Successional Sequences of Microbial Colonization on Three Species of Rhodophycean Macroalgae

The successional sequences of microbial colonization of Centrocerasclavulatum, Bryocladia cuspidata, and Gelidium crinale wereobserved by SEM. Colonization was initiated by filamentous andsmaller rod or coccoid bacteria, and these microbes were replacedby diatom populations in a successional pattern on Centrocerasand Bryocladia. Gelidium was colonized primarily by bacteria.The spatial fouling patterns on each host plant could be correlatedwith plant shape. Differences in epiphyte attachment morphologiescould be correlated in some species with the host plant or withthe position of the epiphyte on the host plant. Diatoms, epiphytes, fouling, microbial colonization, periphyton     

Phytosterols and Other Lipids as Survival Factors for Tetrahymena at 0–5 C

SYNOPSIS. Tetrahymena pyriformis syngen 1, mating type II, (optimal growth temperature ∼ 37 C) ordinarily dies out in 5-14 days at 0-5 C. Dying cells were lumpy, suggesting membrane damage. By supplying crude soy lecithin, survival at 0-5 C was prolonged (after growth in peptone-yeast-dextrin) to at least 22 weeks. Crude soy sterols or sitosterol or stigmasterol, and antioxidant, e.g., Ionox 330 or ascorbylpalmitate, permitted survival of cells in suspension or in growth media for at least 16-22 weeks. These sterols are known to protect against triparanol toxicity, which suggested that triparanol, which blocks cholesterol synthesis in higher animals, might enhance cold-induced injury. Triparanol was more toxic at 0–5 than at 28 C for cell suspensions and cells in growth medium; this toxicity was annulled by crude soy lecithin or β-sitosterol, the only phytosterol tested. The synthetic medium intended as a control on the crude media became toxic at 0–5 C. Protection against cold damage is discussed as a means of elucidating the role of sterols—especially phytosterols—and other lipids in maintaining the integrity of the ciliate cell membrane.     

Physical and Serologic Properties of an Antigen Prepared from Erythrocytes of Horses with Acute Babesiosis*

SYNOPSIS. By means of precipitation with protamine sulfate, a soluble antigen (PS) was obtained from erythrocytes of horses with acute babesiosis due to Babesia caballi and B. equi. This antigen reacted in gel diffusion tests with sera from horses recovered from acute babesiosis. The PS antigen was found to be muco-protein, susceptible to destruction by trypsin and taka-diastase. Analysis of the antigen by paper electrophoresis revealed 2 components which were not present in similar preparations made from erythrocytes of Babesia-free horses. When the PS antigen was heated in boiling water for 30 minutes, a serologically inactive precipitate was formed; however, the supernate remained serologically active and was termed boiled PS (BPS) antigen. This antigen was polysaccharide in nature; its serologic activity was destroyed by taka-diastase. In gel diffusion tests with sera of recovered horses, the PS antigen formed 2 lines of precipitation which coalesced in a single line formed between these sera and the BPS antigen. Both PS and BPS antigens reacted with sera of horses recovered from acute babesiosis in the gel-diffusion test, but not with sera of dogs and rats recovered from acute infection with Babesia canis and Babesia rodhaini, respectively. The serologic specificity of these antigens suggests that they might have application in the serodiagnosis of inapparent Babesia infections of equine animals.     

Colonization factors of diarrheagenicE. coli and their intestinal receptors

WhileEscherichia coli is common as a commensal organism in the distal ileum and colon, the presence of colonization factors (CF) on pathogenic strains ofE. coli facilitates attachment of the organism to intestinal receptor molecules in a species- and tissue-specific fashion. After the initial adherence, colonization occurs, and the involvement of additional virulence determinants leads to illness. EnterotoxigenicE. coli (ETEC) is the most extensively studied of the five categories ofE. coli that cause diarrheal disease, and has the greatest impact on health worldwide. ETEC can be isolated from domestic animals and humans. The biochemistry, genetics, epidemiology, antigenic characteristics, and cell and receptor binding properties of ETEC have been extensively described. Another major category, enteropathogenicE. coli (EPEC), has virulence mechanisms, primarily effacement and cytoskeletal rearrangement of intestinal brush borders, that are distinct from ETEC. An EPEC CF receptor has been purified and characterized as a sialidated transmembrane glycoprotein complex directly attached to actin, thereby associating CF-binding with host-cell response. Three, additional categories ofE. coli diarrheal disease, their colonization factors and their host cell receptors are discussed. It appears that biofilms exist in the intestine in a manner similar to oral bacterial biofilms, and thatE. coli is part of these biofilms as both commensals and pathogens.Abbreviations CF colonization factor - CFA Colonization Factor Antigen - CS coli-surface-associated antigen - EAggEC enteroaggregativeE. coli - ECDD E. coli diarrheal disease - EHEC enterohemorrhagicE. coli - EIEC enteroinvasiveE. coli - EPEC enteropathogenicE. coli - ETEC enterotoxigenicE. coli - Gal galactose - GalNAc N-acetyl galactosamine - LT heat-labile toxin - NeuAc N-acetyl neuraminic acid - PCF Putative colonization factor - RBC red blood cells - SLT Shiga-like toxin - ST heat-stable toxin     

Molecular drift of the bride of sevenless (boss) gene in Drosophila

DNA sequences were determined for three to five alleles of the bride-of- sevenless (boss) gene in each of four species of Drosophila. The product of boss is a transmembrane receptor for a ligand coded by the sevenless gene that triggers differentiation of the R7 photoreceptor cell in the compound eye. Population parameters affecting the rate and pattern of molecular evolution of boss were estimated from the multinomial configurations of nucleotide polymorphisms of synonymous codons. The time of divergence between D. melanogaster and D. simulans was estimated as approximately 1 Myr, that between D. teissieri and D. yakuba as approximately 0.75 Myr, and that between the two pairs of sibling species as approximately 2 Myr. (The boss genes themselves have estimated divergence times approximately 50% greater than the species divergence times.) The effective size of the species was estimated as approximately 5 x 10(6), and the average mutation rate was estimated as 1-2 x 10(-9)/nucleotide/generation. The ratio of amino acid polymorphisms within species to fixed differences between species suggests that approximately 25% of all possible single-step amino acid replacements in the boss gene product may be selectively neutral or nearly neutral. The data also imply that random genetic drift has been responsible for virtually all of the observed differences in the portion of the boss gene analyzed among the four species.      

Evolutionary origin of human and primate malarias: evidence from the circumsporozoite protein gene

We have analyzed the conserved regions of the gene coding for the circumsporozoite protein (CSP) in 12 species of Plasmodium, the malaria parasite. The closest evolutionary relative of P. falciparum, the agent of malignant human malaria, is P. reichenowi, a chimpanzee parasite. This is consistent with the hypothesis that P. falciparum is an ancient human parasite, associated with humans since the divergence of the hominids from their closest hominoid relatives. Three other human Plasmodium species are each genetically indistinguishable from species parasitic to nonhuman primates; that is, for the DNA sequences included in our analysis, the differences between species are not greater than the differences between strains of the human species. The human P. malariae is indistinguishable from P. brasilianum, and P. vivax is indistinguishable from P. simium; P. brasilianum and P. simium are parasitic to New World monkeys. The human P. vivax-like is indistinguishable from P. simiovale, a parasite of Old World macaques. We conjecture that P. malariae, P. vivax, and P. vivax-like are evolutionarily recent human parasites, the first two at least acquired only within the last several thousand years, and perhaps within the last few hundred years, after the expansion of human populations in South America following the European colonizations. We estimate the rate of evolution of the conserved regions of the CSP gene as 2.46 x 10(-9) per site per year. The divergence between the P. falciparum and P. reichenowi lineages is accordingly dated 8.9 Myr ago. The divergence between the three lineages leading to the human parasites is very ancient, about 100 Myr old between P. malariae and P. vivax (and P. vivax-like) and about 165 Myr old between P. falciparum and the other two.      

Oligomeric forms of the membrane-bound acetylcholine receptor disclosed upon extraction of the M(r) 43,000 nonreceptor peptide

Oligomeric forms of the acetylcholine receptor are directly visualized by electron microscopy in receptor-rich membranes from torpedo marmorata. The receptor structures are quantitatively correlated with the molecular species so far identified only after detergent solubilization, and further related to the polypeptide composition of the membranes and changes thereof. The structural identification is made possibly by the increased fragility of the membranes after extraction of nonreceptor peptides and their subsequent disruption upon drying onto hydrophilic carbon supports. Receptor particles in native membranes depleted of nonreceptor peptides appear as single units of 7-8 nm, and double and multiple aggregates thereof. Particle doublets having a main-axis diameter of 19 +/- 3 nm predominate in these membranes. Linear aggregates of particles similar to those observed in rotary replicas of quick-frozen fresh electrolytes (Heuser, J.E. and S. R. Salpeter. 1979, J. Cell Biol. 82: 150-173) are also present in the alkaline-extracted membranes. Chemical modifications of the thiol groups shift the distribution of structural species. Dithiothreitol reduction, which renders almost exclusively the 9S, monomeric receptor form, results in the observation of the 7-8 nm particle in isolated form. The proportion of doublets increases in membranes alkylated with N-ethylmaleimide. Treatment with 5,5’-dithiobis-(nitrobenzoic acid) increases the proportion of higher oligomeric species, and particle aggregates (n=oligo) predominate. The nonreceptor v-peptide (doublet of M(r) 43,000) appears to play a role in the receptor monomer-polymer equilibria. Receptor protein and v-peptide co-aggregate upon reduction and reoxidation of native membranes. In membranes protected ab initio with N- ethylmaleimide, only the receptor appears to self-aggregate. The v-peptide cannot be extracted from these alkylated membranes, though it is easily released from normal, subsequently alkylated or reduced membranes. A stabilization of the dimeric species by the nonreceptor v-peptide is suggested by these experiments. Monospecific antibodies against the v-peptide are used in conjunction with rhodamine- labeled anti-bodies in an indirect immunoflourescence assay to map the vectorial exposure of the v-peptide. When intact membranes, v-peptide depleted and “holey” native membranes (treated with 0.3 percent saponin) are compared, maximal labeling is obtained with the latter type of membranes, suggesting a predominantly cytoplasmic exposure of the antigenic determinants of the v-peptide in the membrane. The influence of the v-peptide in the thiol-dependent interconversions of the receptor protein and the putative topography of the peptide are analyzed in the light of the present results.     

Efficiency of Selenite Cystine and TT Enrichment Broths for the Detection of Salmonella

Comparisons of selenite cystine (SC) and TT enrichment broths for detecting salmonellas were made with pure culture suspensions, with samples of naturally or artificially contaminated foods and with poultry feed. Selenite cystine recovered higher numbers of salmonellas from pure cultures and ground beef, while TT broth recovered higher numbers from pork sausage and poultry feed. Differences in the recovery of salmonellas from other food products appeared to be insignificant. The use of both SC and TT is thus recommended for maximal recovery of these organisms.