Study of color variation in the solitary ascidian Halocynthia roretzi, collected in the Inland Sea of Japan

In the vicinity of Yashiro Island in the Inland Sea of Japan, the solitary ascidian (tunicate) Halocynthia roretzi with tunics of various colors were collected. Samples of these animals were sorted into three groups on the basis of visual observation of tunic color. The red group includes animals with dark-red, light-red, or orange tunics. The pink group includes animals with tunic colors ranging between red and white. The white group includes only animals with completely white tunics. Animals in the white group lacked color internally, with the exception of the hepatopancreas and the gonads in breeding season; the epidermis and gill basket were white. In contrast, animals of both the red group and the pink group were colored internally, with red-orange epidermis and yellow gill basket. Alloreactivity was tested by mixed-hemocyte incubation between different animals belonging to the same color group and between animals belonging to different color groups. Alloreactivity between animals of the white group was 56.3%, between animals of the pink group was 60.0%, and between animals of the red group was 69.3%. The relatively high frequency of compatible combinations among the white animals is discussed.     

Studies on Sudden Fatalities Among Piglets Following Parenteral Iron Therapy

An investigation was carried out in order to clarify whether there is a correlation between the latent iron-binding capacity, UIBG, in the serum of suckling piglets and sudden fatalities occurring among these animals when they are treated with 250 mg trivalent iron in the form of a complex also containing dextrin, sorbitol, citric acid and lactic acid. In all, 97 animals from 9 litters were used. By administering 100 mg oral divalent iron to 22 animals, the iron-binding capacity was saturated or appreciably reduced 3 hrs. after the oral treatment. After this time, the animals were treated with parenteral iron. Seventeen other animals were treated with 100 mg divalent iron and immediately afterwards with parenteral iron. Three hrs. later, the iron-binding capacity of the animals was exceeded. In 32 of the control animals, UIBG was high before the parenteral treatment. No fatalities were observed among the animals treated with parenteral iron. Twenty-three of the animals had a high iron-binding capacity in spite of having diarrhoea. On parenteral treatment of these animals with the iron complex, no fatalities were observed which could be ascribed to the treatment. The mechanism for the sudden fatalities among suckling piglets after parenteral administration of iron is discussed.     

Testicular development, ultrasonographic and histological appearance of the testis in ram lambs immunized against recombinant LHRH fusion proteins

Sixteen native ram lambs weaned at 10 wk of age were divided into two groups. Eight animals were immunized against LHRH with a mixture of two fusion proteins: ovalbumin-LHRH-7 and thioredoxin-LHRH-7. The immunized lambs received a primary immunization plus two booster immunizations at 4 and 12 wks. Animals in the control group (n=8) were not treated. Scrotal measurements and blood samples were taken at 2-week intervals. Beginning at 25 wk of age, semen was collected and sexual behaviour was evaluated on a weekly basis. At 35 and 37 wk of age testes and accessory glands of all animals were subjected to ultrasound scanning. At 37 wk of age animals were slaughtered and testes were evaluated histologically. Serum LHRH antibodies (P<0.01) were detected in animals of the immunized group which had reduced serum testosterone concentrations (P<0.01). Testicular development was suppressed in the immunized animals (P<0.01). Immunized animals exhibited mounting activity 5 wks later than control animals. No mature spermatozoa containing ejaculates were collected from immunized animals. Control animals had moderately echogenic ultrasonographic appearance at 37 wk age, whereas immunized animals had hypoechogenic images. Mean seminiferous tubule diameter in immunized lambs was significantly smaller than that in control lambs. Basal membrane was thickened and hyalinized; there was an increase in peritubular connective tissue. No proliferating spermatogonia or mature spermatozoa were present in the tubules in these animals. There were no differences in the ultrasonographic appearance of prostate and vesicular gland between control and immunized animals. The LHRH recombinant fusion proteins were effective in immunological castration in ram lambs when started at 10 wk of age as noted by differences in serum testosterone, testicular histology and ultrasonographic appearance of testis and weight of accessory sex glands. Determining the effects of immunization on ultrasonographic appearance of the testis related to time after immunization requires further investigations.     

Metabolism of apoprotein B in selectively bred baboons with low and high levels of low density lipoproteins

Very low density lipoprotein (VLDL) and low density lipoprotein (LDL) apoprotein (apo)-B turnover rates were measured simultaneously by injecting 131I-labeled VLDL and 125I-labeled LDL into fasting baboons (Papio sp.) selectively bred for high serum cholesterol levels and having either low or high LDL levels. The radioactivities in VLDL, intermediate density lipoprotein (IDL), LDL apoB, and urine were measured at intervals between 5 min and 6 days. Kinetic parameters for apoB were calculated in each baboon fed a chow diet or a high cholesterol, high fat diet (HCHF). VLDL apoB residence times were similar in the two groups of animals fed chow; they were increased by HCHF feeding in high LDL animals, but not in low LDL animals. Production rates of VLDL apoB were decreased by the HCHF diet in both high and low LDL animals. Most of the radioactivity from VLDL apoB was transferred to IDL. However, a greater proportion of radioactivity was removed directly from IDL apoB in low LDL animals than in high LDL animals, and only about one-third appeared in LDL. In high LDL animals, a greater proportion of this radioactivity was converted to LDL (61.4 +/- 7.2% in chow-fed animals and 49.2 +/- 10.9% in animals fed the HCHF diet; mean +/- SEM, n = 5). Production rates for LDL apoB were higher in high LDL animals than those in low LDL animals on both diets. The HCHF diet increased residence times of LDL apoB without changing production rates in both groups. VLDL apoB production was not sufficient to account for LDL apoB production in high LDL animals, a finding that suggested that a large amount of LDL apoB was derived from a source other than VLDL apoB in these animals.     

Pituitary ACTH responsiveness in the vasopressin deficient rat.

ACTH concentration and the responsiveness of dispersed anterior pituitary cells to hypothalamic extract and to vasopressin were studied in homozygous (DI), heterozygous (HTZ), DI-pitressin treated (DIP) Brattleboro rats, and in control Long-Evans rats.Absolute, but not relative, anterior pituitary weights of HTZ, DI, and DIP animals were significantly smaller than those of controls. The concentration of immunoreactive (I) and bioreactive (B) ACTH in dispersed anterior pituitary cells was greater in DI and DIP than in HTZ or in control animals, although the total amount of ACTH was greater in control than DI or HTZ animals. Media from incubates of pituitary cells derived from DI and DIP animals contained less I and B ACTH than those from HTZ or control animals. Pituitary cells derived from DI animals secreted markedly less ACTH following incubation with hypothalamic extract (NIH-HE-RP-1) than did cells from HTZ animals. The response in DIP animals was intermediate between that of DI and HTZ animals. In contrast, pituitary cells derived from DI and DIP animals were significantly more responsive to vasopressin than those from control or HTZ animals.     

Does immune challenge affect torpor duration?

1. Hibernation may alter the relationship between pathogens and their hosts; low host temperatures can prevent pathogen replication. Therefore, manipulating the timing and duration of torpor bouts could allow animals to gain an advantage over pathogens.
2. Thirty-two Turkish Hamsters ( Mesocricetus brandti ) were placed in short-day, cold conditions. After 10 weeks, 20 animals were challenged with an antigen to simulate a pathogen infection. Ten of these animals were returned to the cold ('cold-challenged'). The other 10 animals were placed in warm conditions ('warm-challenged'). Twelve animals received saline injections and were returned to the cold ('cold-control'). Cold-challenged animals spent significantly more time in torpor than did cold-control animals.
3. After 6·5 weeks, all animals were housed in warm conditions and ceased torpor. Both cold-challenged and warm-challenged animals received a second injection of antigen. There was no correlation between time spent euthermic and level of secondary humoral response of cold-challenged animals. The secondary humoral response of the cold-challenged animals was significantly lower than that of warm-challenged animals.
4. In this study immune status influenced torpor duration, and torpor caused immunosuppression. Hibernators may manipulate body temperature in order to combat pathogens while their own immune systems are suppressed.     

Effects of light on circadian pacemaker development

The effects of raising cockroaches, Leucophaea maderae, in non-24-h light cycles on the response of the circadian system to light was examined. 1. Phase response curves (PRC) were measured for 6-h light pulses for animals raised in LD 11:11 (T22), LD 12:12 (T24), and LD 13:13 (T26). The delay portion of the PRC was found to be significantly reduced in T22 animals (compared to T24 animals) while the advance portion of the PRC was reduced in T26 animals. Compared to T26 animals, phase shifts were more positive at every phase for animals raised in T22. 2. When transferred from constant darkness (DD) to constant light (LL) the freerunning period lengthened significantly less for T22 animals than T24 animals, and in some cases tau in LL was actually shorter than tau in DD in T22 animals. Animals raised in LL were inactive when exposed to LL as adults, and unlike T24 animals, were consistently reset to the beginning of the subjective night (near CT 12) when transferred to DD. 3. Roaches raised in T22 would entrain to LD 6:18, but a few animals exhibited periods of relative coordination indicating that the 24-h light cycle was near the limits of entrainment. These results indicate that the circadian system's responsiveness to light, as well as its freerunning period (Barrett and Page 1989), is dependent on the lighting conditions to which the animals are exposed during development.     

Indirect immunohistochemistry on skin biopsy for the detection of persistently infected cattle with bovine viral diarrhoea virus in Italian dairy herds

Indirect immunohistochemistry (IHC) on skin biopsies for identification of persistently infected (PI) animals has been used as a parallel test to antigen and antibody ELISAs in a bovine viral diarrhoea (BVD) voluntary control program. The aim was to evaluate the reliability and feasibility of IHC on ear skin tissues to detect PI animals in field conditions, including both adult and calves under 6 months of age. In animals over 6 months of age skin biopsy and blood sample were collected at the same time, whereas in young calves blood sampling was performed when animals reached 6 months of age. One hundred and sixty-five animals were tested and immunohistochemical results were compared with those of antigen ELISA. In case of inconclusive results virus isolation and virus neutralization assays were performed. Agreement K value was 0.96. Immunohistochemical staining in positive animals was clearly detectable in the keratinocytes of the epidermis and adnexa.     

Dichloroacetate-induced changes in liver of normal and diabetic rats.

Dichloroacetate (DCA) was administered orally to normal (nondiabetic) and streptozotocin-diabetic rats in a dose of 1000 mg/day/kg rat wt. One group of diabetic animals received DCA both orally and intraperitoneally. DCA therapy lowered the blood glucose values of diabetic animals but did not alter values in nondiabetic rats. The hepatic activity of glucokinase and pyruvate kinase were significantly lower in both DCA-treated nondiabetic and DCA-treated diabetic animals than values observed for untreated animals. However, DCA therapy was accompanied by remarkable increases in the activities of glucose-6-phosphate dehydrogenase and malic enzyme in both nondiabetic and diabetic animals. Glucose-6-phosphate dehydrogenase was 3-fold higher in DCA-treated nondiabetic animals whereas malic enzyme activity was 10-fold higher in the treated animals than observed in the untreated animals. Similar changes, although smaller in magnitude, were observed for these enzymes in the DCA-treated diabetic animals. Although DCA therapy was accompanied by a significant increase in the wet weights of the liver for both nondiabetic and diabetic animals, no morphological changes were seen by light or electron microscopy. Our observations coupled with those of previous investigators suggest that DCA therapy may have an important role in pyruvate metabolism and may lower the blood glucose concentration by inhibiting hepatic gluconeogenesis.     

Change in immunological and biochemical parameters in germ-free animals in response to T-activin

The influence of immunoregulating humoral thymus factor T-activin (fraction AFT-6) on the activity of xenobiotic metabolism enzymes (EMX) and cellular-dependent cytotoxicity was investigated in germ-free guinea-pigs. Germ-free and conventional animals were injected 5 micrograms of T-activin intraperitoneally, on 3 successive days. The control animals were injected a physiologic saline. A day after the last injection the animals were killed, and EMX and K cell activity was measured. It was found that EMX activity in germ-free animals was decreased. T-activin stimulated cytochrome P-450, NADP-cytochrome-C-reductase, glutathione-S-transferase, benzopyrene hydroxylase and epoxyhydrase activity. In conventional animals the activity of these enzymes remained unchanged under the influence of T-activin. K-cell activity in germ-free animals was decreased, as compared to conventional guinea-pigs. Under the influence of T-activin the parameter increased and stood at about 70% of normal values.     

Corynebacterium Pseudotuberculosis Infection in Goats II.

The precalen-ce of caseous lymphadenitis was surveyed in 36 goat herds in Northern Norway. In each herd, information concerning the occurrence of the disease was obtained from the farmer. Adult animals (1 year of age or older) in 35 herds were examined for superficial swellings, and serum samples were collected from most animals in the herds. The sera were examined for antibodies to Corynebacterium pseudotuber-culosis using the bacterial agglutination test (BAT) and the hemolysis inhibition test (HIT). Gaseous lymphadenitis was diagnosed with certainty in 19 herds. Information from the farmers indicated that the disease indeed oc-curred in these herds, and that the majority had been infected with the disease for many years. The herds had apparently become infected through contact with animals from infected herds. Clinical examina-tions were carried out in 18 of these herds and superficial swellings were found in 26 % of the examined animals. The prevalence of ani-mals with lesions varied from 11 to 40 % among the herds. Of the animals in these herds, 81 % were positive in BAT and 84 % in HIT. The prevalence of positive animals varied from 26 to 99 % in BAT and 28 to 99 % in HIT. The prevalence of seropositive animals was lowest in a herd in which animals were kept separately in stalls. Caseous lymphadenitis could not be diagnosed in 16 herds. In-formation from the farmers indicated that the disease indeed seemed to be absent in 14 of these herds. These 14 herds had no history of contact with animals from herds considered to be infected. However, in the remaining 2 herds, the farmers were somewhat uncertain about the occurrence of the disease. One of these 2 herds had a history of contact with infected herds through participation in a goat “breeding circle”. Only a few of the animals were, however, seropositive and all these had low antibody titres. In 1 newly established herd, a single animal showed a high posi-tive titre in BAT only. All the other animals were negative in both tests. This particular herd consisted of animals obtained both from herds with caseous lymphadenitis and from herds in which the disease was not considered to occur.     

The effect of surgical trauma on muscle protein turnover in rats.

The rate of synthesis and catabolism of sarcoplasmic- and myofibrillar-muscle protein was measured in operated, sham-operated and food-restricted rats by using Na2 14CO3. The food-restricted group underwent sham operations and were limited to the food intake of the operated animals. Protein synthesis and catabolism were increased in the sarcoplasmic-muscle fraction in operated rats compared with that in sham-operated or food-restricted rats. The rate of synthesis of the myofibrillar protein decreased in operated animals, but the rate of catabolism was not altered in the myofibrillar-muscle fraction of the operated animals compared with that in food-restricted and sham-operated animals. In the operated animals, there was a net loss of protein from the muscle. Thus the rats that underwent surgery lost muscle protein, primarily as a result of a decrease in synthesis of myofibrillar protein. The changes in protein turnover in operated animals were not due to decreases in food intake, since protein turnover in sham-operated animals that were restricted to the food intake of the operated rats was not different from that in sham-operated rats fed ad libitum.     

Atrial natriuretic peptide (ANP)-granules of auricular cardiocytes in aging Mongolian gerbils.

The atrial natriuretic peptide (ANP)-granules of right auricular cardiocytes were studied in aging Mongolian gerbils by immunohistochemistry and transmission electron microscopy, and analyzed by ultrastructural morphometry. Immunohistochemically, the ANP immunoreactions were weaker in 3 and 4-year-old animals than in 90-day-old, 1- and 2-year-old animals. There was no difference in the reaction among 90-day-old, 1 and 2-year-old animals, or between 3 and 4-year-old animals. Ultrastructurally, the morphological features in 1 and 2-year-old animals were similar to those of 90-day-old animals, but distinct in 3 and 4-year old animals by the presence of a few lysosomal structures in the cell. By ultrastructural morphometry, number and diameter of ANP-granules of 3 and 4-year-old animals were significantly smaller than those in the 90-day-old, 1 and 2-year-old. There was no significant difference in number and diameter among the 90-day-old, 1 and 2-year-old, or between the 3 and 4-year-old.     

Resistance to experimental autoimmune myasthenia gravis in genetically inbred rats. Association with decreased amounts of in situ acetylcholine receptor-antibody complexes

Genetically related susceptibility for experimental autoimmune myasthenia gravis was investigated in nine inbred strains of rats immunized with heterologous acetylcholine (AChR) from Torpedo californica. Wistar Munich and Fischer strain animals consistently developed severe, fatal disease associated with impaired neuromuscular transmission and increased sensitivity to low doses of curare. A lower incidence of disease was induced in Wistar Kyoto, ACI, Brown Norway, Buffalo, and Lewis strain animals. In contrast, Wistar Furth and Copenhagen strain animals were resistant to experimental autoimmune myasthenia gravis, electrophysiologic responses were normal, and animals were insensitive to curare. All strains of animals manifested equivalent amounts of serum antibody to AChR and total muscle AChR was reduced to the same extent in both resistant and susceptible animals. In contrast, the amount of antibody-bound AChR was greater in susceptible Wistar Munich animals than the amount observed in resistant Wistar Furth animals. These data suggest that impaired neurotransmission is correlated with the extent of antibody binding to the AChR. The discordance in the amount of antibody bound to the AChR of resistant and susceptible animals may result from heritable differences in antibody properties. Cross-breeding experiments with Wistar Munich and Wistar Furth animals show that resistance for development of experimental autoimmune myasthenia gravis is recessive and indicate that disease susceptibility is linked to one or two genetic loci.     

Studies of brain myelin in the "quaking mouse"

Myelin was isolated from the brains of "quaking" and littermate control animals and its composition was determined. The brains of quaking animals contained approximately one-fourth as much myelin as the control animals. There were qualitative as well as quantitative differences between the myelin from the two groups. By continuous cesium chloride gradient flotation it was shown that the myelin from the quaking animals consisted solely of a band corresponding to the heavier and smaller of the two bands found in normal controls. Cholesterol and glycolipids were lower and phospholipids (mainly phosphatidylcholine) and protein were higher in quaking animals than in controls. Also, phosphatidal-ethanolamine was decreased, and several consistent differences in the fatty acids (both unsubstituted and hydroxy) and aldehydes of the component lipids were found. In general there were smaller amounts of monounsaturated fatty acids in quaking animals. We suggest from these findings that myelin in the quaking mouse has certain compositional similarities with juvenile myelin, but it may be an abnormal type of myelin.     

The individual postradiation reactions of cyclic nucleotides in the lymphoid cells of the rat spleen]

X irradiation (4.3 Gy) of rats was shown to cause different reactions of the cyclic nucleotide system in differently reactive animals which was determined by the neutrophilic response in the peripheral blood after three-hour immobilization. Changes in the cyclic nucleotide content and in the activity of adenylate cyclase were found to occur in both hyper- and hyporeactive animals 24 h after irradiation. In hyperreactive animals, however, these changes were more pronounced: a relative ability of adenylate cyclase to be activated in hyperreactive animals made 30%, and 52% in hyporeactive animals. The cAMP content in lymphocytes of the spleen of hyporeactive rats was twice as high as that in hyperreactive animals.     

Trypanosoma congolense: thrombocytopenia in experimentally infected cattle

Hereford cattle infected with Trypanosoma congolense developed a thrombocytopenia which was most severe early in the course of infection when parasite levels in peripheral blood were highest. As the disease progressed, parasite levels gradually decreased and a corresponding increase in the number of thrombocytes occurred. In chronically infected animals, the number of thrombocytes was reduced during and shortly after periods of patency. However, during extended remission, thrombocyte values rose to normal or higher levels. Thrombocytes in surviving animals returned to within normal limits. When acutely ill, thrombocytopenic animals were treated with Berenil, a thrombo-cytosis developed rapidly. Accompanying the thrombocytopenia in infected animals was a consistent prolongation of partial thromboplastin times between the 6th and 14th weeks of infection. Plasma protamine paracoagulation tests were positive and fibrinogen levels were decreased in infected animals. No difference were found however in prothrombin times between infected and control animals.     

Effects of chronic ethanol treatment and aging on brain phosphoinositide turnover and adenylate cyclase activity

Inositol phosphate accumulation and adenylate cyclase activity were investigated in the cortex of young and aged ethanol-treated rats. Three months of ethanol treatment of young rats decreased maximal stimulation of inositol phosphate accumulation by carbachol by 26%, from 494 ± 76% of basal turnover in control animals to 396 ± 54% in ethanol-treated animals (mean ± SD). In aged rats ethanol-related changes were no longer observed but age-related changes were evident. EC₅₀ was significantly higher than in young animals and maximal stimulation was significantly lower. Basal adenylate cyclase activity in cortical membranes of all groups of animals was not different. Forskolin-stimulated adenylate cyclase activity was not affected by ethanol treatment, but was higher in aged animals. The activity of forskolin-stimulated adenylate cyclase in the presence of carbachol was higher in both young and aged ethanol-treated animals, when compared to young controls. These results suggest that both ethanol and aging impair the efficiency of receptor/effector coupling.     

Reduction of beta-galactosidase in the ketotic Chinese hamster kidney.

Kidneys from normal, diabetic-nonketotic and ketotic Chinese hamsters were homogenized, fractionated and assayed for beta-glucosidase, and beta-galactosidase activities. The kidneys of the ketotic animals were enlarged but the protein content in each subcellular fraction was similar in all three groups of animals. beta-Glucosidase was found chiefly in the soluble fraction and no difference was observed in these animals. beta-Galactosidase was distributed in both cytoplasmic and particulate fractions; difference in the specific activity of beta-galactosidase between control and ketotic animals was found in nuclear, lysosomal-mitochondrial, microsomal and soluble fractions.     

Acute and chronic dose of alcohol affect the load carrying capacity of long bone in rats

In the present investigation, the effects of acute and chronic dose of alcohol were evaluated on mechanical properties of long bones of Sprague Dawley rats. In "acute study", 18 animals were divided into three groups containing six animals each, i.e. Group A: control animals, normal saline was given to them intraperitoneally for the period of 5 days; Group B: treated animals, given 20% (v/v) absolute alcohol and Group C: treated animals, given 30% (v/v) absolute alcohol, by same route and time duration. In "chronic study", also, 18 animals were divided into three groups containing six animals each, i.e. Group A: control animals, normal saline was given to them intraperitoneally for the period of 6 weeks; Group B: treated animals, given 20% (v/v) absolute alcohol and Group C: treated animals, given 30% (v/v) absolute alcohol by same route and time duration. A significant increase was observed in bone weight of animals taking 20% alcohol but there was decrease in the same for 30% alcohol in case of acute study. For chronic study, there was a decrease in bone weight for both treated groups. During acute study, breaking strength of bone was increased in case of 20% alcohol administration but a slight decrease was shown in the same for 30% alcohol group as compared to control animals. Breaking strength of long bone in the case of chronic study was decreased in case of both groups taking alcohol, i.e. 20% and 30%. The present document is useful in understanding the functional load carrying capacity of bone during alcoholism.