scholarly journals Padronização de um protocolo para assepsia de segmentos nodais de Phalaenopsis para clonagem in vitro

2021 ◽  
Vol 17 (1) ◽  
pp. 10-17
Author(s):  
Fernanda Kelly Mezzalira ◽  
Betty Cristiane Kuhn
Keyword(s):  
Culture Media ◽  
Ornamental Plants ◽  
Lateral Bud ◽  
Cultural Visibility ◽  
Positive Results

Plantbiotechnologysan área ofhighimportancesinceit has forobtainingplantorganismswithcharacteristics superior tothosealreadyonthemarket. Cloningisoneofthetolos that forthisfunction, throughit, organismswithcharacteristicsofinterest are selected and this individual ismultiplied, ensuringthattheregeneratedplants are geneticallyidenticaltothedesiredmatrix, establishing a standardization. Knowingthatthe ornamental plants sector contributessignificantlytotheeconomy and amongthemostesteemed ornamental plantsamongBrazilians, there are orchidswhichhaveacquired cultural visibility and a largenumberofcollectors in recentyears. Theobjectiveofthepresentworkwastoestablishanefficientassepsisprotocolfor lateral meristems and toobtain clones oftheorchidofthegenusPhalaenopsis. Formethodology, aiming at thestandardization o fan asepsisprotocol, 4 treatmentsweredeveloped and testedwithdifferentcombinations (concentration x time) ofagentssuch as sodiumhypochlorite, alcohol, copper, tween and washingtheexplantswithsteriledistilled wáter formeristemsoftheorchidofthegenusPhalaenopsis. Themeristems, alsoknown as lateral bud, were removed fromthestemoftheseedlings, fromtheir floral stems. As forobtained clones, theexperimentcarriedoutconsistedofinoculatingthemeristems, after anasepsisprocess, in Knudson culture media, containingdifferentcombinationsofgrowthregulators, usinganauxin and cytokinin. Theresultsobtained show that, no asepsistreatmentstested in thiswork, presentedsignificant and positive results, so itwasnot posible toestablish, as yet, anasepsisprotocolfor lateral meristemoforchidsofthegenusPhalaenopsis. Sinceallmeristemssufferedcontamination and oxidation. Therefor, itwasnotyet posible toobtain clones, usingthismethodology. Thisworkserves as aninitialbasis, forfutureresearch, regardingcloningthroughmeristems in Phalaenopsis.

scholarly journals Sampling and Characterization of the Environmental Fungi in the Provincial Historic Archive of Pinar Del Río, Cuba

2020 ◽  
Vol 1 (8) ◽  
pp. 404-420
Author(s):  
Sofia Borrego ◽  
Alian Molina ◽  
Tamara Abrante
Keyword(s):  
Respiratory Tract ◽  
Human Health ◽  
Culture Media ◽  
Airborne Fungi ◽  
New Findings ◽  
Del Rio ◽  
Positive Results ◽  
Made In

It has been reported that there is a correlation between indoor airborne fungi and the biodeterioration of valuable documents in archives, libraries and museums, and that these fungi can also cause effects on human health if there are immunological problems or the time of exposure to these environments of low quality is long. The aims of this study were quantifying and characterizing the mycobiota of the indoor air in three repositories of the Provincial Historical Archive of Pinar del Río, Cuba and assessing its impact on the human health. The samplings were made in two different months corresponding to the years 2016 and 2017, one belonging to the rainy season and the other to the season of the little rain using a SAS biocollector and appropriate culture media to isolate fungi. The fungal concentrations and the Indoor/Outdoor (I/O) ratios obtained revealing that the repositories showed good quality environments. In both isolations Cladosporium was the predominant genus followed by Penicillium in the first sampling and Fusarium in the second isolation. The genera Aureobasidium, Sepedonium, Trichaegum and Wallemia were new findings for the Cuban archives. The pathogenic attributes studied showed that 30% of the isolates have spores so small that they can penetrate into the respiratory tract into the alveoli; 10.7% of the taxa obtained in the first isolation and 13.3% of the taxa detected in the second sampling also showed positive results to four virulence tests analyzed "In vitro" (growth at 37°C, hemolytic activity, phospholipase activity and respiratory tract level to which the spores can penetrate). These virulence factors (pathogenic attributes) evidence the risk that environmental fungi represent for the health of personnel in this archive.

scholarly journals Culture media for the multiplication of wild Manihot species

2018 ◽  
Vol 42 (6) ◽  
pp. 598-607 ◽  
Author(s):  
Jucieny Ferreira de Sá ◽  
Emília dos Santos Sampaio ◽  
Maria Inês de Souza Mendes ◽  
Karen Cristina Fialho dos Santos ◽  
Antônio da Silva Sousa ◽  
...  
Keyword(s):  
Plant Height ◽  
Disease Transmission ◽  
Culture Media ◽  
Test Tube ◽  
Nodal Segments ◽  
Randomized Design ◽  
Lateral Bud ◽  
Successive Generations ◽  
Shoot Mass

ABSTRACT The cassava propagation system is slow and favors disease transmission through successive generations. Micropropagation is an alternative to overcome the aforementioned limitations, besides allowing the generation of a larger number of pest- and pathogen-free plants. Therefore, the aim of the present study is to investigate the effect of culture media on the multiplication in vitro of five wild Manihot species. The experiment followed a completely randomized design, at factorial arrangement 5 (wild Manihot species) x 6 (culture media), with 11 repetitions. Explants consisted in nodal segments (91 cm long and one lateral bud) of species Manihot flabellifolia, M. tristis, M. caerulescens, M. chlorosticta and M. jacobinensis, which were extracted in vitro from the collection of wild cassava species. One segment was placed in each test tube added with 10 mL of MS media 0.01, 17N, 12A3, 4E, 8S and WPM, and kept for 90 days in growth room under 30 μmol m-2 s-1irradiance, temperature 27 ± 1 °C and 16h photoperiod. Variables plant height (cm), number of green leaves, number of senescent leaves, number of shoots, number of microcuttings, fresh and dry shoot mass, fresh and dry root mass (mg) and callus mass (mg) were analyzed. Our results showed that the culture medium 12A3 was not responsive to any of the species; however, if one takes into consideration variables plant height and number of microcuttings, this medium can possibly be used in the micropropagation of other wild species belonging to genus Manihot.

scholarly journals In vitro Control of the Fungus Botrytis cinerea Pers. with Plant Extracts

2010 ◽  
Vol 67 (1) ◽  
Author(s):  
Raluca MICLEA ◽  
Carmen PUIA
Keyword(s):  
Botrytis Cinerea ◽  
Plant Extracts ◽  
Culture Media ◽  
Ornamental Plants ◽  
Diffusion Method ◽  
Phytopathogenic Fungus ◽  
Vegetable Crops ◽  
Plant Extraction

The gray mould, caused by Botrytis cinerea Pers., is an economically important disease of plants throughout the world. It is common on vegetable crops, soft fruits, ornamental plants and grapevine, and occurs in greenhouses and in the field as well as in storage and transport. Plant extracts and their purified compounds have antibacterial, antiviral and fungicidal effects both in vitro and in vivo. Their properties were attributed mainly to alkaloids, several flavonoids and phenolic acids. This study was made in order to establish the materials and the methods most used for the in vitro control of the fungus Botrytis cinerea Pers. with plant extracts – types of plant extracts, the most appropriate culture media, methods of inoculation, methods of extraction and methods for testing the antifungal effect of the plant extracts against the phytopathogenic fungus. The most used medium was PDA (potato dextrose agar). Conform to the literature data this medium is the most suitable for the development of this fungus, the growth of the mycelium, the conidia growing and the formation of sclerotia. As plant extraction method, the hydrodistilation is the most ancient method of distillation and the most versatile. Reviewed methods used for testing the efficacy of natural products (plant extracts) were: the agar diffusion method, the poisoned food technique, the radial growth test, the conidial germination assay, the microathmosphere method, the technique of Thompson.

Structural integrity after cold storage of human epidermal model in hypothermosol: A correlative ultrastructural and fluorescent assay

1994 ◽  
Vol 52 ◽  
pp. 270-271
Author(s):  
Henry H. Eichelberger ◽  
John G. Baust ◽  
Robert G. Van Buskirk
Keyword(s):  
Cold Storage ◽  
Structural Integrity ◽  
Culture Media ◽  
Cell Structure ◽  
Natural State ◽  
Sodium Cacodylate ◽  
Ruthenium Tetroxide ◽  
Cacodylate Buffer ◽  
Before And After

For research in cell differentiation and in vitro toxicology it is essential to provide a natural state of cell structure as a benchmark for interpreting results. Hypothermosol (Cryomedical Sciences, Rockville, MD) has proven useful in insuring the viability of synthetic human epidermis during cold-storage and in maintaining the epidermis’ ability to continue to differentiate following warming.Human epidermal equivalent, EpiDerm (MatTek Corporation, Ashland, MA) consisting of fully differentiated stratified human epidermal cells were grown on a microporous membrane. EpiDerm samples were fixed before and after cold-storage (4°C) for 5 days in Hypothermosol or skin culture media (MatTek Corporation) and allowed to recover for 7 days at 37°C. EpiDerm samples were fixed 1 hour in 2.5% glutaraldehyde in sodium cacodylate buffer (pH 7.2). A secondary fixation with 0.2% ruthenium tetroxide (Polysciences, Inc., Warrington, PA) in sodium cacodylate was carried out for 3 hours at 4°C. Other samples were similarly fixed, but with 1% Osmium tetroxide in place of ruthenium tetroxide. Samples were dehydrated through a graded acetone series, infiltrated with Spurrs resin (Polysciences Inc.) and polymerized at 70°C.

Pertumbuhan Dan Perkembangan Anggrek Dendrobium anosmum Pada Media Kultur In Vitro Dengan Beberapa Konsentrasi Air Kelapa

Agrologia ◽  
2018 ◽  
Vol 1 (1) ◽  
Author(s):  
S. Tuhuteru ◽  
Meity L Hehanussa ◽  
Simon H.T Raharjo
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Water Concentration ◽  
Medium Composition ◽  
Coconut Water ◽  
Orchid Species ◽  
Randomized Design ◽  
Wet Weight ◽  
Completely Randomized Design

Dendrobium anosmum is one of natural orchids in Indonesia. Optimization of medium composition for orchid propagation through in vitro culture is necessary to enhance propagule multiplication capabilities and quality. This study was aimed to study the influence of concentration of coconut water in culture medium on in vitro growth and development of D. anosmum orchid species and to determine the optimal coconut water concentration in culture media.  The experiment were arranged in a Completely Randomized Design with four treatments and eight replications. The treatments consisted of the addition of coconut water with concentrations: 0 ml•l -1 (control), 50 ml•l-1, 100 ml•l-1 and 150 ml•l-1. The results showed that addition of coconut water in culture medium gave different effect on shoot growth and multiplication of D. anosmum orchids.  Coconut water concentration of 100 ml•l-1 was the best concentration for growth and multiplication of D. anosmum orchids, based on both shoots and roots growth, plantlet height and wet weight.

Methods of experimental polyploidization and their use in apple breeding

2020 ◽  
Vol 62 ◽  
pp. 85-90
Author(s):  
L. V. Tashmatova ◽  
O. V. Matsneva ◽  
T. M. Khromova ◽  
V. V. Shakhov
Keyword(s):  
Exposure Time ◽  
Cell Walls ◽  
Prolonged Exposure ◽  
Ornamental Plants ◽  
Colchicine Treatment ◽  
Apple Trees ◽  
Open Ground ◽  
High Concentrations ◽  
Diploid Gametes

The article presents methods of experimental polyploidy of fruit, berry and ornamental plants. The purpose of this review is to highlight the problems and prospects of polyploidization of plants in the open ground and in vitro culture and the possibility of their application for apple trees. For the purpose of obtaining apple tetraploids as donors of diploid gametes, seed seedlings were treated with a solution of colchicine in concentrations of 0.1-0.4 % for 24 and 48 hours. Colchicine concentrations of 0.3 % and 0.4 % at 48 hours of treatment had a detrimental eff ect on their development. As a result, tetraploids and chimeras were obtained from seeds from free pollination of the varieties Orlik, Svezhest, Kandil Orlovsky, as well as from seeds obtained from crossing the varieties Svezhest×Bolotovskoe, Moskovskoe Оzherel’e×Imrus, Girlyanda×Venyaminovskoe. The optimal concentration of colchicine was 0.1 %. Methods of colchicine treatment have been studied: 1) adding to the nutrient medium, colchicine concentration: 0.01%, 0.02%, exposure time 24h-19 days; 2) applying amitotic solution to the growth point, colchicine concentration: 0.1 %, 0.2 %, exposure time 24h-7 days. To increase the penetration of colchicine through the cell walls, a 0.1 % dimexide solution was used. Studies have shown that high concentrations and prolonged exposure to colchicine reduce the viability of explants.

scholarly journals TO010A NEW IMMUN-TOXICOLOGICAL TEST TO DETECT POLYSULFONE HYPERSENSITIVITY IN HEMODIALYSIS PATIENTS

2020 ◽  
Vol 35 (Supplement_3) ◽  
Author(s):  
Joachim Beige ◽  
Ralph Wendt ◽  
Despina Rüssmann ◽  
Karl-Peter Ringel
Keyword(s):  
False Negative ◽  
False Negative Rate ◽  
Lymphocyte Transformation ◽  
Clinical Problem ◽  
Lymphocyte Transformation Test ◽  
Laboratory Research ◽  
Humoral Immune ◽  
Before And After ◽  
Positive Results

Abstract Background and Aims Incompatibility of dialysis procedure due to hypersensitivity against dialyzer material which currently is mainly based on polysulfone and derivatives can not be assessed by routine laboratory tests. Although the frequency of such symptoms is suspected to be low (below 2%) such resembles an important clinical problem because dialysis procedures are frequently accompanied by symptoms of non-tolerability with reasons not being entirely clear while circulatory reasons are suspected to play a major role. Method To enlighten the role of polysulfone hypersensitivity, we adapted known standardized material immune-toxicological tests (lymphocyte transformation test, basophil degranulation test) to the specific conditions of dialysis and polysulfone material sensitivity. We developed a method of polysulfone micronisation and measured humoral immune response of isolated patient´s lymphocytes when incubated with polysulfone dispersion. Results 39 samples from 103 patients with suspected polysulfone hypersensitivity showed positive results for type 1 (n=19), type 4 (n=18) or both type (n=2) reactions. There were no significant differences in the level of stimulation measured for DI, SI and lymphogenesis before and after dialysis (average delta -0.4; -0.28; - 1.74, p = 0.71; 0.34; 0.37) and with different dialyzer materials (Tab. 1). Patients with pos. type 4 results (LTT and lymphogenesis) showed highly correlated results in either LTT or lymphogenesis test (Fig. 1, R=0.87, p<0.0001). 8 out of 8 samples from patients with repeated test on different PS showed positive results on either PS. One patient tested positive on PS showed no hypersensitivity with another non-PS (PMMA) material. Conclusion This is the first methodological report showing plausible in-vitro results of patients samples concerning polysulfone intolerance. On the first superficial view, a “false-negative” rate of 60% looks rather disappointing, because all samples derived from patients with suspicion of PS hypersensitivity. However, due to the clinical variability of intolerance symptoms and the high prevalence of any problems after HD initiation, mainly of circulatory origin after initiating extracorporeal circuit, this rate may obviously express the true frequency of isolated PS material hypersensitivity in suspected patients. Alternative pathophysiological pathways of material sensitivity like complement activation, remain to be elucidated and incorporated into a comprehensive future testing panel. Further clinical and laboratory research is needed to define true polysulfone hypersensitivity and to enlighten the field of hypothetic subclinical material incompatibility in patients with impaired dialysis tolerability.

Nephrotoxicity Testing In Vitro: The Current Situation

1997 ◽  
Vol 25 (5) ◽  
pp. 497-503
Author(s):  
Jean-Paul Morin ◽  
Marc E. De Broe ◽  
Walter Pfaller ◽  
Gabriele Schmuck
Keyword(s):  
Proximal Tubule ◽  
Culture Media ◽  
Task Force ◽  
Primary Cultures ◽  
Phenotypic Expression ◽  
Transepithelial Transport ◽  
Specific Cell ◽  
Proximal Tubule Cells ◽  
Tubule Cells

An ECVAM task force on nephrotoxicity has been established to advise, in particular, on the follow-up to recommendations made in the ECVAM workshop report on nephrotoxicity testing in vitro. Since this workshop was held, in 1994, there have been several improvements in the techniques used. For example, the duration of renal slice viability, and the maintenance of functional activities in slices, have been improved by using dynamic incubation systems with higher oxygen tensions and more-appropriate cell culture media. Highly differentiated primary cultures of pig, human and rabbit proximal tubule cells have been established by using specific cell isolation procedures and/or selective culture media. To date, the most comparable phenotypic expression and transepithelial transport capacities to proximal tubules in vivo have been obtained with primary cultures of rabbit proximal tubule cells which are grown on bicompartmental supports; in this system, transepithelial substrate gradients are generated and the transepithelial transport of both organic anions and cations is highly active. This in vitro system has been selected by ECVAM for further evaluation and prevalidation. Industrial needs in the area of nephrotoxicity testing have been identified, and recommendations are made at the end of this report concerning possible future initiatives.

scholarly journals Replacement of Albumin by Preovulatory Oviductal Fluid in Swim-Up Sperm Preparation Method Modifies Boar Sperm Parameters and Improves In Vitro Penetration of Oocytes

Animals ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1202
Author(s):  
Sergio Navarro-Serna ◽  
Evelyne París-Oller ◽  
Ondrej Simonik ◽  
Raquel Romar ◽  
Joaquín Gadea
Keyword(s):  
Calcium Concentration ◽  
Preparation Method ◽  
Culture Media ◽  
Penetration Rate ◽  
Sperm Parameters ◽  
High Concentration ◽  
Sperm Preparation ◽  
Boar Sperm ◽  
Oviductal Fluid

More suitable and efficient methods to protect gametes from external harmful effects during in vitro handling can be achieved by adding preovulatory porcine oviductal fluid (pOF) to in vitro culture media. The objective of this study was to assess the swim-up procedure’s suitability as a sperm selection method using a medium supplemented with 1mg/mL BSA, 1% preovulatory pOF (v/v), 1% v/v pOF plus 1mg/mL BSA, and 5mg/mL BSA. After selection, various sperm parameters were studied, such as sperm recovery rate, sperm morphology, motility (by CASA), vitality, acrosome status and intracellular calcium (by flow cytometry) and ability to penetrate oocytes in vitro. Around 2% of sperm were recovered after swim-up, and the replacement of BSA by pOF showed a beneficial reduction of motility parameters calcium concentration, resulting in an increased penetration rate. The combination of albumin and oviductal fluid in the medium did not improve the sperm parameters results, whereas a high concentration of BSA increased sperm morphological abnormalities, motility, and acrosome damage, with a reduction of calcium concentration and penetration rate. In conclusion, the replacement of albumin by preovulatory oviductal fluid in the swim-up sperm preparation method modifies boar sperm parameters and improves the in vitro penetration of oocytes.

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