James O. Norman

Biological Control of Salmonella typhimurium in Young Chickens

The effect of dietary lactose and anaerobic cultures of cecal microflora of mature chickens on the colonization of young broiler chickens by Salmonella typhimurium was evaluated. Newly hatched chicks were given either no treatment (controls), anaerobic cecal cultures, lactose (2.5%) in the drinking water, or both anaerobic cultures and lactose. Chicks were challenged per os at 3 days of age with either 10(6) or 10(8) S. typhimurium resistant to nalidixic acid and novobiocin. On day 10, the cecal contents of the chicks were examined for S. typhimurium, pH, short-chained volatile fatty acids (VFAs), undissociated VFAs, and lactic acid. Chicks given either lactose alone or cecal anaerobes alone had significantly (P less than 0.05) fewer S. typhimurium recovered from their ceca than the controls. Chicks given the combination of dietary lactose and cecal anaerobes had significantly fewer S. typhimurium recovered from their ceca than the chicks given dietary lactose or cecal anaerobes alone. Chicks given lactose had significant (P less than 0.05) increases in the lactic acid concentration of their cecal contents. Increased lactic acid concentrations were directly correlated to decreased cecal pH values and caused a reduction in the total concentration of VFAs but a significant (P less than 0.05) increase in the undissociated form of some VFAs.

Cytotoxic effect of T-2 mycotoxin on cells in culture as determined by a rapid colorimetric bioassay

We developed a colorimetric assay for determining metabolic activity (viability) of cells exposed to toxic agents. This system is based on the ability of mitochondrial enzymes in viable cells to modify a tetrazolium salt into a blue formazan product that can be detected spectrophotometrically at 570 nm. The assay works equally well for mammalian and insect cell lines and at 48 hr color formation is linear over a cell input range of 1.56-50 X 10(4) cells/ml. The inhibitory effects of T-2 mycotoxin on tetrazolium cleavage in L929 cells is comparable to that observed for protein and DNA synthesis (50% inhibition = 6-8 ng/ml). Using this system to analyze the lethal effect of T-2 toxin on cells from various animal species, it was found that bovine cells were the most sensitive (50% inhibition at 2.2 ng/ml) while hamster cells were the most resistant (50% inhibition at 26.2 ng/ml). Murine cells exhibited intermediate sensitivity (50% inhibition at 10.9 ng/ml). Variable toxin susceptibility was also observed among different cell types. Lymphocytes were 3-fold more sensitive to the T-2 inhibitory effects than comparable tissue culture cell lines. These data indicate that the colorimetric assay system could have broad applications in toxicological studies. Further, the observed differences in species sensitivity may provide insight into the primary mechanism of the T-2 toxin-cell interaction that ultimately leads to cell death.

Polychlorinated biphenyl uptake and transport by lymph and plasma components.

Abstract The uptake and vascular transport of ingested Aroclor 1242, an isomeric mixture of polychlorinated biphenyls (PCB), was investigated in experimental animals. High concentrations of ingested PCB were found in the chylomicron fraction of thoracic duct lymph. When the lymph flow was exteriorized PCB were not subsequently found in the vascular circulation. When lymph was not exteriorized plasma PCB concentrations reached maximal levels 6 hr after ingestion. Less than 1% of total plasma PCB was detected in cellular fractions of blood over a 10-hr period following ingestion. Chylomicrons contained 31% of total plasma PCB 30 min after ingestion, decreasing to less than 6% at 4 hr. A maximum of 10% of plasma PCB at 1 hr, and <5% at 6 hr, after ingestion was associated with very low density lipoproteins (VLDL) or low density lipoproteins (LDL). Although PCB enter the vascular circulation with the chylomicron fractions of lymph, delipoproteinated plasma contained 52% of the total PCB in blood collected 30 min after ingestion. This level increased to 78% after 2 hr, and remained constant at about 80% for an additional 8-hr period. High performance liquid chromatographic (HPLC) examinations of delipoproteinated plasma from blood taken 6 hr after PCB ingestion showed elution of >95% of plasma PCB to coincide with the albumin peak. Electrophoretic examinations of delipoproteinated plasma showed the association of PCB with albumin to be noncovalent. The results suggest that apolar PCB are absorbed into intestinal epithelial cells from which they are secreted into the lymphatic drainage sequestered within the apolar core of chylomicrons, that these PCB transit the thoracic duct and enter the vascular circulation within chylomicrons and are metabolized or otherwise released from chylomicrons during hepatic chylomicron clearance, and that resulting PCB or PCB derivatives circulate in association with plasma albumins.

Reduction of Salmonella typhimurium concentration in broiler chickens by milk or whey.

Whey (5%) in the feed of chicks for the first 10 days of life reduced the mean log10 number of viable S. typhimurium from 5.68 in control chickens to 3.38 in whey-fed chickens. Lactose in drinking water or reconstituted dry milk (5% wt: vol) in drinking water reduced the mean log10 number of S. typhimurium to 2.60 and 2.11, respectively. Milk (5% wt: wt) in feed was not effective in reducing S. typhimurium colonization. The lack of effect of milk in the feed is believed to be because not enough lactose was provided at the 5% (wt: wt) concentration. Lactose in whey or nonfat dried milk offers alternatives to the use of pure lactose in preventing or lowering S. typhimurium numbers in young broiler chickens.

Interaction of phosphatidylinositol-4-monophosphate with a low activity form of dna polymerase alpha: a potential mechanism for enzyme activation

1. DNA polymerase alpha isolated from Norman murine myxosarcoma exhibited two isozyme forms, one with low specific activity and low DNA binding affinity (A1), and one with high specific activity and high DNA binding affinity (A2). 2. DNA polymerase alpha A1, but not A2, showed a significant increase in specific activity after treatment with phosphatidylinositol, ATP and phosphatidylinositol kinase, or with phosphatidylinositol-4-monophosphate. 3. Treatment of DNA polymerase alpha A1 with the phospholipase C hydrolysis product of phosphatidylinositol-4-monophosphate, inositol-1,4-bisphosphate, was sufficient to effect the transient increase in activity of polymerase A1 to a form not chromatographically distinguishable from isozyme form A2.

Effects of aflatoxin B, on distribution of Fe, Cu, Zn, and Mn in rat tissues

The role of aflatoxin B/sub 1/ in trace element metabolism in animals is investigated. Two experiments were conducted using atomic absorption for determining the distribution of Fe, Cu, Zn, and Mn in liver, kidney, spleen, and pancreas. The results indicated body weight loss during aflatoxicosis is dose related and may be due in part to reduced body fat. It remains to be determined whether the redistribution of the metals, especially the massive influx of Fe into the liver, is the actual toxic factor in aflatoxicosis that leads to sickness and death. The preliminary results reported indicate that the effect is a complex problem involving not only a redistribution of the metals but possibly some hormonal action as well. (CAJ)

Benzo[a]pyrene Uptake by Serum Lipids: Correlation with Triglyceride Concentration

Abstract An in vitro study of the relationship between benzo[a]pyrene (BaP) association with serum lipoproteins (LP) and LP composition was conducted using human subjects. BaP partitioning into different serum LP ranged from 53 to 71% of available BaP. Efficiency of BaP partitioning was examined for the relationship with lipid components of different sera. The data indicate that triglyceride (TG) concentrations were more directly correlated with BaP uptake than were concentrations of other LP components. Adjusting sera to a uniform TG concentration (96.5 mg/dl) resulted in the same BaP uptake for each serum type, while adjusting sera to contain a uniform cholesteryl ester concentration (104.6 mg/dl) did not result in similar BaP uptake among serum types. Analysis of serum LP composition suggested that marked differences in both BaP uptake and serum TG concentrations among the subjects were due mainly to differences in serum very low density lipoprotein (VLDL) concentrations. A correlation study using 14 human subjects showed that serum TG concentration was the best predictor (r = 0.973, P < 0.001) for BaP uptake by serum, followed by phospholipid (r = 0.658, P < 0.01) and total cholesterol (r = 0.514, P < 0.05) concentrations. The results indicate that serum TG concentration (typically VLDL-TG) may be the primary factor affecting BaP uptake by serum LP, and suggest that a small change in serum TG concentration could cause a significant increase in BaP uptake by serum LP, contributing to an increased level of circulating carcinogen.

Detection of Phototoxic Activity in Plant Specimens Associated with Primary Photosensitization in Livestock using a Simple Microbiological Test

Ingestion of mushrooms is infrequently observed by owners and may be omitted from the initial history. When mushroom poisoning is suspected on the basis of histopathological lesions, additional history and tests may help to establish the diagnosis. Collection and identification of mushrooms from the animal’s environment will establish the types of mushrooms that the dog was potentially exposed to. Examination of vomitus, gastrointestinal contents, and/or feces for mushroom spores may provide definitive evidence of mushroom ingestion and possible identification of the genus and species. In human patients, early diagnosis is occasionally achieved by detection of amanitins in urine or other body fluids.” Due to delayed onset of clinical signs, patients usually receive only supportive and palliative treatment. The treatment for Amanita mushroom poisoning has recently been reviewed.” Unless treatment is initiated promptly after exposure and before significant amounts of toxin are absorbed, the final outcome of the case is dependent on the amount and type of mushroom ingested.

Purification of norman murine sarcoma DNA polymerase alpha forms with different DNA template primer binding affinity and different specific activity

1. DNA polymerase alpha was isolated from Norman Murine Myxosarcoma cells using ion exchange, immunoaffinity, and DNA affinity chromatography, showing two distinct enzyme forms designated A1 and A2. 2. Chromatographic analysis of polymerase alpha forms A1 and A2 indicate a charge difference and a difference in affinity of binding to DNA between polymerase alpha forms which were equally reactive to anti-DNA polymerase alpha monoclonal IgG. 3. Polymerase A1 specific activity was about 3600 U/mg while A2 specific activity was about 40,000 U/mg.

Effects of Sesquiterpene Lactones on the Growth of Bacillus Thuringiensis

Two sesquiterpene lactones, hymenovin and tenulin, were tested for their effect on growth of two strains of Bacillus thuringiensis. Growth of both strains was 98% inhibited by 0.6 mg of tenulin per ml, but only 15 to 20% was inhibited by the same level of hymenovin. Hymenovin appeared to have a mutagenic effect on B. thuringiensis cultures resulting in production of several variant strains. Some of the variant strains had lost their ability to form spores and crystals. Hymenovin also induced B. thuringiensis to produce significant levels of bacteriophage. All variant strains were as susceptible to phage as the parent strain. Although the bactericidal activity of tenulin was two times as great as of hymenovin, tenulin did not appear to have a mutagenic effect on the bacteria. Images