Corticotypic Technics in Tetrahymena Taxonomy*

SYNOPSIS. Strains of Tetrahymena pyriformis have a cryptic polymorphism reflected in the production of individuals with different numbers of ciliary rows (meridians). Within a strain certain cortical features—particularly the numbers and positions of contractile vacuole pores, the numbers of postoral meridians, and the stability of the cell type-vary in a systematic way with meridianal multiplicity. A preliminary survey of strains representing 10 genetically isolated groups reveals several patterns of correlation of cortical elements. All the groups examined appear to be distinctive with the possible exception of the strains from syngens 6 and 8, which are known to be genetically compatible under certain conditions. These studies encourage a belief that the heterogeneous assembly of genetic species now sheltered under the taxonomic umbrella of T. pyriformis can soon be given independent morphologic identification and appropriate specific designations.     

Range of Variation in the Somatic Infraciliature and Contractile Vacuole Pores of Tetrahymena pyriformis

SYNOPSIS. Meridian ranges and contractile vacuole pore patterns were determined for 36 strains of Tetrahymena pyriformis, including most of the “classic” amicronucleate and micronucleate strains used by many investigators, plus a dozen other strains not previously characterized with respect to these two morphological features. Uniform axenic culturing and staining techniques were employed. Meridian ranges and modes are analyzed with strain-by-strain comparisons to the reports of previous investigators. The overall inclusive range of the 36 strains was 15–25 meridians. Modes for individual strains varied from 16 to 23. Broader ranges and, especially, higher modes and higher limits are recorded than those reported by Corliss (1953) for strains grown in bacterized hay cultures. Two postoral meridians are characteristic, although a single individual having only one and a few having three postorals were observed. Three postorals were found only in those strains having the higher number of total meridians. A few individuals, showing an anterior half or a posterior half of a bipolar meridian missing, indicate a possible explanation of how a single dividing individual could produce a proter and an opisthe differing in the total number of meridians. Meridian ranges of individual strains do not overlap each other in all cases, but they do constitute a progressively overlapping series which does not warrant taxonomic separation of strains at either extreme from each other. Contractile vacuole pores varied from 1 to 3, with 2 pores (1 each in meridians 5 and 6) being characteristic of 22 of the strains. The 2-pore 6-7 pattern was modal for 8 of the strains. Presence of 3 pores (in 5, 6 and 7) was typical of only 2 strains, but 3 pores were found occasionally. Another pattern of 3 pores (in 6, 7 and 8) was observed occasionally in 10 strains. Double pores (in 6 and 7 or 7 and 8) and the triple pore patterns were observed only in the strains having the highest total number of meridians. These variations are not considered adequate justification for taxonomic separation. Strains Ki and Aq are affirmed to be T. pyriformis. Strains V¹ PP and V, formerly regarded as strains of T. vorax, are considered to be T. pyriformis, supporting the view of Shaw and Williams (1963).     

Electrophoretic Isoenzyme Patterns of Entamoeba histolytica and Entamoeba chattoni in a Primate Survey1

Stocks of Entamoeba histolytica grown in a monoxenic culture system from the feces of nonhuman primates are compared with the eleven zymodemes of E. histolytica so far demonstrated from man. In a similar fashion, Entamoeba chattoni has also been grown and identified. Both E. histolytica and E. chattoni have been demonstrated in keepers of the primate collections. Comparisons have been made using the electrophoretic patterns of three enzymes: glucosephosphate isomerase [(GPI) E.C.5.3.1.9], phosphoglucomutase [(PGM) E.C.2.7.5.1], and L-malate—NADP⁺ oxidoreductase (oxaloacetate-decarboxylating) [(ME) E.C. 1.1.1.40]. Enzyme patterns of E. histolytica from the apes were found to be identical with three of those already demonstrated from man. The enzyme pattern of E. chattoni was distinctly different from that of any of the E. histolytica zymodemes. Other protozoa found in the single fecal sample examined from each subject are also listed.     

Patterns of Cortical Stability in Tetrahymena*

SYNOPSIS. A simplified technic is presented for determining clonal stability patterns for numbers of ciliary rows in Tetrahymena. The technic involves plotting the variance of meridian number against the mean number of meridians. By this procedure strains of different syngens, difficult to discriminate on other grounds, may sometimes be shown to be distinctive.     

Terebrospira chattoni sp. n., a Parasite of the Endocuticle of the Shrimp Palaemonetes pugio Holthuis*

SYNOPSIS. Terebrospira chattoni sp. n. may be a species in transition between ectosymbiosis and endosymbiosis. It penetrates the epicuticle of Palaemonetes pugio and feeds on the endocuticle by dissolving galleries through it and absorbing the products of dissolution. Although similar in some respects to species of the endosymbiotic apostome genus Synophrya, T. chattoni is clearly related to ectosymbiotic apostomes that feed on exuvial fluid. However, instead of stimulating the metamorphosis of a phoront to a trophont, the host's ecdysis stimulates T. chattoni to metamorphose from a dedifferentiated trophont with 13 meridional kineties to a protomont, a predivision stage with 10 spiralled kineties. The protomont encysts and dedifferentiates to the division stage, an orthotomont with 13 kineties, either on the new exoskeleton before ecdysis, within a chamber in the endocuticle of the old exoskeleton, or on a substrate away from the host and the molt. The site of division determines the product of the division. On the new exoskeleton, the tomites in the reproductive cyst secrete walls around themselves thereby forming the lenticular, compartmented cysts characteristic of Terebrospira. Each daughter tunnels out of its compartment into the endocuticle. Although its infraciliature remains undifferentiated like that of the orthotomont, the ciliate in the gallery is the trophont, the only feeding stage in the life cycle. Daughters originating from the division of the orthotomont in a chamber in the endocuticle swim out of the exoskeleton at ecdysis. encyst on the substrate, and presumably form tomites. When the protomont itself leaves the molt, it encysts on the substrate and divides to form daughter cells with a rosette and the pattern of ciliature of the conventional tomite of other apostome genera. Tomites carry the infection to new hosts while compartmented cysts insure that the original host retains the infection. Terebrospira chattoni is always astomatous, although a dedifferentiated oral ciliature appears in the orthotomont and persists in the trophont. Terebrospira chattoni sp. n. is separated from T. lenticularis Debaisieux the other species in the genus because the latter species may divide outside a cyst and because T. chattoni has an extra reproductive stage in its life cycle.     

The Fine Structure of the Cytostome of the Apostomatous Ciliate Hyalophysa chattoni*

SYNOPSIS. The fine structure of the organelles concerned with the ingestion of exuvial fluid by the trophont of the apostome ciliate, Hyalophysa chattoni, has been examined. One of the taxonomic characteristics of the order Apostomatida is that cytostomes of ciliates within the taxon are reduced and evolving toward astomy. When examined by electron microscopy the cytostome of H. chattoni appears as a small region of active pinocytosis which is continuous with a very large cortical area, the extended cytostome. The fine structure of the extended cytostome resembles that of the cytostomes of ciliates from other orders in that it is covered by a single membrane underlain with microtubular ribs. Beneath the extended cytostome are accumulations of peculiar organelles that may represent stored membrane for recycling during food vacuole formation. Associated with the site of pinocytosis is a complex fiber that may be contractile.     

Comparative Corticotype Analyses in Tetrahymena*

SYNOPSIS. Strains of Tetrahymena pyriformis, including amicronucleate strain GL and representatives of 9 syngens, have been examined to determine the patterns whereby cortical features vary with numbers of ciliary meridians. The characteristics scored were the positions of the contractile vacuole pores (CVP's), the extent of the area within which CVP's develop, the incidence of supernumerary CVP's, and the number of postoral meridians. Intrasyngenic comparisons were possible in 6 syngens and permitted an assessment of intrasyngenic variation for these characteristics. Only the CVP positions appear to be reasonably constant within syngens in the strains examined. On the basis of this criterion the syngens can be arrayed in an approximate order of 1, 3, 7, 6, 8, 9, 2, 5, GL and 4; the angle formed between the central axis, the stomatogenic meridian and the CVP's is most acute in syngen 1.     

Mating Type Inheritance in Glaucoma*

SYNOPSIS. Mating was observed in collections of Glaucoma from 2 localities in Illinois. The collections could be divided into 2 syngens on the basis of mating type reactions. Corticotype analysis showed that syngen 1 was intermediate in meridian number between Glaucoma chattoni and Glaucoma scintillans, while syngen 2 had the same range as Glaucoma scintillans (Lee, unpublished). The major features of mating type inheritance in syngen 1 are that exconjugant clones usually have identical mating types and that mating types I and II appear in an approximately 1:1 ratio in successive generations. This is the result expected in genic mating type inheritance if one of the original parents was a homozygote and the other a heterozygote at the mating type locus. No significant immaturity period was found in these strains. Only 2 viable pairs were obtained from syngen 2 crosses, but results from these suggest that mating type inheritance may be epigenetic and a longer immaturity period may characterize this syngen.     

Factors Influencing the Growth of Glaucoma chattoni in a Chemically Defined Medium*

SYNOPSIS. Glaucoma chattoni strain A has been grown in the chemically defined medium previously used for Colpidium campylum and Tetrahymena pyriformis. It was necessary to modify the amino acid, nucleotide and other components of the medium in order to obtain optimum growth. The modified medium contained 17 amino acids, nucleic acid components, fatty acids, stigmasterol, sodium acetate, a mixture of B-vitamins and several inorganic salts.     

Hyalophysa lynni n. sp. (Ciliophora,Apostomatida), a new pathogenic ciliate and causative agent of shrimp black gill in penaeid shrimp

The parasitic ciliate causing shrimp black gill (sBG) infections in penaeid shrimp has been identified. The sBG ciliate has a unique life cycle that includes an encysted divisional stage on the host’s gills. The ciliature of the encysted trophont stage has been determined and is quite similar to that of the closely related apostomes Hyalophysa bradburyae and H. chattoni. Hyalophysa bradburyae is a commensal ciliate associated with freshwater caridean shrimp and crayfish, while H. chattoni is a common commensal found on North American marine decapods. Based on 18S rRNA gene sequence comparisons, the sBG ciliate is more closely related to the marine species H. chattoni than to the freshwater species H. bradburyae. The unique life cycle, morphology, 18S rRNA gene sequence, hosts, location, and pathology of the sBG ciliate distinguish this organism as a new species, Hyalophysa lynni n. sp.     

Phylogenetic relationships within the class Oligohymenophorea,phylum Cilophora,inferred from the complete small subunit rRNA gene sequences ofColpidium campylum,Glaucoma chattoni,andOpisthonecta henneguyi

Summary Phylogenetic relationships within the class Oligohymenophorea, phylum Ciliophora, were investigated by determining the complete small subunit rRNA (SSrRNA) gene sequences for the hymenostomesColpidium campylum, Glaucoma chattoni, and the peritrichOpisthonecta henneguyi. The affiliations of the oligohymenophoreans were assessed using both distance matrix (DM) and maximum parsimony (MP) analyses. Variations do exist in the phylogenies created by the two methods. However, the basic tree topologies are consistent. In both the DM and MP analyses the hymenostomes (C. campylum, G. chattoni, and the tetrahymenas) all form a very tight group associated with the peritrichO. henneguyi. TheTetrahymena lineage was monophyletic whereasColpidium andGlaucoma were more closely related to each other than either was to the tetrahymenas. The monophyly of the genusTetrahymena in the present analysis supports the phylogenies determined from morphological data and molecular sequence data from the histone H3II/H4II region of the genome. The perplexing and controversial phylogenetic position of the peritrichs is once again depicted in the present analysis. The distinctiveness of the peritrichOpisthonecta from both hymenostome and nassophorean ciliates based on evolutionary distances suggests that the elevation of the peritrichs to a higher taxonomic rank should be reconsidered.     

Phylogenetic relationships amongTetrahymena species determined using the polymerase chain reaction

Summary The species of theTetrahymena pyriformis complex present a conundrum with regard to their highly conservative morphology and widely divergent molecular characteristics. We have investigated the phylogenetic relationships among these species using the nucleotide sequences from the histone H3II/H4II region of the genome. This region includes portions of the two histone coding sequences, as well as the intergenic region. The DNA sequences of these regions were amplified by the polymerase chain reaction (PCR) and the sequence of each was determined. Nucleotide substitutions and insertions/deletions within this set of sequences were compared to determine the phylogenetic relationships among the species of the complex. These data yield phylogenetic trees with identical topologies when different tree-building routines are used, indicating that the data are very robust.Glaucoma chattoni was used as an outgroup to root the trees for this analysis. The genome organization ofG. chattoni and the divergence of its histone H3II/H4II region sequence relative to those of the complex clearly indicate that this species has diverged considerably from the complex. These results show that PCR amplification analysis is feasible over considerable evolutionary distances. However, DNA-DNA hybridization may be more useful than sequence analysis in resolving the relationships among the closely related species in the complex.     

GWAS研究大肠杆菌和金黄色葡萄球菌种间互作进化机制

【目的】通过实验室培养模拟自然环境微生物相互作用,进而找到影响细菌基因型和表型的基因。【方法】将大肠杆菌和金黄色葡萄球菌在实验室条件下进行单独培养和两两混合培养并连续转接,通过得到的数量表型与最大生长速率表型做全基因组关联分析(GWAS),对得到的与表型相关的SNP进行注释与分析。【结果】162个SNP位点影响到大肠杆菌原始菌株与共培养菌株的生长,36个SNP位点影响大肠杆菌菌株在单独培养和共同培养的生长。总共有85个SNP位点影响金黄色葡萄球菌的原始菌株与单独培养。其中5个基因在之前文献中已有报道。对影响不同时间点细菌数量变化形状的SNP位点进行功能注释,大肠杆菌中有706个与生长性能相关。金黄色葡萄球菌中,129个和不同的生长性能相关。大肠杆菌SNP位点的13个基因在之前的研究中已有报道。【结论】混合培养和单独培养都检测到与生长相关的显著基因,本研究表明了GWAS在研究细菌互作进化机制方面的潜力。     

Phylogenetic Relationships Among Anuran Trypanosomes as Revealed by Riboprinting

Twenty trypanosome isolates from Anura (frogs and toads) assigned to several species were characterized by riboprinting–restriction enzyme digestion of polymerase chain reaction amplified small subunit ribosomal RNA genes. Restriction site polymorphisms allowed distinction of all the recognized species and no intraspecific variation in riboprint patterns was detected. Phylogenetic reconstruction using parsimony and distance estimates based on restriction fragment comigration showed Trypanosoma chattoni to be only distantly related to the other species, white T. ranarum and T. fallisi appear to be sister taxa despite showing non-overlapping host specificities.     

Interdependence of Cell Cycle Events in Tetrahymena thermophila: Formation of Contractile Vacuole Pore and Fission Gap in Ciliary Meridians*

SYNOPSIS The formation of the contractile vacuole pore (CVP) in Tetrahymena thermophila. genotype mol^b/mol^b is temporally and spatially associated with the formation of the fission gap in the CVP meridian. New CVPs arise when fission gaps appear in CVP meridians, the new pores being found anterior to the gaps. When, however, CVP meridians are rotated 180°, the fission gaps develop late. In more than 1/3 of the 180°-rotated CVP meridians, the new CVPs are formed before the appearance of the fission gap. Evidently, the appearance of the gap is not a prerequisite for CVP formation. Nevertheless, mutants exist in which the absence of fission gap and CVP are correlated in some cases and in which the presence of supernumerary fission gap and CVP are correlated in other instances. It is suggested that the 2 developmental events, although not causally related to each other, may be controlled by a common morpho-genetic signal. This commits a certain site (mid-body) along a ciliary meridian to develop the fission gap as well as the CVP; however, after this step of commitment, the appearance of the fission gap is delayed in 180°-rotated CVP meridians.     

New records of Rotifera from India

Fifteen species of monogonont Rotifera are recorded from India for the first time. These are Lecane acanthinula (Hauer), L. aspasia Myers, L. monostyla (Daday), L. ruttneri (Hauer), L. simonneae Segers, Lepadella costatoides Segers, L. cf. favorita Klement, L. minuta (Montet), L. triba Myers, Notommata pachyura (Gosse), N. saccigera Ehrenberg, Sinantherina semibullata (Thorpe), Trichocerca chattoni (De Beauchamp), T. kostei Segers and Taphrocampa selenura (Gosse). Taxonomic and/or zoogeographic notes are added, when appropriate.     

Characterization of Further Erwinia amylovora Strains and the Application of the API 20E System in Diagnosis

The former phenotypic study of Erwinia amylovora (VANTOMME et al. 1982) was extended with a collection of 54 Erwinia amylovora strains from a broad plant and geographic origin. From the 85 phenotypic features studied, 72 (85%) were present in at least 90% of the strains. Only 49 (58%) of the features were shared by all strains. Thirty-eight strains were also examined by the API 20E system. The API 20E code numbers for E. amylovora are unique and, combined with an immature, (green) pear test, may be used for an accurate identification of Erwinia amylovora.     

An Ultrastructural Investigation of 180°-Rotated Ciliary Meridians in Tetrahymena pyriformis*

SYNOPSIS. An ultrastructural investigation has been carried out on 180°-rotated ciliary meridians (inverted meridians) in Tetrahymena pyriformis temperature-sensitive mutant (mol^b/mol^b), syngen 1, strain B. The longitudinal, transverse and postciliary microtubular bands, the kinetodesmal fiber, and the parasomal sac, are shown to be disposed at a 180° angle to their normal positions or orientations. Other abnormalities are as folows: the first 2 basal bodies of the inverted meridian fail to organize into “couplets” and the inverted meridian intrudes into the anterior pole region; an extra longitudinal microtubular band is found in one of the cell lines.     

Anti-mutagenic effect of ultraviolet light on spontaneous tyrosine tRNA ochre suppressor mutations in Escherichia coli

Summary The numbers of tyrosine tRNA ochre suppressor mutations arising spontaneously or after UV irradiation in different strains of Escherichia coli K12 are considered. The DNA sequence change requisite for this type of mutation would be a transversion at a cytosine between two purines, where pyrimidine-pyrimidine photoproducts could not form. We find that UV mutagenesis does not produce these tyrosine tRNA ochre suppressor mutations. With lexA51 recA441 defective cells, the spontaneous yield of these mutations is elevated and UV irradiation produces a significant decrease in the numbers of this particular mutation. As explanation we suggest that the spontaneous appearance of these mutations reflects mutation at apurinic sites, the efficiency of which is elevated in lexA51 recA441 cells (with derepressed SOS functions and an activated form of RecA protein). The addition of UV damage in the DNA of these cells cannot further stimulate the positive functions that are required for the production of these mutations and are typically associated with UV mutagenesis (induction of SOS functions, activation of RecA protein and introduction of a targeting photoproduct) but apparently can have a negative effect on mutagenesis, hitherto not realized.     

Comparative physiology of nitrogenase activity and vesicle development for Frankia strains CpI1, ACN1 ag,EAN1pec and EUN1f

The relationship between nitrogen fixation and development of a specialized cell structure, called the vesicle, was studied using four Frankia isolates. Nitrogenase activity was repressed in all four strains during growth with ammonia. Strain CpI1 formed no vesicles during NH₄ growth. Strains ACN1 ^ag , EAN1_pec and EUN1_f produced low numbers of vesicles in the presence of ammonia. Following transfer to nitrogen-free media, a parallel increase in nitrogenase activity and vesicle numbers occurred with all four isolates. Appearance of nitrogenase activity was more rapid in those strains that possessed some vesicles at the time of shift to N₂ as a nitrogen source. The ratio of vesicle numbers to level of nitrogenase activity varied widely among the four strains and in response to different growth conditions and culture age of the individual strains. Optimum conditions of temperature, carbon and energy source, nitrogen source and availability of iron and molybdenum were different for each of the four strains. Those conditions that significantly reduced nitrogenase activity were always associated with decreased numbers of vesicles.