Ultrastructural distribution of peroxidase in thrombocytes of mammals and submammals

The localization and distribution of peroxidase (PPO) activity were studied ultracytochemically in thrombocytes from lampreys, carps, frogs, snakes, tortoises, rabbits, sheep, dogs, and monkeys. PPO activity was not detectable in the thrombocytes of lampreys, carps, frogs, and snakes. However, this enzyme activity was demonstrated in the nuclear envelope and endoplasmic reticulum of tortoise thrombocytes. Dog and monkey thrombocytes (blood platelets) exhibited PPO activity in the dense tubular system, but this enzyme activity was not detectable in rabbit and sheep thrombocytes. Our observations are interpreted to suggest that thrombocytes from animals lower than amphibia are peroxidase negative. Furthermore, it can be said that thrombocytes from animals higher than reptiles are generally positive, although there are exceptions. PPO activity was localized in the endoplasmic-reticulum system, but not in the cytoplasmic granules of thrombocytes common to submammals and mammals. In this study, we also compared the distribution of peroxidase activity in thrombocytes, neutrophils, and eosinophils and conclude that these are significant differences in the distribution of PPO and myeloperoxidase.     

Ultrastructural distribution of peroxidase in thrombocytes of mammals and submammals

Summary The localization and distribution of peroxidase (PPO) activity were studied ultracytochemically in thrombocytes from lampreys, carps, frogs, snakes, tortoises, rabbits, sheep, dogs, and monkeys. PPO activity was not deteetable in the thrombocytes of lampreys, carps, frogs, and snakes. However, this enzyme activity was demonstrated in the nuclear envelope and endoplasmic reticulum of tortoise thrombocytes. Dog and monkey thrombocytes (blood platelets) exhibited PPO activity in the dense tubular system, but this enzyme activity was not detectable in rabbit and sheep thrombocytes. Our observations are interpreted to suggest that thrombocytes from animals lower than amphibia are peroxidase negative. Furthermore, it can be said that thrombocytes from animals higher than reptiles are generally positive, although there are exceptions. PPO activity was localized in the endoplasmic-reticulum system, but not in the cytoplasmic granules of thrombocytes common to submammals and mammals. In this study, we also compared the distribution of peroxidase activity in thrombocytes, neutrophils, and eosinophils and conclude that these are significant differences in the distribution of PPO and myeloperoxidase.     

Glucose and free amino acids in the blood of lampreys (Lampetra fluviatilis L.) and frogs (Rana temporaria L.) under prolonged starvation

The content variation dynamics of glucose and free amino acids in blood plasma was followed for lampreys and frogs from autumn till spring, when the exogenous feeding is switched off. In October, the glucose level is rather high (8-10 mM) in blood plasma of both lampreys and frogs. It falls by 50% during winter and falls to a critically low level (1-2 mM) during spring. The lamprey plasma amino acid levels increase by 74% from November to April and thus reach the lower limit known for mammals. The amount of free amino acids in frog plasma decreases by 40% by spring in comparison with the values in autumn. More intensive proteolysis in lamprey tissues compared with that in frog tissues has been confirmed by quantitatively determining leucine, isoleucine, and valine in the blood of these animals. Besides these three amino acids, alanine, glycine, lysine, threonine and, in certain periods, tyrosine have been found to be quantitatively significant in the plasma of both animals.     

Comparative study of lipids and fatty acids in blood plasma of river lamprey Lampetra fluviatilis and brown frog Rana temporaria at the periods of elimination of exogenous feeding

Lipids of blood plasma of lampreys and frogs are composed of phospholipids, triglycerides, free fatty acids (FA), cholesterol, cholesterol esters, and waxes. The lipids content in plasma of frogs is markedly lower as compared with that of lampreys. However, the percentage of lipid components is represented by close values. Fluidity of triglycerides and phospholipids in lampreys is determined predominantly by monoenic acids and polyenoic acids of the ω-3-type, whereas that in frogs—by monoenic acids and polyenic acids of the ω-6-type. Free FA are represented mainly by saturated and monoenic acids.     

Comparative physiological characteristics of the action of pentagastrin on gastric secretion in fish, frogs, turtles and chickens]

The same preparation of pentagastrin, which is highly effective in dogs (threshold dose 0.3 mug/kg), has been tested on fishes, frogs, tortoises and hens. In fishes (rays), the threshold dose was equal to 50 mug/kg; the drug only increased juice secretion, but did not decrease pH of the juice. In frogs, the effective dose was equal to 25, in tortoises -- to 50 mug/kg, the drug increasing the secretion of both the juice and HCl. The effect of the drug exhibits seasonal variations, being absent in winter period. In hens, the threshold dose is equal to 5 mug/kg; the drug increases both the volume of the juice and HCl production. In frogs and hens, similar to that in dogs (tortoises were not studied in this respect), pentagastrin also increases the production of proteases of gastric juice. It is suggested that in the animals studied pentagastrin is a specific stimulator of gastric secretion.     

Free amino acids in the blood plasma of lampreys, frogs and rats

Free amino acids in the blood plasma of lampreys, frogs and rats were determined by HPLC. In spite of quantitative differences in the total pool of free amino acids, the specific content of physiologically important amino acids is quite similar in representatives of different classes of vertebrates, from cyclostomes to mammals.     

Neurotrophic regulation of the restorative processes in muscle autografts in vertebrates

In frogs, tortoises and rats autotransplantation of the whole gastrocnemius muscle has been performed into its own bed at sewing the nerve to the autotransplant and at its denervation. Fragments of the skeletal muscles are also transplanted into the brain and into the spinal cord in frogs and rats. By means of light and electron microscopy, restorative processes are studied in the transplanted muscles. Acceleration of histomorphogenesis of the muscular and nervous tissues is stated from the lower toward the higher Vertebrata. When the transplants are deprived of the connection with the nervous system, the cause of the form organization process in all the animals investigated is disturbed, however, the nervous-trophic regulation is of the greatest importance in the mammalia. The experiments reveal mutual regulation of the plastic activity in the nervous and muscular tissues, that becomes more intensive in the course of evolution.     

Natriuretic reaction of the vertebrate kidney

After injection of a 10% solution of NaCl, the kidney of rats, and to a lower extent that of hens, rapidly excretes the excess sodium. Effective sodium excretion in homoiothermic animals is based on high level of glomerular filtration and proximal reabsorption. After injection of a 10% solution of NaCl to the Lamprey Lampetr fluviatilis and frog Rana temporaria, sodiumuretic reaction of the kidney was less evident. During water diuresis, cells of renal tubuli of lampreys, frogs, hens and rats produce high concentration gradient between the blood serum and urine, preventing loss of sodium. In evolution, the kidney was formed presumably as an organ which prevents sodium loss from the organism; this function remains highly effective in all vertebrates, whereas in homoiothermic animals the kidney attains also the capacity for rapid extrusion of excessive sodium.     

The effect of dehydration on the functional activity of the interrenal gland in adenohypophysectomized Rana catesbeiana frogs. A preliminary report

A significant increase of the content of corticosterone in the blood collected from intravenous cannula or by intracardiac punction has been detected using radioimmunoassay in non-operated and adenohypophysectomized frogs Rana catesbeiana subjected to dehydration in 6.2% mannitol solution during 24 hours. The osmolality of the blood plasma of these animals also increases although less significantly than the growth of plasma corticosterone content. There is a tendency to substantial increase of plasma arginine-vasotocin level prior to the growth of corticosterone level, already after 6 hours of dehydration. Based on the present results and literature data, it is suggested that in adenohypophysectomized frogs lacking endogenous ACTH just the increase of blood arginine-vasotocin level results in a substantial activation of corticosteroid-producing cells of the interrenal gland and in the growth of plasma content of corticosterone.     

Western blot can distinguish natural and acquired antibodies to Mycoplasma agassizii in the desert tortoise (Gopherus agassizii)

Mycoplasma agassizi has been identified as a cause of upper respiratory tract disease (URTD) in the threatened Mojave population of the desert tortoise (Gopherus agassizii), and anti-M. agassizii antibodies have been found by ELISA in as many as 15% of these animals across their geographic range. Here we report that a cohort of 16 egg-reared desert tortoises never exposed to M. agassizii had ELISA antibody titers to this organism that overlapped with titers obtained from some M. agassizii-infected tortoises. These natural antibodies were predominantly of the IgM class. Western blots of plasma from these non-infected tortoises produced a characteristic banding pattern against M. agassizii antigens. A group of 38 wild-caught desert tortoises was tested by ELISA, and although some of these tortoises had antibody titers significantly higher than the non-infected tortoises, there was considerable overlap at the lower titer levels. However, Western blot analysis revealed distinct banding patterns that could readily distinguish between the non-infected tortoises and tortoises with acquired antibodies, regardless of ELISA antibody titers. We conclude that desert tortoises have natural antibodies to M. agassizii that can compromise the determination of infection status by ELISA. However, the Western blot technique can distinguish between natural and acquired antibody patterns and can be used to confirm the diagnosis of M. agassizii infections in the desert tortoise.     

Shifting patterns of nitrogen excretion and amino acid catabolism capacity during the life cycle of the sea lamprey (Petromyzon marinus)

The jawless fish, the sea lamprey (Petromyzon marinus), spends part of its life as a burrow-dwelling, suspension-feeding larva (ammocoete) before undergoing a metamorphosis into a free swimming, parasitic juvenile that feeds on the blood of fishes. We predicted that animals in this juvenile, parasitic stage have a great capacity for catabolizing amino acids when large quantities of protein-rich blood are ingested. The sixfold to 20-fold greater ammonia excretion rates (J(Amm)) in postmetamorphic (nonfeeding) and parasitic lampreys compared with ammocoetes suggested that basal rates of amino acid catabolism increased following metamorphosis. This was likely due to a greater basal amino acid catabolizing capacity in which there was a sixfold higher hepatic glutamate dehydrogenase (GDH) activity in parasitic lampreys compared with ammocoetes. Immunoblotting also revealed that GDH quantity was 10-fold and threefold greater in parasitic lampreys than in ammocoetes and upstream migrant lampreys, respectively. Higher hepatic alanine and aspartate aminotransferase activities in the parasitic lampreys also suggested an enhanced amino acid catabolizing capacity in this life stage. In contrast to parasitic lampreys, the twofold larger free amino acid pool in the muscle of upstream migrant lampreys confirmed that this period of natural starvation is accompanied by a prominent proteolysis. Carbamoyl phosphate synthetase III was detected at low levels in the liver of parasitic and upstream migrant lampreys, but there was no evidence of extrahepatic (muscle, intestine) urea production via the ornithine urea cycle. However, detection of arginase activity and high concentrations of arginine in the liver at all life stages examined infers that arginine hydrolysis is an important source of urea. We conclude that metamorphosis is accompanied by a metabolic reorganization that increases the capacity of parasitic sea lampreys to catabolize intermittently large amino acid loads arising from the ingestion of protein rich blood from their prey/hosts. The subsequent generation of energy-rich carbon skeletons can then be oxidized or retained for glycogen and fatty acid synthesis, which are essential fuels for the upstream migratory and spawning phases of the sea lamprey's life cycle.     

Translocation as a conservation tool for Agassiz's desert tortoises: Survivorship,reproduction, and movements

We translocated 120 Agassiz's desert tortoises to 5 sites in Nevada and Utah to evaluate the effects of translocation on tortoise survivorship, reproduction, and habitat use. Translocation sites included several elevations, and extended to sites with vegetation assemblages not typically associated with desert tortoises in order to explore the possibility of moving animals to upper elevation areas. We measured survivorship, reproduction, and movements of translocated and resident animals at each site. Survivorship was not significantly different between translocated and resident animals within and among sites, and survivorship was greater overall during non-drought years. The number of eggs produced by tortoises was similar for translocated and resident females, but differed among sites. Animals translocated to atypical habitat generally moved until they reached vegetation communities more typical of desert tortoise habitat. Even within typical tortoise habitat, tortoises tended to move greater distances in the first year after translocation than did residents, but their movements in the second or third year after translocation were indistinguishable from those of resident tortoises. Our data show that tortoises translocated into typical Mojave desert scrub habitats perform well; however, the large first-year movements of translocated tortoises have important management implications. Projects that employ translocations must consider how much area will be needed to contain translocated tortoises and whether roads need fencing to prevent the loss of animals. © 2012 The Wildlife Society.     

Poikilotherms as reservoirs of Q-fever (Coxiella burnetii) in Uttar Pradesh.

Water snakes (Natrix natrix), rat snakes (Ptyas korros), cobras (Naja naja), pythons (Python molurus), tortoises (Kachuga sp.), plankton fish (Cirrhina mrigala), frogs (Rana tigrina), toads (Bufo sp.) and monitors (Varanus indicus) were screened for evidence of Q-fever infection by the capillary agglutination test on sera to detect antibodies and/or by attempts to demonstrate Coxiella burnetii in spleen and liver samples. Sero-reactors were observed among water and rat snakes, pythons and tortoises. The organism was isolated from the spleen and liver of the monitor, tortoise and python.     

Seroepidemiology of upper respiratory tract disease in the desert tortoise in the western Mojave Desert of California

Several factors have combined with an upper respiratory tract disease (URTD) to produce declines on some population numbers of desert tortoises (Gopherus agassizii) in the western USA. This study was designed to determine the seroepidemiology of URTD in a population of wild adult tortoises at the Desert Tortoise Research Natural Area (DTNA) study site in Kern County (California, USA). Prior to initiation of the study, there was a dramatic decline in the number of individuals in this population. At each individual time point, samples were obtained from 12 to 20 tortoises with radiotransmitters during winter, spring, summer, and fall from 1992 through 1995. During the course of the study, 35 animals were sampled at one or more times. Only 10 animals were available for consistent monitoring throughout the 4 yr period. Specific antibody (Ab) levels to Mycoplasma agassizii were determined for individual tortoises by an enzyme-linked immunosorbent assay (ELISA) test. Specific Ab levels were not influenced by the gender of the tortoise. Levels of Ab and distribution of ELISA+, ELISA- and suspect animals were not consistently affected by season within a single year or for a season among the study years. Significantly more tortoises presented with clinical signs in 1992 and 1995. The profile of ELISA+ animals with clinical signs shifted from 5% (1992) to 42% (1995). In 1992, 52% of tortoises lacked clinical signs and were ELISA-. In 1995, this category accounted for only 19% of tortoises. Based on the results of this study, we conclude that URTD was present in this population as evidenced by the presence of ELISA+ individual animals, and that the infectious agent is still present as evidenced by seroconversion of previously ELISA- animals during the course of the study. There is evidence to suggest that animals may remain ELISA+ without showing overt disease, a clinical pattern consistent with the chronic nature of most mycoplasmal infections. Further, there are trends suggesting that the clinical expression of disease may be cyclical. Continued monitoring of this population could provide valuable information concerning the spread of URTD in wild tortoise populations.     

Western blot can distinguish natural and acquired antibodies to Mycoplasma agassizii in the desert tortoise (Gopherus agassizii)

Mycoplasma agassizi has been identified as a cause of upper respiratory tract disease (URTD) in the threatened Mojave population of the desert tortoise (Gopherus agassizii), and anti-M. agassizii antibodies have been found by ELISA in as many as 15% of these animals across their geographic range. Here we report that a cohort of 16 egg-reared desert tortoises never exposed to M. agassizii had ELISA antibody titers to this organism that overlapped with titers obtained from some M. agassizii-infected tortoises. These natural antibodies were predominantly of the IgM class. Western blots of plasma from these non-infected tortoises produced a characteristic banding pattern against M. agassizii antigens. A group of 38 wild-caught desert tortoises was tested by ELISA, and although some of these tortoises had antibody titers significantly higher than the non-infected tortoises, there was considerable overlap at the lower titer levels. However, Western blot analysis revealed distinct banding patterns that could readily distinguish between the non-infected tortoises and tortoises with acquired antibodies, regardless of ELISA antibody titers. We conclude that desert tortoises have natural antibodies to M. agassizii that can compromise the determination of infection status by ELISA. However, the Western blot technique can distinguish between natural and acquired antibody patterns and can be used to confirm the diagnosis of M. agassizii infections in the desert tortoise.     

Effect of glutaraldehyde on hemoglobin: functional aspects and M?ssbauer parameters

Glutaraldehyde is widely used for the cross-linking of hemoglobin for blood substitute research or for technological purposes. The effects of this reagent on the biochemical properties of hemoglobin were correlated with M?ssbauer data. Human hemoglobin was cross-linked by glutaraldehyde as soluble polymers and insoluble particles. Effects of cross-linking on oxygen affinity, oxidation-reduction potential, autoxidation kinetics, and thermal stability were studied. Stability of cross-linked hemoglobin was specifically studied by M?ssbauer spectroscopy. Oxygen affinity is increased, redox potential is decreased, autoxidation rates are increased, and stability towards thermal denaturation is increased. The regeneration of partially denatured hemoglobin by glutaraldehyde cross-linking is shown. Effects of cross-linking on biochemical properties are explained by the hypothesis of the opening of the heme pocket on the distal-histidine side and the concomitant charge transfer from the iron to the oxygen.     

The Dominance of Introduced Plant Species in the Diets of Migratory Galapagos Tortoises Increases with Elevation on a Human‐Occupied Island

The distribution of resources and food selection are fundamental to the ecology, life history, physiology, population dynamics, and conservation of animals. Introduced plants are changing foraging dynamics of herbivores in many ecosystems often with unknown consequences. Galapagos tortoises, like many herbivores, undertake migrations along elevation gradients driven by variability in vegetation productivity which take them into upland areas dominated by introduced plants. We sought to characterize diet composition of two species of Galapagos tortoises, focussing on how the role of introduced forage species changes over space and the implications for tortoise conservation. We quantified the distribution of tortoises with elevation using GPS telemetry. Along the elevation gradient, we quantified the abundance of introduced and native plant species, estimated diet composition by recording foods consumed by tortoises, and assessed tortoise physical condition from body weights and blood parameter values. Tortoises ranged between 0 and 429 m in elevation over which they consumed at least 64 plant species from 26 families, 44 percent of which were introduced species. Cover of introduced species and the proportion of introduced species in tortoise diets increased with elevation. Introduced species were positively selected for by tortoises at all elevations. Tortoise physical condition was either consistent or increased with elevation at the least biologically productive season on Galapagos. Santa Cruz tortoises are generalist herbivores that have adapted their feeding behavior to consume many introduced plant species that has likely made a positive contribution to tortoise nutrition. Some transformed habitats that contain an abundance of introduced forage species are compatible with tortoise conservation.     

Effect of hypophysectomy on plasma electrolytes and adrenal mineralocorticoid secretion in the terrestrial chelonian Testudo hermanni

Na, K, ACTH, corticosterone and aldosterone plasma levels were studied in Testudo hermanni after hypophysectomy and dexamethasone administration. Hypophysectomy as well as dexamethasone treatment caused a deep decrease in the plasma levels of electrolytes, corticosterone and aldosterone when compared to sham-operated or normal controls. ACTH plasma level was markedly lower in hypophysectomized or dexamethasone-treated tortoises when compared to the controls but never reached undetectable levels. Administration of ACTH significantly increased plasma electrolytes and hormone levels in hypophysectomized and dexamethasone-treated tortoises. The present data seem to indicate that the pituitary plays a significant role on electrolyte homeostasis and on the regulation of plasma mineralocorticoids level in terrestrial chelonians.     

Demographic dynamics of the afro-tropical pig-nosed frog, <Emphasis Type="Italic">Hemisus marmoratus</Emphasis>: effects of climate and predation on survival and recruitment

We studied the population ecology of the West African pig-nosed frog, Hemisus marmoratus, to understand the relative contributions of adult survival and recruitment to population growth rate in savannah frogs using mark-recapture modelling. We marked a total of 821 adult frogs over 6 years and recaptured 74 at least once between years. Between-year adult survival was sex-specific and varied between 0.06 and 0.53 for males and 0.07–0.41 for females. Adult survival was significantly associated with annual rainfall and is cause for concern if rainfall declines further in the study region as predicted by changes in the global climate. There was a significant interaction between rainfall and sex with dry weather having a stronger negative effect on males than females. Pig-nosed frogs experienced boom and bust years with a single decline more dramatic than increases. Recruitment (in situ and immigration; 0.67–0.88) was substantially more important than adult survival (0.12–0.33) in determining realised population growth. In situ recruitment was highly variable between years with 1–36% of eggs and tadpoles released by females into the pond surviving to metamorphosis. Years of low tadpole survival were associated with high numbers of predatory tortoises. Thus, like other pond-breeding anurans, pig-nosed frogs showed highly variable juvenile recruitment, low adult survival and density-independent effects on population growth by predators and weather.     

Effect of vasotocin on plasma free fatty acid level in the migrating anadromous sea lamprey.

The effect of injections of arginine vasotocin (AVT) on plasma free fatty acid (FFA) levels was studied in anadromous sea lampreys collected in the St. John River, New Brunswick, during their upstream spawning migration. Plasma FFA was significantly higher in lampreys injected with a single dose of 1 000 mU vasotocin/kg body weight than in those receiving only the vehicle solution, the difference being the greater at 90 than at 30 min post-injection. The significance of AVT in migration is discussed.