Carbohydrate utilization by Trichomonas gallinae: effects on growth and metabolic activity under nongrowth conditions.

Trichomonas gallinae used 13 of 29 carbohydrates for growth. Quantitative relationships between final populations, acid production, and cellular glycogen contents varied depending on the substrate. The effect of growth on different carbohydrates on the subsequent utilization of carbohydrates by cells under nongrowth conditions was studied by measuring carbohydrate uptake, changes in cellular glycogen content, and gas production. Two major utilization patterns were found. Cells grown on maltose or starch used these substrates well, but cells grown on other sugars did not. All cells used glucose, fructose, galactose, and mannose, but cells grown on maltose or starch did not use them as well as cells grown on other sugars. All cells used ribose slightly but not xylose or arabinose. Turanose, a disaccharide yielding high populations in growth medium, was not used under nongrowth conditions.

Electrophoretic analysis of soluble proteins and esterase, superoxide dismutase and acid phosphatase isoenzymes of members of the protozoan family trichomonadidae.

Polyacrylamide gel electrophoresis was used to compare the proteins and isoenzymes of esterase, superoxide dismutase, and acid phosphatase in soluble, whole-cell extracts of four strains of Trichomonas vaginalis, two strains of Trichomonas gallinae, and one strain each of Tritrichomonas foetus, Tritrichomonas augusta, Tetratrichomonas gallinarum, and Pentatrichomonas hominis. Intraspecific, interspecific, and intergeneric differences were found in protein and isoenzyme profiles. At least four to seven isoenzymes were detected among the ten strains for each of the three enzymes studied. Each strain usually contained one or two isoenzymes of both esterase and acid phosphatase, and two or three isoenzymes of superoxide dismutase.

The gene for the large subunit of ribulose-1,5-bisphosphate carboxylase in Euglena gracilis chloroplast DNA: location, polarity, cloning, and evidence for an intervening sequence.

The gene for the large subunit (LS) of ribulose-1,5,-bisphosphate carboxylase of Euglena gracilis Z chloroplast DNA has been mapped by heterologous hybridization with DNA restriction fragments containing internal sequences from the Zea mays and Chlamydomonas reinhardii LS genes. The Euglena LS gene which has the same polarity as the Euglena rRNA genes has been located with respect to Pst I, Pvu I, and HindIII sites within the Eco RI fragment Eco A. The region of Euglena chloroplast DNA complementary to an 887 bp internal fragment from the Chlamydomonas chloroplast LS gene is interrupted by a 0.5-1.1 kbp non-complementary sequence. This is the first chloroplast protein gene located on the Euglena genome, and the first evidence for an intervening sequence within any chloroplast protein gene.

Treponema innocens lipids and further description of an unusual galactolipid of Treponema hyodysenteriae.

The lipids of Treponema innocens, type strain B256, formerly considered a nonpathogenic isolate of T. hyodysenteriae, have been analyzed and compared with the lipids of T. hyodysenteriae. The lipids of T. innocens comprised 16% of the cell dry weight. Polar lipids amounted to about two-thirds of the total lipids and consisted of 61.9% phospholipids and 38.1% glycolipid. Neutral lipids consisted mainly of sterols. The phospholipids were principally phosphatidylglycerol, phosphatidylcholine, and cardiolipin. Minor amounts of lysophosphatidylcholine, sphingomoyelin, and a relatively nonpolar, unidentified phospholipid were present. The latter lipid has not been detected in T. hyodysenteriae. The glycolipid fraction of T. innocens contained a single component, monoglucosyldiglyceride, in contrast to the occurrence in T. hyodysenteriae of two components: monogallactosyldiglyceride and a less-polar glycolipid tentatively identified as acylmonogalactosyldiglyceride (the additional acyl moieties being 86.6% acetyl, 11.6% propionyl, and 1.6% n-butyryl groups). Alk-1-enyl ether analogs comprised 24.6% of the total phospholipids and glycolipid of T. innocens, or about one-third of the amount in T. hyodysenteriae. The acyl and alk-1-enyl moieties of T. innocens consisted of greater than or equal to 92% of 14:0, iso-15:0, and 16:0 chains. In contrast to T. hyodysenteriae, anteiso-15:0 moieties were not detected, and a reversed distribution of 14:0 and iso-15:0 alk-1-enyl moieties occurred in the two species.

Alk-1-enyl ether phospholipids (plasmalogens) and glycolipids of Treponema hyodysenteriae. Analysis of acyl and alk-1-enyl moieties.

The lipids of Treponema hyodysenteriae B78, the etiologic agent of swine dysentery, comprised 16.4% of the cell dry weight, and consisted of 37.4% glycolipids, 28.6% phospholipids, and 34.0% neutral lipids. Monogalactosyldiacylglycerol, a major lipid in all Treponema except Treponema pallidum, comprised 80% of the glycolipids. An unidentified galactolipid less polar than monogalactosyldiacylglycerol was also detected. Phosphatidylglycerol (19.5% of the total lipids) was the major phospholipid. Phosphatidylcholine, characteristically the major phospholipid of treponemes, comprised 6.1% of the total lipids. Cardiolipin and lysophosphatidylcholine were minor components. The alk-1-enyl ether forms of both the phospholipids (plasmalogens) and glycolipids predominated. The alk-1-enyl ether forms of monogalactosyldiacylglycerol, the unidentified galactolipid, phosphatidylglycerol, cardiolipin, and phosphatidylcholine were 88.3, 96.4, 74.8, 60.6, and 6.3%, respectively. The acyl and alk-1-enyl chains of the organism were qualitatively similar and differed dramatically from those of the medium indicating a capability for fatty acid synthesis that most Treponema do not possess. Saturated C14, C15, and C16 chains comprised more than 95% of the acyl and alk-1-enyl groups. About 25% of the chains were iso-15:0, anteiso-15:0, and other branched moieties.

Unique lipid composition of Treponema pallidum (Nichols virulent strain).

The lipid composition of Treponema pallidum (Nichols virulent strain) was determined after purification of the organisms from the infected testes of corticosteroid-treated rabbits by differential centrifugation, filtration through Nuclepore membranes, and sedimentation in Hypaque density gradients. The total lipids were comprised of 32.2% neutral lipids, mainly cholesterol, and 67.8% phospholipids consisting of phosphatidylcholine (32.1%), sphingomyelin (14.8%), cardiolipin (13.0%), phosphatidylethanolamine (6.2%), phosphatidylinositol-serine (1.2%), and lysophosphatidylcholine (0.4%). Monoglycosyldiglyceride, a glycolipid comprising 25 to 50% of thetotal lipid of all Treponema previously examined, was not detected. The fatty acid composition was similar but quntitatively distinct from that of the infected testes tissue.

Effects of fatty acids on motility retention by Treponema pallidum in vitro.

Treponema pallidum (Nichols virulent strain) was incubated under 75% N2 + 20% H2 + 5% CO2 in prereduced serum-free modified Eagle-Richter medium supplemented with different concentrations of various long-chain fatty acids complexed with fatty acid-free bovine serum albumin. Motility retention was greater in medium with oleic acid containing 15 rather than 2 mg of albumin per ml. Palmitic, stearic, oleic, or linoleic acid alone caused rapid loss of motility at concentrations as low as 5 microgram/ml. Elaidic acid (92 microgram/ml) alone had no effect on motility. Various combinations of saturated plus unsaturated fatty acids did not inhibit motility retention or were less inhibitory than either of the individual fatty acid components. The combination of palmitic plus oleic acids was least toxic. Rapid loss of motility occurred with pairs of unsaturated or saturated fatty acids, or with Tween 40, 60, or 80, alone or combined. Autoxidation of oleic acid resulted in decreased toxicity for T. pallidum but increased toxicity for baby hamster kidney cells.

Unsustained multiplication of treponema pallidum (nichols virulent strain) in vitro in the presence of oxygen.

Treponema pallidum (Nichols virulent strain) was incubated with or without oxygen using a modified medium supplemented with reduced glutathione and a variety of nutrients (PRNF10-B). Two- to fourfold increases in treponemal numbers were observed in cultures without mammalian cells within 96 h of incubation under 5 to 6% oxygen. Treponemal motility and multiplication were maintained more satisfactorily in cultures that were diluted and transferred daily, using an equal volume of fresh medium. Treponemes incubated without oxygen did not significantly increase in number. Virulent microorganisms were detected for at least 96 h in the cell-free system. In the presence of 3 to 4% oxygen, two- to fivefold increases in treponemal numbers were observed in the supernatant fluids of cultures containing human prepuce cells after 48 to 120 h at 35 degrees C. Without oxygen, treponemal numbers rarely approached a threefold increase. Virulent treponemes were detected by the rabbit skin lesion test after at least 120 h in vitro. Regardless of the system of incubation, increases in treponemal numbers could not be sustained for longer than 120 h, and treponemal virulence decreased as a function of time in vitro.

Evidence against gastrointestinal pseudoparasitism by the land planarian, Bipalium kewense Moseley 1878.

The possibility of gastrointestinal pseudoparasitism by the free-living land planarian, Bipalium kewense, was tested by feeding and survival experiments. The intestinal tracts of three dogs were negative for B. kewense after the individual dogs were fed eight, three, and four large worms and autopsied at 12 h, 3 h, and 45 min, respectively. Survival of the worms in Warburg flasks, under N2, AT 27 C, was 2 h or less.. In Gas Pak jars (CO2 + H2) at 37 C, survival was less than 60 min. Aerobically, at 37 C, survival varied from 45 to 60 min. Attra-tion of the worms to stool material was examined by placing planaria inside square whose boundries were constructed of fecal smears. Bipalium kewense exposed to canine feces showed strong avoidance reactions. Urea, a nitrogen end product of these worms, was also shown to be a negative stimulus for B. kewense. Failure to establish even short term passage in the digestive tract, lethality of 37 C and anaerobic environments, and sensitivity to feces makes gastrointestinal pseudoparasitism unlikely in these organisms.

Metabolic changes in Trichomonas gallinae resulting from growth in various carbohydrates.

The influence of the type of growth carbohydrate on the subsequent metabolic activity of Trichomonas gallinae was investigated. Washed suspensions of cells collected from CPL-glucose, CPL-maltose, CPL-galactose, and CPL-glucose-maltose media were examined in the warburg respirometer for their ability to utilize glucose, maltose, and galactose. Comparisons of the metabolic parameters of substrate consumption, changes in glycogen content, and CO2 and H2 production were made. The pattern of utilization of the sugars, both qualitatively and quantitatively, depended upon the type of carbohydrate in the CPL medium used to culture the cells and upon the time of exposure of the cells to a particular sugar in the medium.