The linear alometric relationship between total metabolic energy per life span and body mass of poikilothermic animals

A linear relationship exists between total metabolic energy per life span PT(ls) (kJ) and body mass M (kg) of 54 poikilothermic species (Protozoa, Nematoda, Mollusca, Asteroidae, Insecta, Arachnoidae, Crustacea, Pisces, Amphibia, Reptilia and Snakes): PT(ls) = A(ls*)M(1.0838), where P (kJ/day) is the rate of metabolism and T(ls) (days) is the life span of animals. The linear coefficient A(ls*) = 3.7 x 10(5) kJ/kg is the total metabolic energy, exhausted during the life span per 1 kg body mass of animals. This linear coefficient can be regarded as relatively constant metabolic parameter for poikilothermic organisms, ranging from 0.1 x 10(5) to 5.5 x 10(5) kJ/kg, in spite of 17-degree differences between metabolic rate and body mass of animals. A linear relationship between total metabolic energy per life span and body mass of only 48 poikilothermic multicellular animals (without Protozoa) is: PT(ls) = A(ls*)M(0.9692) with linear coefficient A(ls*) = 2.34 x 10(5) kJ/kg. Since a power relationship exists between the rate of metabolism and body mass of animals of the type: P = aMk (a and k are the alometric constants), an empiric rule could be formulated, that life span is a time interval for which the total metabolic energy per life span becomes proportional to body mass of animals and power coefficient k becomes approximately 1.0.     

Analysis of the polyploidization kinetics of the parenchymal cells in the rat liver

Methodological approaches to kinetics of cell polyploidization in the rat liver parenchyma are discussed. Different ways of hepatocyte polyploidization in the course of postnatal liver growth have been assessed. The intensities of hepatocyte transitions from one ploidy class to another were determined. On the basis of literary experimental data the following is summarized: With the increase in the animal age, there is a decrease in hepatocyte transition from one ploidy class to and ther; in young animals the intensity of formation of tetraploid hepatocytes through the stage of binuclear cells (2c----2c X 2----4c) is 0.39-0.55 within two weeks, the intensity of direct transitions (2c----4c) being 0.00-0.19 within the same time. The intensity of entering to DNA synthesis is reduced with the increase in hepatocyte ploidy levels; in this case the coefficient of the reducing of mitotic activity is calculated as 0.10-0.22, and 0.01-0.05 for 4c- and 8c-hepatocytes, resp. The factors stimulating proliferation in the liver increase the intensity of the direct cell transition (2c----4c) by several times which can exceed the intensity of transition through the binuclear cell stage.     

Vitamin A transport in plasma of the non-mammalian vertebrates: isolation and partial characterization of piscine retinol-binding protein

Studies were conducted to explore vitamin A transport in the non-mammalian vertebrates, especially Pisces, Amphibia, and Reptilia, and to isolate and partially characterize piscine retinol-binding protein. Retinol-containing proteins in fresh plasma obtained from bullfrogs and a turtle exhibited similar properties to those found in mammalian and chicken plasma: i.e., molecular weight of about 60,000-80,000 as estimated by gel filtration and binding affinity to prealbumin on human prealbumin-Sepharose affinity chromatography. In sharp contrast, vitamin A-containing proteins in plasma from larvae of bullfrogs as well as three fishes (carp, blue sharks, and young yellowtails) appeared to be present in plasma as monomeric retinol-binding proteins without any affinity to human prealbumin. On the other hand, plasma vitamin A in the lamprey (Cyclostomes) was found to exist exclusively as an ester form in association with the lipoproteins of hydrated density less than 1.21 g/ml. Piscine retinol-binding protein was isolated from pooled plasma of young yellowtails and was converted (1000-fold purification) to a homogeneous component by a procedural sequence that included gel filtration on Sephadex G-100, chromatography on SP-Sephadex, gel isoelectric focusing, and, finally, polyacrylamide gel electrophoresis. Purified piscine retinol-binding protein showed physico-chemical properties distinctly different from the mammalian and chicken retinol-binding proteins examined, i.e., a smaller molecular weight of approximately 16,000, a lower isoelectric point of 4.3, a prealbumin mobility on analytical polyacrylamide gel electrophoresis, and a lack of binding affinity for human prealbumin; however, it displayed similar characteristics in two ways: a 1:1 molar complex with retinol, and a high content of tryptophan (four residues). These results strongly suggest that the piscine retinol-binding protein is a prototype of the specific vitamin A-transporting protein in plasma of the vertebrates, being modified later in evolution, during phylogenetic development of the vertebrates, to acquire a binding site for prealbumin on the molecule.     

Some aspects of element turnover in living organisms

The net uptake and loss of any element by a living organism can be described as the quotient of the total amount of the element, present in the organism, and its residence time in the organism. Theoretically it can be derived that the residence time tau i, equals Vi1-b/kappa, in which b, the morphometric coefficient, is related to size and shape of the organism (volume Vi); kappa, the turnover coefficient, is related to its metabolic activity. The net uptake or excretion, phi i, of an element then follows from phi = kappa Ci Vib, Ci being the (average, whole animal) concentration of the element. Residence times, derived from the quotient of body volume (weight) and daily food intake of various animal species of different sizes, show that, independent of animal or element species, the morphometric coefficient and turnover coefficient have values of b = 0.735 and k = 71.4 dm. year-1 respectively. The turnover coefficient may show some variation, related to the metabolic activity of the organism. The value of the morphometric coefficient implies that residence times may vary with body size; in small animals the residence times of the elements may be in the order of several hours, whereas in the largest organisms residence times may be as long as several years. The uptake of Na, Mg, Cl and P calculated from the above equation (phi i = k Ci Vib) corresponds very well with data in the literature on the mineral requirements of domestic animals; for Ca the calculated values were lower, for I higher, but they show the same weight-dependence as predicted from the equation. This suggests that the equation may be used tentatively to assess dietary requirements for elements for which up till now no such information exists. Values derived for the uptake and excretion of various elements by the whole biosphere correspond roughly to those estimated for their oceanic fluxes (from data compiled by Brewer, 1975). This supports the hypothesis that the global turnover of (many) elements is closely related to biological (metabolic) processes.     

Inhibitory serpins from rye grain with glutamine as P1 and P2 residues in the reactive center

Six of seven serpins detected in grains of rye (Secale cereale) were purified and characterized. The amino acid sequence close to the blocked N-terminus, the reactive center loop sequence and the second order association rate constant (k(a)') for irreversible complex formation with chymotrypsin were determined for each serpin. Three of four serpins containing the unusual reactive center P2-P1' QQ/S and one with P2-P1' PQ/M were equally efficient inhibitors of chymotrypsin (k(a)' approximately 10(5) M(-1) s(-1)). One serpin with P2-P1' PY/M was a faster inhibitor (k(a)' approximately 10(6) M(-1) s(-1)). Similar but differently organized glutamine-rich reactive centers were recently found in grain serpins cloned from wheat [Ostergaard et al. (2000) J. Biol. Chem. 275, 33272] but not from barley. The prolamin storage proteins of cereal grains contain similar sequences in their glutamine-rich repeats. A possible adaption of hypervariable serpin reactive centers late in Triticeae cereal evolution as defence against insects feeding on cereal grains is discussed.     

Evolution of posture in amniotes–Diving into the trabecular architecture of the femoral head

Extant amniotes show remarkable postural diversity. Broadly speaking, limbs with erect (strongly adducted, more vertically oriented) posture are found in mammals that are particularly heavy (graviportal) or show good running skills (cursorial), while crouched (highly flexed) limbs are found in taxa with more generalized locomotion. In Reptilia, crocodylians have a “semi-erect” (somewhat adducted) posture, birds have more crouched limbs and lepidosaurs have sprawling (well-abducted) limbs. Both synapsids and reptiles underwent a postural transition from sprawling to more erect limbs during the Mesozoic Era. In Reptilia, this postural change is prominent among archosauriforms in the Triassic Period. However, limb posture in many key Triassic taxa remains poorly known. In Synapsida, the chronology of this transition is less clear, and competing hypotheses exist. On land, the limb bones are subject to various stresses related to body support that partly shape their external and internal morphology. Indeed, bone trabeculae (lattice-like bony struts that form the spongy bone tissue) tend to orient themselves along lines of force. Here, we study the link between femoral posture and the femoral trabecular architecture using phylogenetic generalized least squares. We show that microanatomical parameters measured on bone cubes extracted from the femoral head of a sample of amniote femora depend strongly on body mass, but not on femoral posture or lifestyle. We reconstruct ancestral states of femoral posture and various microanatomical parameters to study the “sprawling-to-erect” transition in reptiles and synapsids, and obtain conflicting results. We tentatively infer femoral posture in several hypothetical ancestors using phylogenetic flexible discriminant analysis from maximum likelihood estimates of the microanatomical parameters. In general, the trabecular network of the femoral head is not a good indicator of femoral posture. However, ancestral state reconstruction methods hold great promise for advancing our understanding of the evolution of posture in amniotes.     

Frequency of the stages in the cycle of the seminiferous epithelium in the rat

This study determined the optimum number of tubules to be counted per testis cross section, and the number of animals per treatment group, when changes in stage frequencies in the cycle of the seminiferous epithelium are criteria for assessing effects of treatment on spermatogenesis. A data base of 9,672 observed and staged tubules was collected from testicular cross sections of 15 Sprague-Dawley rats. A significant variation between animals was found for the frequencies of Stages I, II, IV, VI, VIII, and XIII. Computer simulation was used to randomly select different combinations of animal and tubule numbers from the observed data. Stage frequency means from each simulation experiment were compared statistically to observed mean frequencies. A model that used data from all 14 stages was analyzed. The following conclusions were made: a) a minimum of 200 tubule cross sections/testis is recommended for estimating stage frequencies; b) for a fixed number of tubules scored, the number of animals sampled is more important than the number of tubules per animal in reducing variance; c) to detect a difference of 2 standard deviations from the mean with a 2% error rate and examining 200 tubules/testis, at least 12 animals must be used per group when assessing all 14 stages; d) when individual stages are examined using 10 animals per group, only Stage VII has 80% or greater power of test (alpha = 0.05) to detect a frequency difference; e) pooling stages into 3-4 groups is recommended to improve the power of detecting a treatment difference.     

Isolation and partial characterization of IgM-like immunoglobulins from the serum of carp (Cyprinus carpio L.), frog (Rana ridibunda Pall.) and tortoise (Testudo graeca Pall.)

1. IgM-like immunoglobulin fractions have been isolated by combinations of precipitation procedures and gel-filtration from the serum of three vertebrate species, representative of the classes of Pisces (Cyprinus carpio L.), Amphibia (Rana ridibunda Pall.) and Reptilia (Testudo graeca Pall.). 2. The homogeneity of the isolated fractions has been confirmed electrophoretically and immunoelectrophoretically. 3. Electrophoretic mobility of IgM, similar for the three species studied has been observed in polyacrylamide gel; in agar gel immunoelectrophoresis carp and frog IgM mobilities are almost identical, in contrast to the markedly slower cathodic migration of tortoise IgM.     

The ontogeny and maintenance of adult symmetry properties in the ctenophore, Mnemiopsis mccradyi

Ctenophores are biradially symmetrical animals. The body is composed of four identical quadrants which are organized along an oral-aboral axis. Most species have eight comb rows, two tentacles, and an apical organ (located on the aboral surface). During embryogenesis there is a fixed pattern of cleavage, a precocious specification of blastomere developmental potential, and an inability to regulate for portions of the embryo that have been removed. When blastomeres are separated at the two-cell stage each blastomere develops into a "half-animal" with four comb rows, one tentacle, and half an apical organ. In contrast, adult ctenophores regenerate readily. When an adult ctenophore is cut in half to produce "half-animals," in most cases each half regenerates the missing half. In some cases, however, bisected animals remain as "half-animals" which repair the wound site but do not replace all of the missing structures. When animals are cut in half along the tentacular or esophageal axis at different stages of embryogenesis a transition period is detected when the capacity for adult regeneration begins. This transition occurs at the time when the formation of the apical organ is complete and comb row function becomes coordinated. Embryos bisected prior to this time remain as "half-animals" even after growing to large reproductive sizes, while animals bisected after the transition period usually regenerate the missing structures within 2-3 days. When adult "half-animals" (produced by bisection either before or after the transition period) are cut into "quarter-pieces," the pieces regenerate to form either "half-animals" or whole animals. Thus, "half-animals" produced prior to the transition period--although they failed to undergo embryonic regulation--have not irreversibly lost the capacity to form whole animals if challenged to regenerate during adult stages. When aboral blastomeres destined to form the apical organ, tentacles, and comb rows are removed from early cleavage stages (prior to the transition period), the embryo does not form these structures at the appropriate time. However, the resulting deficient adults spontaneously form these structures from remaining blastomere lineages soon after hatching. These experiments suggest that as long as some quadrant-specific cells of the oral pole are present at the time of the transition period, the structures of that quadrant will be spontaneously replaced during the adult period.(ABSTRACT TRUNCATED AT 400 WORDS)     

The evolution of multicellularity and early animal genomes

Several independent molecular datasets, including complete mtDNA sequence, indicate that Choanozoa are most closely related to multicellular animals. There is still confusion concerning basal animal phylogeny, although recent data indicate that Placozoa are not degenerate cnidarians and hence (along with sponges) occupy a pivotal position. The transition in evolution from diploblast to bilaterian animals is becoming better understood, with gene expression data arguing that cnidarians have forerunners of the anteroposterior and dorsoventral body axes, and even a putative homologue of mesoderm. The homeobox and kinase gene families have been further analysed in basal animals, although more data are required to enable detailed comparison with Bilateria.     

Immunity to Litomosoides carinii in Mastomys natalensis. II. Effects of chemotherapeutically abbreviated and postpatent primary infections on challenges with various stages of the parasite

Naive Mastomys natalensis, Litomosoides carinii-infected M. natalensis at a postpatent stage of the infection and L. carinii-infected M. natalensis treated chemotherapeutically with furazolidone (FUR), FUR and diethylcarbamazine (FUR/DEC) or amoscanate (AMOS) were challenged by either injection or implantation of 40 third stage larvae (L3, s.c.), 40 fourth stage larvae (L4, 16 days old, i.p.), 20 male and 20 female preadult worms (36 days old, i.p.), 12 adult female worms (i.p.) or 6 X 10(6) microfilariae/kg (i.v.). Microfilaraemia in animals challenged at a postpatent stage (independent of the kind of challenge), was either totally suppressed or at least greatly reduced. Necropsy of L3-challenged animals showed that neither the length of the worms nor their content of morphologically intact, intrauterine stages was affected. Infected, treated animals challenged with developing stages (L3, L4 and preadult worms) showed reduced levels of microfilaraemia (by up to 75%). Dissection of AMOS-treated, L3-challenged animals showed that both the developmental rate and the fertility of the worms were affected. Microfilaraemia was also reduced after implantation of adult worms into treated animals. This was independent of the interval between treatment and challenge (44-150 days) except in animals challenged 10 days after AMOS-treatment, which showed no difference from naive controls. However, infected, treated M. natalensis, cotton rats and gerbils did not develop immunity against intravenously injected blood microfilariae.     

The regularities in connection between fecundity with body weight and growth rate in fishes

The absolute and relative fecundity of freshwater, anadromous and marine fishes (102 species and subspecies from 33 families) and its dependence on body mass and growth rate were analyzed on the basis of published data. According to the spawning type all studied fishes were divided into species with short-term and single spawning and fishes with extended or long-term spawning. The equations of dependence of absolute fecundity (E) on body mass (W) were calculated: E = 1.033 W0.578 (the first group) and E = 0.792 W0.74 (the second group). If W < 177 g the equations don't differ significantly and one may use the equation E = 1.34(-0.742) for both groups. The body mass of females at age of maturity expressed as a portion of maximal definitive body mass equals 0.22 +/- 0.044 for many different species with 0.95 probability. The relative fecundity (a1) of some species negatively correlates with maximal body mass of adult individuals (Wmax). This dependence is expressed by equations: a1 = 3.033 Wmax-0.549 (for the first group) and a1 = 1.726 Wmax-0.351 (for the second group). Value of ratio Wov/W of different fish species changes irregularly from 0.054 to 0.32 and its average is 0.150 +/- 0.012 for the first group and 0.156 +/- 0.007 for the second one. In such a way, single reproduction effort of fishes is approximately 0.15. Comparison of data on Pisces, Crustacea, Amphibia, Reptilia, and Mammalia revealed that reproduction effort of different aquatic and terrestrial invertebrates and vertebrates varies within rather narrow limits (from 0.05 to 0.44). Average values of this index varies even less--from 0.097 to 0.238, on average 0.162, i.e. approximately 15-18% of animals' body mass falls on their reproduction constituent.     

A simple procedure for estimation of total body surface area and determination of a new value of Meeh's constant in rats

Precise calculation of total body surface area (TBSA) or premarked surface areas (P-MSAs) is of great importance in many biomedical applications. The aim of the paper was to present a simple procedure of measurement of P-MSAs in small animals and to determine a more accurate Meeh's constant (k), for a commonly used weight range of laboratory rats. A series of 30 Wistar rats, weighing 195-240 g, were anaesthetized and weighted. The TBSA of each animal was measured using a clear pocket and a planimeter. The data obtained were entered into the Meeh's formula (TBSA = kW(2/3)), the most commonly used for small experimental animals, so that a k value for each animal as well as a mean k value (9.83) were obtained. The TBSA of the animals was also calculated using the aforementioned mean k value and compared with that obtained using k values reported in previous studies. According to our findings, the new mean k value, determined with the use of our procedure of surface area measurement, ensured greater accuracy in the determination of the TBSA of experimental rats of a specific weight range. We also suggest a new procedure of surface area measurement which is easy, accurate and does not require animal sacrifice.     

Three tiers of genome evolution in reptiles

Characterization of reptilian genomes is essential for understanding the overall diversity and evolution of amniote genomes, because reptiles, which include birds, constitute a major fraction of the amniote evolutionary tree. To better understand the evolution and diversity of genomic characteristics in Reptilia, we conducted comparative analyses of online sequence data from Alligator mississippiensis (alligator) and Sphenodon punctatus (tuatara) as well as genome size and karyological data from a wide range of reptilian species. At the whole-genome and chromosomal tiers of organization, we find that reptilian genome size distribution is consistent with a model of continuous gradual evolution while genomic compartmentalization, as manifested in the number of microchromosomes and macrochromosomes, appears to have undergone early rapid change. At the sequence level, the third genomic tier, we find that exon size in Alligator is distributed in a pattern matching that of exons in Gallus (chicken), especially in the 101-200 bp size class. A small spike in the fraction of exons in the 301 bp-1 kb size class is also observed for Alligator, but more so for Sphenodon. For introns, we find that members of Reptilia have a larger fraction of introns within the 101 bp-2 kb size class and a lower fraction of introns within the 5-30 kb size class than do mammals. These findings suggest that the mode of reptilian genome evolution varies across three hierarchical levels of the genome, a pattern consistent with a mosaic model of genomic evolution.     

Functional implication of the basal metabolic rate in homeothermic animals

Data on energetic profile of many species Passeriformes and Non-Passeriformes show that the basal metabolic rate (BMR) is strongly correlated with potential energy (MPE) and potential productive energy (PPE) (MPE is about 4BMR in all homeothermic animals). BMR is the minimal power of animal and strongly correlated with the daily work output, which may be determined by measuring total animal activity. Hence, BMR is the fundamental scale of power which determines the intensity of the actual interaction of an individual with the environment. The increase in BMR of a particular animal should rise the potential energy (MPE), potential productive energy (PPE), and the level of daily work output. BMR in Passerines birds is 1.3-1.5 fold higher than that in Non-Passerines and Mammals. Origin ofendothermy in the course of evolution should be associated with needs of general activity rather than with the requirements of thermoregulation.     

A metabolic derivation of tritium transfer coefficients in animal products

Tritium is a potentially important environmental contaminant originating from the nuclear industry, and its behaviour in the environment is controlled by that of hydrogen. Animal food products represent a potentially important source of tritium in the human diet and a number of transfer coefficient values for tritium transfer to a limited number of animal products are available. In this paper we present an approach for the derivation of tritium transfer coefficients which is based on the metabolism of hydrogen in animals. The derived transfer coefficients separately account for transfer to and from free (i.e. water) and organically bound tritium. A novel aspect of the approach is that tritium transfer can be predicted for any animal product for which the required metabolic input parameters are available. The predicted transfer coefficients are compared to available independent data. Agreement is good (R ²=0.97) with the exception of the transfer coefficient for transfer from tritiated water to organically bound tritium in ruminants. This may be attributable to the particular characteristics of ruminant digestion. We show that tritium transfer coefficients will vary in response to the metabolic status of an animal (e.g. stage of lactation, diet digestibility etc.) and that the use of a single transfer coefficient from diet to animal product is inappropriate. It is possible to derive concentration ratio values from the estimated transfer coefficients which relate the concentration of tritiated water and organically bound tritium in an animal product to their respective concentrations in the animals diet. These concentration ratios are shown to be less subject to metabolic variation and may be more useful radioecological parameters than transfer coefficients. For tritiated water the concentration ratio shows little variation between animal products ranging from 0.59 to 0.82. In the case of organically bound tritium the concentration ratios vary between animal products from 0.15 (goat milk) to 0.67 (eggs). Received: 28 May 2001 / Accepted: 20 August 2001     

Computing numerator relationships between any pair of animals

We describe a simple method to compute the numerator relationship between any or all pairs of animals in the numerator relationship matrix. The method depends on output of the MTDFNRM program from the MTDFREML set of programs. An option of the MTDFNRM program creates a file that includes the inbreeding coefficient for each animal. The method also makes use of how the inbreeding coefficient is traditionally calculated: one-half of the relationship between the animal's parents. To obtain the numerator relationship between any pair of animals, the original pedigree file is augmented with a dummy animal with an identification number (ID) greater than for any animal in the original pedigree file. The ID of the pair of animals for which the relationship is wanted is included as parents. MTDFNRM is then run with the option to create a file of ordered and original IDs for animals and their parents along with the inbreeding coefficients. We then multiply the inbreeding coefficient for a dummy animal by two to obtain the numerator relationship between the two animals designated as parents.     

The long-term effect of cadmium exposure through food on the postnatal development of the bank vole (Clethrionomys glareolus Schreber, 1780)

Cadmium is well known for its toxicity to the animal body. However, its effect on pregnancy and the development of young animals is still not well understood. This study examined such effects, using bank voles captured from the wild to make the results closer to those which could be expected in the natural environment. One group of animals was fed 7 microg g(-1) cadmium in the food, a second 35 microg g(-1), and a third no cadmium, as a control. The concentrations of cadmium in the whole bodies of young bank voles were determined on the 3rd, 5th, or 10th day of life. The cadmium level in the bodies of animals exposed to 35 microg g(-1) of cadmium was significantly higher than in those from either the control group or the group receiving 7 microg g(-1) of cadmium, which did not differ from each other. The cadmium level did not change with animal age in any of the study groups. Concentrations of Zn, Cu, and Fe were also determined in the whole body of young animals, as cadmium is known to disturb the metabolism of these essential metals through antagonistic activity. Both Cu and Fe levels were negatively correlated with cadmium concentrations, while a positive correlation was found between zinc and cadmium in the young animal bodies. Also found was higher offspring mortality in the group receiving 35 microg g(-1) of cadmium in food. There was no difference in young animal body weight between the study groups.