Differential proliferation of erythroid and granuloid spleen colonies following sublethal irradiation of the bone marrow donor

Spleen colonies produced by sublethally irradiated mouse bone marrow cells were compared to those produced by unirradiated marrow cells in lethally irradiated mice. Sublethally irradiated marrow cells gave rise to many fewer spleen colonies. At seven days of colony age, the ratio of erythroid colonies to granuloid colonies was lower (< 1) than for colonies formed by unirradiated marrow (2 to 3 or more). Delay of harvest of colonies to day 10 or 12 resulted in 6 to 11 fold increase in the ratio of erythroid to granuloid colonies due largely to the belated appearance of erythroid colonies.     

The determinants of queen size in a socially polymorphic ant

In social animals, body size can be shaped by multiple factors, such as direct genetic effects, maternal effects, or the social environment. In ants, the body size of queens correlates with the social structure of the colony: colonies headed by a single queen (monogyne) generally produce larger queens that are able to found colonies independently, whereas colonies headed by multiple queens (polygyne) tend to produce smaller queens that stay in their natal colony or disperse with workers. We performed a cross‐fostering experiment to investigate the proximate causes of queen size variation in the socially polymorphic ant Formica selysi. As expected if genetic or maternal effects influence queen size, eggs originating from monogyne colonies developed into larger queens than eggs collected from polygyne colonies, be they raised by monogyne or polygyne workers. In contrast, eggs sampled in monogyne colonies were smaller than eggs sampled in polygyne colonies. Hence, eggs from monogyne colonies are smaller but develop into larger queens than eggs from polygyne colonies, independently of the social structure of the workers caring for the brood. These results demonstrate that a genetic polymorphism or maternal effect transmitted to the eggs influences queen size, which probably affects the social structure of new colonies.     

Transmission of Vairimorpha invictae (Microsporidia: Burenellidae) infections between red imported fire ant (Hymenoptera: Formicidae) colonies

Red imported fire ant, Solenopsis invicta, colonies were successfully infected with the microsporidium Vairimorpha invictae by introducing live larvae, pupae, or dead adults from V. invictae-infected field colonies collected in Argentina. Introductions with 4th instar larvae or non-melanized pupae obtained from infected field colonies, resulted in infection of 40% of the inoculated colonies. Introductions of 4th instars or melanized pupae produced from colonies that were initially infected in the laboratory, resulted in infections of 83% of the colonies, thus perpetuating the infection in other colonies. Infection was detected in 2 of 6 colonies after introducing adult worker caste ants that had died with V. invictae. The average number of adults and the volume of immature ants per colony were significantly lower in the infected than in the control colonies. Infected colonies had 86% fewer adults per colony and 82% less immature ants than the controls. A portion of the 16S rRNA gene of the V. invictae identified from these studies was amplified, cloned, and sequenced; the 1251 nucleotide amplicon was 100% identical to the 16S rRNA gene sequence recorded previously in the GenBank database, thus verifying the species as V. invictae. This is the first report of the artificial transmission of this pathogen to uninfected ant colonies, and demonstration of its ability to hinder growth in individual colonies.     

Colony formation in agar by multipotential hemopoietic cells.

Agar cultures of CBA fetal liver, peripheral blood, yolk sac and adult marrow cells were stimulated by pokeweed mitogen-stimulated spleen conditioned medium. Two to ten percent of the colonies developing were mixed colonies, documented by light or electron microscopy to contain erythroid, neutrophil, macrophage, eosinophil and megakaryocytic cells. No lymphoid cells were detected. Mean size for 7-day mixed colonies was 1,800-7,300 cells. When 7-day mixed colonies were recloned in agar, low levels of colony-forming cells were detected in 10% of the colonies but most daughter colonies formed were small neutrophil and/or macrophage colonies. Injection of pooled 7-day mixed colony cells to irradiated CBA mice produced low numbers of spleen colonies, mainly erythroid in composition. Karyotypic analysis using the T6T6 marker chromosome showed that some of these colonies were of donor origin. With an assumed f factor of 0.2, the mean content of spleen colony-forming cells per 7-day mixed colony was calculated to vary from 0.09 to 0.76 according to the type of mixed colony assayed. The fetal and adult multipotential hemopoietic cells forming mixed colonies in agar may be hemopoietic stem cells perhaps of a special or fetal type.     

Genetics of Ustilago violacea. XXXV. Transposition in Haploid and Diploid Sporidia and Germinating Teliospores

Ustilago violacea sporidia of the white (w) MAD strain (a-2 w lys-3 ino-1 thi) incubated on minimal medium containing 100 mM KClO3 (potassium chlorate) produced only colonies with the pink phenotype. Sporidia from these colonies retained their pink color on complex medium. Sporidia of the diploid D1 strain (a-1 y nic-1 thi/a-2 w met-1 arg-f Chl70 thi) and of the diploid D2 strain (a-1 y his-1 glu-1 thi/a-2 w lys-3 ino-1 thi) produced pink colonies on complex medium. Streaks of diploid D1 sporidia from the pink colonies were stable on complex medium. In contrast, streaks of diploid D2 sporidia, which are heterozygous for the MAD strain, were unstable, initially producing pink colonies on complex medium but then, with continued incubation, producing white termini. Sporidia from the white termini with diploid morphology continued to yield white colonies. Teliospore colonies from three crosses with the MAD strain as a common parent were uniformly pink or had a pink sector instead of the expected uniformly white colonies or colonies having a white sector. Four of 20 and 13 of 20 teliospore colonies, respectively, from two of the three crosses had both a-1 and a-2 sporidia, and the remaining colonies had only a-1 or only a-2 sporidia. All 40 teliospore colonies from the third cross had only a-1 or only a-2 sporidia. All of these observations indicated that the MAD strain may have two autonomous, transactive transposable elements in different chromosomes and that insertional mutations in at least two haplolethal loci were responsible for the teliospore colonies with only a-1 or only a-2 mating type. Crossing over between a haplolethal locus and the centromere would account for teliospore colonies with both a-1 and a-2 sporidia.     

A population genetic analysis of migration: the case of the noctule bat (Nyctalus noctula)

Although rarely assessed, the population genetics of hibernating colonies can help to understand some aspects of population structure, even when samples from nursery or mating colonies are not available, or in studies of migration when both types of samples are available and can be compared. Here we illustrate both points in a survey of mitochondrial DNA (mtDNA) control region sequences used to study the population genetics of hibernating colonies of a migrating species, the noctule bat (Nyctalus noctula). Lacking samples from Scandinavian nursery colonies, we use a North European hibernacula to suggest that Scandinavian populations are isolated from Central and East European colonies. Then, we compare genetic diversities of nursery and hibernating colonies. We find a significantly higher haplotype diversity in hibernacula, confirming that they consist of individuals from different nursery colonies. Finally, we show that pairwise comparisons of the haplotype frequencies of nursery and hibernating colonies contain some information on the migration direction of the noctule bat.     

Effect of estradiol on splenic repopulation by endogenous and exogenous hemopoietic cells in irradiated mice

Estradiol treatment of irradiated mice during repopulation of their spleens by endogenous hemopoietic cells reduced the number of myelocytic colonies and increased the numbers of erythropoietic and undifferentiated colonies. The inhibitory effects of the hormone on myelopoiesis were not dependent on stimulation of erythropoiesis, since they occurred in the absence of erythropoiesis in mice made polycythemic by hypertransfusion. Treatment of bone marrow donors with estradiol reduced the ability of their marrow cells to form spleen colonies, particularly reducing the proportion of myelopoietic colonies relative to the total number of colonies of all types. Conversely erythropoietic colonies, though reduced in absolute number, were increased in relative number. Such treatment also decreased the volume and cell content of the marrow cavity through stimulation of endosteal bone formation. Estradiol treatment of lethally irradiated recipient mice did not detectably alter the total numbers or types of hemopoietic spleen colonies formed in these animals from transplanted marrow cells; however, without estradiol treatment, myelopoietic colonies were so few and erythropoietic colonies so numerous that the effects of the hormones may have been undetectable by the methods employed. The sex of the donor or recipient mouse did not affect the numbers or types of colonies formed, suggesting that endogenous levels of estradiol were too low to exert effects dectectable by the methods used. However, since our mice were only 8 weeks old, the data do not exclude the possibility that older female mice, with higher levels of estradiol, would have differed from males in relative numbers of myelopoietic as compared with erythropoietic colonies.     

Effects of hyperbaric oxygen on the growth and properties of Pseudomonas aeruginosa

Growth of Pseudomonas aeruginosa in 2 atmospheres absolute of 100% oxygen at 37 degrees C produced two types of abnormal colonies--stunted, rough colonies, termed dwarfs, and large, domed, mucoid colonies, termed giants. The occurrence of these variants depended upon the partial pressure of oxygen and the inoculum size. Subculture of dwarf or giant colonies produced a mixture of both colony types after incubation in hyperbaric oxygen, and colonies of normal appearance after incubation in air. Electron micrographs of ultrathin sections showed that cells from dwarf colonies had a more clearly defined envelope region than cells from normal colonies. Giant colony and normal colony-derived cells were of similar appearance. Whole cells from giant colonies contained more carbohydrate, readily extractable lipid, neutral lipid and free fatty acid than cells from normal colonies; the two cell types showed similar contents of 2-keto,3-deoxyoctonic acid and total phospholipid, but different proportions of individual phospholipids. Cells from dwarf, giant and normal (air-grown) colonies were incubated in air on nutrient agar containing either polymyxin, tetracycline or phenoxyethanol. Relative to cells from normal colonies, cells from dwarf colonies showed enhanced resistance to all three agents and cells from giant colonies showed enhanced resistance to polymyxin and tetracycline only. The resistance of cells from variant colonies was lost following a single subculture in air in the absence of antibacterial agents. It was concluded that the envelopes of cells from dwarf and giant colonies differed both from each other and from those of normal cells. These differences, and the formation of variant colonies, appeared to result from bacterial adaptation to hyperbaric oxygen rather than from mutation.     

Possible demonstration of multipotential nature of embryonic neural retina by clonal cell culture.

The possible multipotential nature of the neural retina of early chick embryos was examined by the technique of clonal cell culture. Cultures were prepared from cells dissociated from freshly excised neural retinas of 3.5-day-old chick embryos or from cells harvested from primary highdensity cultures. The following four colony types were obtained: colonies differentiating into “lentoid bodies”; colonies with pigment cells; colonies with both “lentoid bodies” and pigment cells; and colonies comprised entirely of unidentifiable cells. Neuronal differentiation occurred frequently in the early stages of culture (up to about 10 days). In some of these neuronal colonies, “lentoid bodies” and, rarely, both “lentoid bodies” and pigment cells differentiated after a further culture period of up to 30 days. Secondary colonies established from primary colonies after 9–10 days demonstrated that these original colonies fell into four different categories: those giving rise to secondary colonies containing only “lentoid bodies,” those giving rise to pigmented colonies only, those developing both lentoid and pigmented colonies, and finally those which gave rise to secondary colonies of all three types, lentoid, pigmented, and mixed colonies. When primary pigmented colonies were recloned at about 30 days after inoculation, the differentiated pigment cells transdifferentiated into lens. Whether multispecific colonies were really of clonal origin or not is discussed. The possible presence of a multipotent progenitor cell able to give rise to multispecific clones in the neural retina of 3.5-day-old chick embryos is suggested. A sequence of differentiation starting from multipotent neural retinal cells to be terminated with lens through the differentiation of neuronal and pigment cells is hypothetically proposed.     

Reinvasion and colony expansion of Coptotermes formosanus (Isoptera: Rhinotermitidae) after areawide elimination

The population recovery of Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae) colonies were monitored after an areawide elimination of all detectable colonies from September 2003 to August 2005 in Louis Armstrong Park, New Orleans, LA. Six colonies reinvaded the vacant niche created by the full elimination. These colonies expanded their territories throughout the study period. This represented 43% of the original number of colonies present in the park before the elimination. To determine the mode of the reinvasion, nuptial pair establishment was monitored during the C. formosanus dispersal flight seasons. Nuptial pairs were discovered up to 1 yr after the elimination. Morphological and genetic data were collected from field colonies before the full elimination in 2002 and again in 2005 after the reinvasion of these territories by new colonies. These data were used to estimate the relative age of reinvading colonies as compared with their predecessors. It is proposed that the first three reinvading colonies detected were smaller colonies that were undetectable before the full elimination, or were older, established colonies present outside of the park, that expanded their foraging territories into the park in the absence of competition from the eliminated populations. The subsequent three colonies to reinvade seemed to be small colonies founded during or just before the study period by an imago pair after a dispersal flight into the park from outlying areas. The implications of this study on subterranean termite areawide integrated pest management strategies are discussed.     

Influence of pollen diet in spring on development of honey bee (Hymenoptera: Apidae) colonies

The effects of changes in spring pollen diet on the development of honey bee, Apis mellifera L. (Hymenoptera: Apidae), colonies were examined in a 3-yr study (2002-2004). Pollen-supplemented and pollen-limited conditions were created in colonies every spring, and brood rearing and honey yields were subsequently monitored throughout the summer. In all 3 yr, colonies that were supplemented with pollen or a pollen substitute in the spring started rearing brood earlier than colonies in other treatment groups and produced the most workers by late April or early May. In 2002, these initial differences were reflected by a two-fold increase in annual honey yields by September for colonies that were pollen-supplemented during the spring compared with pollen-limited colonies. In 2003 and 2004, differences between treatment groups in the cumulative number of workers produced by colonies disappeared by midsummer, and all colonies had similar annual honey yields (exception: in one year, productivity was low for colonies supplemented with pollen before wintering). Discrepancies between years coincided with differences in spring weather conditions. Colonies supplemented with pollen or a substitute during the spring performed similarly in all respects. These results indicate that an investment in supplementing the pollen diet of colonies would be returned for situations in which large spring populations are important, but long-term improvement in honey yields may only result when spring foraging is severely reduced by inclement weather. Beekeepers should weigh this information against the nutritional deficiencies that are frequently generated in colonies by the stresses of commercial management.     

A comparison of stem cell assays using early or late spleen colonies

The distribution of spleen colony diameters was determined 5.5, 8.0, 10.5 and 13.0 days after injection of normal bone marrow cells to lethally irradiated recipients. A relative lack of small colonies on day 8.0, as compared with days 5.5, 10.5 and 13.0, argued against a time continuum in colony appearance. The spleen colonies observed after 10 days or more probably represented a mixture of colonies which developed from the originally transplanted CFU-S and those arising from secondary CFU-S. Thus, late appearing spleen colonies may not necessarily identify a different, less mature, population of CFU-S. Administration of increasing amounts of bone marrow cells was used for comparing the linearity of the CFU-S assay for colonies observed after 8 days or after 12 to 13 days. The influence of overlapping colonies on the results was considerably augmented if large spleen colonies were observed after 12 or 13 days. Subsequently the CFU-S assay lost much of its quantitative character. We believe that some previously published data might have been misinterpreted by neglecting the important differences between 'early' and 'late' CFU-S assays.     

Cell kinetic study on GM colonies using autoradiography and collagen gel culture with special reference to unique proliferation of eosinophils

Using a newly developed technique of autoradiography and collagen gel culture, a kinetic study on human GM colonies was attempted. Colonies of immature cells appeared first on day 5. The number of mixed colonies (mixture of immature cells, neutrophils, and/or monocyte-macrophages) and neutrophil colonies attained a maximum on days 8 to 10 and a broad peak of monocyte-macrophage colonies was observed on days 11 to 16. Eosinophil colonies appeared first on day 12, reached a maximum on day 18, and then gradually decreased. A detailed analysis of the order of appearance of the colonies suggests that mixed, neutrophil and monocyte-macrophage colonies originate from immature cell colonies or clusters, while eosinophil colonies do not. An autoradiographic study was designed to study the proliferation characteristics of each colony. Labeling indices (LI) with 3H-TdR of the cells in immature cell colonies were always high. LI of the cells in differentiated colonies such as neutrophil, monocyte-macrophage, and mixed colonies were low throughout the observation period. In contrast, LI of the cells in eosinophil colonies were constantly high regardless of the size of cell aggregates and the duration of the culture period. Both mitotic indices and mean grain counts on the nuclei of eosinophils were similar to those of immature cells. These results suggest that eosinophil colonies develop from their own small clusters and that eosinophils retain a fairly good proliferative capacity even when differentiated to the level in which specific granules appear in the cytoplasm.     

A comparison of stem cell assays using early or late spleen colonies

Abstract The distribution of spleen colony diameters was determined 5.5, 8.0, 10.5 and 13.0 days after injection of normal bone marrow cells to lethally irradiated recipients. A relative lack of small colonies on day 8.0, as compared with days 5.5, 10.5 and 13.0, argued against a time continuum in colony appearance. The spleen colonies observed after 10 days or more probably represented a mixture of colonies which developed from the originally transplanted CFU-S and those arising from secondary CFU-S. Thus, late appearing spleen colonies may not necessarily identify a different, less mature, population of CFU-S. Administration of increasing amounts of bone marrow cells was used for comparing the linearity of the CFU-S assay for colonies observed after 8 days or after 12 to 13 days. The influence of overlapping colonies on the results was considerably augmented if large spleen colonies were observed after 12 or 13 days. Subsequently the CFU-S assay lost much of its quantitative character. We believe that some previously published data might have been misinterpreted by neglecting the important differences between 'early'and 'late'CFU-S assays.     

Honey supplementation and its developmental consequences: evidence for food limitation in a paper wasp, Polistes metricus

ABSTRACT.

• 1 Polistes metricus Say (Hymenoptera: Vespidae, Polistinae) field colonies were supplemented with dilute honey during the pre-emergence and early post-emergence phases of colony development.
• 2 Supplementation did not increase number of nest cells constructed or rates of loss to predation or foundress disappearance compared with controls.
• 3 Colonies receiving honey supplementation produced first offspring earlier in the season than control colonies. The difference is due to a shorter time span between founding and first emergence.
• 4 Foundresses of supplemented and control colonies did not differ in wing length or per cent body fat.
• 5 Offspring of both supplemented and control colonies had shorter wing lengths than did foundresses of supplemented colonies. Offspring of control colonies had shorter wing lengths than did foundresses of control colonies but not foundresses of supplemented colonies.
• 6 The per cent body fat of offspring from control colonies was lower than that of all foundresses and of offspring from supplemented colonies.
• 7 The per cent body fat of offspring from supplemented colonies was higher than that of all foundresses.

     

Banding,age and growth in the calcareous hydrozoan Millepora complanata Lamarck

Prominent bands or lines were present on colonies of Millepora complanata on coral reefs at Barbados, West Indies. These bands appeared as regular, dark and light horizontal zones or stripes on vertically growing plates and blades making up the colonies. Bands are formed by wave-like undulations or currugations of the skeletal surface. The number of bands correlates with colony height and mean band width equals the mean annual vertical growth of measured colonies. Variation in annual growth and differences in growth rates of colonies between different reefs were determined from band width measurements. Bands provide a rapid and non-destructive means for measuring growth and ageing colonies of M. complanata.     

Effect of formic acid formulations on honey bee (Hymenoptera: Apidae) colonies and influence of colony and ambient conditions on formic acid concentration in the hive

The interaction between the effects of varroa, Varroa destructor Anderson & Trueman, and formic acid treatments on colonies of honey bees, Apis mellifera L., were examined in two field experiments. In experiment 1, colonies with low varroa levels were exposed to two different slow-release formulations and compared with untreated colonies. In experiment 2, colonies inoculated with varroa and uninoculated colonies were exposed to a slow-release formulation, a pour-on formulation, or were left untreated. The effects of treatments, hive temperature, and hive relative humidity on formic acid concentration in hive air also were examined. Slow-release formic acid application improved colony development in colonies that had been inoculated with varroa. However, in uninoculated colonies where the mean abundance of varroa was low, slow-release formic acid application suppressed colony development. The pour-on application did not have a negative impact on worker population growth in uninoculated colonies, but also it was not as effective as the slow-release treatment in improving population growth in varroa-inoculated colonies. Equivalent volumes of acid applied in pour-on and slow-release formulations provided the same cumulative dose in hive air but differed in the daily pattern of formic acid release. Colonies that were not inoculated with varroa had higher concentrations of formic acid in hive air than colonies that were inoculated with varroa on three of the five pour-on application dates. The data suggest that reductions in worker population and/or activity caused by varroa can interact with ambient conditions to affect the volatilization or sorption of formic acid in the hive.     

STUDIES OF HEMATOPOIESIS IN MURINE SPLEENS: SIGNIFICANCE OF HEMATOPOIETIC COLONIES FORMED ON THE SURFACE OF SPLEENS

Conies of hematopoietic tissue are formed in spleens of lethally irradiated mice by the injection of small numbers of hematopoietic cells. Some of these colonies appear as surface colonies, others can be identified only in serial sections of the spleen. The present studies have related the number and cellular composition of total hematopoietic colonies in the murine spleen to their visual recognition on the splenic surface. These studies demonstrate that only 50% of the total colonies in a spleen are recognized as surface colonies and that of those colonies on the surface, approximately 80% contain erythroid elements. At least four factors play important roles in the recognition of hematopoietic colonies as splenic surface colonies: (1) dose of repopulating cells or hematopoietic stem cells injected into the irradiated animal; (2) location of colonies within the spleen; (3) size of colonies; and (4) cellular content of the colonies. These studies demonstrate that surface colony formation reflects primarily erythropoiesis and not total hematopoiesis.     

Detection of multiple-strain carriers: the value of re-sampling

Bacteria may colonize a carrier with more than one strain of a species at any one time. Attempts to determine multiple colonization are labour intensive because of the large number of colonies per carrier which need to be tested. A possible solution -- in which only 3 colonies per carrier are initially tested and only multiple-strain carriers are re-sampled -- was recently described. We evaluated the accuracy of the "re-sampling method" devised by Cespedes et al. with 500,000 stochastic simulations per scenario. Re-sampling is acceptable where > or = 5 colonies are initially tested or where one strain predominates over others in colony counts. The method introduces bias towards overestimation which decreases with the number of available carriers, increases with the proportion of truly multiple carriers, with decreasing number of colonies available for testing, and with decreasing number of colonies tested per carrier. Initial testing of 5-8 colonies tested with re-sampling are adequate for a large study (>100 carriers), or a small study where it is suspected that no strain predominates over the other in colony counts. Testing 9-20 colonies with re-sampling is necessary for small studies where one strain predominates over others. Re-sampling is unnecessary where >20 colonies are tested.     

Tetrazolium staining by optical scanning overestimates colony size and number of colonies counted

We measured the effect that staining with 2-(P-iodophenyl)-3-(p-nitrophenyl)-5-phenyl tetrazolium chloride (INT) had on the number and size distribution of tumor colonies counted using an optical image analyzer (FAS II). Staining increased the number of tumor colonies counted. By using opaque tumor cells or pigmented melanoma cells and measuring colony growth kinetics, we demonstrated that the use of INT staining to assist in counting tumor colonies artificially increased the size of viable tumor cell aggregates by adding a red precipitate to the outside surface of the cells. Laboratories that are using the INT method for drug screening are probably measuring colonies down to and below 42 microns in diameter. These small colonies could result from as few as one or two divisions. Thus, potentially useful drugs may be missed in the screen because of the presence of abortive colonies: i.e., lethally damaged cells completing only one or two divisions.