Novel Multi-Slit Large-Volume Air Sampler

Scientific investigators who are interested in the various facets of airborne transmission of disease in research laboratories and hospitals need a simple, continuous, high-volume sampling device that will recover a high percentage of viable microorganisms from the atmosphere. Such a device must sample a large quantity of air. It should effect direct transfer of the air into an all-purpose liquid medium in order to collect bacteria, viruses, rickettsia, and fungi, and it should be easy to use. A simple multi-slit impinger sampler that fulfills these requirements has been developed. It operates at an air-sampling rate of 500 liters/min, has a high collection efficiency, functions at a low pressure drop, and, in contrast to some earlier instruments, does not depend upon electrostatic precipitation at high voltages. When compared to the all-glass impinger, the multi-slit impinger sampler collected microbial aerosols of Serratia marcescens at 82% efficiency, and aerosols of Bacillus subtilis var. niger at 78% efficiency.     

STUDIES AT OYSTER BAY IN JAMAICA,WEST INDIES. IV. OBSERVATIONS ON THE MORPHOLOGY AND ASEXUAL CYCLE OF PYRODINIUM BAHAMENSE PLATE12

The morphology and asexual cycle of the luminescent armoured dinoflagellate, Pyrodinium bahamense Plate, were examined in situ and in vitro. Several previously unreported aspects of the cycle are described. Attempts to induce the formation of these alternate life stages by imposing new conditions were unsuccessful.     

Modification of the zona-free hamster ova bioassay of boar sperm fertility and correlation with in vivo fertility

These studies were designed to evaluate the ability of the zona-free hamster ova bioassay to detect differences in fertility of boar sperm. In the first study, sperm from two previously infertile boars were compared to sperm from seven previously fertile boars. The percentage of zona-free hamster ova penetrated by sperm from the previously infertile boars was significantly lower than the percentage of ova penetrated by sperm from previously fertile boars (18% of ova penetrated vs. 83%, P < .001). In the 14 ejaculates from the previously infertile boars that had ejaculate motilities of 50% or greater, the percentage of zona-free hamster ova penetrated continued to be lower than in ejaculates from the fertile boars. One of the two previously infertile boars consistently had a normal semen analysis. The only two observed manifestations of his reduced fertility were his zero conception rate and the limited ability of his sperm to penetrate zona-free hamster ova. In the second study, females were inseminated with equal numbers of sperm from two previously fertile males and the paternity of offspring determined at birth. The experiment was replicated with four combinations of six boars. A high correlation was observed between the percentage of offspring sired and the ability to penetrate zona-free hamster ova (R = .89). Neither morphology nor the ability of the sperm to undergo an acrosome reaction during in vitro incubation was correlated with fertility in the competitive mating situation. These results suggest the zona-free hamster ova bioassay can improve the in vitro fertility assessment of fresh boar semen.     

Pathogenesis of hemosiderosis in lemurs: Role of dietary iron,tannin, and ascorbic acid

Clinical disease associated with excess iron deposition (hemosiderosis) in the duodenum, liver, and spleen occurs in captive lemurs. In this report we review the occurrence of hemosiderosis and related disease in the Zoological Society of San Diego lemur collection; we then define and describe potential pathogenic factors with the goal of establishing rational husbandry methods to limit or prevent the disease. At the San Diego Zoo, all 49 lemurs necropsied since 1968 were hemosiderotic, the severity increasing with increasing age; liver and kidney disease were common. Our review of iron metabolism, current knowledge on the pathogenesis of hemosiderosis in humans, and the diets of captive and wild lemurs reveals several key dietary substances that may contribute to lemur hemosiderosis: iron, tannins, and ascorbic acid. In captivity, excess dietary iron (commercial monkey chow) and high levels of ascorbic acid (citrus fruits) lead to enhanced iron uptake and increased toxicity of stored iron due to free radical formation. In the wild, lemurs have an unusual preference for leaves, fruits, and bark high in tannin, a polyphenolic secondary plant compound that rapidly chelates iron, protein, and minerals in the gastrointestinal (GI) tract, preventing their absorption. These findings suggest that hemosiderosis in captive lemurs results from a diet high in iron, high in ascorbic acid, and lacking in tannin. Immediate correction of captive diets may limit hemosiderosis in lemurs in the future.     

Ortholinea alata n. sp. (Myxosporea: Ortholineidae) in the northern butterfly fish Chaetodon rainfordi

Ortholinea alata n. sp. is described from the northern butterfly fish, Chaetodon rainfordi collected at Heron Island, Great Barrier Reef, Australia. Spherical, disporous trophozoites (10-15 micrometers) and spores were observed in the lumina of kidney tubules and collecting ducts. Spores are broadly triangular with two short, broad processes that extend dorsoventrad from the posterior end of each of the two spore valves. Valves are bisected by a suture in the plane of the polar capsules. Spores are 12.6 micrometers (length) x 9.6 micrometers (width) x 9.9 micrometers (length), and at the anterior end contain two spherical, divergent polar capsules measuring 4.6 (4.1-5.1) micrometers. Sporogenesis is similar to that of renal Sphaerospora spp.; the intraluminal trophozoites of O. alata n. sp. correspond to pseudoplasmodia described for Sphaerospora spp. and no large, multinucleated plasmodia are formed. No significant histopathological changes were observed in the kidneys of infected fish.     

Multiple T and B cell epitopes in the S1 subunit ("A"-monomer) of the pertussis toxin molecule

The immunogenicity and reactogenicity of Bordetella pertussis vaccine are mediated in part by the S1 subunit of pertussis toxin (PT). To identify the immune epitopes in the S1 subunit of PT, synthetic peptides were prepared and tested for their capacity to induce antibodies in mice with different MHC genotypes. In BALB/c mice, peptides corresponding to sequences 1-17, 70-82 and 189-199 generate T cell proliferative responses, induce the production of antibodies capable of neutralization of the toxin in the Chinese hamster ovary-cell assay, and protect mice from a shock-like syndrome caused by alternate injections of BSA and PT. Protection and neutralization correlated with the ability of these peptides to elicit high anti-PT titers. Different B cell epitopes were detected in other inbred mouse strains. The antibody reactivity against synthetic peptides from two infants vaccinated with pertussis vaccine was tested. These infants had antibodies reactive to a variety of epitopes in the S1 subunit, including peptides 1-17, 70-82, 99-112, 135-145, and 189-199. Thus, it appears that there are multiple T and B cell epitopes in the S1 subunit of PT.     

Comparison of [125I]Iodolysergic Acid Diethylamide Binding in Human Frontal Cortex and Platelet Tissue

The human platelet contains a functional 5-hydroxytryptamine (5-HT) receptor that appears to resemble the 5-HT2 subtype. In this study, we have used the iodinated derivative [125I]iodolysergic acid diethylamide ([125I]iodoLSD) in an attempt to label 5-HT receptors in human platelet and frontal cortex membranes under identical assay conditions to compare the sites labelled in these two tissues. In human frontal cortex, [125I]iodoLSD labelled a single high-affinity site (KD = 0.35 +/- 0.02 nM). Displacement of specific [125I]iodoLSD binding indicated a typical 5-HT2 receptor inhibition profile, which demonstrated a significant linear correlation (r = 0.97, p less than 0.001, n = 17) with that observed using [3H]ketanserin. However, [125I]iodoLSD (Bmax = 136 +/- 7 fmol/mg of protein) labelled significantly fewer sites than [3H]ketanserin (Bmax = 258 +/- 19 fmol/mg of protein) (p less than 0.001, n = 6). In human platelet membranes, [125I]iodoLSD labelled a single site with affinity (KD = 0.37 +/- 0.03 nM) similar to that in frontal cortex. The inhibition profile in the platelet showed significant correlation with that in frontal cortex (r = 0.96, p less than 0.001, n = 16). We conclude that the site labelled by [125I]iodoLSD in human platelet membranes is biochemically similar to that in frontal cortex and most closely resembles the 5-HT2 receptor subtype, although the discrepancy in binding capacities of [125I]iodoLSD and [3H]ketanserin raises a question about the absolute nature of this receptor.     

Ineffectiveness of ethylene biosynthetic and action inhibitors in phenotypically reverting theEpinastic mutant of Tomato (Lycopersicon esculentum mill.)

Five-day-old, dark-grown seedlings of theEpinastic (Epi) tomato mutant (Lycopersicon esculentum Mill.) and its parent, cultivar VFN8, were used as a system for assessing the role of ethylene in theEpi phenotype. The distinguishing features ofEpi seedlings are an increase in hypocotyl diameter and reduced hypocotyl length. Treatment of VFN8 seedlings with 0.5 l/liter ethylene closely mimicked theEpi phenotype. The rate of ethylene production by 5-day-old, dark-grownEpi seedlings was double that of VFN8 seedlings. Nevertheless, treatment ofEpi seedlings with inhibitors of ethylene biosynthesis (aminoethoxyvinylglycine or Co²⁺) or ethylene action (silver thiosulfate or norbornadiene) failed to normalize theEpi phenotype.Epi seedlings grown in sealed jars containing ethylene and CO₂ adsorbants also expressed the characteristicEpi phenotype. The results indicate that the physiological lesion resulting from theEpi gene mutation is not simply an overproduction of ethylene.