Contributing factor toMycoplasma pneumoniae produced stimulation of rhinovirus-RNA synthesis

It has been demonstrated thatRhinovirus ribonucleic acid (RNA) synthesis was greater inMycoplasma pneumoniae inoculated KB (Human Carcinoma of Nasopharynx) cells than in PPLO-free tissue systems (Milligan III & Fletcher, 1969). In this study,Mycoplasma pneumoniae was grown on glass to eliminate the PPLO medium. However, if PPLO medium was added to theMycoplasma pneumoniae inoculum, stimulation of viral-RNA synthesis was greater than in the presence of PPLO alone. Rhinovirus-RNA synthesis, measured by uridine-³H uptake, in the presence ofMycoplasma pneumoniae andMycoplasma pneumoniae plus medium was enhanced 1.2-fold and 1.4-fold respectively, above virus synthesis in untreated KB cell systems. PPLO medium did not appear to significantly stimulate viral RNA synthesis. However, a 5-fold increase in the concentration of the medium or its components resulted in greater viral RNA replication. The medium components, yeast extract and PPLO broth (Difco), stimulated viral RNA synthesis 4-fold and 2-fold, respectively. Our findings also demonstrated that other components, agar, phenol red, and dextrose, inhibited viral replication, whereas horse serum appeared to have no effect. This work was supported by the Life Sciences Division of the U.S. Army Research Office under contract/grant No. DAHC-20-67C-0122/DAHC 19-69-G-0011.     

Identification of Phosphocholine-Containing Glycoglycerolipids Purified from Mycoplasma fermentans-Infected Human Helper T-Cell Culture as Components of M. fermentans

Previously, we have reported the occurrence of novel phosphocholine-containing glycoglycerolipids (GGPLs: GGPL-I and GGPL-III) in human helper T-cell culture (MT-4 cell line) (Matsuda et al, Glycoconjugate J. 10: 340). However, the GGPLs disappeared from the MT-4 cells after treatment with an antimycoplasma agent. This disappearance suggested the involvement of microorganisms in the GGPL expression. In this paper, we show that the novel lipids are components of Mycoplasma fermentans itself. The supernatant fluid of the antimycoplasma agent-untreated MT-4 cell culture produced mycoplasma-like colonies on PPLO agar plates, and PCR and immunological methods revealed the presence of M. fermentans. GGPLs were expressed again in the treated MT-4 cells after infection with the isolated M. fermentans. The isolated M. fermentans had glycoglycerolipids corresponding to GGPL-I and GGPL-III. Thin-layer chromatography-mass spectrometry and immunological analyses showed that these glycoglycerolipids which were derived from the isolated M. fermentans were identical with GGPL-I and GGPL-III previously obtained. This is the first report that shows mycoplasma has phosphocholine-containing glycoglycerolipids.     

Assessment of aphid colonies by hoverflies. I maple aphids andEpisyrphus balteatus (de Geer) (Diptera: Syrphidae)

In the syrphid hoverflyEpisyrphus balteatus (de Geer), females in search of oviposition sites are assessing the qualitative and quantitative value of aphid colonies for securing the successful development of their offspring. They select small, young and “promising” aphid colonies consisting of nymphs and/or fundatrices as their oviposition sites, and neglect large, older ones including winged adults. Such “buy-futures” tactics will benefit the offspring in finding suitable and sufficient food.     

The Impact of Recent Queenloss and Colony Pheno-type on the Removal of Small Hive Beetle (Aethina tumida Murray) Eggs and Larvae by African Honeybee Colonies (Apis mellifera capensis Esch.)

The removal of small hive beetle [=SHB] eggs and larvae was studied in queenright and recently queenless Cape honeybee, Apis mellifera capensis, colonies over a range of phenotypes. The overall removal efficiency was not influenced by phenotypes or queenstate, because all introduced eggs and larvae were removed within 24 hours. Queenless colonies removed them merely slower than queenright ones. The latter ones rejected up to 300 larvae within one hour. However, colonies undergoing preparation for absconding did not completely remove SHB offspring, suggesting that removal efficiency was reduced. Since even small and recently queenless colonies effectively removed immature SHB, and no differences in the overall efficiency was found compared to A. m. scutellata we conclude that this defense behavior is well developed in African honeybees.     

Untersuchungen über die Morphologie und die Vermehrung der pleuropneumonieähnlichen Organismen und der L-Phase der Bakterien

Zusammenfassung Lichtmikroskopische Untersuchungen der pleuropneumonieähnlichen Organismen und der L-Phase vonBacterium proteus undVibrio cholerae zeigten bei beiden 0,5–1,0 große, runde Gebilde, die vielfach zu traubenoder kettenförmigen Verbänden vereinigt sind. Im Gegensatz zu den PPLO mit relativ regelmäßigen Teilchen zeigen die L-Phasen auch sehr große Körperchen mit Durchmessern bis zu mehreren . Außerdem tritt bei den L-Phasen häufig Vacuolisierung auf. Ähnliche große Gebilde konnten bei den PPLO nur auf ungünstigen Nährböden erhalten werden. Auch die Kolonieform war bei den PPLO stark von der Konsistenz des Nährbodens abhängig.Im Phasenkontrastmikroskop wurde die Vermehrung von PPLO und L-Phasen an wachsenden Mikrokulturen laufend beobachtet und einzelne Stadien photographisch festgehalten. Dabei ergab sich, daß die einzige beobachtbare Vermehrungsweise eine multi- oder unipolare Knospung ist, die je nach der Nährbodenbeschaffenheit zu verschiedenen Formen führt. Häufig kommt es zu perlschnurartigen Ketten, die aber nicht als Mycel bezeichnet werden können, wie es andere Autoren wiederholt taten.Auf die große Ähnlichkeit zwischen PPLO und großen Viren wird hingewiesen.Herrn Ministerialdirigent i. R. Professor Dr.Gustav Seiffert in Verehrung und Dankbarkeit zum 70. Geburtstag gewidmet.     

Serological study of a pleuropneumonia-like organism and an associatedCorynebacterium species

Summary The complement fixation test was employed to study the possible serological relationship between PPLO 4387 andCorynebacterium C4387. No cross reactions between the two organisms were demonstrated by the methods employed. The limitations of the complement fixation test in this study were discussed. Because of these limitations and the accumulated evidence from other types of studies, the authors concluded that the results obtained are not necessarily proof of the absence of a relationship between the PPLO and theCorynebacterium sp. This work was supported by a PHS Research Grant E-2332 from the National Institute of Allergy and Infectious Diseases, U.S. Public Health Service. This paper is part of a thesis presented to the Graduate School of the University of Maryland in partial fulfillment of the requirements for the M.S. degree in Microbiology. Mailing address: Department of Bacteriology, Walter Reed Army Institute of Research, Washington 12, D.C.     

The crystal structure of Pichia pastoris lysyl oxidase

Pichia pastoris lysyl oxidase (PPLO) is unique among the structurally characterized copper amine oxidases in being able to oxidize the side chain of lysine residues in polypeptides. Remarkably, the yeast PPLO is nearly as effective in oxidizing a mammalian tropoelastin substrate as is a true mammalian lysyl oxidase isolated from bovine aorta. Thus, PPLO is functionally related to the copper-containing lysyl oxidases despite the lack of any significant sequence similarity with these enzymes. The structure of PPLO has been determined at 1.65 A resolution. PPLO is a homodimer in which each subunit contains a Type II copper atom and a topaquinone cofactor (TPQ) formed by the posttranslational modification of a tyrosine residue. While PPLO has tertiary and quaternary topologies similar to those found in other quinone-containing copper amine oxidases, its active site is substantially more exposed and accessible. The structural elements that are responsible for the accessibility of the active site are identified and discussed.     

Experimental subcutaneous granulomas in sheep with Dermatophilus-like microorganisms from porcine tonsil

The pathogenicity of the Dermatophilus-like microorganisms from porcine tonsil and the light and electron microscopic findings were studied with adult ewes. The early lesions were abscessess and advanced ones were granulomas after the subcutaneous inoculation. The granulomas were composed of the central bacterial colonies and the layers of the neutrophils, epithelioid cells and giant cells, and peripheral connective tissues. Epithelioid cells and giant cells were identified by the large euchromatic nuclei, abundant organelles and interdigitation of the blunt pseudopods in the ultrastructure. The lesions were very similar to subcutaneous granulomas in sheep and cattle due to Dermatophilus congolensis (atypical dermatophilosis), actinomycosis and nocardiosis.     

Fungi in healthy and diseased sea fans (<Emphasis Type="Italic">Gorgonia ventalina)</Emphasis>: is <Emphasis Type="Italic">Aspergillus sydowii</Emphasis> always the pathogen?

Caribbean corals, including sea fans (Gorgonia spp.), are being affected by severe and apparently new diseases. In the case of sea fans, the pathogen is reported to be the fungus Aspergillus sydowii, and the disease is named aspergillosis. In order to understand coral diseases and pathogens, knowledge of the microbes associated with healthy corals is also necessary. In this study the fungal community of healthy Gorgonia ventalina colonies was contrasted with that of diseased colonies. In addition, the fungal community of healthy and diseased tissue within colonies with aspergillosis was contrasted. Fungi were isolated from healthy and diseased fans from 15 reefs around Puerto Rico, and identified by sequencing the nuclear ribosomal ITS region and by morphology. Thirty fungal species belonging to 15 genera were isolated from 203 G. ventalina colonies. Penicillum and Aspergillus were the most common genera isolated from both healthy and diseased fans. However, the fungal community of healthy fans was distinct and more diverse than that of diseased ones. Within diseased fans, fungal communities from diseased tissues were distinct and more diverse than from healthy tissue. The reduction of fungi in diseased colonies may occur prior to infection due to environmental changes affecting the host, or after infection due to increase in dominance of the pathogen, or because of host responses to infection. Data also indicate that the fungal community of an entire sea fan colony is affected even when only a small portion of the colony suffers from aspergillosis. An unexpected result was that A. sydowii was found in healthy sea fans but never in diseased ones. This result suggests that A. sydowii is not the pathogen causing aspergillosis in the studied colonies, and suggests several fungi common to healthy and diseased colonies as opportunistic pathogens. Given that it is not clear that Aspergillus is the sole pathogen, calling this disease aspergillosis is an oversimplification at best. Communicated by Biology Editor Dr Michael Lesser     

Long-term Effects of <Emphasis Type="Italic">Lupinus lepidus</Emphasis> on Vegetation Dynamics at Mount St. Helens

The nitrogen-fixing legume Lupinus lepidus is the most abundant herb on new volcanic surfaces at Mount St. Helen. We compared vegetation structure in 30 Lupinus colonies in three age classes (old, mature, or young based on known years of their establishment) to adjacent sites that were sparsely populated by Lupinus. Our goals were to determine if the age of colonies affected either species composition or vegetation structure and if colonies altered vegetation structure of a site. Species richness increased with site age and colonies had significantly greater richness. Mature colonies had lower cover and frequency of other species, while old colonies had significantly higher cover of other species, particularly mosses. Frequencies in colonies were higher than in adjacent sites. Diversity measures were highest in old colonies and least in mature ones. ANOVA showed that density effects of Lupinus were stronger than age effects on richness, cover, frequency, dominance, and diversity. The floristic similarity between colonies and adjacent sites declined with colony age, and sites without Lupinus became heterogeneous with age. Detrended correspondence analysis demonstrated floristic change as colonies aged and that the differences between colonies and sparse sites increased with age. Regressions of Lupinus frequency with measures of structure showed weak second order relationships that suggested that low densities of Lupinus inhibited other species, while higher densities promoted abundance and diversity. Together these results suggest that Lupinus is a key element that alters floristic successional trajectories and accelerates succession. However, its effects are complicated by the wide ranges of Lupinus density, the sequence of invasion, and by herbivores.     

Efficient integrative transformation of the phytopathogenic fungus Alternaria alternata mediated by the repetitive rDNA sequences

An attempt was made to transform Alternaria alternata protoplasts using a plasmid vector, pDH25, bearing the Escherichia coli hygromycin B (Hy) phosphotransferase gene (hph) under the control of the Aspergillus nidulans trpC promoter. Transformants arose on a selective medium containing 100 μg Hy/ml. There were two types of transformants, forming large and small colonies on the selective medium. Transformation with one μg of the vector produced an average of 4.5 large colonies and 600 small ones. In large-colony transformants, the vector often integrated into the recipient chromosome in the form of highly rearranged tandem arrays. To increase transformation efficiency, fragments of the highly repetitive ribosomal RNA gene cluster (rDNA) of A. alternata were used to construct four new vectors for homologous recombination system. Use of these vectors gave higher transformation efficiency than the original plasmid. The best vector, pDH25r1a, gave rise to large-colony transformants at a frequency 20 times higher than pDH25. Transformation events in A. alternata with pDH25r1a occured by homologous recombination as a single crossover between the plasmid-borne rDNA segment and its homologue in the chromosome, often giving rise to tandemly repeated vector DNA.     

Ascospore Dimorphism and Mating Type in Chromocrea spinulosa (Fuckel) Petch n. comb

The asci contain four large and four small ascospores, eachtwo-celled, arranged in the six patterns expected if spore sizewere controlled by a pair of allelic genes, the locus showing65 per cent, second-division segregation. The small ascosporesproduce sterile colonies, the large ones moderately fertilecolonies whose asci again show segregation for spore size. Fertilityis stimulated where colonies from large and small spores meet.In crosses of various mutants with the wild type, all asci inperithecia developed along the line of junction show segregationfor the mutant as well as for spore size. Evidently Chromocreais heterothallic, the spore-size difference being a pleiotropicexpression of mating type. One allele, that governing largespores, occasionally mutates to the other allele, resultingin fertility of colonies from large spores.     

Clonal propagation by the azooxanthellate octocoral Dendronephthya hemprichi

 The azooxanthellate octocoral Dendronephthya hemprichi (Octocorallia, Alcyonacea) is the most abundant benthic organism inhabiting the under-water surfaces of oil jetties at Eilat (Red Sea); however, it is very rare on Eilat’s natural reefs. This soft coral exhibits a newly discovered mode of clonal propagation that results in autotomy of small-sized fragments (2–5 mm in length). They possess specialized root-like processes that enable a rapid attachment onto the substrata. An autotomy event is completed within only 2 days; large colonies can bear hundreds of pre-detached fragments. Temporal fluctuations in the percentage of fragment-bearing colonies indicate that autotomy is stimulated by exogenous factors, probably flow-related events. Recruitment of fragments onto PVC plates placed horizontally adjacent to D. hemprichi colonies, was immediate and remarkably high. Attached fragments were observed 2 days after placement of plates; after 52 days, densities of more than one recruit per cm² were recorded. The negative buoyancy of fragments causes them to land on horizontal surfaces rather than on vertical ones. However, their survivorship on vertical surfaces is much higher. This pattern corresponds with dominance of D. hemprichi on vertical substrata at the oil jetties and on natural vertical reefs of the northern Red Sea. Such a mode of clonal propagation provides an efficient mean for genets of D. hemprichi to exploit food resources within a zooxanthellate-dominated reef community. Accepted: 2 February 1996     

Growth or reproduction: Intrasexual competition in a colonial damselfishStegastes nigricans

Intrasexual competition within colonies of the damselfishStegastes nigricans was investigated. Both males and females held individual territories, and these territories were adjacently distributed, forming distinct conspecific colonies with different size compositions. In both large and small colonies, only relatively large individuals participated in reproduction. Breeding males and females left their territories to court, spawn and aggregate more frequently than non-breeders of the same colonies. Breeders were less frequently attacked by other fish during these excursions than were non-breeders. Most attacks were conducted by breeders of the same sex. This intrasexual aggression may suppress the reproduction of non-breeders at least in large colonies. Non-breeders in large colonies had smaller gonads than breeders of both large and small colonies, but they grew faster than breeders of the same body size in small colonies. This suggests that non-breeders in large colonies allocate energy to growth in order to be reproductively competitive in the future, although only a few of them remained in the same colonies upon maturity. Colony size thus has an effect on the reproductive life histories of individual damselfish.     

Foraging decisions of individual workers vary with colony size in the greenhead ant Rhytidoponera metallica (Formicidae, Ectatomminae)

Summary. The ability of worker ants to adapt their behaviour depending on the social environment of the colony is imperative for colony growth and survival. In this study we use the greenhead ant Rhytidoponera metallica to test for a relationship between colony size and foraging behaviour. We controlled for possible confounding ontogenetic and age effects by splitting large colonies into small and large colony fragments. Large and small colonies differed in worker number but not worker relatedness or worker/brood ratios. Differences in foraging activity were tested in the context of single foraging cycles with and without the opportunity to retrieve food. We found that workers from large colonies foraged for longer distances and spent more time outside the nest than foragers from small colonies. However, foragers from large and small colonies retrieved the first prey item they contacted, irrespective of prey size. Our results show that in R. metallica, foraging decisions made outside the nest by individual workers are related to the size of their colony.Received 23 March 2004; revised 3 June 2004; accepted 4 June 2004.     

Effect of external environmental factors on the morphology of Spiroplasma citri.

Spiroplasma citri was examined by electron microscopy for morphological changes when maintained under a variety of conditions. PPLO serum fraction maintained spiral and helical morphology of S. citri at pH values of 8.0, 7.5, and 7.0, but only partially at pH 6.0 and 5.0. The absence of PPLO serum fraction resulted in round, deteriorated cells at all pH values tested. Bovine serum albumin (BSA), Phytone, soluble starch, potato starch, spermine, lipid-extracted PPLO serum fraction, and lipid-extracted BSA could substitute for PPLO serum fraction in maintaining spiral and helical morphology at pH 7.5. At pH 5.0, only BSA, lipid-extracted BSA, and lipid-extracted PPLO serum fraction were effective. Only BSA supported growth of S. citri for more than two transfers, whereas all other substitutes could not support growth longer than two transfers.     

Ontogenetic changes in forager polymorphism and foraging ecology in the leaf-cutting ant Atta cephalotes

In the leaf-cutting ant Atta cephalotes (L.) small colonies produce a relatively narrow size-range of small workers, whereas large colonies produce a much wider size-range of workers. In this study, I compared the foraging of four small A. cephalotes colonies (fewer than 5000 workers) with published data on foraging of large colonies to examine how colony size and worker size-range may be related to foraging ecology in leaf-cutting ants. I found that the foraging ecology of small A. cephalotes colonies is very different from that of large colonies. In small colonies, a relatively narrow size-range of foragers (1.4–6.7 mg, mean 3.3 mg) cut primarily herbs (ferns, grasses, and other small herbaceous plants) located within 7 m of the nest. In contrast, in large colonies, a broader size-range of workers (1.4–30 mg, mean 7.3 mg) participate in foraging, generally harvesting from trees 20–80 m from the nest, with larger workers cutting on trees with thicker and tougher leaves. Small colonies' dependence on small herbaceous plants near the nest may have a profound impact on distribution of A. cephalotes. A. cephalotes colonies are rarely found in primary forest, where the low occurrence of small herbaceous plants in the understory may preclude the establishment of young colonies.     

Facultative polygyny in Ectatomma tuberculatum (Formicidae, Ectatomminae)

Summary. Polygyny, the presence of several mated queens within the same colony, is widespread in insect societies. This phenomenon is commonly associated with ecological constraints such as limited nest sites. In habitats where solitary nest foundation is risky, monogynous colonies can reintegrate young daughter queens (secondary polygyny). We studied the reproductive structure (i.e. queen number) of the ectatommine ant Ectatomma tuberculatum from Bahia State, Brazil. This species was found to present facultative polygyny: out of a total of 130 colonies collected, 39.2% were monogynous, while 43.8% were polygynous. Polygynous colonies had significantly more workers than monogynous ones. Queen number in polygynous colonies ranged from 2 to 26, with an average of 4 ± 4 queens per colony. All nestmate queens were egg-layers with no apparent dominance hierarchy or agonistic behavior. Non-nestmate queens were adopted by monogynous colonies suggesting that polygyny is secondary, originating through queen adoption. This species is characterized by an open recognition system, which probably allows a switch from monogynous to polygynous colonies. The behavioral acts of queens showed that resident queens remained frequently immobile on or near the brood, contrarily to alien or adopted queens and gynes. In addition, monogynous queens showed no behavioral or physiological (i.e. by ovarian status) differences in comparison with polygynous ones. Secondary or facultative polygyny, probably associated with queen adoption, may have been favored in particular environmental conditions. Indeed, by increasing colony productivity (i.e. number of workers) and territory size (by budding and polydomy), polygyny could uphold E. tuberculatum as a dominant species in the mosaic of arboreal ants in Neotropical habitats.Received 7 April 2004; revised 11 November 2004; accepted 15 November 2004.     

Comparative Investigation of Mesenchymal Stem Cells Isolated from the Bone Marrow and Fetal Liver of Mouse and Rat

We studied the properties of cells forming fibroblast colonies from the bone marrow and fetal liver of mouse and rat. Bone marrow and fetal liver cells formed colonies in vitro including fibroblasts as well as a considerable proportion of macrophages. The colonies formed from bone marrow and hepatic cells of rat differed from the murine ones by a higher proportion of fibroblasts. Most colonies derived from the bone marrow of both mouse and rat included a fraction of cells expressing alkaline phosphatase, and hence, capable of osteogenic differentiation; the colonies derived from the fetal liver included low proportions of such cells. The cell layers derived from the colony-forming fibroblasts of both bone marrow and fetal liver of mouse maintained hematopoiesis in the peritoneal cavity of irradiated mice, which indicated that these progenitor cells can form hematopoietic microenvironment.     

Evidence of prey depletion around lesser kestrel Falco naumanni colonies and its short term negative consequences

Predation may reduce prey numbers in such extent that prey may be depleted, which has negative effects on predator populations. Prey depletion would be more likely when the number of predators increase and/or concentrate their activity in a certain area, as is the case of colonial birds. As a matter of fact, food depletion due to intraspecific competition is considered a major cost of coloniality, and several studies have shown indirect evidence of this. However, no direct measures of food depletion have been provided along with its consequences for the fitness of the colony inhabitants. We carried out a field study with the lesser kestrel Falco naumanni, a raptor that breeds in colonies ranging from two to dozens of pairs. During the nestling period we sampled the main prey of the kestrels around different sized colonies at increasing distances. At the same time, we recorded hunting distances and prey delivery rates to the nest. In addition, we monitored the reproductive success in colonies of different sizes. Lesser kestrels feed their nestlings mainly with grasshoppers and these prey became depleted through the season in the surroundings of the large colonies. Prey depletion made kestrels fly longer distances to forage and prey delivery rates to the nest decreased. Lower feeding rates were not compensated by bringing larger prey, hence, the net amount of energy provided to the chicks decreased with the date in large colonies. By contrast, none of this occurred around small colonies, where both prey abundance and hunting distance remained constant throughout the season. As a consequence, the seasonal decline in the reproductive success (number of fledglings and fledgling body condition) was greater the larger the colony. Thus, these results evidence that food depletion and its fitness costs are related to colony size, as they are suffered by the kestrels breeding in large colonies but not by those settled at small ones. Finally, the consequences of prey depletion on the demographic dynamics and the regulation of colony size are discussed.