Initial Colonies Research Articles

Costs of Dust Collection by Trichodesmium: Effect on Buoyancy and Toxic Metal Release

AbstractThe marine cyanobacterium Trichodesmium has the remarkable ability to interact with and utilize air‐borne dust as a nutrient source. However, dust may adversely affect Trichodesmium through buoyancy loss and exposure to toxic metals. Our study explored the effect of desert dust on buoyancy and mortality of natural Red Sea puff‐shaped Trichodesmium thiebautii. Sinking velocities and ability of individual colonies to stay afloat with increasing dust loads were studied in sedimentation chambers. Low dust loads of up to ∼400 ng per colony did not impact initial sinking velocity and colonies remained afloat in the chamber. Above this threshold, sinking velocity increased linearly with the colony dust load at a slope matching prediction based on Stoke's law. The potential toxicity of dust was assessed with regards to metal dissolution kinetics, differentiating between rapidly released metals, which may impact surface blooms, and gradually released metals that may impact dust‐centering colonies. Incubations with increasing dust concentrations revealed colony death, but the observed lethal dose far exceeded dust concentrations measured in coastal and open ocean systems. Removal of toxic particles as a mechanism to reduce toxicity was explored using SEM‐EDX imaging of colonies incubated with Cu‐minerals, yet observations did not support this pathway. Combining our current and former experiments, we suggest that in natural settings the nutritional benefits gained by Trichodesmium via dust collection outweigh the risks of buoyancy loss and toxicity. Our data and concepts feed into the growing recognition of the significance of dust for Trichodesmium's ecology and subsequently to ocean productivity.

First report of leaf spot disease caused by Alternaria alternata on Polygonatum cyrtonema Hua in Hunan Province of China.

Polygonatum cyrtonema Hua, a perennial plant of the Asparagaceae family, is an important herb in Chinese medicine and is mainly grown in the Chinese provinces of Guizhou, Hunan, Yunnan, Anhui, and Zhejiang (Chen et al. 2021). In June 2021, a new case of leaf spot disease was detected in an 80 m2 plantation of P. cyrtonema on Xuefeng Mountain, Huaihua City, Hunan Province (27°17'30″N, 110°24'20″E). It infected almost 40% of the total planted area. Initially, irregular light brown spots appeared on the leaves, gradually turning dark brown and coalescing to form large necrotic areas, after which the affected plant turned yellow and eventually died. Ten disease samples were collected from ten plants in the plantation area. The leading edge of necrotic tissues were rinsed with sterile water and then disinfected with 3% hydrogen peroxide for 30 s, followed by 75% ethanol for 90 s, and rinsed three times with sterile water. Samples were then placed on water agar plates and incubated in the dark in a constant temperature incubator at 28 ℃ for 3-5 days. After mycelial growth was observed in the media, the hyphae were transferred to potato dextrose agar plates and incubated for 3-5 days at 28 ℃ in the dark. Ultimately, 12 purified fungal isolates were obtained, some of which were morphologically similar, including 10 that were Alternaria (83.3% isolation rate). Three representative isolates (HJYB1, HJYB2, and HJYB3) were selected for further study. The initial colonies were grayish green with white fluffy mycelia on the surface and a prominent white rim, which became brown with dense, cottony aerial mycelia as the colonies matured. The conidia were obpyriform or ellipsoidal, pale to dark brown, with 0-4 transverse and 0-3 longitudinal septa, some with a short cylindrical beak at the tip. They measured 11.826-28.873 × 6.231-26.018 μm (n = 100). To further confirm the identity of the isolates, their rDNA internal transcribed spacer region (ITS), β-microtubulin (TUB2) and translation elongation factor-1 (TEF-1) genes were amplified and sequenced using the ITS4/ITS5, TUB2F/R and EF-526F/1567R primers, respectively (Hong et al. 2006). The sequences were submitted to GenBank (ITS: OR513924, OR513964, OR519874; TUB2: OR526928, OR533421, OR526929; TEF: OR526926, OR533420, OR526927). A concatenated phylogenetic tree of the three genes showed that the isolate clustered significantly with Alternaria alternata. Based on morphological identification and phylogenetic tree analysis, the isolate was identified as A. alternata. We carried out pathogenicity tests on four uniformly growing P. cyrtonema plants. Three of these plants were used as experimental plants and one as a control. For each plant, three young leaves were selected and inoculated with 6 × 6 mm PDA blocks, while sterile PDA blocks were used as controls. The treated plants were subjected to 10 days of stable temperature in a climatic chamber set at 28°C, 80% constant relative humidity and 12 hours of light per day. The pathogenic lesions appeared and the pathogens re-isolated from the diseased leaves showed similar morphological characteristics to representative isolates and were confirmed as A. alternata by DNA sequencing, thus fulfilling Koch's postulates. A. alternata is the major causal agent of leaf spot on P. sibiricum (Zou et al. 2023) and Agrimonia pilosa (Jiang et al. 2023). As far as we know, leaf necrosis caused by A. tenuissima has been found on P. cyrtonema (Li et al. 2020). To our knowledge, this is the first report of A. alternata causing leaf spot disease in P. cyrtonema. These findings form the basis for the management of this leaf spot disease.

A live-cell image-based machine learning strategy for reducing variability in PSC differentiation systems

The differentiation of pluripotent stem cells (PSCs) into diverse functional cell types provides a promising solution to support drug discovery, disease modeling, and regenerative medicine. However, functional cell differentiation is currently limited by the substantial line-to-line and batch-to-batch variabilities, which severely impede the progress of scientific research and the manufacturing of cell products. For instance, PSC-to-cardiomyocyte (CM) differentiation is vulnerable to inappropriate doses of CHIR99021 (CHIR) that are applied in the initial stage of mesoderm differentiation. Here, by harnessing live-cell bright-field imaging and machine learning (ML), we realize real-time cell recognition in the entire differentiation process, e.g., CMs, cardiac progenitor cells (CPCs), PSC clones, and even misdifferentiated cells. This enables non-invasive prediction of differentiation efficiency, purification of ML-recognized CMs and CPCs for reducing cell contamination, early assessment of the CHIR dose for correcting the misdifferentiation trajectory, and evaluation of initial PSC colonies for controlling the start point of differentiation, all of which provide a more invulnerable differentiation method with resistance to variability. Moreover, with the established ML models as a readout for the chemical screen, we identify a CDK8 inhibitor that can further improve the cell resistance to the overdose of CHIR. Together, this study indicates that artificial intelligence is able to guide and iteratively optimize PSC differentiation to achieve consistently high efficiency across cell lines and batches, providing a better understanding and rational modulation of the differentiation process for functional cell manufacturing in biomedical applications.

A SERIES OF SIMPLE DECONTAMINATION METHODS OF BACTERIAL FLORA FOUND ON MUSICAL WIND INSTRUMENTS

Objective: This study was aimed to compare the efficacy of cleaning techniques using hot water treatment soap containing 2% triclosan and chemical antiseptics in reducing the bacterial contamination observed on shared musical wind instruments.
 Methods: The trumpet, mellophone, trombone, and tuba were evaluated in this study. To count the initial bacterial colonies on the instrument, the total amount of bacteria adhered to it was extracted using the swab procedure. The mouthpieces were immersed in hot water at a temperature of 100 °C for 5 min and then were soaked in soap that contained 2% triclosan to achieve the effect of decontamination. Then the survival colonies were counted. As a series of decontamination technique, this study also examined the disinfection ability of phenol, chloroxylenol, povidone-iodine, and 70% alcohol utilizing the Rideal Walker method.
 Results: When compared to liquid soap (50.30-91.67%), the cleaning procedure that uses immersion in hot water of 100 °C for 5 min greatly lowers the quantity of bacteria (91.85-99.91%). However, due to their huge surface area, tuba mouthpieces were the most straightforward to sterilize using both techniques. The highest phenol coefficient value was shown by chloroxylenol; however, all tested disinfectants showed stronger antibacterial activity than 1% phenol.
 Conclusion: The mouthpieces of shared wind instruments can be cleaned quickly, easily, and effectively by immersing them in hot water at a temperature of 100 °C for 5 min. However, chloroxylenol has the strongest ability to eradicate bacteria from the instrument's mouthpiece.

First Report of Paramyrothecium breviseta Causing Leaf Spot Disease of Coffea canephora in China.

Coffee is a tropical plant with two widely cultivated species, namely Coffea arabica and Coffea canephora. A leaf spot disease causing brownish and necrotic lesions was broken out on the C. canephora coffee seedlings in a nursery in Ruili County, Yunnan Province, China, during 2018 to 2019. The incidence of the disease was 15% ~ 20%. Ten diseased leaf samples from five diseased plants were collected for pathogen isolation by tissue separation method. Leaf pieces were cut from the margin of the necrotic lesions (4 × 6 mm), surface-sterilized for 30 s in 75% ethanol, followed by 0.1% arsenic mercury solution for 15 s, then washed 3~4 times with sterilized distilled water and transferred onto potato dextrose agar (PDA) medium in petri plates. Four morphologically similar isolates were obtained from lesions and cultivated on PDA at 25°C. Initial colonies of isolates were round, neat edge, white, floccose mycelium and developed dark green-to-black concentric rings that were sporodochia bearing viscid spore masses after 5~7 days. Conidia were acetates, hyaline and cylindrical with both rounded ends and 4.8 to 6.4 µm long × 1.6 to 2.6 µm wide. Koch's test were conducted on three healthy plants leaves of original source variety C. canephora No.2 and C.arabica Catimor CIFC7963 (control plants) with spore suspension (1 × 106/mL), respectively. Meanwhile, equal numbers of healthy plants were inoculated with water as controls. After inoculation, the plants were transferred into an incubator at 25℃ with saturated humidity. After 10 days of inoculation, all the tested plants presented similar typical symptoms with the diseased leaves under natural conditions; whereas the controls remained healthy. Koch's postulates were performed by re-isolating the fungus from the inoculated leaves and verifying its colony and morphological characters. Two single spore isolates cultured on PDA medium were selected for DNA extraction. The ribosomal internal transcribed spacer (ITS) was PCR amplified by using primers ITS1 and ITS4 (White et al., 1990), β-tubulin gene by Bt2a and Bt2b (Glass and Donaldson, 1995), the RNA polymerase II second largest subunit (rpb2) by RPB2-5F2 and RPB2-7cR (O'Donnell et al, 2007), calmodulin (cmda) gene by CAL-228F and CAL2Rd (Groenewald et al., 2013). The sequences of ITS (MT853067 ~ MT853068), β-tubulin (MT897899 ~ MT897900), rpb2 (MW256264~ MW286265) and cmda (MT897897~ MT897898) were deposited in GenBank databases. BLAST analysis revealed that the representative isolates sequences shared 99.31%~99.65% similarities to the ITS sequence of Paramyrothecium breviseta (Accession Nos. NR_155670.1), 99.43% similarities to the β-tubulin sequence of P. breviseta (Accession Nos. KU846406.1), 98.98% similarities to the rpb2 sequence of P. breviseta (Accession Nos. KU846351.1), and 98.54%~98.71% similarities to the cmda sequence of P. breviseta (Accession Nos. KU846262.1). As it shown in the phylogenetic tree derived from combined ITS, β-tubulin, rpb2, and cmda gene sequences, the two representative isolates were clustered together with P. breviseta CBS 544.75 with 98% strong bootstrap support, which confirmed that P. breviseta is the causal agent of leaf spot of Coffea canephora. To our knowledge, this is the first report of a leaf spot disease caused by P. breviseta on C. canephora in China, which raised the caution that P. breviseta is also pathogenic to Coffea Arabica.

Poly(lactic-co-glycolic) Acid-Lipid Hybrid Microparticles Enhance the Intracellular Uptake and Antibacterial Activity of Rifampicin.

An urgent demand exists for the development of effective carrier systems that systematically enhance the cellular uptake and localization of antibiotic drugs for the treatment of intracellular pathogens. Commercially available antibiotics suffer from poor cellular penetration, restricting their efficacy against pathogens hosted and protected within phagocytic cells. In this study, the potency of the antibiotic rifampicin against intracellular small colony variants of Staphylococcus aureus was improved through encapsulation within a strategically engineered cell-penetrant delivery system, composed of lipid nanoparticles encapsulated within a poly(lactic-co-glycolic) acid (PLGA) nanoparticle matrix. PLGA-lipid hybrid (PLH) microparticles were synthesized through the process of spray drying, whereby rifampicin was loaded within both the polymer and lipid phases, to create a nanoparticle-in-microparticle system capable of efficient redispersion in aqueous biorelevant media and with programmable release kinetics. The ability of PLH particles to disintegrate into nanoscale agglomerates of the precursor nanoparticles was shown to be instrumental in optimizing rifampicin uptake in RAW264.7 macrophages, with a 7.2- and 1.6-fold increase in cellular uptake, when compared to the pure drug and PLGA microparticles (of an equivalent initial particle size), respectively. The enhanced phagocytosis and extended drug release mechanism (under the acidic macrophage environment) associated with PLH particles induced a 2.5-log reduction in colony forming units compared to initial colonies at 2.50 μg/mL rifampicin dose. Thus, the ability of PLH particles to reduce the intracellular viability of S. aureus, without demonstrating significant cellular toxicity, satisfies the requirements necessary for the safe and efficacious delivery of antibiotics to macrophages for the treatment of intracellular infections.

Abstract 5095: Concurrent Dnmt3a haploinsufficiency partially abrogates competitive disadvantage in Smc3 haploinsufficient myeloid cells

Abstract SMC3 encodes a subunit of cohesin associated with recurrent heterozygous mutations in acute myeloid leukemia (AML) and other myeloid malignancies. About one-third of these mutations are predicted loss-of-function (nonsense and splice-site mutations) and the rest are missense with unclear function. To understand whether these missense mutations might have dominant-negative effects or phenocopy loss-of-function effects, we compared the consequences of Smc3 deficient and haploinsufficient mouse models. We found that both embryonic and somatic deletion of homozygous Smc3 alleles led to complete hematopoietic failure. Furthermore, in competitive transplantation, following engraftment and subsequent tamoxifen treatment, the Smc3fl/fl/ERT2-Cre+/- BM cells were rapidly outcompeted, with earliest cell loss in the Gr1+ myeloid compartment. Hence, Smc3 is indispensable for hematopoiesis and AML-associated SMC3 missense mutations are unlikely to have dominant-negative effects. We examined the consequences of somatic Smc3 haploinsufficiency using Smc3fl/+/ERT2-Cre+/-. We found that Smc3 haploinsufficiency did not lead to increased number of initial methylcellulose colonies, nor did these cells replate beyond two weeks. In competitive transplantations, we again observed a significant competitive disadvantage in the Smc3fl/+/ERT2-Cre+/- BM cells, most pronounced in the Gr1+ myeloid cells. Competitive disadvantage is counter-intuitive for a leukemia-associated mutation. SMC3 mutations frequently co-occur with DNMT3A mutations. We, therefore, asked whether Smc3 haploinsufficiency might lead to a competitive advantage if it occurred in the background of Dnmt3a haploinsufficiency. With the addition of Dnmt3a haploinsufficiency, the severe myeloid competitive disadvantage was partially ameliorated, but the significant competitive disadvantage in other lineages remained intact. In summary, our data demonstrate that homozygous Smc3 deletion was incompatible with embryonic or adult hematopoiesis, with the greatest defects noted within myeloid compartment, indicating that AML-associated SMC3 mutations are unlikely to have dominant-negative effects. Smc3 haploinsufficiency resulted in a myeloid competitive disadvantage, which was partially abrogated in the presence of concurrent Dnmt3a haploinsufficiency, suggesting lineage-specific interactions between the mutations. Citation Format: Tianjiao Wang, John S. Welch. Concurrent Dnmt3a haploinsufficiency partially abrogates competitive disadvantage in Smc3 haploinsufficient myeloid cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5095.

A growing microcolony can survive and support persistent propagation of virulent phages

Bacteria form colonies and secrete extracellular polymeric substances that surround the individual cells. These spatial structures are often associated with collaboration and quorum sensing between the bacteria. Here we investigate the mutual protection provided by spherical growth of a monoclonal colony during exposure to phages that proliferate on its surface. As a proof of concept we exposed growing colonies of Escherichia coli to a virulent mutant of phage P1. When the colony consists of less than [Formula: see text]50,000 members it is eliminated, while larger initial colonies allow long-term survival of both phage-resistant mutants and, importantly, colonies of mostly phage-sensitive members. A mathematical model predicts that colonies formed solely by phage-sensitive bacteria can survive because the growth of bacteria throughout the colony exceeds the killing of bacteria on the surface and pinpoints how the critical colony size depends on key parameters in the phage infection cycle.

Initial clonogenic potential of human endothelial progenitor cells is predictive of their further properties and establishes a functional hierarchy related to immaturity

Endothelial progenitor cells (EPCs) generate in vitro Endothelial Colony Forming Cells (ECFCs) combining features of endothelial and stem/progenitor cells. Their angiogenic properties confer them a therapeutic potential for treating ischemic lesions. They may be isolated from umbilical cord blood (CB-ECFCs) or peripheral adult blood (AB-ECFCs). It is generally accepted that CB-ECFCs are more clonogenic, proliferative and angiogenic than AB-ECFCs. Nevertheless, only a few studies have focused on the functional heterogeneity of CB-ECFCs from different individuals. Moreover, AB-ECFC loss of function is yet to be precisely described. We have focused on these two issues that are critical for clinical perspectives.The detailed clonogenic profile of CB-ECFCs and AB-ECFCs was obtained and revealed a high inter individual heterogeneity and the absence of correlation with age. Most CB-ECFCs yielded initial colonies and had functional properties similar to those of AB-ECFCs. Conversely, a high clonogenicity was associated with an enhanced proliferative and angiogenic potential and stemness gene overexpression, confirming that immaturity, lost by AB-ECFCs, was a prerequisite to functionality. We thus demonstrated the importance of selecting CB-ECFCs according to specific criteria, and we propose using the initial clonogenicity as a relevant marker of their potential efficacy on vascular repair.

Cooperation and profit allocation in two-echelon logistics joint distribution network optimization

Collaborative two-echelon logistics joint distribution network can be organized through a negotiation process via logistics service providers or participants existing in the logistics system, which can effectively reduce the crisscross transportation phenomenon and improve the efficiency of the urban freight transportation system. This study establishes a linear optimization model to minimize the total cost of two-echelon logistics joint distribution network. An improved ant colony optimization algorithm integrated with genetic algorithm is presented to serve customer clustering units and resolve the model formulation by assigning logistics facilities. A two-dimensional colony encoding method is adopted to generate the initial ant colonies. Improved ant colony optimization combines the merits of ant colony optimization algorithm and genetic algorithm with both global and local search capabilities. Finally, an improved Shapley value model based on cooperative game theory and a cooperative mechanism strategy are presented to obtain the optimal profit allocation scheme and sequential coalitions respectively in two-echelon logistics joint distribution network. An empirical study in Guiyang City, China, reveals that the improved ant colony optimization algorithm is superior to the other three methods in terms of the total cost. The improved Shapley value model and monotonic path selection strategy are applied to calculate the best sequential coalition selection strategy. The proposed cooperation and profit allocation approaches provide an effective paradigm for logistics companies to share benefit, achieve win–win situations through the horizontal cooperation, and improve the negotiation power for logistics network optimization.

Differences of Streptococcus mutans adhesion between artificial mouth systems: a dinamic and static methods

Background: Various materials have been used for treating dental caries. Dental caries is a disease that attacks hard tissues of the teeth. The initial phase of caries is a formation of bacterial biofilm, called as dental plaque. Dental restorative materials are expected for preventing secondary caries formation initiated by dental plaque. Initial bacterial adhesion is assumed to be an important stage of dental plaque formation. Bacteria that recognize the receptor for binding to the pellicle on tooth surface are known as initial bacterial colonies. One of the bacteria that plays a role in the early stage of dental plaque formation is Streptococcus mutans (S. mutans). Artificial mouth system (AMS) used in bacterial biofilm research on the oral cavity provides the real condition of oral cavity and continous and intermittent supply of nutrients for bacteria. Purpose: This study aimed to compare the profile of S. mutans bacterial adhesion as the primary etiologic agent for dental caries between using static method and using artificial mouth system, a dinamic. method (AMS). Method: The study was conducted at Faculty of Dentistry and Integrated Research and testing laboratory (LPPT) in Universitas Gadjah Mada from April to August 2015. Composite resin was used as the subject of this research. Twelve composite resins with a diameter of 5 mm and a width of 2 mm were divided into two groups, namely group using static method and group using dynamic method. Static method was performed by submerging the samples into a 100µl suspension of 1.5 x 108 CFU/ml S. mutans and 200µl BHI broth. Meanwhile AMS method was carried out by placing the samples at the AMS tube drained with 20 drops/minute of bacterial suspension and sterile aquadest. After 72 hours, five samples from each group were calculated for their biofilm mass using 1% crystal violet and read by a spectrofotometer with a wavelength of 570 nm. Meanwhile, one sample from each group was taken for its surface image using Scanning Electron Microscope (SEM). Result: The results showed that S. mutans biofilm mass in the group using static method was 0.34, while in the group using AMS method was 0.09. The results of the statistical analysis then showed that there was a significant difference (p=0.02) in the formation of bacterial biofilm mass between those groups. SEM image in the group using static method also showed that the attachment of S. mutans was more numerous and had a longer chain than in the group using AMS method. Conclusion: There is a difference in the profile of S. mutans bacterial adhesion between using AMS method and static method.

Post-Fire Effect of Savannah Vegetation on the Establishment of New Colonies ofAtta sexdens rubropilosa(Hymenoptera: Formicidae)

Establishing their initial colony is probably the most critical moment in the life of leaf-cutting ants. The non-establishment is connected to abiotic and biotic factors, and the high mortality rates of initial colonies are possibly associated with entomopathogenic or antagonistic microorganisms to the symbiotic fungus present in the soil, hosted by these ants. Fire in the vegetation, depending on the intensity, is known to cause significant changes to the soil physical, chemical, and microbiological properties. The impact of a fire in savannah vegetation (Cerrado) on the establishment of early colonies of Atta sexdens rubropilosa Forel (Hymenoptera: Formicidae) was evaluated. For this end, two areas were selected, one where there had been an accidental fire, and a contiguous one with the same size and vegetation characteristics without burning. In these areas and in soil collected in the same areas and stored in the laboratory, females recently fertilized in the nuptial flight were placed to excavate the soil and establish their colonies. Post-fire changes in the soil chemical and microbiological properties were quantified and correlated successfully in the establishment of new colonies of this leaf-cutting ant. Under field conditions, the females of A. sexdens rubropilosa did not show preference for which areas to excavate: the ones that had been burned or the ones that were unburned; under this condition, no colony survived according to the evaluation performed 120 d after the nuptial flight. Under laboratory conditions, the majority of the females excavated the soil, whether it had been burned or not. However, the establishment of initial colonies was significantly higher in soils collected far from the surface and in areas that had not directly been affected by the fire, showing a negative effect of fire on colony establishment under laboratory conditions. ResumoEstabelecer sua colonia inicial provavelmente seja o momento mais critico da vida das formigas-cortadeiras. O nao-estabelecimento esta ligado a fatores abioticos e bioticos e, possivelmente, as altas taxas de mortalidade de colonias iniciais estejam associadas aos microrganismos entomopatogenicos ou antagonicos ao fungo simbionte cultivado por essas formigas presentes no solo. Sabe-se que o fogo na vegetacao, dependendo de sua intensidade, provoca alteracoes significativas nas propriedades fisicas, quimicas e microbiologicas do solo. Assim, neste trabalho avaliou-se o impacto de um incendio em vegetacao de savana (cerradao) no estabelecimento de colonias iniciais de Atta sexdens rubropilosa Forel (Hymenoptera: Formicidae). Para isso, foram selecionadas areas onde ocorreu uma queima acidental e outra, de mesmo tamanho, com mesmas caracteristicas vegetacionais e contigua sem queima. Nessas areas e em solo coletados nessas mesmas areas e acondicionados em laboratorio foram colocadas femeas recem copuladas no dia da revoada para perfurarem o solo e fundarem suas colonias. Alteracoes nas propriedades quimicas e microbiologicas do solo pos-fogo foram quantificadas e correlacionadas com sucesso no estabelecimento de novas colonias dessa formiga-cortadeira. Em condicoes de campo, as femeas de A. sexdens rubropilosa nao apresentaram preferencia para perfurar o solo de areas queimadas ou nao e, nessa condicao, nenhuma colonia sobreviveu, em avaliacao realizada 120 dias depois da revoada. Em condicoes de laboratorio, a maioria das femeas perfurou o solo, independentemente de o solo onde elas foram acondicionadas ter sido queimado ou nao. Entretanto, o estabelecimento inicial de colonias foi significativamente maior em solos coletados distantes da superficie e em areas que nao sofreram acao direta do fogo, o que mostrou efeito negativo do fogo no estabelecimento de colonias no laboratorio. View this article in BioOne

Social structure of the polygynous ant, Crematogaster osakensis

The colony structures of social insects vary greatly among species. In ant societies, the number of queens within a colony is variable during colony maturation. We investigated the social structure of a polygynous ant Crematogaster osakensis in a series of field and laboratory experiments. First, the initial colonies headed by multiple queens were found in the field. In laboratory experiments, queens that were artificially cohabited after their nuptial flight harmoniously co-existed even in the presence of newly emerged workers, suggesting that mated queens of this species can establish their colony cooperatively as primary polygynous colonies. In addition, the mating frequency of a queen was typically more than one, estimated from the sperm number stored in field-collected males and queens, and from genetic relatedness among daughters of lab-reared monogynous colonies. Second, our assessment of genetic relatedness in a mature field colony of this species revealed that dealated queens, as well as workers and alates, were relatives. The number of developed oocytes identified reproductive skew in two of five field-collected nests. Moreover, under laboratory conditions, the most fertile egg layer altered over 3 months of observation. Based on these lines of evidence, we propose that in C. osakensis, polygynous foundresses might either be unrelated and subsequently be replaced by daughter queens of particular foundresses, or be related in the first place.

Initial development and production of CO2 in colonies of the leaf-cutting ant Atta sexdens during the claustral foundation

Queens in the genus Atta are solely responsible for fungus cultivation and for care of herself and her offspring. Only few studies have investigated their nests in the claustral foundation and it is unknown the production rate of expelled carbon dioxide and/or of oxygen consumption in the initial colonies of leaf-cutting ants. Thus, we have studied the development of 50 initial colonies of Atta sexdens, and production of expelled carbon dioxide under laboratory conditions. The number of eggs was counted one week after nest foundation on the seventh day, the larvae counted on day 28, and the pupae between days 42 and 49. The workers emerged on the 63rd and 70th day. The CO2 concentration increased steeply in the 42nd days (20.60 ± 8.36%) and 49th days (15,37 ± 13,11 %), at 42nd days, and subsequently returned to lower values, for example, 3.35±2.84% at week seven. The present study is the first to present CO2 emission data in initial nests, in their claustral foundation under laboratory conditions.

Variation in honey yield per hive of Africanized bees depending on the introducing time of young queens

ABSTRACT: The objective of this research was to evaluate the honey production per hive and the egg laying rates of queens produced in 2007, 2008 and 2010. Thirty colonies initiated with a queen per colony at each climatic season were used during the three years. The years, started on January (summer), April (autumn), July (winter) and October (spring) and ended 12 months later, at the same periods related to each season of the later years. Honey supply were weighed before and after centrifugation to evaluate the quantity of the stored honey. Colonies with queens introduced during autumn and winter in the three years produced 57.2±6.0kg and 60.7±7.5kg of honey, respectively. In the first year of production activity, after the introduction of queens in the initial colonies, values were significantly higher than those obtained in colonies with queens introduced in the summer (39.3±7.6kg) and spring (41.8±3.7kg). Egg laying rates of queens were higher in spring (98.2±3.9%) and summer (88.4±7%), indicating greater food flow (flowerings) in these seasons compared to the averages in autumn (30.3±8.1%) and winter (24.5±7.2%). Produce and introduce queens of Africanized Apis mellifera in colonies initiated during autumn and winter was found to be economically feasible. Honey production of colonies initiated in these periods were higher and they had greater population stability in times of scarcity of flowerings.

Improvement of urban lake water quality by removal of Escherichia coli through the action of the bivalve Anodonta californiensis.

High levels of fecal indicator bacteria, such as Escherichia coli, can be indicative of poor water quality. The use of shellfish to reduce eutrophication has been proposed, but application of bivalves to reduce bacterial levels has not been extensively reported. Removal of E. coli by the native freshwater mussel Anodonta californiensis was studied using laboratory batch systems and field-based flow-through systems. Batch systems were utilized to determine the fate and inactivation of E. coli after uptake by the mussel. Batch experiments demonstrated that uptake patterns followed first order kinetics and E. coli was inactivated with less than 5% of the initial colonies recoverable in fecal matter or tissue. Flow-through systems located at an urban impaired lake in San Francisco, CA were utilized to determine uptake kinetics under environmentally relevant conditions. The bivalves maintained a 1-log removal of E. coli for the duration of exposure. The calculated uptake rates can be used in conjunction with hydrologic models to determine the number of bivalves needed to maintain removal of E. coli in different freshwater systems. The outcomes of this study support the use of native freshwater bivalves to achieve the co-benefits of rehabilitating a freshwater ecosystem and improving water quality via reduction of E. coli in contaminated freshwater systems.

In Vitro Toxicity Test of Strichnos johnsonnii (Loganiaceae) on a Strain of Staphylococcus aureus

The aim of this study was to test the in vitro toxicity of an extract of Strychnos johnsonii (Loganiaceae) on a strain of Staphylococcus aureus. The tests were performed in the bacteriology unit of the Douala General Hospital biology laboratory, dealing with an extract from the bark of stem of Strychnos johnsonii (Loganiaceae) harvested at Etome village, South West Cameroon and authenticated by a botanist. The plant extract was obtained by maceration in 300 mL of ethanol for 120 hours. The filtrate obtained was evaporated under vacuum, at 50° C, 250 mbar of pressure and at a speed of 125 rounds per minute. The residual solvent was eliminated in an incubator at 37° C for one week to give dry extract. Selected bacterial strain came from pus collected from an in-patient. By its biochemical and enzymatic characters, this strain showed 90.9% homology with the Staphylococcus aureus ATCC 29213-reference strain. No bacterial growth were observed on Mannitol Salt, EMB and Sabouraud-Chloramphenicol agar plates after 48 hours of incubation, evidence that the extract contained no germs before the test. The number of initial colonies for the time t0 averaged 225. The point of intersection between the inhibition curve and the x-axis as the MIC corresponds to 0,04 g/mL. The smallest concentration of the extract for which the growth of Staphylococcus aureus is zero on the Mannitol salt agar was 0.04 g/mL. Therefore, the MIC amounts to MBC. The results obtained showed a bactericidal effect, which could be, attributed to the presence of indole alkaloids in the plant.

First Report of Myrothecium roridum Causing Leaf Spot on Zantedeschia aethiopica in China

Zantedeschia aethiopica (L.) Spreng. (calla lily), belonging to family Araceae, is a popular ornamental plant in China. In the summer of 2010, leaves of calla lily with typical symptoms of necrotic lesions were observed in a commercial glasshouse in Beijing, China (116°20' E, 39°44' N). The initial symptoms were circular to subcircular, 1 to 3 mm, and dark brown lesions on the leaf lamina. Under high humidity, lesions expanded rapidly to 5 to 10 mm with distinct concentric zones and produced black sporodochia, especially on the backs of leaves. Later, the infected leaves were developing a combination of leaf lesions, yellowing, and falling off; as a result, the aesthetic value of the plant was significantly impacted. Leaf samples were used in pathogen isolation. Symptomatic leaf tissues were cut into small pieces and surface sterilized with 70% ethanol for 30 s and then in 0.1% mercuric chloride solution for 1 to 3 min. After being washed in sterile distilled water three times, the pieces were plated on potato dextrose agar (PDA) and incubated at 25°C in darkness for 7 days (5). Initial colonies of isolates were white, floccose mycelium and developed dark green to black concentric rings that were sporodochia bearing viscid spore masses after incubating 5 days. Conidiophores branched repeatedly. Conidiogenous cells were hyaline, clavate, and 10.0 to 16.0 × 1.4 to 2.0 μm. Conidia were hyaline, cylindrical, both rounded ends, and 6.0 to 8.2 × 1.9 to 2.4 μm. Morphological characteristics of the fungus were consistent with the description of Myrothecium roridum Tode ex Fr. (3,4). To confirm the pathogenicity, three healthy plants of calla lily were inoculated with a conidial suspension (1 × 106 conidia per ml) brushed from a 7-day-old culture of the fungus. Control plants were sprayed with sterile water. The inoculated plants were individual with clear plastic bags and placed in a glass cabinet at 25°C. After 7 days, all inoculated leaves developed symptoms similar to the original samples, but control plants remained disease free. Re-isolation and identification confirmed Koch's postulates. For molecular identification, genomic DNA of a representative isolate (MTL07081001) was extracted by modified CTAB method (1), and the rDNA-ITS region was amplified by using primers ITS1 (5-TCCGTAGGTGAACCTGCGG-3) and ITS4 (5-TCCTCCGCTTATTGATATGC-3). The 465-bp amplicon (GenBank Accession No. KF761293) was 100% identity to the sequence of M. roridum (JF724158.1) from GenBank. M. roridum has an extensive host range, covering 294 host plants (2). To our knowledge, this is the first record of leaf spot caused by M. roridum on calla lily in China.

A Critical View of Colony Losses in Managed Mayan Honey-Making Bees (Apidae: Meliponini) in the Heart of Zona Maya

This research considered native Mayan stingless bees, Melipona beecheii, with special attention to decrease in their managed colonies. From a total of 155 beekeepers located in 60 communities, 58 were randomly selected to survey in 2011. Their experience ranged from less than one to 50 years, and initial colonies from one to 100. Both structured and open interviews were conducted. Participants generally reported they believed bees were obtaining less food, which could produce colony loss. The present and a previous survey in the Zona Maya show colony loss averages 4–5% each year. In this study, during an average of 10 years, 27 beekeepers lost none, 9 lost all, and the remainder lost 44% of their colonies. Further analysis revealed colony loss had no association with relative habitat disturbance, presumed Africanized honey bee abundance, or beekeeping experience. However, those initially with more colonies in a meliponary lost them at a greater rate, indicating competition for food. Initial colony number was near 11, but currently is near 4 per meliponary. Little colony propagation (husbandry) was the norm until recently, when initiatives including meliponiculture workshops stimulated more husbandry. Twenty-six percent of beekeepers had less than one year experience and they began meliponaries using wild colonies. Because established meliponicultors were found to very seldom rely on new wild colonies, increased husbandry efforts are necessary to offset natural mortality of managed colonies. Five meliponicultors increased their colonies over 300% in two years (40 to 123 colonies), whereas a 34% loss in nine years (480 to 206 colonies) was found among the individuals randomly surveyed.

To clone or not to clone? Induced pluripotent stem cells can be generated in bulk culture.

Induced pluripotent stem cells (iPSCs) are usually clonally derived. The selection of fully reprogrammed cells generally involves picking of individual colonies with morphology similar to embryonic stem cells (ESCs). Given that fully reprogrammed cells are highly proliferative and escape from cellular senescence, it is conceivable that they outgrow non-pluripotent and partially reprogrammed cells during culture expansion without the need of clonal selection. In this study, we have reprogrammed human dermal fibroblasts (HDFs) with episomal plasmid vectors. Colony frequency was higher and size was larger when using murine embryonic fibroblasts (MEFs) as stromal support instead of HDFs or human mesenchymal stromal cells (MSCs). We have then compared iPSCs which were either clonally derived by manual selection of a single colony, or derived from bulk-cultures of all initial colonies. After few passages their morphology, expression of pluripotency markers, and gene expression profiles did not reveal any significant differences. Furthermore, clonally-derived and bulk-cultured iPSCs revealed similar in vitro differentiation potential towards the three germ layers. Therefore, manual selection of individual colonies does not appear to be necessary for the generation of iPSCs – this is of relevance for standardization and automation of cell culture procedures.