Internal ultrastructure of the planktonic larva of the coral Porites panamensis (Anthozoa: Scleractinia) / Ultraestructura interna de la larva planctónica del coral Porites panamensis (Anthozoa: Scleractinia)

Abstract Introduction: The scleractinian coral life cycle includes planktonic larvae that settle on the benthos, allowing the primary polyp to clone and build a sexually reproducing adult colony. The larval physiology and ecology of Eastern Tropical Pacific scleractinians needs the exploration of basic aspects such as the internal morphology of planulae. Objective: To describe histological and cytological characteristics of Porites panamensis larvae. Methods: During August-July 2019, at Islas Marias Biosphere Reserve, Central Mexican Pacific, we made 14 collections of coral larvae and identified the species with cytochrome oxidase subunit 1 gene. We used a scanning electron microscope and other techniques. Results: The ectoderm was composed by heterogeneous, mono-ciliated, columnar epithelial cells. Nematocysts were clustered at the oral pole of the ectoderm, and cells were evident in the aboral pole of the ectoderm gland. The endoderm had secretory cells, lipids and symbionts. Conclusions: The abundance of secretory cells and nematocysts in the aboral pole suggests their importance in substrate exploration and larval settlement. Our results support previous descriptions of larval ultrastructure in other coral species.

Resumen Introducción: El ciclo de vida del coral escleractinio incluye larvas planctónicas que se asientan en el bentos, lo que permite que el pólipo primario se clone y construya una colonia de adultos con reproducción sexual. La fisiología y ecología larvaria de los escleractinios del Pacífico Tropical Oriental necesita la exploración de aspectos básicos como la morfología interna de las plánulas. Objetivo: Describir las características histológicas y citológicas de las larvas de Porites panamensis. Métodos: Durante agosto-julio 2019, en la Reserva de la Biosfera Islas Marías, Pacífico Central Mexicano, realizamos 14 recolectas de larvas de coral e identificamos las especies con el gen citocromo oxidasa subunidad 1. Utilizamos un microscopio electrónico de barrido y otras técnicas. Resultados: El ectodermo está compuesto por células epiteliales columnares heterogéneas, monociliadas. Los nematocistos se agrupan en el polo oral del ectodermo, mientras que en el polo aboral son visibles células glandulares. El endodermo presentó células secretoras, lípidos y simbiontes. Conclusiones: La abundancia de células secretoras y nematocistos en el polo aboral sugiere su importancia en la exploración del sustrato y asentamiento larvario. Nuestros resultados respaldan las descripciones previas de la ultraestructura de las larvas en otras especies de coral.

Spatio-temporal variability in the Cladocera assemblage of a subtropical hypersaline lagoon / Variabilidade espaço-temporal da assembleia de Cladocera de uma lagoa hipersalina subtropical

Cladocera represent an important zooplankton group because of their seasonal prominence in terms of abundance and their contribution in controlling primary production (phytoplankton). On a global scale, there are few studies on Cladocera in hypersaline environments. The present work aims to evaluate the spatio-temporal variation of the Cladocera assemblage across a salinity gradient in the habitats of the Araruama Lagoon. Samples were collected in random months over a period of four years at 12 fixed stations in the Araruama Lagoon using a WP2 plankton net equipped with a flow meter. Our results do not reveal significant influence of the tide and seasonal variation as factors affecting the Cladocera assemblage. Five Cladocera species were found in the Araruama Lagoon, only in stations 11 and 12 where they reached an average of 1,799 ± 3,103 ind. m-3. The mean of the Shannon Diversity Index was 0.45 ± 0.2. The species that stood out in terms of frequency and abundance were: Penilia avirostris (frequency of occurrence: 71%), followed by Pseudevadne tergestina (41%). The same species also stood out in terms of relative abundance, Penilia avirostris (87%) and Pseudevadne tergestina (11%). The absence of Cladocera in the innermost parts of the lagoon suggests that their entrance to these locations is possibly inhibited by the salinity and temperature gradient of the lagoon, being the main factors influencing the dynamics of the Cladocera assemblages.

Os cladóceros representam um importante grupo de zooplâncton, devido ao seu destaque sazonal em termos de abundância e à sua contribuição no controlo da produção primária (fitoplâncton). Em escala global, há poucos estudos com Cladocera em ambientes hipersalinos. O presente trabalho tem como objetivo avaliar a variação espaço-temporal da assembleia de Cladocera em um gradiente de salinidades da Lagoa de Araruama. As amostras foram coletadas em meses aleatórios, durante um período de quatro anos, em 12 estações fixas na Lagoa de Araruama, utilizando uma rede de plâncton WP2 equipada com um fluxômetro. Os nossos resultados não revelam uma influência significativa da maré e da variação sazonal como fatores que afetam a assembleia de Cladocera. Cinco espécies de Cladocera foram encontradas na Lagoa de Araruama. A assembleia apresentou maior densidade absoluta estações 12 e 11, atingindo uma a média de 1.799 ± 3.103 inds-3. A média do Índice de Diversidade de Shannon foi de 0,45 ± 0,2. As espécies que se destacaram em termos de frequência e abundância foram: Penilia avirostris (frequência de ocorrência: 71%), seguida de Pseudevadne tergestina (41%). As mesmas espécies também se destacaram em termos de abundância relativa Penilia avirostris (87%) e Pseudevadne tergestina (11%). A ausência de Cladocera nas partes mais interiores da lagoa sugere que a sua entrada nestes locais é possivelmente inibida pela salinidade e gradiente de temperatura da lagoa, sendo os principais fatores que influenciam a dinâmica da assembleia de Cladocera.

Comparison of Application of MD-VF-Auto SEM Method and Plankton Gene Multiplex PCR System in the Diagnosis of Drowning / 法医学杂志

OBJECTIVES@#To compare the application effect of microwave digestion - vacuum filtration - automated scanning electron microscopy (MD-VF-Auto SEM) method and plankton gene multiplex PCR system in the diagnosis of drowning.@*METHODS@#Lung, liver and kidney tissue of 10 non-drowning cases and 50 drowning cases were prepared for further MD-VF-Auto SEM method analysis and plankton gene multiplex PCR system analysis. The positive detection rate of the two methods in each tissue was calculated.@*RESULTS@#The positive rate of the MD-VF-Auto SEM method detecting diatoms in drowning cases was 100%, and few diatoms were detected in the liver and kidney tissues of 6 non-drowning cases. By using the plankton gene multiplex PCR system, the diatom positive rate of drowning cases was 84%, and all the non-drowning cases were negative. There were significant differences in the positive rate of the liver, kidney tissues between MD-VF-Auto SEM method and plankton gene multiplex PCR system (P<0.05), as well as the total positive rate of cases. However, no significant differences were found in the positive rates of lung tissues (P>0.05).@*CONCLUSIONS@#MD-VF-Auto SEM method is more sensitive than plankton gene multiplex PCR system in diatom test. But the plankton gene multiplex PCR system can also detect plankton other than diatoms. Combination of the two methods can provide a more reliable basis for the diagnosis of drowning.

Daily vertical distribution of zooplankton in two oligo-mesotrophic north Patagonian lakes (39° S, Chile) / Distribuição vertical diária do zooplâncton em dois lagos oligo-mesotróficos ao norte da Patagônia (39º S, Chile)

Abstract The zooplankton communities often exhibit daily vertical migrations to avoid natural ultraviolet radiation and/or fish predation. However there is no information on this topic in Chilean North Patagonian lakes up to date. Therefore, this study deals with a first characterization of plankton crustacean daily vertical migration in two temperate, oligotrophic lakes (Villarrica and Panguipulli lakes, 39°S) in Southern Chile. Zooplankton were collected at different depths intervals (0-10m, 10-20 m, 20-30m, 30-40m) at early morning, middle day, evening and night in the studied site. The results revealed that zooplankton species (Daphnia pulex, Ceriodaphnia dubia, Neobosmina chilensis, Mesocyclops araucanus, and Tropocyclops prasinus) are abundant in surface zones at night, early morning and evening, whereas at middle day the zooplankton abundances are high at deep zones. The results agree with observations for Argentinean and North American lakes where these daily migration patterns in crustacean zooplankton species were reported due mainly natural ultraviolet radiation exposure, whereas for northern hemisphere lakes the vertical migration is due to combined effect of natural ultraviolet radiation and fish predation exposure.

Resumo As comunidades zooplanctônicas frequentemente exibem migrações verticais diárias para evitar a radiação ultravioleta natural e/ou a predação de peixes. No entanto, não há informações sobre esse tema em lagos chilenos no norte da Patagônia até a presente data. Portanto, este estudo trata de uma primeira caracterização da migração vertical diária de crustáceo planctônico em dois lagos temperados e oligotróficos (lagos Villarrica e Panguipulli, 39º S) no sul do Chile. O zooplâncton foi coletado em diferentes profundidades (0-10 m, 10-20 m, 20-30 m e 30-40 m) no início da manhã, ao meio-dia, à tarde e à noite no local estudado. Os resultados revelaram que as espécies de zooplâncton (Daphnia pulex, Ceriodaphnia dubia, Neobosmina chilensis, Mesocyclops araucanus e Tropocyclops prasinus) são abundantes nas zonas de superfície à noite, de manhã cedo e à tarde, enquanto, ao meio-dia, as abundâncias do zooplâncton são altas nas zonas de profundidade. Os resultados expostos corroboram as observações para outros lagos argentinos e da América do Norte, onde foram reportados esses padrões de migração diária em espécies crustáceas de zooplâncton por causa, principalmente, da exposição à radiação ultravioleta natural, enquanto, para lagos do hemisfério norte, a migração vertical se dá em razão do efeito combinado da radiação ultravioleta natural e exposição à predação.

Profile of host cell responses to exposure to stressed bacteria in planktonic; dislodged, and intact biofilm mode

Abstract The host defense response to microbial challenge emerging from the root canal system leads to apical periodontitis. The aim of this study was to evaluate the expression of inflammatory cytokines and Nitric Oxide (NO) by macrophages after interaction with Enterococcus faecalis in the: plankton and dislodged biofilm mode; intact biofilm mode stimulated by calcium hydroxide (CH), CH and chlorhexidine (CHX) or Triple Antibiotic Paste (TAP). For this purpose, culture of macrophages from monocytes in human peripheral blood (N=8) were exposed to the different modes of bacteria for 24 hours. Subsequently, the cytokines, such as, Tumor Necrotic Factor- alfa (TNF-α), interleukin (IL)-1β, IL-6, IL-10; and NO were quantified by Luminex xMAP and Greiss reaction, respectively. In addition to the potential therapeutic effects of the intracanal medication, their antimicrobial activity against Enterococcus faecalis biofilm were also tested in vitro by confocal microscopy. The experiments` data were analyzed by the Kruskal-Wallis test with the Dunn post hoc test (α < 0.05). Bacteria in dislodged biofilm mode were shown to be more aggressive to the immune system than bacteria in plankton mode and negative control, inducing greater expression of NO and TNF-α. Relative to bacteria in intact biofilm mode, the weakest antimicrobial activity occurred in Group CH. In Groups CH/CHX and TAP the percentage of dead bacteria was significantly increased to the same extent. Interestingly, the biofilm itself did not induce the release of pro-inflammatory cytokines - except for NO - while the biofilm treated with TAP and CH based pastes enhanced the levels of IL-6 and TNF-α; and IL-1 β, respectively. In contrast, the levels of a potent anti-inflammatory (IL-10) were increased in Group TAP.

Resumo A resposta de defesa do hospedeiro ao desafio microbiano que emerge do sistema de canais radiculares leva à periodontite apical. Os objetivos deste estudo foram avaliar a expressão de citocinas pró e anti-inflamatórias e Óxido Nítrico (NO) por macrófagos após interação com Enterococcus faecalis no modo: planctônio e de biofilme desalojado; biofilme intacto estimulado por hidróxido de cálcio (CH), CH e clorexidina ou Pasta Tri Antibiótica (TAP). Para isto, a cultura de macrófagos originados de monócitos do sangue periférico de humanos (N=8) foi exposta aos diferentes tipos de bactéria por 24 horas. Então, a quantificação da produção de of Fator de Necrose Tumoral- alfa (TNF-α), interleucina (IL)-1β, IL-6, IL-10 e NO por macrófagos se deu por meio do Luminex xMAP e reação de Greiss, respectivamente. Além dos potenciais efeitos terapêuticos desses compostos, sua atividade antimicrobiana contra E. faecalis também foi testada através microscopia confocal. Os dados dos experimentos foram analisados através do teste de Kruskal-Wallis com Dunn`s post hoc (α < 0.05). Bactéria em modo de biofilme desalojado se mostrou mais agressivo ao sistema imune que as bactérias planctônicas e controle negativo induzindo a maior excreção de NO e TNF-α. Em relação ao biofilme intacto, a atividade antimicrobiana mais fraca ocorreu no grupo de CH. Os grupos CHX e TAP aumentaram significativamente a porcentagem de bactérias mortas na mesma extensão. Interessantemente, o biofilme por ele mesmo não induziu a liberação de citocinas pro-inflamatórias - exceto por NO - enquanto que o biofilme tratado com TAP ou pastas a base de CH aumentaram os níveis de IL-6; e TNF-α e IL-1 β respectivamente. Em contraste, os níveis da potente citocina anti-inflamatória (IL-10) foram aumentados pelo grupo TAP.

Problems and puzzles in echinoderm demography / Problemas y acertijos en la demografía de los equinodermos

Introduction: There are problems and puzzles in understanding reproduction, growth and mortality in echinoderm life cycles. Objective: Explore problems and puzzles in life cycles that are important and challenging. Methods: The literature is used to elucidate problems associated with all life stages. Results: Sources of larvae that settle at a site are explored using oceanographic modelling and genetic methods. There are few studies that have estimated larval mortality in the plankton under field conditions and results differ from experimental results or patterns of settlement. In a small number of studies, mortality rate of newly settled larvae appears to change rapidly as individuals grow. There are problems measuring size, and measurement bias that interferes with many tagging methods used to estimate growth. There also are problems with the use of natural growth lines and commonly used software to estimate both growth and mortality from size-frequency data. An interesting puzzle is that echinoderms may show negative senescence with mortality rate decreasing with size. There is a problem in fertilization success based on density so there should not be rare species where sexes are separate with free spawning of gametes yet there seem to be rare echinoderms. Conclusions: All parts of echinoderm life cycles provide problems and puzzles that are important and challenging.

Introducción: Existen problemas y acertijos en la comprensión de la reproducción, el crecimiento y la mortalidad en los ciclos de vida de los equinodermos. Objetivo: Explorar los problemas y acertijos en los ciclos de vida que son importantes y desafiantes. Métodos: La literatura es usada para dilucidar los problemas asociados con todas las etapas de vida. Resultados: Las fuentes de larvas que se asientan en un sitio se exploran usando modelos oceanográficos y métodos genéticos. Existen pocos estudios que han estimado la mortalidad larval del plancton bajo condiciones de campo y los resultados difieren de los resultados experimentales o los patrones de asentamiento. En un número pequeño de estudios, la tasa de mortalidad de las larvas recién asentadas parece cambiar rápidamente a medida que los organismos crecen. Existen problemas para medir tamaños y el sesgo de medición interfiere con el uso de muchos métodos de marcado para estimar el crecimiento. También hay problemas con el uso de las líneas de crecimiento natural y con los programas comúnmente usados para estimar tanto el crecimiento como la mortalidad a partir de datos de frecuencia de tamaño. Un acertijo interesante es que los equinodermos pueden mostrar senescencia negativa con una tasa de mortalidad que disminuye con el tamaño. Existe un problema con el éxito de la fertilización basado en la densidad, por lo que no debería haber especies raras cuando los sexos están separados y existe un desove libre de gametos, sin embargo, parece haber equinodermos raros. Conclusiones: Todas las partes de los ciclos de vida de los equinodermos proveen problemas y acertijos que son importantes y desafiantes.

Spatio-temporal Distribution Pattern of Oyster Larvae in Guaratuba Bay, Paraná State - Brazil

HIGHLIGHTS Crassostrea oysters larvae patterns in estuarine environments. Larvae distribution patterns in Guaratuba Bay (Paraná State - Brazil).

Abstract Crassostrea oysters present planktonic/planktotrophic larval development. Thus, it is essential knowing larvae distribution patterns in estuarine environments presenting oyster farms and spat collection. The aim of the current research is to investigate the spatio-temporal distribution of oyster larvae in Guaratuba Bay (Paraná State - Brazil) based on monthly samplings conducted with a plankton net (50 cm diameter and 225 µm mesh size) from June 2003 to June 2004 in three different points (Point I - bay´s entrance; Point II - cultivation park; Point III - median sector of the bay). Plankton samples were preserved in 4% buffered formaldehyde. They were concentrated and quantified in the laboratory. The total number of larvae in the sample from each net was counted and corrected to a standard larvae/m3 collection tow. Data about sea water temperature, salinity, transparency and pluviosity were also collected. Based on results about the spatial distribution of larvae, mean density was 33.30 ±42.73 larvae/m³ in Point I; 17.84 ±16.88 larvae/m³ in Point II and 55.53 ±78.31 larvae/m³ in Point III, during the studied period. Spring and Summer were the seasons recording the most expressive mean number of larvae; the largest concentration of them was found in Point III, in the middle section of the bay.

Application of Chlorophyte ChlB Gene and Cyanophyte NIES Gene in the Detection of Drowning-Related Plankton / 法医学杂志

Objective To construct a polymerase chain reaction-capillary electrophoresis （PCR-CE） detection method using ChlB gene and NIES gene, investigate the method's specificity and sensitivity, and to evaluate its application value in drowning diagnosis. Methods The specific primers ChlB and NIES were designed for the conserved sequence of chlorophyte ChlB gene and cyanophyte NIES gene in GenBank to construct PCR-CE detection method; 50 species of standard DNA samples were amplified; the sensitivity was determined by gradient concentration detection of positive standard samples; 25 actual cadaver lung tissue samples （drowned： 20, natural death： 5） were detected, and the simultaneous detection results of microwave digestion-vacuum filtration-automated scanning electron microscopy （MD-VF-Auto SEM） were simultaneously compared. Results The minimum DNA detection concentration of primers ChlB and NIES was 0.161 ng and 0.109 ng, respectively, which could specifically amplify chlorophyte （Chlorella pyrenoidosa） and cyanophyte ［Microcystis aeruginosa （producing and not producing toxin）］ widespread in water. The product fragments were 156 bp and 182 bp, respectively. The results of non-drowning tissues were negative. Conclusion This method has high sensitivity and specificity. It can be applied to the detection of plankton related to drowning and combined with MD-VF-Auto SEM method, can increase the detection range of plankton related to drowning and improve the evidence power of drowning diagnosis.

Heterotrophic flagellates (Amorpha and Diaphoretiches) in phytotelmata bromeliad (Bromeliaceae) / Flagelados heterotróficos (Amorpha e Diaphoretiches) em fitotelmata de bromélia (Bromeliaceae)

Abstract Many plants may accumulate rainwater, forming phytotelmata, aquatic microhabitats inhabited by various organisms. The aim of this study was to conduct an inventory of heterotrophic flagellates associated with phytotelmata of the bromeliad Aechmea distichantha Lem., found in rocky cliffs on the Upper Paraná River. The bromeliads were removed manually from the rocky wall, the water was removed and cultures of organisms of each plant were mounted in Petri dishes. Sixteen species of heterotrophic flagellate were recorded, drawn and described, among them one species belonging to the Amorpha Domain and 15 species to the Diaphoretiches Domain. The groups with most species were Euglenida and Kinetoplastea. The low diversity of heterotrophic flagellates recorded in this study, compared to the plankton of lakes and reservoirs, is probably related to the fact that phytotelmata are habitats with extreme environmental conditions, thus selecting organisms tolerant to these environments.

Resumo Em sua superfície externa, muitas plantas podem acumular água da chuva formando fitotelmata, ou seja, micro-habitat aquáticos colonizados por diversos organismos, entre eles, os flagelados protistas. Fez-se um inventário de flagelados heterotróficos associados ao fitotelmata da bromélia Aechmea distichantha Lem., encontrada em paredões rochosos em um trecho da bacia do alto rio Paraná. As bromélias foram retiradas manualmente de paredão rochoso e, em laboratório, a água foi retirada dos tanques de cada planta, e foram montadas culturas dos organismos em placas de Petri. Os flagelados heterotróficos foram, então, observados in vivo sob microscópio óptico Olympus BX51, com sistema de contraste de interferência diferencial (DIC) acoplado. Dezesseis espécies flagelados heterotróficos foram registradas, desenhadas e descritas, sendo uma espécie pertencente ao Domínio Amorpha e 15 espécies ao Domínio Diaphoretiches. Os grupos mais especiosos foram Euglenida e Kinetoplastea. A diversidade relativamente reduzida de flagelados heterotróficos, quando comparada ao plâncton de lagos e reservatórios, pode estar relacionada ao fato de os fitotelmata serem habitats limitados, com condições ambientais extremas, selecionando, assim, organismos tolerantes a esses ambientes.

Antibacterial effect of different concentrations of Galla Chinensis extract on cariogenic bacteria in a biofilm model / 대한구강보건학회지

OBJECTIVES: Galla chinensis inhibited the adherence of planktonic oral bacteria and acid production by cariogenic bacteria. However, little is known about the relevant conditions of Galla Chinensis extract (GCE) exposure time and concentration and the effect of GCE on the structural and functional activity of cariogenic bacteria. The antibacterial effects of natural G. Chinensis extract on S. mutans , S. sanguinis, and S. oralis biofilms were evaluated in vitro.METHODS: Biofilms formed on glass surfaces were treated with different concentrations of GCE at different exposure times. The effects were assessed by examining the bactericidal activity, acidogenesis, minimum inhibitory concentration, and morphology.RESULTS: There was a statistically significant difference in the bacterial growth inhibition depending on the concentration of the GCE, with bacterial growth being inhibited as the concentration of GCE increased. A concentration of 1.0 mg/ml GCE had similar bactericidal effects against S. mutans and S. oralis biofilms to those produced by 2.0 mg/ml CHX. In the 1.0 mg/ml GCE group, incomplete septa were also observed in the outline of the cell wall, together with disruption of the cell membrane. In addition, there was also a slight exudation of the intracellular content from the bacteria in the 1.0 mg/ml GCE and 2 mg/ml CHX groups.CONCLUSIONS: These results indicate that GCE inhibits the growth of S. mutans, S. sanguinis, and S. oralis with increasing concentrations. It alters the microstructure of S. mutans biofilms. These results suggest that GCE might be a useful anti-bacterial agent for preventing dental caries.

Antibacterial effects of sodium tripolyphosphate against Porphyromonas species associated with periodontitis of companion animals

Porphyromonas species are closely associated with companion animal periodontitis which is one of the most common diseases in dogs and cats and leads to serious systemic diseases if left untreated. In this study, we evaluated the antimicrobial effects and mode of action of sodium tripolyphosphate (polyP3, Na5P3O10), a food additive with proven safety, using three pathogenic Porphyromonas species. The minimum inhibitory concentrations (MICs) of polyP3 against Porphyromonas gulae, Porphyromonas cansulci, and Porphyromonas cangingivalis were between 500 and 750 mg/L. PolyP3 significantly decreased viable planktonic cells as well as bacterial biofilm formation, even at sub-MIC concentrations. PolyP3 caused bacterial membrane disruption and this effect was most prominent in P. cangingivalis, which was demonstrated by measuring the amount of nucleotide leakage from the cells. To further investigate the mode of action of polyP3, high-throughput whole-transcriptome sequencing was performed using P. gulae. Approximately 30% of the total genes of P. gulae were differentially expressed by polyP3 (> 4-fold, adjusted p value < 0.01). PolyP3 influenced the expression of the P. gulae genes related to the biosynthesis of thiamine, ubiquinone, and peptidoglycan. Collectively, polyP3 has excellent antibacterial effects against pathogenic Porphyromonas species and can be a promising agent to control oral pathogenic bacteria in companion animals.

Susceptibility of Mutans streptococci in the Planktonic and Biofilm State to Erythrosine / 대한소아치과학회지

The aim of this study was to investigate the susceptibility of Mutans streptococci in both planktonic and biofilm states to erythrosine.S. mutans was cultured in brain-heart infusion (BHI) broth. Erythrosine was diluted in BHI broth and prepared at a concentration range of 0.02 – 10000 µg/L. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were measured using the microdilution method. After forming biofilms on 96-well plates, the minimum biofilm inhibitory concentration (MBIC) and minimum biofilm eradication concentration (MBEC) were measured.S. mutans was susceptible to erythrosine in both planktonic and biofilm states. MIC and MBC values were both 19.5 µg/L for the planktonic state, while MBIC and MBEC values were 313 µg/L and 2500 µg/L, respectively, for the biofilm state.Erythrosine (19.5 µg/L) exhibited a bactericidal effect on S. mutans (killing 99.9%) in the planktonic state. For biofilms, erythrosine inhibited biofilm growth and eradicated 99.9% of biofilm bacteria at higher concentrations than MIC and MBC. These MBIC and MBEC concentrations are much lower than known noxious doses, and the MIC, MBC, and MBIC values were even lower than clinical concentrations.

Dinámica del plancton en la primera milla náutica frente a la provincia de El Oro, Ecuador / Plankton dynamics in the first nautical mile of El Oro province, Ecuador

Resumen El Oro incluye diferentes sistemas costeros y estuarinos y es parte del Golfo de Guayaquil. Estas áreas proporcionan diferentes servicios ecosistémicos, pero con el aumento de la población y de las diversas actividades económicas, los recursos naturales locales poco estudiados pueden verse afectados negativamente. Es por ello que esta investigación tuvo como objetivo describir la dinámica del plancton, productividad y ecológica de esta área durante la estación seca de 2012, para lo cual se establecieron 17 estaciones para estudiar la distribución espacial y variación mensual de los nutrientes inorgánicos, temperatura, oxígeno disuelto, salinidad, zona fótica, abundancia y composición del plancton y larvas de peces, con métodos estándar. Se establecieron patrones espaciales y temporales mediante análisis de conglomerados y de ordenamiento multidimensional. Los resultados obtenidos de las variables bióticas y abióticas analizadas determinaron un patrón característico de un ecosistema estuarino productivo, así como también la existencia de áreas específicas asociadas a condiciones naturales y antrópicas, que son: a) interior del Archipiélago de Jambelí y frente a la isla Puná, influenciada por las actividades productivas y la hidrografía del área; b) una zona, conformada por las estaciones ubicadas en el estuario exterior donde la influencia oceánica es evidente y c) el canal de Jambelí, caracterizado por procesos de mezcla producto del flujo de mareas. Lo que se evidenció en la presencia de fitoplancton oceánico, nerítico y estuarino; copépodos con distribución uniforme en el área de estudio y dominancia de la familia Engraulidae representativa en este tipo de ecosistema acuático en lo referente a larvas de peces. Temporalmente, mayo registró precipitaciones que excedieron los valores normales, lo que habría determinado las altas concentraciones de nutrientes, la disminución de salinidad y de la zona fótica, registradas en este mes. Sin embargo la disponibilidad de nutrientes existente permitió a lo largo de todo el período de estudio mantener la composición y abundancia del fitoplancton con alternancia de especies representativas y que el zooplancton registre periodos de mayor (junio a agosto) y menor (octubre a diciembre) abundancia, lo que sería resultado de patrones poblacionales específicos de las especies presentes e influencia de la marea y flujos del río aportante; comportamiento similar registró el ictioplancton resultado de las concentraciones de Anchoa macrolepidota, especie característica de este ecosistema. Estos patrones de distribución espacio-temporal, permiten establecer el desarrollo de múltiples interacciones abióticas y bióticas que determinan diferentes estrategias para mantener la productividad del área, por lo que se recomienda desarrollar esta investigación durante la estación lluviosa en el Ecuador.

Abstract El Oro includes different coastal and estuarine systems and the important Gulf of Guayaquil. These areas provide different ecosystem services, but with the increase of population and of the varied economic activities, the barely known local natural resources may be negatively impacted. Thus this research aimed to study plankton dynamics, productivity and ecological balance for the area during the dry season of 2012. For this, we established 17 stations and studied the monthly variation and spatial distribution of inorganic nutrients, temperature, dissolved oxygen, salinity, photic zone, plankton and fish larvae abundance and composition with standard methods, while spatial and temporal patterns where identified using cluster and multidimentional scaling analyses. Obtained results of biotic and abiotic parameters defined a pattern characteristic of productive estuarine zones as well as the existence of three specific areas associated with the natural and anthropic conditions of the area, such as: a) inner part of the Jambelí Archipelago and in front of Puná island influenced by productive activities and the hydrography of the area; B) an area, formed by the stations located in the outer estuary where the oceanic influence is evident and c) the Jambelí channel, characterized by mixing processes resulting from tides flow. This was supported by the presence of oceanic, neritic and estuarine phytoplankton; copepods with uniform distribution in the study area, and dominance of Engraulidae fish larvae, representative of this type of aquatic ecosystem. Temporally, rainfall records in May exceeded normal values, which would have influenced the high concentrations of nutrients, decrease of salinity and of the photic zone registered during this month. However, the availability of nutrients throughout the study period, allowed to maintain the composition and abundance of phytoplankton, with changes in representative species, recorded periods of greater (June to August) and lower (October to December) zooplankton abundance, could be the result of specific population patterns of present species, and the influence of the tide and the flows of the contributing river basin. A similar behavior was recorded for ichthioplankton, as a result of Anchoa macrolepidota concentrations, species, characteristics of this type of ecosystem. These patterns of spatio-temporal distribution, allowed the development of multiple abiotic and biotic interactions that determine different strategies to maintain the productivity of the area. We recommend new studies during the rainy season in Ecuador. Rev. Biol. Trop. 66(2): 836-847. Epub 2018 June 01.

Spatial distribution and habitat use by early fish stages in a dammed river basin, Southern Brazil / Distribución espacial y uso de hábitat de peces en estadios tempranos de una represa del sur de Brasil

Abstract Fish diversity loss is threatened by the construction of dams as they prevent the regular natural dispersal among populations. Thus, conservation of key riverine habitats for fish reproduction may be essential for the recruitment of new native species of fish. The present study aimed to identify key habitats for fish spawning and early development in the Paranapanema River basin, as well as to determine the taxonomic composition, reproductive and life-history strategy, and to report spatial distribution of eggs, larvae and juveniles. The importance of lagoons, tributaries, and sub-tributaries was evaluated in the Paranapanema River basin between October 2012 and March 2013. Eggs and larvae samples were collected at dawn and dusk with conical plankton nets (0.5 mm mesh size), whereas juveniles were captured during the day with seine and sieve (0.5 cm mesh size). A total of 547 eggs, 904 larvae and 1 228 juveniles were captured. We observed that 2 larvae and 288 juveniles of non-migratory species, parental care, and equilibrium life-history strategy, predominated in lagoons and tributaries. On the other hand, 13 larvae and 60 juveniles of short migratory distance, no parental care, and periodic life-history strategy predominated in sub-tributaries. The highest densities of eggs were recorded in tributaries and sub-tributaries (Tukey's test, P = 0.001 and P = 0.03, respectively), and the highest densities of larvae were recorded for lagoons and tributaries (P = 0.005 and P = 0.0001, respectively). Captures of eggs and larvae were higher at night; while the highest catches per unit effort of juveniles were recorded for tributaries and sub-tributaries. Fish species that adopt different life-history strategies can use diverse types of habitats during the early stages. Lagoons, tributaries and sub-tributaries of the Paranapanema River play different roles in the reproductive success of fish fauna in a heavily modified basin. The preservation of spawning and nursery areas trapped between reservoirs is necessary for Neotropical fish species recruitment and survival. Rev. Biol. Trop. 66(2): 605-621. Epub 2018 June 01.

Resumen Los riesgos de pérdida de diversidad de peces aumentan con la construcción de represas, ya que impide la dispersión de individuos entre las poblaciones. Por lo tanto, la conservación del habitat ribereño clave para la reproducción de peces puede ser esencial para el reclutamiento de nuevos individuos de especies nativas. El presente estudio tiene como objetivo identificar hábitats clave para el desove y el desarrollo temprano de peces en la cuenca del río Paranapanema, así como determinar la composición taxonómica, la estrategia reproductiva y la historia de vida, y reportar la distribución espacial de huevos, larvas y juveniles. Se evaluó la importancia de lagunas, afluentes y subafluentes en la cuenca del río Paranapanema entre octubre de 2012 y marzo de 2013. Se recogieron muestras de huevos y larvas al amanecer y al atardecer con redes de plancton cónico (tamaño de malla de 0,5 mm), mientras que los juveniles fueron capturados durante el día con redes de cerco y tamiz (tamaño de malla de 0,5 cm). Se capturaron un total de 547 huevos, 904 larvas y 1 228 juveniles. Como resultado fue observado que 2 larvas y 288 juveniles de especies no migratorias, cuidado parental y estrategia de vida de equilibrio son predominantes en lagunas y afluentes. Por otro lado, 13 larvas y 60 juveniles de espécies migratorias de corta distancia, sin cuidado parental y estrategia de vida periódica predominaron en subafluentes. Se registraron las densidades más altas de huevos para afluentes y subafluentes (Prueba de Tukey, P = 0.001 y P = 0.03, respectivamente), y se registraron las densidades más altas de larvas para las lagunas y afluentes (P = 0.005 y P = 0.0001, respectivamente). Las capturas de huevos y larvas eran más altas por la noche. Las mayores capturas por unidad de esfuerzo de juveniles se registraron para los afluentes y subafluentes. Las especies de peces que adoptan diferentes estrategias de historia de vida pueden utilizar diversos tipos de hábitats durante las primeras etapas. Lagunas, afluentes y subafluentes del río Paranapanema desempeñan diferentes papeles en el éxito reproductivo de la fauna de peces en una cuenca fuertemente modificada. La preservación de áreas de desove y cría atrapadas entre el embalse es necesaria para el reclutamiento de especies de peces neotropicales.

Influence of Helicobacter pylori culture supernatant on the ecological balance of a dual-species oral biofilm

Abstract Dental caries is a chronic progressive disease occurring in the tooth hard tissue due to multiple factors, in which bacteria are the initial cause. Both Streptococcus mutans and Streptococcus sanguinis are main members of oral biofilm. Helicobacter pylori may also be detected in dental plaque, playing an important role in the development of dental caries. Objective The aim of this study was to investigate the effect of H. pylori culture supernatant on S. mutans and S. sanguinis dual-species biofilm and to evaluate its potential ability on affecting dental health. Material and methods The effect of H. pylori supernatant on single-species and dual-species biofilm was measured by colony forming units counting and fluorescence in situ hybridization (FISH) assay, respectively. The effect of H. pylori supernatant on S. mutans and S. sanguinis extracellular polysaccharides (EPS) production was measured by both confocal laser scanning microscopy observation and anthrone-sulfuric acid method. The effect of H. pylori supernatant on S. mutans gene expression was measured by quantitative real-time PCR (qRT-PCR) assays. Results H. pylori supernatant could inhibit both S. mutans and S. sanguinis biofilm formation and EPS production. S. sanguinis inhibition rate was significantly higher than that of S. mutans. Finally, S. mutans bacteriocin and acidogenicity related genes expression were affected by H. pylori culture supernatant. Conclusion Our results showed that H. pylori could destroy the balance between S. mutans and S. sanguinis in oral biofilm, creating an advantageous environment for S. mutans, which became the dominant bacteria, promoting the formation and development of dental caries.

Effect of γ-lactones and γ-lactams compounds on Streptococcus mutans biofilms

Abstract Considering oral diseases, antibiofilm compounds can decrease the accumulation of pathogenic species such as Streptococcus mutans at micro-areas of teeth, dental restorations or implant-supported prostheses. Objective To assess the effect of thirteen different novel lactam-based compounds on the inhibition of S. mutans biofilm formation. Material and methods We synthesized compounds based on γ-lactones analogues from rubrolides by a mucochloric acid process and converted them into their corresponding γ-hydroxy-γ-lactams by a reaction with isobutylamine and propylamine. Compounds concentrations ranging from 0.17 up to 87.5 μg mL-1 were tested against S. mutans. We diluted the exponential cultures in TSB and incubated them (37°C) in the presence of different γ-lactones or γ-lactams dilutions. Afterwards, we measured the planktonic growth by optical density at 630 nm and therefore assessed the biofilm density by the crystal violet staining method. Results Twelve compounds were active against biofilm formation, showing no effect on bacterial viability. Only one compound was inactive against both planktonic and biofilm growth. The highest biofilm inhibition (inhibition rate above 60%) was obtained for two compounds while three other compounds revealed an inhibition rate above 40%. Conclusions Twelve of the thirteen compounds revealed effective inhibition of S. mutans biofilm formation, with eight of them showing a specific antibiofilm effect.

Antimicrobial Effect on Streptococcus mutans in Photodynamic Therapy using Different Light Source / 대한소아치과학회지

In a photodynamic therapy, the difference of antibacterial capacity was compared according to the type of source of light when the same quantity of energy is irradiated.After S. mutans is formed in planktonic state and biofilm state, erythrosine diluted to 40 µM was treated for 3 minutes, and as the type of light source, Halogen, LED, and Plasma arc were used, which were irradiated for 30 seconds, 15 seconds and 9.5 seconds, respectively.After the completion of the experiment, CFU of each experiment arm was measured to compare the photodynamic therapeutic effects according to each condition.The CFU of each experiment arm had no statistically significant difference.Under the same quantity of energy, the photodynamic therapeutic effect can be said to be the same regardless of types of light source, which is a useful result in the clinical field with various light irradiators.

Plankton crustaceans in bays with different trophic status in Llanquihue lake (41° S Chile) / Crustáceos planctónicos crustáceo em baías com estado trófico diferente no lago Llanquihue (41° S Chile)

Abstract The Llanquihue lake is included in the called Araucanian or Nord Patagonian lakes located between 38-41° S. These lakes are characterized by their oligo-mesotrophic status due to human intervention which takes to the increase in nutrients inputs from industries and towns. Effects on zooplankton assemblages are observed with marked increase of daphnids abundance. The aim of the present study is to analyze the trophic status and zooplankton relative abundance in different bays of Llanquihue lake. It was found direct associations between chlorophyll a with daphnids percentage, total dissolved nitrogen with reactive soluble phosphorus nitrogen/phosphorus molar radio with cyclopoids percentage, and an inverse relation between daphnids and calanoids percentages. The occurrence of three kinds of microcrustacean assemblages and environmental conditions was evidenced: the first one with high calanoids percentage, low species number and low chlorophyll and nutrients concentration, a second with moderate chlorophyll and nutrients concentration and moderate daphnids percentage; high species number and a third site with high chlorophyll and nutrients concentration, high daphnids percentage and high species number. Daphnids increase under mesotrophic status, agree with similar results observed for southern Argentinean and New Zealand lakes.

Resumo O lago Llanquihue está incluído nos chamados lagos araucana ou Nord Patagônia localizado entre 38-41° S. Estes lagos são caracterizados pela condicao oligo-mesotrofica debido a intervencao humana, com aumento da carga de nutrientes provenientes de industrias y areas urbanas com efeitos sobre as assembleias zooplantonicas sao observadas, com aumento acentuado de dafnideos. O objetivo do presente estudo é analisar o estado trófico a abundancia relative do zooplancton em diferentes compartimentos do lago Llanquihue. Foram encontradas associações diretas entre clorofila a com percentual de dafinídeos, nitrogênio total dissolvido com fósforo solúvel reativo molares razao molar nitrogênio / fósforo com percentual de ciclopóides, e uma relação inversa entre percentuais de dafinídeos e calanóides porcentagens. A ocorrência de três tipos de assembleias de microcrustáceos e as condições ambientais fora: a primeira com alta porcentais de calanóides, baixo número de espécies e baixa clorofila e a nutrientes, uma segunda com concentracoes moderadas de clorofila e nutrientes percentual moderado de daphnideos e alto número de espécies; e uma terceiro local com alta concentração de clorofila e nutrientes, alta abundância dafinídeos e número elevado de espécies. Resultados similares com aumento de dafnideos em condicoes mesotroficas também foram observados para lagos da Argentina e Nova Zelândia do sul.

Influence of Eichhornia crassipes and Lemna sp. on the natural diet of Astyanax altiparanae Garutti & Britski, 2000 in the initial stages / Influência de Eichhornia crassipes e Lemna sp. na alimentação natural de Astyanax altiparanae Garutti & Britski, 2000 na fase inicial

Aiming to study the changes in the diet of Astyanax altiparanae in the early developmental stages, in the presence of Eichhornia crassipes (EC) and Lemna sp. (LM), larvae of Astyanax altiparanae were stocked in eight tanks of 900 L (500 ind. tank-1), four with EC and four with LM covering 50% surface. Tanks were inoculated with plankton and received 3.0 g inorganic fertilizer (NPK-7:14:8) and 1.5 g every week. At every three days, three larvae were taken from each tank, fixed in 4% formaldehyde and grouped into four age classes (I: 6 to 12 days, II:15 to 21 days, III:24 to 30 days; IV:33 to 39 days). The analysis of dominance and frequency of occurrence (FO) of food items indicated that, at this stage, A. altiparanae is generalist. In class I, there was a greater dominance and FO of Lecane bulla and Alona sp. in digestive tracts in the presence of both EC and LM, respectively; and larvae started to consume larger organisms, such as Chironomidae, in other age classes with EC. In the presence of LM, Alona sp. was dominant in class II; Chironomidae, in class III and Centropyxis sp., in class IV. Diatoms have greater FO both in EC and in LM. In class III, with EC, Chironomidae had higher FO, as well as Arcella sp. and Scenedesmus spp. with LM. The highest FO in class IV was found for L. bulla and Scenedesmus spp., with EC and LM, respectively. It can be concluded that EC and LM influenced the diet of Astyanax altiparanae, with alterations according to the availability of organisms and increase in age.

Objetivou-se estudar mudanças da dieta nos estágios iniciais de desenvolvimento de Astyanax altiparanae, na presença de Eichhornia crassipes (EC) e Lemna sp. (LM). Larvas de Astyanax altiparanae foram estocadas em oito tanques de 900 L (500 ind. tanque-1), sendo quatro com EC e quatro com LM cobrindo 50% da superfície destes. Os tanques foram inoculados com plâncton e receberam 3,0 g de fertilizante inorgânico (NPK-7:14:8) e 1,5 g semanalmente. A cada três dias foram coletadas três larvas de cada tanque, fixadas em formol a 4% e agrupadas em quatro classes de idade (I: 6 a 12 dias, II: 15 a 21 dias, III: 24 a 30 dias e IV:33 a 39 dias). Relacionando a dominância e frequência de ocorrência (FO) dos itens alimentares verificou-se que, nesta fase, o A. altiparanae é generalista. Na classe I, houve maior dominância e FO de Lecane bulla e de Alona sp. nos tratos digestórios na presença de EC e LM, respectivamente, passando a consumir organismos maiores como Chironomidae nas demais classes com EC. Na presença de LM, Alona sp. foi dominante na classe II, Chironomidae na III e Centropyxis sp. na IV. As diatomáceas tiveram maior FO tanto em EC quanto em LM. Na classe III, com EC Chironomidae teve maior FO, assim como Arcella sp. e Scenedesmus spp. com LM. A maior FO na classe IV foi de L. bulla e Scenedesmus spp. com EC e LM, respectivamente. Concluiu-se que EC e LM influenciaram a dieta de A. altiparanae, alterando-a de acordo com a disponibilidade dos organismos e com o aumento da idade.