A comparison of intrinsic and extrinsic tracer methods for estimating calcium bioavailability to rats from dairy foods.

Dairy products doubly labeled with 45Ca and 47Ca were used to evaluate an extrinsic labeling procedure for calcium bioavailability determination. Nonfat milk, yogurt, and fresh cheese curd were prepared from caprine milk that was intrinsically labeled with 45Ca. The products were then labeled extrinsically with 47Ca and administered to rats by gavage. The 47Ca to 45Ca ratio in bone and teeth averaged about 1.00 with either milk, yogurt, or CaCl2, but the ratio was about 1.04 when dosed with cheese curd. Ca absorption, determined by whole-body counting of 47Ca, was lower (P less than 0.05) in cheese curd (59%) than in either milk (69%), yogurt (72%), or CaCl2 (72%). Expressed as percent of dose, the absorption of 47Ca was highly correlated with bone 47Ca (r = 0.973) and with bone 45Ca (r = 0.946). Correlation between tibia 47Ca and tibia 45Ca was r = 0.923. For the dairy products tested, our results indicated that extrinsic 47Ca was absorbed similarly to intrinsic 45Ca. Moreover, the percent of radioactive dose retained in bone appears to be a valid indicator of relative bioavailability of food Ca.

Endotoxin of Escherichia coli and permeability of the mammary gland of goats.

Serial collections of milk were used to determine where in the mammary gland endotoxin of Escherichia coli was effective in altering the transfer of selected milk components into blood and blood components into milk. Lactating goats had half the gland infused with 1 microgram of endotoxin and the other half served as a control. Sodium-24 and 42K or [14C] lactose were included with 141Ce in the infusate in some experiments, whereas in others 99mTc-labelled albumin or 24Na and 42K were given intravenously 2 h after the endotoxin infusion. Milk was collected 3 h after endotoxin infusion. Endotoxin increased the loss of 24Na, 42K, and [14C] lactose from the mammary gland and increased the transfer of 24Na and 99mTc-albumin into the gland. The transfer in of 42K was reduced compared with control halves. Movement of stable Na and K was in accord with the movement of the 24Na and 42K. Endotoxin was effective in all parts of the gland but particularly from the mid-portion upward to the alveoli. For the control halves there was evidence that some 24Na and 42K crossed the ductal or cisternal epithelium into blood outside of the alveoli, whereas only 42K provided evidence for transfer from blood to milk in these same regions. There was no demonstrable transfer of lactose and albumin in regions other than the alveoli.

Passive transfer of mucosal antibody to Streptococcus equi in the foal.

Passive transfer of mucosal antibody to Streptococcus equi was studied in foals during the first 2 months of life. Immunoglobulin G (IgG) and IgA antibodies were found in sera and nasal secretions of foals shortly after colostrum intake. Titers were highest 2 days after birth; IgG predominated in sera, and IgA predominated in nasal washes. Intragastrically administered 99mTc-labeled IgA was transported from the bloodstream to the nasal mucosa of a newborn foal within a few hours of colostrum intake. Western blot analysis of the specificities of colostral and serum antibodies showed that selective transfer of immunoglobulins of defined specificity did not occur. Antibodies from milk samples taken a month or more into lactation had different specificities than those of colostrum or serum samples. Acid-extracted M protein fragments of S. equi recognized by milk antibodies were the same as those recognized by IgG and IgA from nasopharyngeal mucus of horses recently recovered from strangles. We postulate that passive antibody protection of the foal is derived both by secretion of colostral immunoglobulins onto the nasopharyngeal mucosa and by immunoglobulins ingested in milk that directly coat the upper respiratory and oral mucosa during the first months of life.

In vitro radioisotopic labeling of the L-3 stage of Brugia malayi.

Ten radioisotopes (133Ba, 207Bi, 109Cd, 51Cr, 59Fe, 203Hg, 54Mn, 125Sb and 75Se) were used to label the L-3 stage of Brugia malayi. The in vitro uptake at 4 hours in a buffered salt solution was greatest for 207Bi and exceeded that of 203Hg, by a factor of 6 and the rest of the radionuclides by factors of 26 or more. Dimethylsulfoxide (DMSO) (1%) in the medium only slightly enhanced the uptake of 207Bi but increased 203Hg uptake by a factor of 2.5. After incubation in non-radioactive medium only 2% of the 207Bi was retained; for 203Hg the retention was 70% or better. Increasing the labeling time increased total uptake and retention of the radionuclides. The retention of the 203Hg injected intraperitoneally into jirds (Meriones unguiculatus) in the form of labeled L-3s was followed over a 16 day period by counting the jirds in a whole animal NaI crystal detector. One L-4 stage of B malayi was recovered at the end of this period.

Effects of endotoxin on mammary secretion of lactating goats.

The objectives were to describe the magnitude and time course of changes in milk pH, Na, K, lactose, and somatic cells and to determine if paracellular pathways were altered after infusion of Escherichia coli endotoxin (serotype #0128:AB12) to produce inflammation in one-half of the udder of the goat. Intramammary infusion of endotoxin increased pH, number of somatic cells, and Na and decreased K and lactose in milk. Sodium and number of somatic cells were increased by as little as .1 microgram of endotoxin; .25 microgram produced changes in most of the other parameters; maximal effect was elicited by 1 microgram of endotoxin. The gland response peaked from 5 to 7 h after infusion of endotoxin with an increase in milk cellularity as the only significant effect noted in the control gland. Infusion of [14C]lactose into the gland and [99mTc]albumin into the blood demonstrated that large molecules were more able to cross into and out of udder halves after endotoxin treatment. It is suggested that ion interchange rather than bulk flow across paracellular paths is responsible for changes. In addition, endotoxin appeared to reduce lactose secretion and synthesis.

203Hg and other gamma-emitting radio-isotopes as labels for Dirofilaria immitis microfilariae.

The in vitro uptake of gamma-emitting radionuclides by microfilariae of Dirofilaria immitis was investigated. Radionuclides tested were 133Ba, 207Bi, 82Br, 109Cd, 51Cr, 60Co, 59Fe, 203Hg, 125I, 54Mn, 32P, 125Sb, 46Sc, 75Se and 65Zn. Only 207Bi, 59Fe, 203Hg, 54Mn and 46Sc showed more than 2% of the available radioactivity to bind to the microfilariae. When tested for retention in vitro only 203Hg showed retention levels approaching 90%. Moreover, when dimethyl-sulphoxide was incorporated into the medium at levels of 1% (v/v) the uptake of 203Hg could be increased by 3-5 times; no other radio-isotope tested responded in this manner. The uptake of 203Hg was directly related to temperature and time of incubation. Mercury, as mercuric chloride, was toxic to the microfilariae and represents an impediment to the incorporation of high levels of 203Hg in microfilariae.

External monitoring of 203Hg-labelled Dirofilaria immitis microfilariae in mice and dogs.

Microfilariae of Dirofilaria immitis were labelled with 203Hg2+ in vitro and injected into irradiated mice and Beagle dogs. With irradiated mice it was possible to demonstrate microfilariae present in the blood and to detect 203Hg by external counting as long as 28 days after dosing. The 203Hg2+ label had a half-time of 4-5 days; the amount of stable mercury in the labelling medium strongly influenced the survival of microfilariae in vivo. In dogs, external counting showed the lungs to be a major location of the microfilariae soon after reinjection into the host. Evidence was obtained that labelled microfilariae can circulate; however, the detection of dispersed microfilariae is difficult because of the relative insensitivity of the detecting system. For radiomercury the accumulation of the inorganic form in the liver and kidneys limits the long-term usefulness of 203Hg2+ as a label if the organism being studied also accumulates in these organs.

Isolation, purification and characterization of bovine spleen ferritin.

Ferritin was isolated from bovine spleen and used to prepare apoferritin and reconstituted ferritin. The mol. wt of bovine ferritin was 464,000 with monomer subunits about 18,000-19,500. Gel electrophoresis showed three bands each for ferritin, apoferritin and reconstituted ferritin; all stained for protein and carbohydrate. Only apoferritin failed to stain for iron. Bovine ferritin had higher concentrations of proline, threonine, and valine than equine or human ferritin. The iron:protein ratio of bovine ferritin was 0.161 and of equine ferritin was 0.192. After iron uptake by the apoferritins the iron:protein ratios were 0.186 and 0.278 for the bovine and equine ferritins, respectively.

Isolation of sheep spleen ferritin.

Ferritin was isolated from the spleen of sheep and found to contain an iron/protein content of 21 per cent. Electrophoretic behaviour on 6 per cent polyacrylamide gel revealed three protein bands of Rf values of 5.7, 19.3 and 76.1. Sheep ferritin has a molecular weight of about 475,000 daltons and is high in glutamic acid, leucine and aspartic acid and low in methionine and cysteine. Sheep ferritin is not crystallisable by 10 per cent cadmium sulphate and has an ultraviolet-visible spectrum similar to that of bovine ferritin. Anti (sheep) ferritin antibodies raised in rabbits showed a 57.1 per cent binding at a serum dilution of 1:800 and 30 per cent binding at a 1:12,800 dilution, in a titre determination using double antibody assay procedure.

Effect of low and high ambient temperatures on metabolism of radioiodine by the lactating goat.

Two lactating goats given an oral dose of iodine-125 excreted 16.8, 52.5, and 9.8% of the dose into the milk, urine, and feces when the ambient temperature was 33 C. The thyroids contained about .7% of the oral dose. At 5 C only 2.6% of the radioiodine was in the milk, but 71.2% was in the urine, 18.1% in the feces, and about 10% in the thyroid. The reduced competition by the thyroid plus enhanced transfer of radioiodine from blood to milk at 33 C was responsible for the large transfer of radioiodine into milk at 33 C.