Larval stages of Paradasygyius depressus (Bell, 1835) (Crustacea : Decapoda : Brachyura : Majidae) and a phylogenetic analysis for 21 genera of Majidae

All larval stages and the first crab instar of Paradasygyius depressus (Bell) were obtained in laboratory culture. Larval development consists of two zoeal stages, followed by the megalopa. Each larval stage is described in detail. Beginning with the first zoea, the duration of each stage was 4--7 (4.5 +/- 0.7), 4-5 (4.5 +/- 0.5), and 7 days, the megalopa and first crab instar appearing 11 +/- 1 and 15 days after hatching, respectively. A phylogenetic analysis of 21 genera of Majidae is provided based on 34 zoeal and three megalopal characters. The phylogenetic analysis resulted in four equally parsimonious trees 173 steps long (CI = 0.66, RI = 0.71, and RC = 0.47) supporting the monophyly of Oregoniinae, Majinae, and Inachinae (with the exclusion of Macrocheira de Haan incertae sedis). Based on general agreement of sister-group hypotheses, we provide sets of larval characters that define Oregoniinae, Majinae, and Inachinae. Our phylogenetic hypothesis suggests that Oregoniinae is the most basal clade within the Majidae, and Majinae and the clade (Epialtus H. Milne Edwards + Inachinae [excluding Macrocheira incertae sedis]) are sister taxa. Within Inachinae, all trees suggest that Inachus Weber and Macropodia Leach are sister taxa nested as the most derived clade, followed by Achaeus Leach, Pyromaia Stimpson, Paradasygyius Garth, Anasimus A. Milne-Edwards, and the most basal Stenorhynchus Lamarck. The sister-group relationships of the clade (Pisa Leach (Taliepus A. Milne-Edwards + Libinia Leach)), Mithrax Latreille and Microphrys H. Milne Edwards remained unresolved.

Utilização de meios alternativos e diferentes tipos de recipientes no cultivo de Ankistrodesmus gracilis (Reinsch) Korshikov (Chlorophyceae) em laboratório

A laboratory culture of Ankistrodesmus gracilis algae was evaluated by studying the biology of the species and its chemical composition in a traditional medium (CHU 12) and in two alternative culture media, NPK (20-5-20) and macrophyte (Eichhornia crassipes) + NPK, in three different types of recipients (fiberglass, carboy and plastic bag). First peak in the growth curve of Ankistrodesmus gracilis occurred on the ninth day in macrophyte + NPK medium (74.16 x 10 5 cells mL -1) in a fiberglass recipient. However, highest density (p < 0.01) was reported in medium CHU 12 (122.87 x 10 5 cells mL -1) in a plastic bag on the twelfth day. Cell density was over 70 x 10 5 cells mL -1 starting on the twelfth day. Growth rate of A. gracilis was similar (p > 0.05) in culture media in the three recipients. Protein and fiber were similar (p > 0.05) in the treatments, but lipids were higher (p < 0.05) in NPK. Nitrate, ammonia, total phosphorus and orthophosphate contents were over 1 mg L -1 in NPK (p < 0.01). Results show that alternative media, such as NPK and macrophyte + NPK, are possible for large-scale culture of A. gracilis cultured in three types of recipients. Costs are low, occupying less space when cultured in plastic bags and in the laboratory.

Evaluation of Palmistichus elaeisis Delvare & LaSalle (Hymenoptera: Eulophidae) as parasitoid of the Sarsina violascens Herrich-Schaeffer (Lepidoptera: Lymantriidae)

This is the first report on the parasitoid Palmistichus elaeisis, genus Eulophidae, found in the field parasitizing pupae of defoliating eucalyptus. Lepidopterous pests occur in eucalyptus plantations in Brazil, reaching high population levels. Due to the complexity of pest control in eucalyptus forests, alternative control methods have been proposed, for instance biological control through use of parasitoids. Natural enemies play an important role in regulating host populations because their larvae feed on the eggs, larvae, pupae or adults of other insects. The parasitic Hymenoptera are important agents in biological control programs against forest pests, and may provide economic and environmental benefits. The generalist endoparasitoid Palmistichus elaeisis Delvare and LaSalle, 1993 (Hymenoptera: Eulophidae) can develop in its host’s pupae, which overcome the host’s physiology and can therefore be used for biological control of agricultural and forest pests. This study aimed to evaluate the impact of P. elaeisis as a pupal parasitoid of S. violascens in providing a potential alternative to chemical control of the pest and creation of an alternative host. The experiment was developed in the Laboratory for Biological Control of Forest Pests, Universidade Estadual Paulista "Julio Mesquita Filho”. Parasitoids used in this test were originally collected on pupae of E. eucerus. (Lepidoptera: Riodinidae) in eucalyptus plantations at Lençois Paulistas, São Paulo state, Brazil, in 2011. Thereafter, a laboratory culture has been maintained, using pupae of Spodoptera frugiperda (J. E. Smith) (Lep.: Noctuidae) as hosts. S. violascens eggs were collected in a eucalyptus clonal plantation in Sao Paulo state (Brazil). Larvae were reared under ambient conditions on Eucalyptus urophylla S.T. Blake (Myrtaceae) leaves. The following parameters were determined: parasitism level, numbers of emerged and non-emerged parasitoids and duration of egg-adult cycle. The S. violascens pupae were dissected to evaluate the non-emerged parasitoids. The parasitism level reached 100%, with a 100% emergence rate. It was verified that 113.2±0.8 parasitoids emerged per individual pupa versus only 0.7±0.1 that did not emerge. The P. elaeisis egg-adult cycle was 20.3±0.6 days in S. violascens pupae. This opens new perspectives for utilizing this parasitoid in biological control programs against caterpillars important to forestry. Sarsina violascens in Brazil.

Large scale laboratory cultures of Ankistrodesmus gracilis (Reisch) Korsikov (Chlorophyta) and Diaphanosoma biergei Korinek, 1981 (Cladocera)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Population growth and development of two species of Cladocera, Moina micrura and Diaphanosoma birgei, in laboratory

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Comparison between qualitative and semiquantitative catheter-tip cultures: laboratory diagnosis of catheter-related infection in newborns

Este estudo prospectivo avaliou os métodos semiquantitativo e qualitativo de cultura de cateter para o diagnóstico de infecção relacionada a cateter (IRC) em recém-nascidos (RN). Foram incluídas pontas de cateteres provenientes de recém-nascidos internados na Unidade Neonatal do Hospital das Clínicas da Faculdade de Medicina de Botucatu, UNESP. Foram utilizadas as técnicas semiquantitativa e qualitativa de cultura de cateter. Para o diagnóstico de IRC, os microrganismos isolados das culturas de cateteres e de hemoculturas periféricas foram identificados e submetidos ao teste de sensibilidade a antimicrobianos. O padrão ouro correspondeu ao diagnóstico de certeza de IRC, com o isolamento do mesmo microrganismo (espécie e perfil de sensibilidade a antimicrobianos) isolado em hemocultura periférica. Foram estudados 85 cateteres provenientes de 63 RN. A cultura semiquantitativa, embora tenha apresentado menor sensibilidade (90%), apresentou uma maior especificidade (71%) em comparação à sensibilidade de 100% e especificidade de 60% encontradas na cultura qualitativa. Através da identificação dos microrganismos obtidos nas culturas de cateteres, observou-se uma predominância de espécies de Estafilococos coagulase-negativa (ECN). A espécie Staphylococcus epidermidis foi a prevalente (77,5%) nos cateteres com culturas semiquantitativas positivas. Dos 11 episódios de IRC diagnosticados, 8 (72,7%) foram associados a espécies de ECN, dos quais 6 eram da espécie S. epidermidis. Também foram detectados dois casos de IRC por S. aureus e um caso por Candida parapsilosis. O método de cultura semiquantitativo cateter apresentou vantagens para o diagnóstico de IRC em RN quando comparado com o método qualitativo tradicional.

Intraspecific and interspecific pre-adult competition on the neotropical region colonizer Zaprionus indianus (Diptera: Drosophilidae) under laboratory conditions

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

A comparative study of different diets to optimize cultivation of Hyalella azteca in the laboratory

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Tuberculosis diagnosis after bleach processing for early stage tuberculosis laboratory capacity building

The diagnosis of tuberculosis is seriously hampered in the absence of standard biosafety laboratory facilities for specimen concentration and Mycobacterium tuberculosis culture. Within a laboratory twinning arrangement, heat-fixed direct smear and sediment from 74 bleach-processed and 20 non-processed specimens from Cumura Hospital, Guinea-Bissau, were sent to Lisbon for molecular evaluation of rifampicin resistance. Sequence analysis of a 369 base-pair ppoB locus detected 3.2% (3/94) resistant specimens. To our knowledge, this represents the first report on the molecular analysis of M. tuberculosis from bleach-processed sputum, an alternative to current diagnostic practice in low-resource settings.

Behavior of Atta sexdens rubropilosa (Hymenoptera: Formicidae) workers during the preparation of the leaf substrate for symbiont fungus culture

The behavioral repertory of Atta sexdens rubropilosa Forel (Hymenoptera: Formicidae) workers marked by size category was studied during the preparation of the leaf substrate in the laboratory. The workers were marked according to three physical castes, i.e., minima, generalist and forager. Seven types of behavioral acts were recorded for each caste, with the following frequencies: licking leaf fragments (64.6%), holding fragments on the surface of the fungus garden (16.4%), shredding the fragments (6.0%), chewing and crimping the edges of the fragments (9.0%), incorporating the fragments (2.7%), touching the surface of the fungus with their mandibles and other mouthparts after incorporation (0.3%), and depositing fecal fluid (0.1%). The minima workers were found to be more specialized in the activities related to the preparation of the leaf substrate, which represented 52% of the total number of tasks performed. The generalists performed 40.3% of these tasks, and the foragers 7.9%. Licking the substrate was the behavior most frequently recorded and performed for a longer period of time. In this way, the workers may feed and at the same time eliminate microorganisms that are harmful to the symbiont fungus. The smaller castes, minima and generalists, are those most responsible for the preparation of the leaf substrate and predominate within a colony. From a practical viewpoint, with the introduction of toxic bait containing insecticides, for example, these size categories will be those most intensely intoxicated, especially through the behavior of licking bait pellets. On the basis of the data obtained about these behaviors, we may raise the hypothesis that trophallaxis in not the major factor triggering contamination with an insecticide among the workers of a colony.

Effects of some parameters in upscale culture of Haematococcus pluvialis Flotow

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Isolation, Culture and Differentiation of Buffaloes Bone Marrow Mesenchymal Stem Cells Obtained from the Coxal Tuberosity

Currently, much attention has been devoted to the renewal of knowledge about Stem Cells and Cell Therapy in domestic species. In this sense, the present work aimed to develop a methodology for collecting, processing and cultivation of mesenchymal stem cells obtained from bone marrow of coxal tuberosity in buffaloes. The collection was performed using a Komiyashiki needle, which was introduced in the coxal tuberosity and the bone marrow aspirated into a heparinized syringe with the aid of negative pressure. Directly after collection samples were processed at the laboratory at FMVZ - UNESP. The samples took approximately 32 days to reach 80% confluence, when the first passage and differentiation was performed. To confirm the mesenchymal origin, cells were induced to differentiate into adipogenic and osteogenic lineages. Samples showed morphological changes during differentiation protocol, but not all presented production of extracellular deposits of calcium or intracellular fat droplets, observed after staining with Alizarin Red and Oil Red respectively. Compared with the material obtained from other species and processed in the same laboratory, the primary culture was longer. Therefore, more studies are needed to standardize the age of animals used and to test other inducers of cell differentiation.

Diagnostic accuracy of semi-quantitative and quantitative culture techniques for the diagnosis of catheter-related infections in newborns and molecular typing of isolated microorganisms

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)