Effects of a charcoal powder-wood vinegar compound solution in piglets for raw pigeon pea seed meal

Histological intestinal villus alterations were studied in piglets fed a raw pigeon pea meal (PM) diet including a powder mixture of amorphous charcoal carbon and wood vinegar compound solution (CWVC). Twenty-eight male castrated piglets were divided into seven dietary groups of four piglets each. The control group was fed raw PM supplemented to the basal diet (178 g/kg crude protein, 4.23 kcal/g gross energy) at 0 g/kg (CONT), 200 g/kg (PM200) and 400 g/kg (PM400). The treatment groups were fed CWVC in both PM200 and PM400 diet groups at levels of 10 g/kg and 30 g/kg (PM200 + CWVC10, PM200 + CWVC30, PM400 + CWVC10 and PM400 + CWVC30). With increasing dietary PM levels, daily feed intake tended to increase. In contrast, daily body-weight gain tended to decrease, significantly in the PM400 group (P < 0.05), resulting in a significant decrease of feed efficiency in PM groups (P < 0.05). Body-weight gain and feed efficiency were higher in the CWVC groups compared with the PM groups. The duodenum and ileum were longer (P < 0.05) in the PM400 group than in CONT, but were similar to CONT in CWVC groups. The liver was heavier (P < 0.05), whereas the weights of the heart, kidney and stomach were decreased in the CWVC groups than in other groups. Most values for the intestinal villus height, cell area and cell mitosis number were lower in PM groups than those in CONT (P < 0.05) for each intestinal segment; however, these values were higher in CWVC groups than in PM groups (P < 0.05). The epithelial cells on the duodenal villus surface of the PM200 group showed cell morphology almost similar to CONT. However, the PM400 group had a smooth villus surface due to the presence of flat cells. The epithelial cells of the CWVC groups were protuberated, resulting in a much rougher surface than CONT. The current growth performance and histological intestinal alterations in piglets fed PM and PM + CWVC diets demonstrate that the intestinal features might be atrophied by feeding PM, resulting in decreased growth performance. CWVC might prevent the harmful effects of PM dietary toxins on intestinal function, resulting in a normal growth performance.     

酶解乳蛋白对初生仔猪小肠组织学结构及隐窝细胞增殖的影响(英文)

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The measurement of villus cell population size in the mouse small intestine in normal and abnormal states: a comparison of absolute measurements with morphometric estimators in sectioned immersion-fixed material

From a functional viewpoint, the most important part of the small intestinal mucosa is the villus epithelium; in the experimental study of intestinal adaptation it is often necessary to estimate the size of the population. For these reasons, a systematic study of methods of measuring the size of the villus cell population was undertaken in villi of normal morphology from control animals, and in villi of abnormal morphology as produced by treatment of mice with cytosine arabinoside (Ara-C). The villus cell population can be measured with precision and accuracy in both normal and abnormal mucosal states, with a relative standard error usually less than 5%, with good reproducibility, and with very low counting errors. In the mouse a practically linear relationship between the villus cell population size and position in the small intestine can be shown. In normal, control animals, linear estimates of villus population, for example the villus height, the villus core height, and the villus row count measured in sections of immersion-fixed material, were found to be rough approximations as indicators of the villus cell population, and were highly correlated with it. Product variables, comprising the products of a height with a width measurement, show a large improvement over the linear variables as estimators of villus cell population. In populations of abnormally shaped villi produced by Ara-C, both linear and product variables showed a considerable decrease in correlation with the villus cell population. Consequently, indirect, linear measurements of the villus population size do not accurately reflect the size of the villus population in abnormal villi. Finally, a method comparing theoretical and measured surface: volume indices was used to indicate that the most appropriate shape for normal mouse villi is a simple cylinder; this method would also be applicable for other villus cell populations.     

The measurement of villus cell population size in the mouse small intestine in normal and abnormal states: a comparison of absolute measurements with morphometric estimators in sectioned immersion-fixed material

Abstract From a functional viewpoint, the most important part of the small intestinal mucosa is the villus epithelium; in the experimental study of intestinal adaptation it is often necessary to estimate the size of the population. For these reasons, a systematic study of methods of measuring the size of the villus cell population was undertaken in villi of normal morphology from control animals, and in villi of abnormal morphology as produced by treatment of mice with cytosine arabinoside (Ara-C).
The villus cell population can be measured with precision and accuracy in both normal and abnormal mucosal states, with a relative standard error usually less than 5%, with good reproducibility, and with very low counting errors. In the mouse a practically linear relationship between the villus cell population size and position in the small intestine can be shown. In normal, control animals, linear estimates of villus population, for example the villus height, the villus core height, and the villus row count measured in sections of immersion-fixed material, were found to be rough approximations as indicators of the villus cell population, and were highly correlated with it. Product variables, comprising the products of a height with a width measurement, show a large improvement over the linear variables as estimators of villus cell population. In populations of abnormally shaped villi produced by Ara-C, both linear and product variables showed a considerable decrease in correlation with the villus cell population. Consequently, indirect, linear measurements of the villus population size do not accurately reflect the size of the villus population in abnormal villi.
Finally, a method comparing theoretical and measured surface: volume indices was used to indicate that the most appropriate shape for normal mouse villi is a simple cylinder; this method would also be applicable for other villus cell populations.     

A fluorescence-based demonstration of intestinal villi and epithelial cell in chickens fed dietary silicic acid powder including bamboo vinegar compound liquid

This study investigates the combined effect of silicic acid and bamboo vinegar compound liquid (SPV) on the growth and intestinal histological alterations in poultry. Forty-eight 7-day-old male Sanuki Cochin chickens were fed a commercial mash diet supplemented with SPV at 0, 0.1, 0.2, and 0.3% level ad libitum for 112 days. Body weight gain tended to improve with increased concentrations of dietary SPV, although these results were not statistically significant (P<0.1). Tissue observation by light microscopy revealed that the jejunal villus height (P<0.01) and duodenal and jejunal villus area (P<0.05) increased in the 0.2 and 0.3% SPV groups, respectively, compared with the control. Cell mitosis within the duodenum and jejunum also increased in the 0.2 and 0.3% SPV groups. Scanning electron microscopy revealed a prominent increase in the number of protuberant cells on the villus apical surface of the duodenum and jejunum for the 0.2 and 0.3% SPV groups compared with the control. Poultry in the 0.3% SPV group had the highest body weight gain and hypertrophied histological alterations of intestinal villi. Fluorescent microscopic images of cell mitosis and protuberant cells in the duodenal crypt clearly confirmed positive reactions for the activator protein 2α (AP-2α) and proliferating cell nuclear antigen (PCNA), compared with the control. The present results indicate that dietary SPV stimulates adsorption by the epithelial cells, which activate cell proliferation and self-renewal and regulate the expression of cell cycle regulators AP-2α and PCNA, resulting in higher body weight gain. Thus, we can conclude that a concentration of 0.3% dietary SPV is ideal for promoting growth in poultry.     

Effects of fermented soybean meal on carbon and nitrogen metabolisms in large intestine of piglets

Fermented soybean meal (FSM), which has lower anti-nutritional factors and higher active enzyme, probiotic and oligosaccharide contents than its unfermented form, has been reported to improve the feeding value of soybean meal, and hence, the growth performance of piglets. However, whether FSM can affect the bacterial and metabolites in the large intestine of piglets remains unknown. This study supplemented wet-FSM (WFSM) or dry-FSM (DFSM) (5% dry matter basis) in the diet of piglets and investigated its effects on carbon and nitrogen metabolism in the piglets’ large intestines. A total of 75 41-day-old Duroc×Landrace×Yorkshire piglets with an initial BW of 13.14±0.22 kg were used in a 4-week feeding trial. Our results showed that the average daily gain of piglets in the WFSM and DFSM groups increased by 27.08% and 14.58% and that the feed conversion ratio improved by 18.18% and 7.27%, respectively, compared with the control group. Data from the prediction gene function of Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) based on 16S ribosomal RNA (rRNA) sequencing showed that carbohydrate metabolism function families in the WFSM and DFSM groups increased by 3.46% and 2.68% and that the amino acid metabolism function families decreased by 1.74% and 0.82%, respectively, compared with the control group. These results were consistent with those of other metabolism studies, which showed that dietary supplementation with WFSM and DFSM increased the level of carbohydrate-related metabolites (e.g. 4-aminobutanoate, 5-aminopentanoate, lactic acid, mannitol, threitol and β-alanine) and decreased the levels of those related to protein catabolism (e.g. 1,3-diaminopropane, creatine, glycine and inosine). In conclusion, supplementation with the two forms of FSM improved growth performance, increased metabolites of carbohydrate and reduced metabolites of protein in the large intestine of piglets, and WFSM exhibited a stronger effect than DFSM.     

RADIATION EFFECTS ON CELL POPULATIONS IN THE INTESTINAL EPITHELIUM OF MICE AND ITS THEORY

Mice were exposed to 1000 R of X-rays to their trunks and sacrificed every day up to the tenth day after exposure. Cell counts were made on histological sections of the duodenum. The cell counts in the crypts were reduced to about 50% of the control value on the first day after exposure. The cell counts began to recover on the third day and an overshoot of 170% was observed on the fourth day; thereafter the crypt cell counts tended to return to the control level. The cell counts on the villi reached their minimum value on the third day after exposure. Following an overshoot on the sixth day, the villus cell counts returned to the control level by the tenth day. The above experimental results were analysed using a two-compartment model with a feedback term. A logistic proliferation was assumed for the proliferative crypt cells, while for the postmitotic villus cells the compartment was assumed to be a first in-first out type. The calculated results with this model are in general consistent with the experimental ones. The model seems to possess some essential features of the dynamics of cell renewal in the intestinal mucosa.     

Intestinal development and growth performance of early-weaned piglets fed a low-threonine diet

High dietary threonine extraction by the digestive tract suggests that threonine contributes to maintain gut integrity. The aims of this study were to investigate the intestine development and the growth performance of early-weaned piglets pair-fed either a control well-balanced (C: 9.3 g threonine/kg diet) or a low-threonine diet (LT: 6.5 g threonine/kg diet) for 2 weeks. As expected, LT piglets presented lower plasma free threonine compared with C piglets (118 v. 356 ± 12 μmol/l, P < 0.001). Dietary threonine supply altered neither growth performance nor growth of the intestine and of the other portal-drained viscera (stomach, spleen and pancreas). Nevertheless, villus height was reduced in the ileum of the LT piglets compared with C piglets (446 v. 714 ± 74 μm, P < 0.05). This was also associated with a decrease in crypt width (P < 0.05) and villus height-to-crypt depth ratio (P < 0.05). Whereas maltase and lactase activities did not change between the two groups, aminopeptidase nitrogen activity was decreased in the ileum of LT piglets (269 v. 374 ± 27 IU/mg protein, P < 0.05). The number of mucin-containing goblet cells was not modified in the ileum and in the proximal part of the large intestine of the LT piglets compared with the C piglets. In conclusion, despite no alteration of intestinal growth, villus hypotrophy associated with a reduction of aminopeptidase nitrogen activity suggest an alteration of the structure of the ileum in early-weaned piglets fed a diet supplying inadequate dietary threonine.     

Intestinal intraepithelial lymphocyte gamma delta-T cell-derived keratinocyte growth factor modulates epithelial growth in the mouse

Keratinocyte growth factor (KGF) promotes intestinal epithelial growth. To understand the relevance of intraepithelial lymphocyte (IEL)-derived KGF expression on epithelial growth, we used a mouse model of villus atrophy by the administration of total parenteral nutrition, and a model of villus hypertrophy by the creation of a short bowel syndrome. KGF expression was confined to gammadelta-TCR(+) IELs. IEL-derived KGF expression was highest in the crypts, somewhat less in the lower portion of villi, and markedly lower in the upper portion of villi. Total parenteral nutrition administration was associated with a down-regulation of IEL-derived KGF expression, and short bowel syndrome was associated with an up-regulation of IEL-derived KGF expression. In the absence of gammadelta-TCR(+) IEL, using gammadelta(-/-) mice, intestinal epithelial cell proliferation decreased in control, and in both mucosal atrophy (22% decline) and mucosal hypertrophy (14%) models. These results show that KGF from IELs is an important factor for maintenance of intestinal epithelial cell proliferation and villus growth.     

Morphological effects of a large single dose of cycloheximide on the intestinal epithelium of the rat.

Adult male rats received 15 mg/kg cycloheximide and the subsequent morphological effects at three and six hours after injection were evaluated using histometry, light and electron microscopy, histological demonstration of terminal web and acid phosphatase, and radioautography with tritiated thymidine. Rapid atrophy of the villi took place, progressing from the villus tip by premature exfoliation of epithelial cells. The crypts also diminished by random exfoliation of many crypt cells and by partial or complete disintegration. Mitosis and epithelial cell migration were absent. By six hours, the area occupied by the villi and the crypts per unit length of histological section was decreased by about 70-90% in most of the small intestine but only by about 40-60% in the duodenum and the terminal ileum. In the upper half of the villi, the epithelium was strongly positive for acid phosphatase and contained large numbers of round bodies resembling primary lysosomes. In the lower half, the microvillous border and terminal web were found to be disrupted. Animals receiving only 5 mg/kg cycloheximide also showed the atrophy of villi and crypts, and the round bodies resembling lysosomes. Evidence from several sources has indicated that protein synthesis in normal villus epithelial cells subsides toward the villus tip and becomes minimal at exfoliation. At exfoliation, proteins responsible for epithelial cohesion probably fail because they are no longer replenished. Cycloheximide appears to accelerate this process.     

CHANGES IN GROWTH KINETICS OF JEJUNAL EPITHELIUM IN MICE MAINTAINED ON AN ELEMENTAL DIET

Changes in the kinetics of the intestinal epithelium were observed in mice maintained on an elemental diet containing hydrolysed protein and medium chain triglycerides. An increase in the length of the villi seen shortly after commencement of the diet was followed by a reduction in the rate of proliferation in the crypt. After 7 days on the diet, an equilibrium state was reached with the cellularity of the villi being 120% that of control while the number of proliferative cells/crypt was reduced by 35%. The proliferative response of the crypt following irradiation occurrred 16 hr later in diet-fed mice than in controls. It was postulated that, because of the increased cellularity of the villus compartment in diet-fed mice, additional time was required to reduce the number of villus cells to a critical level at which a proliferative response is induced in the crypt.     

Heat shock protein 70 is upregulated in the intestine of intrauterine growth retardation piglets

The objective of this study is to investigate the expression and distribution of heat shock protein 70 (Hsp70) in the intestine of intrauterine growth retardation (IUGR) piglets. Samples from the duodenum, prejejunum, distal jejunum, ileum, and colon of IUGR and normal-body-weight (NBW) piglets were collected at birth. The results indicated that the body and intestine weight of IUGR piglets were significantly lower than NBW piglets. The villus height and villus/crypt ratio in jejunum and ileum of IUGR piglets were significantly reduced compared to NBW piglets. These results indicated that IUGR causes abnormal gastrointestinal morphologies and gastrointestinal dysfunction. The mRNA of hsp70 was increased in prejejunum (P < 0.05), distal jejunum (P < 0.05), and colon in IUGR piglets. However, the hsp70 mRNA in ileum of piglets with IUGR was decreased. Similar to hsp70 mRNA, the protein levels of Hsp70 in prejejunum (P < 0.05), distal jejunum, and colon (P < 0.05) in IUGR piglets were higher than those in NBW piglets. These results indicated that the expression of Hsp70 in the intestinal piglets was upregulated by IUGR, and different intestinal sites had different responses to stress. Meanwhile, the localization of Hsp70 in the epithelial cells of the whole villi and intestinal gland rather than in the lamina propria and myenteron suggested that Hsp70 has a cytoprotective role in epithelial cell function and structure.     

Changes in growth kinetics of jejunal epithelium in mice maintained on an elemental diet.

Changes in the kinetics of the intestinal epithelium were observed in mice maintained on an elemental diet containing hydrolysed protein and medium chain triglycerides. An increase in the length of the villi seen shortly after commencement of the diet was followed by a reduction in the rate of proliferation in the crypt. After 7 days on the diet, an equilibrium state was reached with the cellularity of the villi being 120% that of control while the number of proliferative cells/crypt was reduced by 35%. The proliferative response of the crypt following irradiation occurred 16 hr later in diet-fed mice than in controls. It was postulated that, because of the increased cellularity of the villus compartment in diet-fed mice, additional time was required to reduce the number of villus cells to a critical level at which a proliferative response is induced in the crypt.     

Effects of Weaning on Intestinal Upper Villus Epithelial Cells of Piglets

The intestinal upper villus epithelial cells represent the differentiated epithelial cells and play key role in digesting and absorbing lumenal nutrients. Weaning stress commonly results in a decrease in villus height and intestinal dysfunction in piglets. However, no study have been conducted to test the effects of weaning on the physiology and functions of upper villus epithelial cells. A total of 40 piglets from 8 litters were weaned at 14 days of age and one piglet from each litter was killed at 0 d (w0d), 1 d (w1d), 3 d (w3d), 5 d (w5d), and 7 d (w7d) after weaning, respectively. The upper villus epithelial cells in mid-jejunum were isolated using the distended intestinal sac method. The expression of proteins in upper villus epithelial cells was analyzed using the isobaric tags for relative and absolute quantification or Western blotting. The expression of proteins involved in energy metabolism, Golgi vesicle transport, protein amino acid glycosylation, secretion by cell, transmembrane transport, ion transport, nucleotide catabolic process, translational initiation, and epithelial cell differentiation and apoptosis, was mainly reduced during the post-weaning period, and these processes may be regulated by mTOR signaling pathway. These results indicated that weaning inhibited various cellular processes in jejunal upper villus epithelial cells, and provided potential new directions for exploring the effects of weaning on the functions of intestine and improving intestinal functions in weaning piglets.     

Epidermal growth factor improves intestinal morphology by stimulating proliferation and differentiation of enterocytes and mTOR signaling pathway in weaning piglets

Epidermal growth factor(EGF) has been shown to improve piglet intestinal morphology and epithelial recovery. In an attempt to further understand the mechanisms behind these improvements, this study tested the hypothesis that dietary EGF may affect intestinal morphology by stimulating the proliferation and differentiation of enterocytes in weaning piglets. In piglets receiving200 μg kg–1 EGF, crypt depth and villus height increased(P0.05). Adding 400 μg kg–1 EGF increased villus height-to-crypt depth ratio(P0.05), but reduced crypt depth(P0.05). Dietary supplementation with 200 μg kg–1 EGF significantly increased the number of Ki67-positive cells(P0.01) and tended to increase the mRNA level of proliferating cell nuclear antigen(P0.10).However, this supplementation decreased the expression level of intestinal fatty acid-binding protein(P0.05). Piglets fed with400 μg kg–1 EGF had an increased mRNA level of intestinal alkaline phosphatase(P0.05). The phosphorylation of m TOR(mammalian target of rapamycin) was observed in the 200 μg kg–1 EGF group. These results suggest that dietary supplementation with a low level of EGF improved piglet intestinal morphology through stimulating the proliferation and differentiation of enterocytes, and the mTOR signaling pathway may partly be involved in this process.     

Effects of cold shock and phospholipase A2 on intact boar spermatozoa and sperm head plasma membranes

Head plasma membranes (HPM) isolated from cryopreserved boar spermatozoa show an excessive fluidization, which might be involved in the loss of fertility. The current study assessed the ability of cold shock (5 degrees C) and phospholipase A2 (PA2) to duplicate these effects on membrane structure and to affect 45Ca2+ uptake and gross morphological characteristics of whole, fresh boar-sperm. The HPM from cold-shocked sperm showed a significantly greater rate of fluidization over time than did HPM from control sperm. Addition of PA2 (bee or snake venom, 0.1 or 10.0 ng/ml) to HPM from control sperm caused fluidization similar to cold shocking, but to a lesser degree (P less than 0.05). Cold-shocked intact sperm exhibited severe acrosomal disruption, loss of motility, and increased 45Ca2+ uptake relative to control sperm. Addition of PA2 (bee or snake venom, 0.1, 1.0., 10.0, and 1,000 ng/ml) to control sperm had no effect on gross morphology or motility while maintaining or increasing sperm extrusion of 45Ca2+. Therefore, although PA2 can, to some extent, duplicate the effects of cold shock on HPM molecular organization, its lipid hydrolytic action is insufficient to cause all the gross disruptions of severe thermal shock. Both PA2 and cold shock disrupted HPM structure, but only cold shock increased 45Ca2+ uptake, suggesting that cold shock may be increasing 45Ca2+ uptake in areas other than the head. Cold shock disrupts sperm on three levels; membrane molecular organization, intracellular Ca2+ regulation, and gross morphology/motility.     

The kinetics of villus cell populations in the mouse small intestine

Abstract. The cell population kinetics of the villus epithelium of the mouse have been analysed with respect to the size, flux and time. Microdissection methods were employed to measure the villus cell population size and yielded reproducible, precise results. There was a proximodistal negative size gradient in villus cell population and, in those villi of normal morphology, there was a good correlation with the usual morphometric estimators such as height and row count, although correlation was improved by a product variable consisting of a height multiplied by a width parameter.
Flux onto the villus is the product of the crypt cell production rate, which was measured by a metaphase arrest method using vincristine and crypt microdissection, and the crypt:villus ratio; net villus influx was maximum proximally in the bowel, where the largest villi were found, and decreased distally. The distribution of transit times of labelled cells to the crypt: villus junction and to the villus tip was measured, allowing the measurement of the median villus transit time.
Comparison of the measured villus transit time with the theoretical transit time calculated from the villus influx and population size gave results consistent with a steady state hypothesis. It was found, at each level of the small intestine studied, that the number of epithelial cells on the villus was equivalent to the total number of crypt cells associated with the villus.     

EFFECT OF VILLUS LENGTH ON CELL PROLIFERATION AND MIGRATION IN SMALL INTESTINAL EPITHELIUM

For the interpretation of data supporting the hypothesis of a feedback regulation of proliferative activity in intestinal crypts by the functional villus cell compartment the life span and migration rate of epithelial cells on villi of experimentally reduced length should be known. Autoradiographic studies and scintillation counting of isolated villi at different time intervals after ³H-thymidine labelling were carried out 36, 48 and 60 hr intervals after X-irradiation. The results showed that the life span of epithelial cells in rat small intestine (36–48 hr) is independent of the villus length. In villi of reduced length the migration rate of the epithelial cells was found to be decreased compared with controls. Changes in the migration rate in turn seem to be dependent on the production of epithelial cells in the crypt. Comparative studies on the recovery of crypt and villus epithelium after various doses (300 and 700 R) of X-radiation support the hypothesis that increased proliferative activity in the crypt cell compartment is related to a reduction of the number of functional villus cells below a critical villus length. The importance of these findings in the interpretation of data on (micro) biochemical analyses of certain cell differentiation characteristics during increased proliferative activity is discussed.     

Intestinal villus structure contributes to even shedding of epithelial cells

In the intestinal epithelium, proliferated epithelial cells ascend the crypts and villi and shed at the villus tips into the gut lumen. In this study, we theoretically investigate the roles of the villi on cell turnover. We present a stochastic model that focuses on the duration over which cells migrate the shortest paths between the crypt orifices and the villus tips, where shedding cells are randomly chosen from among those older than the shortest-path cell migration times. By extending the length of the shortest path to delay cell shedding, the finger-like shape of the villus would tightly regulate shedding-cell ages compared with flat surfaces and shorter projections; the villus allows epithelial cells to shed at around the same age, which limits them from shedding early or staying in the epithelium for long periods. Computational simulations of cell dynamics agreed well with the predictions. We also examine various mechanical conditions of cells and confirm that coordinated collective cell migration supports the predictions. These results suggest the important roles of the villi in homeostatic maintenance of the small intestine, and we discuss the applicability of our approach to other tissues with collective cell movement.     

Effects of Enterococcus faecium and Bacillus cereus var. toyoi on the morphology of the intestinal mucous membrane in piglets

Eighty piglets aged 14, 28, 35 and 56 days — weaned at day 28 — were subjected to this investigation. Each age-group consisted of five animals which were fed an Enterococcus faecium NCIMB 10415 (Cylactin®) and Bacillus cereus var. toyoi (Toyocerin®) based diet. Five animals served as controls. Tissue samples were collected immediately after sacrifice at 8.30 h a.m. from duodenum, jejunum, ileum, cecum and colon to examine intestinal morphology and histochemistry. The results showed that with respect to villus height and crypt depth supplementation of probiotics in piglets feed seemed to influence the morphology and enlargement factor not at all or only to a certain extent. With respect to the number of goblet cells, the difference between probiotic fed animals and control animals was generally extremely low. The shape of the villi of the small intestinal segments greatly varied in all age groups of control and probiotic fed animals. However, this morphological variety does not depend on the mode of feeding.