Sex specificity of myocardial damage in mice fed a purified diet

Both sexes of BALB/c and B6C3F1 mice were divided into test groups and fed either a purified diet (AIN-76A) or a natural ingredient diet (NIH-07) containing graded levels of 2-acetylaminofluorine (2-AAF) for 90 days. A large number of dead or moribund B6C3F1 males fed the AIN diet were removed from the study prematurely. AIN-fed B6C3F1 mice removed early as well as some sacrificed at the end of the study showed myocardial damage with hemorrhage. A much smaller number of BALB/c males fed the AIN diet also exhibited these signs while none of the females from either stock were affected. Mice having these lesions were confined largely to 2 of 5 treatment groups. Increased levels of serum aspartate aminotransferase (GOT) (P less than .01) occurred in the AIN-fed B6C3F1 male mice that were sacrificed, supporting the histopathological observation of myocardial damage. There was no other significant difference in the GOT between diets or 2-AAF doses. No environmental factors could be associated with the problem and microbiological and chemical analyses of the diets showed no convincing evidence of specific pathogenic organisms or nutritional deficiencies that might have caused these lesions. Extended storage (up to 4 months) and one batch of feed in particular seemed to be associated with mice having myocardial damage. These associations were highly strain and sex dependent and suggest that great care must be taken in the manufacture and handling of the diet. Furthermore, it seems likely that the diet may be marginally adequate for some strains of mice and may require modification before it will become generally useful.     

The utilization of nitrogen for growth in mice fed blends of purified proteins

Reasons for differences in growth of weanling mice fed different blends of purified proteins in diets apparently adequate in indispensable amino acids were investigated. Dietary nitrogen provided at increasing levels from a mixture of casein, gelatin, and zein, or from these plus gliadin and lactalbumin resulted in similar weight gains and efficiency of feed utilization in weanling mice. These responses were always lower (P less than 0.05) than for mice fed the control diet. When diets were based on true digestible nitrogen and amino acids, weight gains and feed efficiency decreased (P less than 0.05) as the dietary nitrogen level increased. High plasma glycine concentrations and high phenylalanine to tyrosine ratios were associated with high dietary glycine levels provided by gelatin. When a mixture of constant proportions of protein sources, predominantly casein, supplied most of the dietary nitrogen, small increases in weight gains and feed efficiency were observed as the nitrogen level in the diet increased. Varying the proportions of sources, which increased the percentage of gelatin, resulted in decreased gains in response to more dietary nitrogen. Replacing the amino acids found in gelatin by crystalline L-amino acids and replacing glycine by glutamic acid improved gains and feed efficiency (P less than 0.05) compared with gelatin-containing diets. These studies demonstrated that glycine at dietary concentrations of approximately 1-2%, either from gelatin or as the free amino acid, inhibited growth and feed utilization of growing mice.     

Heat stress aggravates viral myocarditis in mice

While a beneficial effect of hyperthermia on viral infection has been hypothesized, there are no data on viral myocarditis in vivo. To investigate whether hyperthermia might attenuate the course or severity of viral myocarditis, we studied the pathological changes in a murine model of viral myocarditis. C3H mice were inoculated i.p. with the encephalomyocarditis virus (500 pfu). They were anesthetized and heated to a body temperature of 42.5+/-0.2 degrees C for 30 min. The latter was performed 4 hr before (n=28, HB) or 4 hr after (n=28, HA) the viral inoculation; results were compared with nonheated, infected controls (n=30, Cont). Cardiac viral titers were recorded on day 3, and the body weight (BW), heart weight (HW) and pathological changes were recorded on days 5 and 10. The incidence of spontaneous mortality on day 10 was significantly higher in the HA group (all deaths occurring by day 7 post-inoculation) as compared with the HB (35%) or Cont (18%) groups. Viral titers in the HA group (n=4) were significantly (P<0.05) higher than those in the Cont (n=7) or HB (n=7) groups (4.11+/-0.54 vs 3.01+/-0.44 and 3.23+/-0.45 LogTCID50/mg, respectively). On day 5, the HW, the BW/HW ratio, and the severity of myocardial necrosis were all significantly higher in the HA than in the Cont and HB groups. To confirm the effect of hyperthermia on the expression of heart shock protein (HSP), immunohistochemical staining was done in the virus-infected hearts. The nucleus and cytoplasm of the injured myocardium in the HA group strongly expressed HSP70, whereas the HB and Cont groups were negative for this protein. In conclusion, induction of hyperthermia after viral inoculation aggravated the viral-induced myocardial necrosis and increased the mortality rate in a murine model of viral myocarditis and induced myocardial heat shock protein 70.     

Iron status in rats fed a purified diet without vitamin A

The effect of vitamin A deficiency on iron status was investigated in rats. After 28 d of feeding either low or high vitamin A diets (0 vs 4000 IU of vitamin A per kg feed), the final body weight was slightly but significantly lowered by the low vitamin A diet. Plasma retinol concentrations were decreased in rats fed diets low in vitamin A. Marginal vitamin A deficiency produced slightly, but significantly lower blood hemoglobin concentrations; it did not clearly affect hematocrit. The concentration of iron in liver was significantly higher when diets low in vitamin A were fed while significantly lower levels were observed in femur.     

Cardiac myosin induces myocarditis in genetically predisposed mice

After infection with coxsackie virus B3 (CB3), H-2 congenic mice on an A- background develop immunologically mediated myocarditis associated with an increased titer of myosin autoantibody, part of which is specific for the cardiac myosin isoform. The present study demonstrates that cardiac myosin itself induces severe myocarditis and high titers of myosin autoantibodies in A/J, A.SW/SnJ, and A.CA/SnJ mice. As in CB3-induced myocarditis, one population of these autoantibodies was specific for cardiac myosin. A.BY/SnJ and B10.A/SgSnJ mice also developed the disease after immunization, but the prevalence and the myosin autoantibody titers were lower. In contrast, C57BL/6J and C57BL/10J mice were resistant to myocarditis induced by cardiac myosin and did not develop increased myosin autoantibodies or cardiac myosin-specific autoantibodies. Immunization with skeletal muscle myosin had no effect compared with controls injected with complete Freund's adjuvant, thereby suggesting that the immunogenic epitopes are unique to the cardiac myosin isoform. Furthermore, we found that susceptibility to myocarditis induced by cardiac myosin is influenced by the major histocompatibility complex and by genes not closely linked to the major histocompatibility complex. Because there are parallels between myocarditis induced by cardiac myosin and that induced by CB3, this new animal model can be used to analyze the pathologic mechanisms in autoimmune heart disease.     

Brain cholecystokinin in fasted and fed mice

Immunoreactive cholecystokinin (iCCK) in brain extracts is reduced to 133 ± 6 ng/g from 274 ± ng/g in 2 to 5 day fasted C₃H mice. Brain weights of fed and fasted mice do not differ although the body weights of fasted mice averaged only about two-thirds of the fed weights. Since the cortex contains over 80% of the brain iCCK in fed mice, it is concluded that food-deprivation is associated with decreased synthesis of cortical iCCK.     

Reduction of chlortetracycline-resistant Escherichia coli in weaned piglets fed fermented liquid feed

We investigated the change in chlortetracycline resistance in 360 Escherichia coli strains separated from the feces of piglets fed fermented liquid feed (FLF) in comparison with those fed dry feed (control). The total amount of lactic acid bacteria in feces was 8.77x10(8)CFU/g DM at weaning, which increased to 1.23x10(12)CFU/g DM (FLF) at 28 days after weaning (P<0.001). As a result of the antibiotic susceptibility test, almost all isolates were chlortetracycline-resistant (CTC(R)) until 14 days after weaning both in the FLF and control groups. At 28 days, the CTC(R)E. coli decreased to 22.2% in the FLF group, while the proportion of resistant bacteria was 88.9% in the control group. In addition, as a result of the gene analysis, it was clarified that there is a relation between the decrease in the minimum inhibitory concentration values and the possession rate of the tetracycline-resistance gene tet(A), tet(B) and tet(D). These results show that FLF caused an increase in the number of lactic acid bacteria in the intestines, and suggested that the feeding of FLF can possibly reduce antibiotic-resistance bacteria.     

Experimental reovirus myocarditis in newborn mice. Electron microscopic observations

Reovirus is a double-stranded RNA-virus which induces myocarditis in newborn mice. Due to the large diameter of the viral particles (70-75 nm) it can be detected by electron microscopy. Subcutaneous inoculation of 0.05 ml reovirus type 3 (TCID50-titer: 10(8.5)/ml) into newborn NMRI-mice (12-18 h after birth) caused a grey-yellow mottling on the ventricular surface first seen on the 5th day after birth. At the same time muscle fiber necrosis was observed which increased with time. Electron microscopic investigations of the diseased heart muscle disclosed a marked interstitial oedema, swelling of the tubular system and sarcoplasmic reticulum, and degenerative changes in the mitochondria of individual myocardiocytes as early as the 2nd post-inoculation day. Simultaneously, an enlarged Golgi-apparatus and an increasing number of lysosomes, partially exhibiting acid phosphatase activity, was detected in the perinuclear region of ventricular myocardiocytes. On the 5th day after infection, viruses were detected either within single membrane vesicles, dispersed in cytoplasm or as aggregated clusters in the perinuclear region. These in vivo electron microscopic findings correspond with observations of virus propagation in cell-culture systems.     

Lactation in mice fed endophyte-infected tall fescue seed

This study was conducted to assess the effects of endophyte-infected Acremonium coenophialum tall fescue (KY-31) seed (80% infected) on lactation in CD-1 dams and suckling performance of pups as measured by pup survival and growth rates. Twenty-four pairs of mature CD-1 mice were randomly allocated to four dietary treatments: 1) 100% mouse chow ad libitum; 2) 40% endophyte-infected tall fescue seed and 60% mouse chow (w/w); 3) reduced intake (100% chow), adjusted daily to the intake level of Treatment 2; and 4) 60% infected tall fescue seed and 40% chow. The mice were preconditioned on their respective diets for 100 d prior to 96 h of cohabitation between pairs of males and females. At parturition the litters were removed, and each dam was given a litter of six pups of equal weight, size and sex ratio to suckle for 15 days. All pups given to all the dams were born to other mice that were not part of the study and had not been exposed to endophyte-containing diets. Dams and litter weights were measured daily for 15 consecutive days. The combined body weight measurements of litters from dams fed the tall fescue containing diets (Treatments 2 and 4) were significantly lower (2.07 +/- 0.41 g/d) than that of litters from dams fed the chow containing diets (Treatments 1 and 3) during the suckling trial (P<0.05). Similarly, nine of ten (90%) dams fed the chow containing diets maintained five or more pups (5.5 +/- 0.2) throughout the study as compared to five of nine (55.6%) dams fed the tall fescue containing diets that maintained less than five pups (4.5 +/- 0.2).