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l -cysteine (L-cys) modified ZnS (LZS) quantum dots (QDs) were synthesized by a two-step way. The effects of LZS QDs on the phenotype, biomass, morphology and extracellular protein content of Aspergillus oryzae (A. oryzae) were studied in detail for the first time. The results illustrated that both uncoated and coated QDs had a cubic blend ZnS structure, and the particle size was about 4.0 and 4.3 nm, respectively. It was found that LZS QDs acted as an activator to stimulate the growth of A. oryzae in both solid and liquid media, and the biomass with 50 μg/mL (25 μg/mL) QDs in solid (liquid) medium was the highest and was about 1.65 (3.74) times higher than that in control medium (without QDs). The data of ultraviolet–visible (UV–Vis) revealed that the protein concentration of A. oryzae was 824.904 and 467.748 μg/mL relating to liquid medium with and without QDs, respectively. This work can provide a new way to increase the production of A. oryzae or microorganisms.

Acanthamoeba spp. is one of the free-living amoebas in the environment that many studies focused on. This parasite is of veterinary and public health importance. The present study aimed to evaluate several available media for Acanthamoeba spp. culture to find a suitable axenic liquid medium. In the present study four culture media, including TYM, RPMI-1640, and hydatid cyst fluid (HCF) as well as non-nutrient agar (NNA) as the control group, were examined to evaluate the proliferation of Acanthamoeba T4 genotype at 24, 48, and 72 h. Based on our results, at 24 and 48 h, the highest number of cysts was observed in TYM medium and the lowest in NNA medium. Moreover, at 72 h, the highest and lowest number of cysts was detected in the HCF and NNA media, respectively. In the present study, the best medium for proliferation of the parasite in the short term was TYM. Furthermore, the parasite proliferated continuously in a considerable number in HCF medium, and this medium can be used if continuative examinations are needed on the parasite.

Effects were assessed for the post-culture liquid medium originating from the cultivation of microorganisms that are present in the ecosystem called symbiotic consortium bacteria and yeast (SCOBY). The effectiveness of protecting Scotch pine wood samples against decomposition caused by the fungus Coniophora puteana was evaluated. The obtained results confirmed that impregnation of wood with post-culture medium reduces the possibility of decomposition of this wood by the test fungus. The potential biocidal effect of the post-culture medium from SCOBY was attributed to the presence the substances of a potential fungicidal nature, which were synthesized in the culture medium during metabolic processes occurring in the culture. The obtained results encourage further studies on the potential use of metabolites obtained from SCOBY breeding to protect wood against biodegradation.

The objective: to study the phenotypic sensitivity of Mycobacterium tuberculosis (MTB) to fluoroquinolones, ofloxacin and moxifloxacin and correlate it with MTB genome mutations associated with resistance to fluoroquinolones to determine the possibility to use these data when compiling chemotherapy regimens IV and V. Subjects and methods. The study included 108 multiple drug resistant Mycobacterium tuberculosis cultures obtained in 2018-2019 from specimens of tuberculosis patients from three regions of Russia. To test drug susceptibility of the isolated cultures, standard methods of the modified method of proportions on a liquid medium by Bactec MGIT 960 were used. Genetic determinants of multiple and extensive drug resistance of tuberculosis mycobacteria were detected using the test system of TB-TEST, OOO Biochip-IMB. Results. Of 66 cultures resistant to ofloxacin, 26 (39.4%) were found to be sensitive to moxifloxacin, while 40 (60.6%) were resistant. Thus, the interpolation of data of ofloxacin drug susceptibility testing to moxifloxacin is unjustified, and using the absolute concentration method as the only phenotypic drug susceptibility testing method in modern laboratories is unacceptable. It was demonstrated that the high level of resistance to moxifloxacin (0.5 μg/ml) was mainly due to the presence of the gyrAD94G, gyrAD94N, and gyrAD94H substitutions in the genome. It was suggested to revise the existing concentrations for moxifloxacin during phenotypic drug susceptibility testing.

Among more than 600 types of micromycetes of clinical importance, fungi of the Candida spp. are the most common pathogen, causing different forms of candidiasis (superficial candidiasis, visceral candidiasis). The aim of this work was to determine the antifungal activity of new derivatives of 4-(het)aryl-2,4-dioxobutanoic acids, in relation to typical strains of Candida yeast and to identify the most active compounds. The study of the antifungal activity of the compounds was carried out by the micromethod of double serial dilutions in a liquid medium. The antifungal activity of 6 compounds in relation to 4 typical strains of Candida spp. was studied; the high antimycotic activity of 4 compounds against the Candida albicans strain was established. A leader compound with high antifungal activity for all the studied strains was revealed. The prospects of studying the modified compounds of this series are shown.

Objectives The aim of this study was to describe the evaluation of the use of MALDI-TOF MS for the identification of non-tuberculous mycobacteria (NTM) and Mycobacterium tuberculosis directly from liquid MGIT cultures from January 2017 to December 2017. Material/methods A total of 155 isolates (mainly respiratory) were analyzed by MALDI-TOF MS (Bruker Daltonics) directly from MGIT liquid medium with a previous extraction procedure. Results MALDI-TOF MS generated acceptable scores for 152 isolates (98.06%). Fifty isolates were identified as M. tuberculosis complex and the remaining 105 as NTM (M. abscessus subsp. abscessus, M. avium, M. celatum, M chelonae, M. chimaera, M. fortuitum, M. gordonae, M. intracellulare, M. kansasii, M. lentiflavum, M. mageritense, M. mucogenicum and M. xenopi). Conclusions These results indicate that MALDI-TOF MS can be useful to identify mycobacteria directly from MGIT cultures and is an accurate, rapid and cost-effective system to be used as a routine method.

In this study, strategies for recovery of ammonia-stressed AD reactors were attempted, by addition of preserved bioaugmentation consortium in gel (BioG), fresh consortium in liquid medium (BioL), woodchip biochar (BW), and straw biochar (BS). In comparison to control group with ammonia, effective treatments, i.e., BioG, BioL, BW and BS raised the maximum methane production rate by 77%, 23%, 35%, and 24%, respectively. BW possibly acted as interspecies electrical conduits for Direct Electron Transfer based on conductivity and SEM analysis. BioG facilitated slow release of bioaugmentation inocula from gel into the AD system, which protected them from a direct environmental shock. According to microbial analysis, both BioG, BioL and BW resulted in increased relative abundance of Methanothermobacter thermautotrophicus; and BS induced selective raise of Methanosarcina thermophila. The increase of methanogens via these strategies led to the faster recovery of the AD process. publishedVersion

The results of comparative experimental studies of identification of nontoxigenic C. diphtheriae strain by three different commercial laboratories are presented. A typical nontoxigenic strain of C. diphtheriae biovar mitis was used. For the studies, three lines of ten-fold dilutions of bacterial culture were prepared, followed by control planting on the medium and counting CFU/ml. In the experiment, tampons were pooled with a 24-hour bacterial culture of a nontoxigenic C. diphtheriae strain. Tampons were provided from three different laboratories - ∑-Transwab® with Ames liquid medium (from the first and second laboratories) and a viscose tampon with coal medium (from the third laboratory). After pooled, tampons were delivered to commercial laboratories. And as a result of the experiment, Corynebacterium spp. was identified in first laboratory (103 CFU/tamp), S. epidermidis (102 CFU/ml) - in second laboratory and nontoxigenic C. diphtheriae biovar gravis - in third laboratory. The study indicates that there is a need to the supervision of bacteriological investigations conducted in various laboratories. This will improve the quality of investigations on diphtheria infection and identify of diphtheria carrier, which is a reservoir of the causative agent of diphtheria, and will contribute to the maintenance of sanitary and epidemiological well-being in our country.

Hazelnut (Corylus avellana L.), contains a valuable medicinal substance known as Paclitaxel®, which is one of the most effective anticancer drugs. The original plants produce negligible amount of paclitaxel; therefore, tissue culture techniques, especially hairy root culture, could be one of the most practical methods to enhance the amount of paclitaxel. The main goal of this study was to assess the induction of hairy roots in C. avellana. The effects of different strains of Agrobacterium rhizogenes including c58c1pRiA4, K599, and 15834, and six culture media, MS (Murashige and Skoog), half-strength MS, quarter-strength MS, WPM (woody plant media), half-strength WPM, and quarter-strength WPM, were evaluated. The results showed that the maximum amounts of the rooted explants were obtained with c58c1pRiA4 strain in quarter-strength WPM medium. The investigations of explant type (leafstalk, petiole, lamina, and stem) and different propagation media (quarter-strength WPM, half-strength MS, and half-strength SH ((Schenk and Hildebrandt) medium) showed that the leafstalk was the most optimal explant for hairy root induction, and half-strength SH was the best culture medium for growth of the hairy roots in liquid medium. HPLC analyses confirmed the presence of paclitaxel (3.2 μg g−1 (DW)) in hairy root extracts.

The aim of this research was to evaluate the redox system of Noc­ardia corallina B-276 in the biotransformation of 1-phenyl-1-propanone ( 1a ), 2-hydro­xy-1-phenylethanone ( 2a ) and methyl (2-chlorophenyl)(oxo)acetate ( 3a ) into 1-phenylpropan-1-ol ( 1b ), 1-phenyl-1,2-ethanediol ( 2b ) and methyl (2-chloro­phenyl)(hydroxy)acetate ( 3b ). The biomass of N. corallina was obtained in a liquid medium with an initial pH of 8.50, but the pH changed during the 96 h of the culture media, the final pH was between 4.74 and 7.62. The N. corallina biomass biocatalyzed the enantioselective reduction of 1a – 3a to the corresponding alcohols. Whereas, during the process of oxidation of the rac -alcohols 1b–3b, 1b was oxidized in enantioselective way, the oxidation of 2b was not selective, but 3b was biotransformed mainly to ( R )- 3b . These results are indicative that N. corallina produced reductases and oxidases, whereby the biocatalytic activity was influenced by the final pH of the culture media, the reaction time and structure of the substrate.

The study about the growth of white oyster mushroom mycelium (Pleurotus ostreatus) (Jacq) (P. Kumm) from liquid and solid inoculum conducted on January until July 2018 in Biotechnology laboratory, Biology Department, Faculty of Math and Science, Tadulako University. This study was aimed to find out the mycelium growth of source of stem inoculum and hood in liquid and solid media. This study was designed by Complete Randomized Design (CRD) which consisted of four treatments and five replications. The treatment were M1 (source of stem inoculum in liquid medium), M2 (source of stem inoculum in solid medium), M3 (source of hood inoculum in liquid medium), and M4 (source of hood inoculum in solid medium). The parameters were (a) incubation time till mycelium fills medium, (b) CFU, and (c) the viability of inoculum on producing medium. The result showed that the faster incubation were M1 and M3 (for 2 days, higher CFU was M3 with an average number 8,2 x 10 10 CFU /ml. The faster viability of mycelium growth were M3 with an average growth of mycelium 6,97 cm/days.

In accordance with the requirements of the State Pharmacopoeia of the Russian Federation, 14th edition, Clostridium novyi 198 test strain is used to determine growth promoting properties of thioglycollate medium when testing sterility of immunobiological medicinal products. The preservation of C. novyi 198 as spores requires a special liquid medium containing insoluble calcium chloride casein (CChC). At present, there is no reference standard available on the Russian market for the quality control of CChC used as a component of the media for preservation of C. novyi 198.The aim of the study was to develop and certify the first batch of calcium chloride casein industry reference standard (IRS) for the quality control of CChC batches and liquid media containing CChC which is used to preserve C. novyi 198 as spores.Materials and methods: thermally denatured CChC which was chemically synthesized from industrial CChC was used as candidate material. The quality control was performed using the test procedures described in procedural guidelines MUK 4.2.2316-08. C. novyi 198 spores were obtained as described in the «Sterility testing» section of MUK 4.2.2316-08 (preparation, inoculation and incubation in the medium containing calcium chloride casein IRS).Results: a list of quality parameters was developed for calcium chloride casein IRS: «Spore formation» (certifiable parameter), «Appearance», «Loss on drying», «Clarity of medium». The first batch of the candidate calcium chloride casein IRS was certified. The value of the «Spore formation» certifiable parameter was found to be (32.6 ± 7.3) %, the IRS shelf life was 5 years when stored in a dry place protected from light at a temperature of 15 to 25 °C. A documentation package for the IRS 4228-444-2018 was prepared and approved. The liquid medium containing CChC reference material (0.3 g/10 mL) allows for preservation of the biological properties of the C. novyi 198 working culture for at least 12 months.Conclusions: the certification results demonstrate that the calcium chloride casein IRS meets the applicable requirements and can be used as reference material to control the quality of new CChC batches and to assess the suitability of liquid medium containing CChC for preservation purposes. The use of the calcium chloride casein IRS makes it possible to standardise and greatly simplify the procedure for preserving the stability of C. novyi 198, to increase the efficiency and reliability of immunobiologicals quality control in terms of Sterility, and to reduce costs when using a freeze-dried strain.

The purpose of the work was to investigate the organoleptic, physico-chemical and microbiological indices of traditional Carpathian cheeses, in particular, brynza, butz and vurda, made directly on the mountain valleys with traditional technology in Rakhiv district of the Transcarpathian region. Of 16 samples of cheese, 13 samples were cheese butz (samples 1, 2, 3, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15); vurda cheese (samples 4 and 16) and cheese of brynza a year ago (sample 14). Experimental studies of organoleptic, physico-chemical and microbiological parameters of cheese samples were carried out in the laboratory of the milk and milk products department and in the department of biotechnology at Rzeszow University (Poland). In samples of cheese, organoleptic parameters were studied in accordance with GOST 7616-85; active acidity – by potentiometric method using the pH meter of the ARN-9 brand; titrated acidity – titrarily (in degrees Turner) according to GOST 3624-67. The mass fraction of salt in the cheese was determined by titration of the extract with nitric oxide silver (GOST 3627-81). The detection of bacteria of the genus Staphylococcus aureus was determined by sowing on a solid Gissa with mannitol, with further detection and confirmation of the belonging of the grown colonies to Staphylococcus aureus (GOST 30347-97). The detection of bacteria of the genus Salmonella was determined by sowing on the Endo solid medium (GOST 31659-2012). The bacteria of the intestinal stem group were determined by sowing on a Kessler liquid medium (GOST 9225-84). Cultivation of microorganisms was carried out at temperature +37 °С for 48 h. According to researches of organoleptic parameters of cheeses it was found that samples of cheese butz were characterized by sour milk, and samples of 3 and 5 sour taste and aroma, a dense humus consistency and from gray to white color. Figure – with single small and large bodies of irregular shape. According to studies of organoleptic parameters of cheese, the taste and aroma of the product should be given – light sour milk, salted, which is confirmed by the salt content – 4.6%. Consistency was crumbly, the color was yellow. Samples of cheese vurda were characterized by delicate butter, sour milk taste and smell, paste, crumbly consistency and white color. Analyzing the acidity of butz cheese it was that it was high particularly in samples 3, 8 and 14, which was reflected in organoleptic parameters. In the samples of vurda titrated acidity was 75 °T in sample 4 (pH 5.7) and 16 °T in sample 16 (pH 6.4), while brynza was characterized by the highest acidity – 260 °T (pH 4.9). According to microbiological parameters, all specimens were characterized by the presence of pathogenic microflora. In particular, 1, 3, 5–9, 11–15 samples showed Staphylococcus aureus, 10 sample of Staphylococcus epidermis, 2 sample of bacteria of the genus Salmonella, 16 sample of Enterobacter aerogenes, and 4 samples of mold.

Background and objectives: Gynura procumbens has been widely used as a traditional medicine in Indonesia. Micropropagation of this plant was important to fulfill the need of pharmaceutical industry. The aims of this study were to increasing the number of G. procumbens shoots in balloon-type bubble bioreactor influenced by sucrose concentration and inoculums density. Materials and Methods: Nodal segments of G. procumbens were used as explants which were grown in balloon type bubble bioreactor in MS liquid medium supplemented with 2 mg LG1 IAA and 4 mg LG1 BAP. Shoot culture were conducted under treatment of various concentration of sucrose (10, 30 and 50 g LG1) and various inoculums density (5, 10 and 15 explants). Results: The highest multiplication of shoots and growth index were obtained at 50 g LG1 sucrose treatment and 5 explants as initial inoculums; there are 9.36 shoots/explants and 13.8 shoots/explants, respectively. Conclusion: The study found that G. procumbens shoots cultured in balloon-type bubble bioreactors increased shoots multiplication significantly.

Discharge of textile dyes in wastewater of textile factories represents a major environment problem threatening the aquatic life. The decolorization of such harmful chemicals is of paramount importance. In this work, Aspergillus niger, a brown rot fungi, isolated from the effluent samples around the textile industry of El- Baragil, Giza, Egypt was evaluated for its efficiency in decolorization of two textile dyes; Remazole blue and Remazol red. Effluent samples were also analyzed for their physiochemical properties. The optimum conditions for decolorization were found to be 4 days incubation, 30 ° C and pH4 in Sabouraud liquid medium supplemented with dextrose (10 g/l) and ammonium sulfate (2 g/l) as a carbon and nitrogen source, respectively. The response of the isolate to the increase in dye concentrations was found to be growth inhibitory. Decolorization extent and facile conditions showed the potential for the A.niger to be used in biological treatment of textile dyes.

Oldenlandia umbellata L. gains importance due to its medicinal properties and the presence of anthraquinones based natural dyes in the roots. Present study describes the effect of Murashige and Skoog’s (MS) liquid medium (full strength) on in vitro regeneration, flower bud induction and ex vitro rooting in O. umbellata. Shoot segments with 2-3 nodes (each node with 2 axillary buds) served as explants for establishment of cultures. The liquid medium augmented with 2.0 mg L-1 6-benzylaminopurine (BAP) with additives (50 mg L-1 of ascorbic acid and 25 mg L-1 each of arginine, adenine sulphate and citric acid) was effective for shoot bud induction (6.4±0.19 shoots per explant within 2-3 weeks). The shoots were further multiplied (89.3±1.07 shoots, 2-3 weeks) when the shoot clusters obtained from the culture initiation directly transferred to the full-strength MS liquid medium incorporated with 1.0 mg L-1 BAP and 0.5 mg L-1 indole-3 acetic acid (IAA) with additives. Flower buds were induced (12.0±0.15 buds per shoot) when the shoots were cultured on 1.0 mg L-1 BAP and kinetin (Kin, 6-furfurylaminopurine) and 0.5 mg L-1 of IAA at 45 µmol m−2 s−1 SFPD (Spectral Flux Photon Density) light intensity for 14/10h (light/dark) photoperiod. The adventitious roots were induced on 1/4 strength MS medium supplemented with 1.5 mg L-1 indole-3 butyric acid (IBA). Ex vitro rooting was achieved (16.0±0.53 roots per shoot) by pulse treatment of the shoots with 300 mg L-1 IBA for 2 min. The in vitro produced plantlets were acclimatized in the greenhouse and finally translocated to the in vivo conditions with 93 % success rate. This is the foremost (use of liquid MS medium) and cost-effective method for large scale multiplication of O. umbellata.

Javanese ginseng (Talinum paniculatum) is an herb and one of the traditional medicinal plants in Indonesia which accumulate saponin in the root organ. However, slow root growth hampers the accumulation of the compound in this plant. Therefore, in vitro culture of adventitious roots offers an alternative way to increase the production of saponin. Furthermore, this study was performed to investigate the effect of methyl jasmonate (MeJA) and salicylic acid (SA) on the growth and saponin content in adventitious root of Javanese ginseng. Adventitious root culture was induced on solid Murashige and Skoog (MS) medium supplemented by 10 µM IBA (indole-3-butyric acid). Subsequently, adventitious roots were subcultured into a half strength MS liquid medium with the addition of different concentrations of MeJA or SA and incubated for 5, 10 and 15 days. Despite inhibition on the root growth, saponin production increased by 1.5 and 1.3 fold upon elicitation with 0.2 mM MeJA and SA for 15 days, respectively. Thus, MeJA and SA elicitations regulate saponin biosynthesis in adventitious root culture of Javanese ginseng as a time and dose-dependent manner. Key words: Adventitious root, elicitation, Javanese ginseng, saponin, Talinum paniculatum.

The development of low-cost, fast, and reliable methods for detecting Mycobacterium tuberculosis (MTB) infection and drug susceptibility is critical for tuberculosis control. The new microscopic examination of liquid drug susceptibility assay (MODS) examines early MTB colonies in a liquid medium, which permits for more convenient diagnosis and testing of drug susceptibility. The sensitivity of MODS (91%) was superior as compared to the sensitivity of different culture methods (92%). The MGIT and MODS were used to monitor tuberculosis-positive sputum samples for isoniazid and rifampin susceptibility. Concordance between MODS and MGIT was found in 89 percent of cases. MODS are the fastest method used for diagnostic and susceptibility testing (median, 10.0 or 9 days). MODS is a fast, low-cost, responsive, and particular method for detecting and testing MTB susceptibility; it is specifically well-suited utilize in growing countries with high infection rates and a growing number of multidrug-resistant cases.

Resumo As pesquisas com fungos em busca de substâncias com potencial atividade biológica ganham cada vez mais destaque por serem fontes sustentáveis de novas moléculas. Diplodia pinea é um fungo fitopatogênico encontrado em espécies florestais, que tem apresentado poucos relatos de moléculas com atividade biológica e pode apresentar potencial biotecnológico promissor. Assim, o objetivo deste estudo foi detectar substâncias produzidas por esse fungo em meio líquido, sob condições de estresse. Um metabólito foi detectado na fração alcoólica e caracterizado por difração de raios-X como D-manitol. Esta é a primeira descrição deste monossacarídeo produzido por D. pinea. Abstract Research with fungi in search of substances with potential biological activity is gaining more prominence since they are sustainable sources of new molecules. Diplodia pinea is a phytopathogenic fungus found in forest species, which has shown few reports of molecules with biological activity and may present promising biotechnological potential. Thus, the objective of this study was to detect substances produced by this fungus in a liquid medium, under stressful conditions. A metabolite was detected in the alcoholic fraction characterized by X-ray diffraction as D-mannitol. This is the first description of this monosaccharide produced by D. pinea. Resumen La investigación con hongos en busca de sustancias con potencial actividad biológica está ganando más protagonismo ya que son fuentes sostenibles de nuevas moléculas. Diplodia pinea es un hongo fitopatógeno que se encuentra en especies forestales, que ha mostrado pocos informes de moléculas con actividad biológica y puede presentar un potencial biotecnológico prometedor. Así, el objetivo de este estudio fue detectar sustancias producidas por este hongo en un medio líquido, en condiciones de estrés. Se detectó un metabolito en la fracción alcohólica caracterizada por difracción de rayos X como D-manitol. Ésta es la primera descripción de este monosacárido producido por D. pinea Made available in DSpace on 2021-07-05T19:00:56Z (GMT). No. of bitstreams: 1 Celso-16424-Article-211106-1-10-20210621.pdf: 233954 bytes, checksum: 30751e54426cfbefcdf2d764b2ddfa00 (MD5) Previous issue date: 2021

SUMMARY Anabaena azollae can be completely eliminated from megasporocarps of Azolla by the removal of the indusium and apical membrane after horizontally cutting the megaspore apparatus beneath the apical membrane. All subsequent sporelings of Azolla are free from Anabaena. These do not fix N and fail to survive on N2-free media. They grow normally, however, in N2-containing liquid medium or in a soil-water system with small amounts of added N and eventually bear sporocarps. Anabaena is again absent in the new megasporocarps and sporelings of Azolla derived from the second generation of megaspores are likewise Anabaena-free.

The availability of high-quality seeds is now a necessity. This is due to a government program to replace oil palm trees in smallholder plantations with high quality seeds. An efficient protocol to produce a large number of embryos is needed. To increase the number of embryogenic callus production, the callus proliferation experiment was carried out through suspension culture. This study aimed to examine the proliferation ability of embryogenic callus from three different oil palm clones, in several repeated subcultures. Liquid MS media added with 1 ppm 2.4-D and 0.1 ppm NAA were used. Embryogenic callus was weighed by 0.1 - 0.2 g, transferred into the liquid media, shaking at 60-80 rpm and 27 ºC for 8 weeks without light. Continues subcultures were repeated up to 7 times. The results showed that the growth rate of embryogenic callus increased in the third and fourth subcultures and then decreased in subsequent subcultures. It also revealed that the entire embryogenic callus from the first subculture up to seventh subculture still has the ability to regenerate into new plants. These results indicate that oil palm embryogenic callus can be proliferated by suspension culture with a limit up to the fourth subculture. Ketersediaan benih kelapa sawit berkualitas saat ini merupakan kebutuhan karena adanya program pemerintah untuk menggantikan tanaman sawit di kebun-kebun petani. Salah satu cara vegetatif yang dapat dilakukan adalah meningkatkan jumlah kalus embriogenik yang dihasilkan melalui pengembangan kultur suspensi. Penelitian ini bertujuan mengkaji kemampuan proliferasi kalus embriogenik dari tiga klon kelapa sawit, pada beberapa kali subkultur yang berulang. Media cair MS dengan penambahan 1 ppm 2,4-D dan 0,1 ppm NAA digunakan untuk memperbanyak 0,1–0,2 g kalus embriogenik, dikocok pada 60-80 rpm dan suhu 27 ºC tanpa cahaya selama 8 minggu. Subkultur berulang dilakukan hingga 7 kali. Hasil percobaan menunjukkan bahwa kemampuan proliferasi kalus dipengaruhi oleh genotip tanaman induk. Rata-rata kalus embriogenik dapat meningkat pada subkultur ke-3 dan ke-4 dan semakin menurun pada subkultur selanjutnya. Kalus embriogenik hasil proliferasi subkultur pertama hingga ke-7 dapat tumbuh menjadi calon tanaman baru. Hasil ini menunjukkan bahwa kalus embriogenik kelapa sawit dapat diperbanyak dengan kultur suspensi pada batas sampai subkultur ke-4.

Nanocellulose fibers bioengineered by bacteria are a high-performance three-dimensional cross-linked network which can confine a dispersed liquid medium such as water. The strong chemical and physical interactions of dispersed water molecules with the entangled cellulosic network allow these materials to be ideal substrates for effective liquid separation. This type of phenomenon can be characterized as green with no equivalent precedent; its performance and sustainability relative to other cellulose-based or synthetic membranes are shown herein to be superior. In this work, we demonstrated that the renewable bacterial nanocellulosic membrane can be used as a stable liquid-infused system for the development of soft surfaces with superwettability and special adhesion properties and thus address intractable issues normally encountered by solid surfaces.

Nematode-trapping fungi (NTF) and nematodes are common and sympatric in nature. The molecular basis that underlies this interkingdom predator-prey interaction remains largely uncharacterized. Both NTF and nematodes can be easily isolated from soil samples. NTF do not form traps in nutrient-rich environments, yet trap morphogenesis can be observed under nutrient-poor conditions and upon simultaneous sensing of the nematode cues. Here, we present protocols for laboratory maintenance and culturing of the model NTF Arthrobotrys oligospora. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Growth of nematode-trapping fungi on solid medium Basic Protocol 2: Growth of nematode-trapping fungi in liquid medium Basic Protocol 3: Collection of conidia from solid medium Support Protocol 1: Preparation of Miracloth filter funnel Basic Protocol 4: Induction of trap morphogenesis Alternate Protocol: Quantitative measurement of trap induction Support Protocol 2: Preparation of synchronized C. elegans L4 Basic Protocol 5: Establishing C. elegans survival rate upon exposure to A. oligospora Basic Protocol 6: Storage of nematode-trapping fungi strains.

Cryptococcus neoformans is an opportunistic fungal pathogen primarily targeting immunosuppressed populations in both resource-rich and resource-limited nations. Successful treatment is limited to a few antifungals that have become compromised by cryptococcal resistance, leading to intensive research seeking new drug candidates. Two distinguishing hallmarks of this species are the ability to develop a polysaccharide capsule and melanization of the fungal cells. These also act as virulence factors, protecting this pathogen in the host as well as in the environment. Here we describe two classic methods to document capsule and melanin. Although initially described and documented several decades ago, these methods remain relevant in spite of the advent of more sophisticated methodology, due in part to their simplicity and cost efficiency. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Capsule visualization by India ink counterstaining Basic Protocol 2: Assessment of melanin on solid media Alternative Protocol: Quantification of melanin production in liquid medium.

In this study, mixed cultures of brown-rot fungus Gloeophyllum trabeum and filamentous fungus Trichoderma viride in decolorizing Methylene Blue (MB) has been investigated. T. viride culture was mixed with G. trabeum culture and then MB was added into the media to achieve final concentration 95.24 mg/L. The results showed single culture of G. trabeum and T. viride can decolorize MB only 31.50% and 53.89%, respectively. While decolorization of MB by mixed cultures of G. trabeum and T. viride reached to 74.72% in a liquid medium of potato dextrose broth (PDB) for 7 days. The results showed mixed cultures had the highest ability to decolorize MB among those variations. Based on the LC- TOF/MS analysis, the peak of MB in mixed cultures treatment had a lower intensity compared to the peak of MB in abiotic control. The metabolites from MB biodecolorization by G. trabeum were C8H10N2SO5, C6H8N2SO3, C16H20N3SO, C6H7NSO3, and C12H10N3S, while the metabolites from MB biodecolorization of T. viride were C16H23N3O3S, C22H26NO2S, and C31H48N3S. On the other hand, the metabolites from the MB biodecolorization by mixed cultures were C16H23N3O3S, C31H48N3S, C31H47N2OS and C17H20N2S. This study indicated that mixed cultures of brown-rot fungus G. trabeum and filamentous fungus T. viride was effective to decolorize MB dye.

As in any other economically important crop, the possibility of producing fully homozygous, doubled haploid lines in cucumber allows for faster and cheaper breeding. At present, the fastest way to doubled haploidy is the production of cucumber haploid plants and duplication of their chromosomes to make them doubled haploid. In this chapter, we describe a complete protocol to successfully produce cucumber doubled haploid plants, including the evaluation of their ploidy level by flow cytometry. Briefly, this protocol involves a first step of anther culture to induce microspores to divide and proliferate forming calli. The calli produced are isolated from anthers and transferred first to a liquid medium and then to a solid medium to induce organogenesis. Organogenic shoots will eventually give rise to entire DH plants.

A method based on reversed phase high-performance liquid chromatography coupled with electrospray ionization high-resolution mass spectrometry (RP-HPLC-ESI-HRMS) for the comprehensive and reliable detection of secondary metabolites of Trichoderma reesei cultured in synthetic minimal liquid medium is presented. A stable isotope-assisted (SIA) workflow is used, which allows the automated, comprehensive extraction of truly fungal metabolite-derived LC-MS signals from the acquired chromatographic data. The subsequent statistical data analysis and a typical outcome of such a metabolomics data evaluation are shown by way of example in a previously published study on the influence of the pleiotropic regulator transcription factor Xylanase promoter binding protein 1 (Xpp1) in T. reesei on secondary metabolism.

Yersinia pseudotuberculosis has been studied for many decades, and research on this microbe has taught us a great deal about host-pathogen interactions, bacterial manipulation of host cells, virulence factors, and the evolution of pathogens. This microbe should not be cultivated at 37°C because this is a trigger that the bacterium uses to sense its presence within a mammalian host and results in expression of genes necessary to colonize a mammalian host. Prolonged growth at this temperature can result in accumulation of mutations that reduce the virulence of the strain, so all protocols need to be modified for growth at room temperature, or 26°C. This article describes protocols for cultivating this microbe and for its long-term storage and its genetic manipulation by transformation and conjugation. © 2020 Wiley Periodicals LLC. Basic Protocol 1: Growth of Y. pseudotuberculosis from a stock Basic Protocol 2: Growth of Y. pseudotuberculosis in liquid medium from a single colony Basic Protocol 3: Freezing Y. pseudotuberculosis in glycerol for long-term storage Basic Protocol 4: Transformation of Y. pseudotuberculosis by electroporation Basic Protocol 5: Tri-parental mating/conjugation.

Identification of mycobacteria by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) requires not only a good protein extraction protocol but also an adequate cutoff score in order to provide reliable results. The aim of this study was to assess the cutoff scores proposed by the MALDI-TOF MS system for mycobacterial identification. A total of 693 clinical isolates from a liquid medium and 760 from a solid medium were analyzed, encompassing 67 different species of nontuberculous mycobacteria (NTM). MALDI-TOF MS identified 558 (80.5%) isolates from the liquid medium and 712 (93.7%) isolates from the solid medium with scores of ≥1.60. Among these, four (0.7%) misidentifications were obtained from the liquid medium and four (0.5%) from the solid medium. With regard to species diversity, MALDI-TOF MS successfully identified 64 (95.5%) different species, while PCR-reverse hybridization (GenoType Mycobacterium CM and AS assays) identified 24 (35.8%) different species. With MALDI-TOF MS scores of ≥2, all isolates were correctly identified, and with scores in the range from 1.60 to 1.99, most isolates were correctly identified, except for Mycobacterium angelicum, M. parascrofulaceum, M. peregrinum, M. porcinum, and M. gastri. In conclusion, MALDI-TOF MS is a useful method for identifying a large diversity of NTM species. A score threshold of 1.60 proved useful for identifying almost all the isolates tested; only a few species required a higher score (≥2.00) to obtain a valid definitive identification.

Vibrio fischeri is a nonpathogenic organism related to pathogenic Vibrio species that can be readily grown and stored with common laboratory equipment. In this article, protocols for routine growth, storage, and phenotypic assessment of V. fischeri, as well as recipes for useful media, are included. Specifically, this article describes procedures and considerations for growth of this microbe in complex and minimal media. It also describes assays for biofilm formation, motility, and bioluminescence, three commonly assessed phenotypes of V. fischeri. © 2020 Wiley Periodicals LLC. Basic Protocol 1: Growth of V. fischeri from frozen stocks Basic Protocol 2: Growth of V. fischeri in rich, undefined liquid medium Alternate Protocol 1: Growth of V. fischeri in minimal medium Basic Protocol 3: Storage of V. fischeri in frozen stocks Basic Protocol 4: Biofilm assay on solid agar Alternate Protocol 2: Biofilm assay in shaking liquid culture Alternate Protocol 3: Biofilm assay in static liquid culture Basic Protocol 5: Motility assay Basic Protocol 6: Luminescence assay.

Tissue culture have been used for plant propagation generally, and the medium has been important role in its growth. Vegetative propagation on Phalaenopsis sp orchids can be through the protocorm like bodies (PLB). Medium of affect on propagation of PLB was carried out on medium type, kind of basal medium and concentrations ratio of naphtaleneacetic acid (NAA) and benzylamino purine (BAP). The experiment used Completely Randomized Factorial Design with 3 replications and continued with the Duncan Multiple Range Test (DMRT) if there were significant differences. The results showed that the best callus formed in a combination of solid medium type and Murashige & Skoog (MS) basal medium was 100%. The most number of PLB produced from a combination of liquid medium types and a concentration ratio of NAA 1 mgL-1 and BAP 5 mgL-1, the most number of plantlet produced from a combination of MS basal medium and the concentrations ratio of NAA 0.1 mgL-1 and BAP 0.1 mgL-1, the number of PLB germination and PLB with leaves were influenced by each single factor.

The Lead [Pb(II)] tolerance and uptake ability of four fungal species, two from the genus Penicillium and two from the genus Talaromyces were investigated in this study. The species were isolated from a polluted tributary and identified to be closest to P. canescens , P. simplicissimum , T. macrosporu s and another Talaromyces sp. via PCR targeting their internal transcribed spacer 1 and 4 sequences. All isolates have tolerances for up to 2000 µg/mL and 3000 µg/mL Pb(II) on solid and liquid medium, respectively. Both Penicillium isolates have increasing removal rates dependent on initial Pb(II) concentration at 500 to 2000 µg/mL, while removal rates of both Talaromyces isolates are not significantly influenced by initial Pb(II) concentrations. The Pb(II) uptake of all isolates increases with increasing Pb(II) concentration but is depressed at 3000 µg/mL, with the exception of T. macrosporus . The recorded total uptake capacities for both Penicillium isolates in this study are higher than in most literature, at 7.0 – 407.4 mg/g and 50.8 – 412.6 mg/g for P. canescens and P. simplicissimum , respectively. The study also reports the exemplary Pb(II) uptake capacities of both Talaromyces isolates at 58.9 – 601.0 mg/g and 60.9 – 402.3 mg/g for T. macrosporus and Talaromyces sp., respectively.

Objective This study aims to determine the optimum liquid medium for the reproduction of Trypanosoma cruzi strains and provide cryopreservation. Methods The reproduction density of T. cruzi strain was evaluated in the following five different commercial liquid culture media: RPMI 1640, Medium 199 (M199), Schneider's Insect Medium (SIM), Nutrient Broth (NB), Brain Heart Infusion Broth (BHIB). Cultures were monitored on every other day for a period of 24 days. Cryopreservation of T. cruzi was also performed and viability was tested after six months. Results Epimastigotes of T. cruzi were not found to be produced in NB and BHIB media. Significant difference was not observed among the reproduction potential of RPMI-1640, M199, and SIM after evaluating the data for the first 10 days. Between days 12 and 24, RPMI-1640 was found to be the best reproduction medium. From the 18th day onwards, parasites transformed amastigotes. On the 24th day, the highest level of amastigote amount was observed, and reproduction was determined to have stopped. As a result of cryopreservation, it was determined that the survival of T.cruzi continued after six months.. Conclusion Thus, the selection of RPMI-1640 medium, followed by M199 and SIM media would be appropriate when studying T. cruzi epimastigotes. Studies using epimastigotes should be planned for up to 18 days and for those using amastigotes, it would be appropriate to plan the studies after the 18th day. Moreover, T. cruzi can be cryopreserved with 15% DMSO and stored for a long time in liquid nitrogen.

To investigate some interesting functional properties of Bacillus spp. in cocoa fermentation processing, 702 strains of Bacillus sp. strains were isolated from six main Ivorian cocoa producer regions. Three mains technological and useful properties for good cocoa beans fermentation were monitored. Among the 386 pectinolytic Bacillus spp. strains, 86.79% of them were confirmed for enzymes production in liquid medium. These 335 pectinolytic Bacillus strains displayed variety of pectinolytic activity with 50.45% of them producing both pectinase and polygalacturonase (PG) enzymes. Among these 169 strains, 70% displayed both the ability to acidify the medium and to degrade the citrate. In mimic conditions of pH and temperature variations as for fermentation heap, 11 strains were able to maintain their activity ranged within 2.12 ± 0.02 to 0.41 ± 0.03 µmol/min/mg of proteins for PG and 2.36 ± 0.03 to 0.25 ± 0.03 µmol/min/mg of proteins for pectinase. Two species (T10T2 and T6HS10) identified as Bacillus subtilus and Bacillus thuringiensis, were selected as the best based on pectinase production level. They were able to continuously produce both pectinolytic enzymes in different conditions of organic acids and ethanol contents. The highlighted properties in this study and the performance of these two strains may make them best candidate as starters for cocoa fermentation control.

The data proving low probability of observing Biofilm Formation (BF) by contemporary clinical strains of M. tuberculosis growing on liquid medium in vitro are discussed. A hypothesis about the role of MDR/XDR development hindering BF production was proposed. It was found that strains capable of producing BF grow on Lewenstein–Jensen medium generated R-form specific colonies shaped as a disk with a convex center, “UFO-colonies”. Sixty seven “UFO”- strains were investigated to BF production, resistance to antibiotics and their belonging to the main epidemics clusters of the Beijing genotype (CC1 and CC2-W148). It was shown that MDR/XDR strains were also capable of BF production that, however, was remarkably more frequent in strains of CC1 and CC2-W148 genotypes. Thus, it was hypothesized that BF production might potentially influence an outcome of chronic forms of TB-infection.

fA study was carried out to screen lovastatin producing ability in Higher Basidiomycetous Fungi isolates from India. For this an extended screening was performed for lovastatin production in Potato Dextrose medium (solid and liquid medium), among a total of 40 basidiomycetous isolates which were obtained from NTCC, Forest Pathology Discipline, Forest Protection Division, Dehradun, Uttarakhand, India). Lovastatin production was evaluated by the disc diffusion method and agar well diffusion method. During the bioassay experiment with Neurospora crassa 159 only six isolates were found to be lovastatin producers viz., Pleurotus ostreatus (NTCC 1253), P. sajor-caju (NTCC 1010), P. floridanus (NTCC 1126), P. eous (NTCC 1264), P. eryngii (NTCC 1070) and P. cystidiousus (NTCC 1006) in liquid and solid growth media with centrifugation and the filtration method.

Dehydrated aerobic granules, if can be sufficiently recovered without significant loss of structural stability and biological activities, presents a promising long-time storage option in practical use. This study dehydrated aerobic granules by six protocols: air drying at 25 or 50 °C, freeze-dry, acetone or ethanol dehydration, and microwave heating, and then recovered them in liquid medium, with the measured characteristics being reported. The granule stability has no correlation with measured settleability, hydrophobicity or extracellular polymeric substances compositions; instead, is correlated with the functional strains presented in the recovered granules. Air dry dehydration minimally damage the functional strains including genus Brevundimonas and genus Comamonas and markedly deteriorated structural breaker such as Acinetobacter of Moraxellaceae to lead to stable and tough recovered granules.

Aim. To determine the variability of the trait of carotenoids biosynthesis in two mutant strains Streptomyces globisporus 1912 after deep cultivation and storage. Relevance. Study of the spontaneous variability of strains-producers of carotenoids after deep cultivation and long-term storage is necessary for understanding the mechanisms of intrapopulation inheritance and the possibility of their control. Methods. Visually the analysis of phenotype of streptomyces colonies, statistical analysis was performed with Windows Software Excel 2007. Results. The appearance of non-productive/non-active variants (yellow, creamy and colorless) in populations of strains S. globisporus 4Lcp-Hp7 and S. globisporus 7Crt after cultivation in corn-soy liquid medium was 10-3 and 10-2, at 1–2 orders of magnitude higher than their standard cleavage with frequent transplantation of subcultures. The variability during storage had high indexes, which were dependent on storage temperature. Non-productive variants of strains S. globisporus 4Lcp-Hp7 and S. globisporus 7Crt at storage temperature of 4 °C were about 50% and 10% of the population, respectively. There were near 10% non-active colonies in the population of strain S. globisporus 4Lcp-Hp7, and 20–30% ones in the case strain S. globisporus 7Crt at storage temperatures of 21 °C and 28 °C. After one-year storage near 52% and 66% colonies of the populations of lyophilized cultures S. globisporus 4Lcp-Hp7 and S. globisporus 7Crt supported the high level of synthesis of corresponding carotenoids. Conclusions. It has been shown, that the trait of carotenoids biosynthesis in the mutant strains S. globisporus 4Lcp-Hp7 and S. globisporus 7Crt was showing variability with the high frequency after deep cultivation (10-3 and 10-2 non-active colonies) and during long storage on slants and in lyophilized state (10–50%).

The aim of this research is to Bioassay of lovastatin from Fusarium spp. obtained from NTCC, Forest Pathology Discipline, Forest Protection Division, Forest Research Institute, Dehradun, India. Lovstatin is an inhibitor of 3-hydroxy-3-methylglutaryl-coenzyme-A- reducatase (HMG-CoA reducatse) enzyme and a competitive inhibitor of the biosynthesis of cholesterol. In the present study, six different fungal species viz., Fusarium oxysporum (NTCC 367), F. moniliforme (NTCC 390), F. avenaceum (NTCC 1056), Fusarium equiseti (NTCC 1142), Fusarium solani (NTCC 1145) and Fusarium semitectum (NTCC 1150) were screened for lovastatin production in liquid and solid growth media. The lovastatin extraction from the fungal colonies was done by centrifugation and filtration method. Different techniques viz., paper disc and well preparation were employed to compare their relative sensitivity and efficacy. All the Fusarium spp. Shows positive result for lovastatin bioassay. According to results it was concluded that in comparison to the liquid medium (Potato Dextrose Broth), the solid medium (Potato Dextrose Agar) was more able to produce lovastatin.

The objective of this work was to evaluate the viability of liquid medium inoculation of Rhizobium etli in the planting furrow and to certify the efficiency of its strain UFLA 02-100 as a potential inoculant for common bean (Phaseolus vulgaris). The treatments consisted of three application rates of liquid inoculant applied in the planting furrows or in common bean seed, besides two controls without inoculation. The inoculant contained 109 cells of Rhizobium etli mL-1. Regardless of the application method, the yield obtained with the inoculation was equivalent to that of N from urea; however, the application in the furrows, at 0.6 L ha-1, is more advantageous due to the operational practicality and reduced costs. Resumo: O objetivo deste trabalho foi avaliar a viabilidade da inoculação líquida de Rhizobium etli no sulco de semeadura e certificar a eficiência da estirpe UFLA 02-100 como inoculante potencial para o feijoeiro-comum (Phaseolus vulgaris). Os tratamentos consistiram de três doses do inoculante líquido aplicadas aos sulcos e uma dose aplicada às sementes de feijão-comum, além de dois controles sem inoculação. O inoculante continha 109células de Rhizobium etli mL-1. Independentemente do método de aplicação, a produtividade obtida com a inoculação foi equivalente à da aplicação de N da ureia, mas a aplicação ao sulco, com 0,6 L ha-1, é mais vantajosa pela praticidade operacional e redução de custos.

The essential oils of Artemisia ifranensis J. Didier (Asteraceae), collected from Timahdite (Moroccan Middle-Atlas) in April and June (2012) (vegetative and early bloom stages, respectively), were obtained by hydrodistillation and then analysed by gas chromatography-flame ionization detector (GC/FID) and GC-mass spectrometry (GC/MS). Moreover, antibacterial activity essential oil was evaluated against pathogen germs isolated from hospital. Disc-diffusion method in solid medium and macrodilution method in liquid medium were, respectively, used to determine inhibition diameters and the antibacterial parameters to knowing, minimal inhibitory and bactericide concentrations (MIC and MBC). Essential oil yields were 0.58 ± 0.1% and 1.11 ± 0.3% (ml/100 g), respectively, in vegetative and blooming stages. For these periods, essential oil analyses revealed that β-Thujone was the most abundant compound in the plant with 50.0 and 41.1%, respectively. Oxygenated monoterpenes and sesquiterpenes are the most important groups among identified compounds. For antibacterial activity, the results proved that essential oil was highly active against the Staphylococcus aureus.

The effect of sucrose concentration on the micropropagation of axillary shoots of willow was investigated. The following factors were examined: the culture system (semisolid medium in glass jars versus liquid medium in temporary immersion bioreactors), the type of explant (apical and basal sections), the frequency of immersion, and CO2 enrichment. Shoots and leaf growth were significantly higher in RITA® bioreactors than in the jars for all the sucrose treatments. Apical or basal sections of willow cultured in bioreactors under high light intensity (150 µmol m−2 s −1 ) and ventilated six times a day with CO2 -enriched air were successfully proliferated without sucrose, whereas shoots cultured in jars did not proliferate well if sucrose concentration was 0.5% or lower. More roots were formed when sucrose was added to the medium. Shoots cultured in bioreactors were successfully acclimatized irrespective of the sucrose treatment and the root biomass when transferred to ex vitro conditions. This is the first report of photoautotrophic willow micropropagation, our results confirm the importance of proper gaseous exchange to attain autotrophy during in vitro propagation., This research was funded by Xunta de Galicia (Spain) (projects IN607A and Contrato Programa 2019-2020), by CYTED (P117RT0522) and by CSIC (COOPB20584).

The aim of this work was to develop and evaluate ready-made dishes of hybrid sorubim fish fillets obtained by the sous vide process. Three treatments were investigated: T1 with brine containing fine-herbs, T2 with brine containing fine-herbs, olive oil, and rosemary; and T3 with the same brine as in T1 but added of 30% of tomato sauce. Results showed the protein content increased according to the type of liquid medium employed, where the lowest value was determined for T1 and the highest value for T3. The sous vide process did not influence the lipid and protein contents of the treatments, be by the addition of olive oil (T2) nor by tomato sauce (T3). Treatments differed for all sensory attributes and presented acceptability indexes above 78.66%. indicating that they are good alternatives for fish consumption. However, T1 had the highest percentage of purchase intention (97.23%), representing the best product for commercialization.

Triazole-resistance in Aspergillus fumigatus has been frequently found in agricultural soils. We determined if triazole-resistance in A. fumigatus can be acquired in soil and earthworm guts after exposure to environmentally relevant concentrations of formulated propiconazole (FPRO) and difenoconazole (FDIF). Acquired triazole-resistance was found in two strains of A. fumigatus after exposure to 2.5 mg/L of FPRO and three strains of A. fumigatus after induction by 0.1–2.5 mg/L of FDIF for 40 d in liquid medium. Resistance development depended upon exposure time and substrate concentration. There were one, three, and two resistant strains of A. fumigatus in soils receiving FPRO, FDIF, and their combination, respectively. Resistance was identified in two strains of A. fumigatus isolated from the guts of earthworms exposed in soils contaminated by FDIF and two strains of A. fumigatus isolated from the guts of earthworms exposed to the combination of FDIF and FPRO. We conclude that normal applications of FPRO, FDIF, and their combination are likely to induce triazole-resistance in A. fumigatus in agricultural fields. Earthworms may act as disseminators of triazole-resistant A. fumigatus.

Developing scale-up system and automation of micropropagation in a bioreactor has been a possible way of cost reduction and intensive manual handling. We report a comparison between the results of experiments aimed at improving carnation micropropagation using new bioreactor according to Temporary Immersion Bioreactor (TIB) and solid culture. By applying different levels of BAP, at the concentration of 3 mg L−1, we observed 14.3 new shoots in TIB, but the number of new shoots on solid medium reached to 5.7 at the same treatment. Our results also showed that with 3 mg L−1 BAP in TIB, the initial fresh weight of plant material increased from 10 g to 450 g after 15 days. It is concluded that TIB showed more than 10 times shoot production of carnation. Shoot elongation and rooting induction was successfully stimulated in TIB by applying 1 mg L−1 IBA. Rooting of proliferated plantlets from TIB and solid culture were successfully happened, and led to highest number of roots (4.6 cm) and highest length of roots (6.87 cm) in TIB. More than 90% of plantlet was acclimatized to ex vitro. Our results suggested that mass production of carnation shoots in our simple TIB, with effective result, can be considered as a critical first step toward large scale production of carnation.

Achieving high drug loading capacity and controlling drug delivery are two main challenges related to drug carriers. In this study, polymeric macroporous sponges with very high pore volume and large porosity are introduced as a new-type of drug carrier. Due to the high pore volume (285 and 166 cm3/g for the sponges with densities of 3.5 and 6.0 mg/cm3, respectively), the sponges exhibit very high drug loading capacities with average values of 1870 ± 114 and 2697 ± 73 mg/g in the present study, which is much higher than the meso and microporous drug carriers (< 1500 mg/g). In order to control the release profiles, an additional poly(p-xylylene) (PPX) coating was deposited by chemical vapor on the drug loaded sponge. Consequently, Artemisone (ART) release in the liquid medium could be retarded, depending on the density of the sponge and the thickness of the coating. In future, the new 3D polymeric sponges would be highly beneficial for drug carriers and release for treatment of chronic diseases.

Agrobacterium rhizogenes is responsible for hairy root induction in infected sensitive plants. Since tissue-specific behaviour and synthesis are maintained in hairy roots, hairy root cultures are suitable for large-scale production of secondary metabolites and other chemicals. T. Murashige has proposed a concept of “artificial seed” as an approach to the mass propagation of elite plant varieties. Artificial seed is prepared by encapsulating somatic embryos or regenerated tissue in polysaccharide gels such as alginate and coating with a membrane to prevent water evaporation. Hairy roots must be cut to produce artificial seeds. To harvest a large quantity of root fragments with apical meristems or lateral roots from a whole root culture, hairy roots can be treated with various concentrations of auxin, then randomly-picked root fragments can be encapsulated and transferred to Murashige and F. Skoog liquid medium in the light.

Enokipodins are antimicrobial sesquiterpenes produced by Flammulina velutipes in a mycelial culture medium. To date, enokipodin production has not been reported in other members of the genus Flammulina. Hence, in this study, the production of enokipodins A, B, C, and D by F. velutipes and F. rossica was investigated. Some strains of F. rossica were confirmed to produce at least one of the four enokipodins in the culture medium. However, some strains of F. velutipes did not produce any of the enokipodins. In an antibacterial assay using liquid medium, enokipodin B showed the strongest growth inhibitory activity against Bacillus subtilis among the four types of enokipodins. Enokipodin B inhibited the spore germination of some plant pathogenic fungi. Enokipodins B and D exerted moderate anti-proliferative activity against some cancer cell lines, and enokipodins A and C inhibited the proliferation of the malarial parasite, Plasmodium falciparum. Productivity of enokipodins of some strains of F. velutipes and F. rossica and antibacterial activity of culture filtrate extract against B. subtilis.

Pollen tubes growing in the transmitting tract are presented with an extracellular matrix rich in a variety of substances. The expression of a multitude of genes for transport proteins in the pollen tube indicates that pollen tubes take up at least some of the components provided by the transmitting tract, for example nutrients, ions, or signaling molecules. FRET (Forster resonance energy transfer)-based nanosensors are perfectly suited to study the uptake of these molecules into pollen tubes. They are genetically encoded and can easily be expressed in Arabidopsis pollen tubes. Furthermore, the method is noninvasive and nanosensors for a wide range of substances are available. This chapter will describe the design of plasmids required to generate stable Arabidopsis lines with a pollen tube-specific expression of nanosensor constructs. We also present a method to germinate Arabidopsis pollen tubes in a flow chamber slide that allows the perfusion of the pollen tubes with liquid medium supplemented with the substrate of the nanosensor. Simultaneous evaluation of the FRET efficiency of the nanosensor by confocal microscopy reveals whether the substance is taken up by the pollen tubes. Together with the great number of available nanosensors this method can generate a detailed picture of the substances that are taken up during pollen tubes growth.

Pseudomonas fragi is the dominant spoilage organism in various foods, especially in spoiled milk, fish, and meats. Its growth can be inhibited by releasing allyl isothiocyanate (AITC) from ground mustard seeds in food packages. This paper aims to investigate the antimicrobial potential of ground mustard seeds against P. fragi growth and the effectiveness of released AITC concentration from mustard seeds on microbial inhibition of the spoilage bacteria growing in the liquid medium. The AITC concentration in the headspace and the liquid medium was measured and the growth of P. fragi in the liquid medium was monitored. Depending on the concentration of AITC, not only growth was inhibited but a reduction of the total count of P. fragi was even observed. The inactivation rate (k) of P. fragi was estimated using first-order inactivation kinetics and the minimum gaseous-released AITC to inactivate P. fragi was determined. Higher AITC concentration in the headspace and liquid medium was observed when using a higher amount of ground mustard seeds and a lower food to headspace ratio. Increasing the amount of ground mustard seeds (&gt, 100 mg per 10 mL liquid medium) led to full inactivation of P. fragi in 48 hours. By using an inhibition sigmoid Emax model, the minimum gaseous-released AITC for inactivation of P. fragi in 48 hours was observed around 15 &micro, g/l headspace. These results indicate that inhibition of the spoilage bacteria and extending the shelf life using ground mustard seeds is only possible by applying a careful design of the packaging system.