabbit

Six female rabbits fitted with a simple glass cannula in terminal ileum and a further 6 non-cannulated rabbits were used to perform digesta flow and transit measurements. The animals received ad libitum a diet based mainly on lucerne meal. Flow and transit measurements were carried out using two particulate markers: ytterbium (Yb) fixed on lucerne meal cell-walls by soaking, and chromium (Cr) fixed by mordancing. Both markers were incorporated in the same diet before pelleting. The digestibility coefficient or total mean retention time (MRTt) in the whole tract was not affected by cannulation. MRT values between cecum and rectum could be obtained in cannulated rabbits: 9.6 h. The MRTt values estimated with Yb were higher than those obtained with Cr because the size of particles labelled with Cr was probably greater and thus the particles were excreted more rapidly in hard feces. Flow measurements were not affected by the choice of marker, confirming indirectly the validity of ileal digesta samples. These first ileal flow measures obtained on cannulated rabbits indicate that about half (49%) of the digestible organic matter and two-thirds of the digestible crude protein apparently disappeared before the cecum. Cell-walls (CW), determined as NDF fraction, were totally recovered at the terminal ileum, although hemicellulose (NDF-ADF) digestion could occur in both the small intestine and the cecocolic parts. These results are discussed in relation to the specificity of the Van Soest method for digesta cell-wall analysis.     

Some generalized stochastic compartment models for digesta flow

Digesta flow models have been based on linear compartment theory that assumes exponential retention times, and on a generalized theory that incorporates nonexponential (Erlang) retention times (Matis, 1987, Journal of Theoretical Biology 124, 371-376). This paper develops a new family of passage models for heterogeneous digesta by mixing the previous models with assumed parametric, usually gamma, mixing distributions. The utility of the resulting models is demonstrated with experimental data on two treatments, namely a chopped and a ground straw, given to each of four cows. Treatment differences are apparent in the preferred model form and in the means of the estimated mean residence times. The models are relatively easy to fit to data using standard estimation procedures, and they should have broad application to other compartment modeling problems with "heterogeneous particles."     

Digestive enzymes of human and nonhuman primates

All living organisms need to consume nutrients to grow, survive, and reproduce, making the successful acquisition of food resources a powerful selective pressure. However, acquiring food is only part of the challenge. While all animals spend much of their daily activity budget hunting, searching for, or otherwise procuring food, a large part of what is involved in overall nutrition occurs once the meal has been swallowed. Most nutritional components are too complex for immediate use and must be broken down into simpler compounds, which can then be absorbed by the body. This process, digestion, is catalyzed by enzymes that are either endogenous or produced by the host's microbial population .¹ Research shows that the nutritional value of food is partially constrained by the digestive abilities of the microbial community present in the host's gut and that these microbes rapidly adapt to changes in diet and other environmental pressures .² An accumulating body of evidence suggests that endogenously produced digestive enzymes also have been, and still are, common targets of natural selection, further cementing their crucial role in an organism's digestive system .^3–5