STUDI BACILLUS FIRMUS  SEBAGAI KANDIDAT PROBIOTIK DALAM MENGHADAPI Aeromonas hydrophila PADA MEDIA BUDIDAYA The Study of Bacillus firmus as Probiotic Candidate in Supressing Aeromonas hydrophila in Culture Media

ABSTRAK Pengendalian penyakit bakterial yang umum dilakukan dengan pemakaian antibiotik atau bahan kimia sudah tidak diperbolehkan lagi karena menimbulkan patogen yang resisten terhadap bahan kimia tersebut, terlebih jika penggunaan tidak sesuai dengan anjuran yang diberikan. Dampak negatif terhadap kesehatan konsumen berupa residu antibiotik juga menjadi pertimbangan yang harus diperhatikan. Manipulasi terhadap populasi mikroba yang berada di perairan guna pencegahan sebelum terjadinya serangan bakteri yang bersifat mematikan perlu dilakukan sebagaimana konsep probiotik sebagai biokontrol. Tujuan penelitian ini adalah menguji kandidat probiotik dalam menekan atau menghambat bakteri patogen Aeromonas hydrophila. Penelitian ini dilaksananakan dalam dua tahap. Tahap pertama adalah tahap pengujian bakteri kandidat probiotik secara in vitro menggunakan metode zona hambat dan kultur bersama pada media agar. Tahap kedua adalah uji tentang bakteri kandidat probiotik dengan patogen pada media budidaya. Hasil terbaik penelitian tahap pertama pada uji kultur bersama antara kandidat probiotik B. firmus dengan A. hydrophila pada skala in vitro adalah dengan penambahan probiotik B. firmus sebanyak 108 cfu/ml. Sedangkan pada penelitian tahap kedua didapatkan hasil berturut-turut perlakuan D dengan tingkat kelangsungan hidup (SR) mencapai 90%, perlakuan C dengan SR 75%, perlakuan A dengan SR 50% dan perlakuan K dengan SR 50%. Kata kunci: Bacillus firmus, probiotik, Aeromonas hydrophila, media budidaya ABSTRACT Controlling bacterial disease with the use of antibiotics or chemicals is no longer allowed as it results in pathogens that are resistant to the chemicals, especially when not in accordance with the recommendations provided. The negative impactsof the antibiotics residues on the consumers’ health also need to be considered. Manipulation of microbial populations present in the waters as preventation before the lethal attack of bacteria needs to be done which is in accordance with the concept of probiotics as biocontrol.The purpose of this study was to test the probiotic candidates in suppressing or inhibiting pathogenic bacteria Aeromonas hydrophila. This study was conducted in two stages. The first stage was to test a candidate probiotic bacteria in vitro using culture methods and inhibition zone on the media together. The second stage wasto test candidate probiotic bacteria to pathogens on the cultivation media. The best results in the first phase of the research is shared culture test between probiotic candidate B. FIRMUS with A. hydrophila on vitro scale is the addition of the probiotic B. FIRMUS 108 cfu / ml. While in the second phase of the research results obtained successively: treatment D with a survival rate (SR) reaches 90%, treatment C with SR 75%, treatment A with SR 50% and treatment K with SR 50%. Keywords: Bacillus FIRMUS, probiotics, Aeromonas hydrophila, media cultivation

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Screening of Probiotic Bacteria from Intestine and Culture Environment of Hoeven’s slender carp Leptobarbus hoeveni Blkr to Control Pathogenic Bacteria

Jurnal Akuakultur Indonesia ◽

10.19027/jai.9.127-135 ◽

2010 ◽

Vol 9 (2) ◽

pp. 127

Author(s):

. Sunarto ◽

. Sukenda ◽

. Widanarni

Keyword(s):

Aeromonas Hydrophila ◽

Pathogenic Bacteria ◽

Probiotic Bacteria ◽

Streptococcus Iniae ◽

Mycobacterium Fortuitum ◽

Fish Pathogen ◽

Culture Environment ◽

Control Disease

The ability of probiotic bacteria to control disease infection has been used in aquaculture. This experiment was conducted to isolate and characterize probiotic bacteria; the competition test its ability probiotic bacteria against pathogenic bacteria; and to improve survival rate of Leptobarbus hoeveni. The bacteria were isolated from Leptobarbus hoeveni and its culture environment, and then tested to know its ability to inhibit bacterial fish pathogen in-vitro. Furthermore, the selected probiotic bacteria were tested in vivo to evaluate their ability to inhibit pathogen of Leptobarbus hoeveni. The result showed that probiotic bacteria inhibit the growth of Streptococcus iniae, Flexibacter columnaris, Mycobacterium fortuitum and Aeromonas hydrophila in vitro. Isolate DD3 was the best of candidate probiotic because of the ability to inhibit pathogen, especially A. hydrophila, the most virulent bacteria in Leptobarbus hoeveni. Key Words : probiotic bacteria, Leptobarbus hoeveni, pathogenic bacteria Abstrak Kemampuan bakteri probiotik untuk mengendalikan penyakit infeksi telah digunakan dalam akuakultur. Tujuan penelitian ini adalah mengisolasi dan mengkarakterisasi bakteri probiotik, menguji kemampuan bakteri probiotik terhadap bakteri patogen, sehingga dapat meningkatkan tingkat kelangsungan hidup ikan jelawat. Bakteri diisolasi dari usus ikan jelawat dan lingkungan budaya, kemudian diuji kemampuannya menghambat bakteri patogen secara in-vitro. Selanjutnya bakteri probiotik yang dipilih diuji secara in vivo untuk mengevaluasi kemampuannya dalam menghambat patogen di dalam tubuh ikan jelawat. Dari hasil penelitian diperoleh bakteri probiotik yang diisolasi dari usus dan lingkungan budaya ikan jelawat menunjukkan penghambatan pertumbuhan terhadap Streptococcus iniae, Flexibacter columnaris, Mycobacterium fortuitum dan Aeromonas hydrophila secara in vitro. Isolat DD3 merupakan kandidat probiotik terbaik, karena mempunyai kemampuan untuk menghambat bakteri patogen, khususnya bakteri A. hydrophila adalah bakteri yang paling viluren bagi ikan jelawat. Kata Kunci: bakteri probiotik, ikan jalawat dan baktri patogen

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Application of Bacillus probiotic to prevent Aeromonas hydrophilla infection in Clarias sp.

Jurnal Akuakultur Indonesia ◽

10.19027/jai.13.105-114 ◽

2015 ◽

Vol 13 (2) ◽

pp. 105 ◽

Cited By ~ 3

Author(s):

Mohammad Faizal Ulkhaq ◽

, Widanarni ◽

Angela Mariana Kusumastuti

Keyword(s):

Aeromonas Hydrophila ◽

Pathogenic Bacteria ◽

Clarias Gariepinus ◽

Positive Control ◽

Negative Control ◽

Probiotic Bacteria ◽

African Catfish ◽

Control Application

ABSTRACT The aim of this study was to test the effectiveness of a probiotic Bacillus for the prevention of motile aeromonad septicemia (MAS) disease caused by Aeromonas hydrophila in African catfish (Clarias gariepinus). The study consisted of the inhibition testing of A. hydrophila by Bacillus (in vitro) and the application of probiotic in African catfish (in vivo). The in vivo test, consisted of five treatments such as the addition of probiotic Bacillus P4I1 RifR, Bacillus P4I2 RifR, Bacillus P4I1 RifR + Bacillus P4I2 RifR (Kom), positive control (K+; only added with A. hydrophila) and negative control (K-; without probiotic nor A. hydrophila addition). African catfish (13.35±2.80 g) was maintained in 15 aquariums (40 L in volume) with 30 fishes each for 30 days. Probiotic bacteria was applied in water once a day, whereas pathogenic bacteria A. hydrophila RifR (103 cfu/mL) were added once in earlier treatment (except for the negative control). The result showed that the optimal concentration of Bacillus to inhibit A. hydrophila on in vitro test was 104 cfu/mL. In vivo test showed that the addition of probiotic in media of cultivation could reduce the number of A. hydrophila, improve immune response, and also increase the survival of African catfish compared to positive control. Application of probiotic P4I1 RifR showed the highest survival (92.23%) of all treatments. Keywords: Bacillus, Clarias gariepinus, motile aeromonad septicemia, probiotic ABSTRAK Penelitian ini bertujuan untuk menguji efektivitas probiotik Bacillus dalam pencegahan penyakit motile aeromonad septicaemia (MAS) yang disebabkan oleh Aeromonas hydrophila pada ikan lele dumbo (Clarias gariepinus). Penelitian terdiri atas pengujian penghambatan bakteri probiotik Bacillus terhadap A. hydrophila secara in vitro, dilanjutkan dengan aplikasi pada budidaya ikan lele dumbo (in vivo). Pada uji in vivo, penelitian terdiri atas lima perlakuan yaitu budidaya ikan lele dumbo dengan penambahan probiotik Bacillus P4I1 RifR, Bacillus P4I2 RifR, kombinasi probiotik Bacillus P4I1 RifR + Bacillus P4I2 RifR (Kom), kontrol positif (K+; hanya ditambahkan A. hydrophila) dan kontrol negatif (K-; tanpa pemberian probiotik dan A. hydrophila). Ikan lele dumbo (13,35±2,80 g) dipelihara pada akuarium volume 40 L dengan kepadatan 30 ekor/akuarium selama 30 hari. Bakteri probiotik ditambahkan pada media pemeliharaan ikan setiap hari, sedangkan bakteri patogen A. hydrophila RifR (103 cfu/ mL) diberikan sekali pada awal pemeliharaan (kecuali pada kontrol negatif). Hasil penelitian menunjukkan bahwa konsentrasi terbaik pada penghambatan in vitro adalah dengan penambahan Bacillus 104 cfu/mL. Hasil uji in vivo menunjukkan perlakuan penambahan probiotik pada media budidaya efektif dapat menekan jumlah bakteri A. hydrophila, memperbaiki respons imun, dan meningkatkan kelangsungan hidup ikan lele dumbo dibanding kontrol positif. Perlakuan probiotik P4I1 RifR memberikan hasil terbaik dengan tingkat kelangsungan hidup tertinggi yaitu 92,23%. Kata kunci: Bacillus, Clarias gariepinus, motile aeromonad septicemia, probiotik

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Antimicrobial efficacy of a new tri-antibiotic combination against resistant endodontic pathogens: An in-vitro study

Brazilian Dental Science ◽

10.14295/bds.2021.v24i1.2186 ◽

2020 ◽

Vol 23 (4) ◽

pp. 8p ◽

Cited By ~ 1

Author(s):

Prasanna T. Dahake ◽

Sudhindra M Baliga

Keyword(s):

Root Canal ◽

Pathogenic Bacteria ◽

Culture Media ◽

In Vitro Study ◽

Vitro Study ◽

New Combination ◽

Antimicrobial Efficacy ◽

E Coli ◽

Background Removal

Background: Removal of all the pathogenic bacteria from the root canal system is of prime importance for the success of endodontic therapy. Objective: The study aimed to determine the antimicrobial efficacy of three antibiotics and their new combination against selected endodontic pathogens. Methods: In this in-vitro study, we used bacterial strains associated with the refractory endodontic condition and determined MIC and MBC of Clindamycin (C), Metronidazole (M), Doxycycline (D) as well as their combination CMD. We cultured Candida Albicans, Pseudomonas Aeruginosa, Escherichia Coli, Enterococcus Faecalis, Streptococcus Mutans, Bacillus Subtilis subsp. spizizenii, Actinomyces Actinomycetemcomitans on selective culture media. We analyzed the data using paired 't' test, one-way ANOVA, and Tuckey's HSD post hoc test. Results: Clindamycin inhibited the growth of C. Albicans (90%) and S. Mutans (90%) significantly and P. Aeruginosa, E. Coli, E. Faecalis, B. Subtilis, and A. Actinomycetemcomitans were resistant to it. Metronidazole did not inhibit any of the bacteria. Doxycycline inhibited C. Albicans (90%), P. Aeruginosa (90%), and S. Mutans (90%) significantly while E. Coli, E. Faecalis, B. Subtilis, and A. Actinomycetemcomitans were resistant to it. The combination of CMD inhibited all the microbes significantly. However, at bactericidal concentrations of CMD, E. Faecalis (p = 0.024), B. Subtilis (p = 0.021) and A. Actinomycetemcomitans (p = 0.041) were eliminated significantly, while C. Albicans (p = 0.164), P. Aeruginosa (p = 0.489), E. Coli (p = 0.106) and S. Mutans (p = 0.121) showed resistance. Conclusion: Combination CMD can be used against resistant endodontic pathogens to achieve predictable endodontic results.KEYWORDSAntimicrobial agents; Clindamycin; Doxycycline; Metronidazole; Root canal therapy.

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Bioactivity of Selected Phenolic Acids and Hexane Extracts from Bougainvilla spectabilis and Citharexylum spinosum on the Growth of Pectobacterium carotovorum and Dickeya solani Bacteria: An Opportunity to Save the Environment

Processes ◽

10.3390/pr8040482 ◽

2020 ◽

Vol 8 (4) ◽

pp. 482 ◽

Cited By ~ 1

Author(s):

Nader A. Ashmawy ◽

Said I. Behiry ◽

Asma A. Al-Huqail ◽

Hayssam M. Ali ◽

Mohamed Z. M. Salem

Keyword(s):

Phenolic Acids ◽

Tannic Acid ◽

Defense Mechanisms ◽

Pathogenic Bacteria ◽

Soft Rot ◽

Inhibition Zone ◽

Pectobacterium Carotovorum ◽

Bacterial Isolates ◽

Dickeya Solani

Phenolic acids and natural extracts, as ecofriendly environmental agents, can be used as bio bactericides against the growth of plant pathogenic bacteria. In this study, isolation trails from infected potato tubers and stems that showed soft rot symptoms in fields revealed two soft rot bacterial isolates and were initially identified through morphological, physiological, and pathogenicity tests. The molecular characterization of these isolates via PCR, based on the 16S rRNA region, was carried out by an analysis of the DNA sequence via BLAST and Genbank, and showed that the soft rot bacterial isolates belong to Pectobacterium carotovorum subsp. carotovorum (PCC1) and Dickeya solani (Ds1). The in vitro results of the tested phenolic acids against the cultured bacterial isolates proved that concentrations of 800, 1600, and 3200 μg/mL were the most effective. Ferulic acid was the potent suppressive phenolic acid tested against the Ds1 isolate, with an inhibition zone ranging from 6.00 to 25.75 mm at different concentrations (25–3200 μg/mL), but had no effect until reaching a concentration of 100 μg/mL in the PCC1 isolate, followed by tannic acid, which ranged from 7.00 to 25.50 mm. On the other hand, tannic acid resulted in a significant decrease in the growth rate of the PCC1 isolate with a mean of 9.11 mm. Chlorogenic acid was not as effective as the rest of the phenolic acids compared with the control. The n-hexane oily extract (HeOE) from Bougainvillea spectabilis bark showed the highest activity against PCC1 and Ds1, with inhibition zone values of 12 and 12.33 mm, respectively, at a concentration of 4000 μg/mL; while the HeOE from Citharexylum spinosum wood showed less activity. In the GC/MS analysis, nonanal, an oily liquid compound, was found ata percentage of 38.28%, followed by cis-2-nonenal (9.75%), which are the main compounds in B. spectabilis bark HeOE, and 2-undecenal (22.39%), trans-2-decenal (18.74%), and oleic acid (10.85%) were found, which are the main compounds in C. spinosum wood HeOE. In conclusion, the phenolic acids and plant HeOEs seem to raise the resistance of potato plants, improving their defense mechanisms against soft rot bacterial pathogens.

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In vitro ovarian follicle growth: a comprehensive analysis of key protocol variables†

Biology of Reproduction ◽

10.1093/biolre/ioaa073 ◽

2020 ◽

Vol 103 (3) ◽

pp. 455-470

Author(s):

Leah E Simon ◽

T Rajendra Kumar ◽

Francesca E Duncan

Keyword(s):

Wildlife Conservation ◽

Culture Media ◽

Ovarian Follicle ◽

Three Dimensional ◽

Ovarian Follicles ◽

Culture Methods ◽

Growth And Survival ◽

Follicle Culture ◽

Culture Techniques

Abstract Folliculogenesis is a complex process that requires integration of autocrine, paracrine, and endocrine factors together with tightly regulated interactions between granulosa cells and oocytes for the growth and survival of healthy follicles. Culture of ovarian follicles is a powerful approach for investigating folliculogenesis and oogenesis in a tightly controlled environment. This method has not only enabled unprecedented insight into the fundamental biology of follicle development but also has far-reaching translational applications, including in fertility preservation for women whose ovarian follicles may be damaged by disease or its treatment or in wildlife conservation. Two- and three-dimensional follicle culture systems have been developed and are rapidly evolving. It is clear from a review of the literature on isolated follicle culture methods published over the past two decades (1980–2018) that protocols vary with respect to species examined, follicle isolation methods, culture techniques, culture media and nutrient and hormone supplementation, and experimental endpoints. Here we review the heterogeneity among these major variables of follicle culture protocols.

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Commercial Biocontrol Agents Reveal Contrasting Comportments Against Two Mycotoxigenic Fungi in Cereals: Fusarium Graminearum and Fusarium Verticillioides

Toxins ◽

10.3390/toxins12030152 ◽

2020 ◽

Vol 12 (3) ◽

pp. 152 ◽

Cited By ~ 2

Author(s):

Lucile Pellan ◽

Noël Durand ◽

Véronique Martinez ◽

Angélique Fontana ◽

Sabine Schorr-Galindo ◽

...

Keyword(s):

Fusarium Graminearum ◽

Culture Media ◽

Fusarium Verticillioides ◽

Inhibition Zone ◽

Culture Conditions ◽

Dual Culture ◽

Growth Inhibition Zone ◽

Mycotoxigenic Fungi ◽

The Impact

The aim of this study was to investigate the impact of commercialized biological control agents (BCAs) against two major mycotoxigenic fungi in cereals, Fusarium graminearum and Fusarium verticillioides, which are trichothecene and fumonisin producers, respectively. With these objectives in mind, three commercial BCAs were selected with contrasting uses and microorganism types (T. asperellum, S. griseoviridis, P. oligandrum) and a culture medium was identified to develop an optimized dual culture bioassay method. Their comportment was examined in dual culture bioassay in vitro with both fusaria to determine growth and mycotoxin production kinetics. Antagonist activity and variable levels or patterns of mycotoxinogenesis inhibition were observed depending on the microorganism type of BCA or on the culture conditions (e.g., different nutritional sources), suggesting that contrasting biocontrol mechanisms are involved. S. griseoviridis leads to a growth inhibition zone where the pathogen mycelium structure is altered, suggesting the diffusion of antimicrobial compounds. In contrast, T. asperellum and P. oligandrum are able to grow faster than the pathogen. T. asperellum showed the capacity to degrade pathogenic mycelia, involving chitinolytic activities. In dual culture bioassay with F. graminearum, this BCA reduced the growth and mycotoxin concentration by 48% and 72%, respectively, and by 78% and 72% in dual culture bioassay against F. verticillioides. P. oligandrum progressed over the pathogen colony, suggesting a close type of interaction such as mycoparasitism, as confirmed by microscopic observation. In dual culture bioassay with F. graminearum, P. oligandrum reduced the growth and mycotoxin concentration by 79% and 93%, respectively. In the dual culture bioassay with F. verticillioides, P. oligandrum reduced the growth and mycotoxin concentration by 49% and 56%, respectively. In vitro dual culture bioassay with different culture media as well as the nutritional phenotyping of different microorganisms made it possible to explore the path of nutritional competition in order to explain part of the observed inhibition by BCAs.

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In Vitro and FTIR Spectroscopy: Local Black Fruit Seed Extract as Antibacterial Aeromonas Hydrophila

International Journal of Biology and Biomedical Engineering ◽

10.46300/91011.2021.15.12 ◽

2021 ◽

Vol 15 ◽

pp. 92-98

Author(s):

Yori Turu Toja ◽

Eddy Suprayitno ◽

Aulanni’am ◽

Uun Yanuhar

Keyword(s):

Antibacterial Activity ◽

Ethyl Acetate ◽

Aeromonas Hydrophila ◽

Antibacterial Properties ◽

Inhibition Zone ◽

Seed Extract ◽

Water Fraction ◽

Natural Treatment ◽

Ft Ir

Indonesia has many local plants with potential as herbal antibacterial properties, one of which is the local black fruit of Wandama. The objective of this study is to determine the antibacterial activity of the extract of black fruit seed against A. Hydrophila by in vitro and FTIR by Tilapia. The benefit of this research is to make black fruit seed extract as a natural remedy against tilapia attacked by Aeromonas Hydrophila. The antibacterial activity of black fruit seed extract against A. Hydrophila is investigated in vitro and using FTIR on tilapia in this research. The aim of this study is to establish black fruit seed extract as a natural treatment for tilapia infected with Aeromonas Hydrophila. The disc approach was used to conduct the in vitro research. Agar medium was poured into a sterile petri dish, and 2 drops of bacteria from the liquid medium were uniformly distributed and cooled. On the press, disc paper with black fruit seed extract was put and incubated for 18-24 hours at 35 C. An FTS 1000 version spectrophotometer system was used for the FTIR test. For the FT-IR study, a fraction of dried black fruit seeds was used. For clear preparation, 100 mg of dry extract is condensed in KBr pellets. A specimen of loaded black fruit seed samples was analyzed using FT-IR spectroscopy with a scanning range of 400-4000 cm-1 and a resolution of 4 cm-1. The results showed that the ethyl acetate fraction produced an inhibition zone (13.65 mm), the water fraction produced an inhibition zone (11.21 mm), and the n-hexane fraction produced the weakest inhibition zone (11.21 mm) (5.31mm). The concentration test results from 125 ppm, 250 ppm, 500 ppm and 1000 ppm obtained a large inhibition zone at a concentration of 1000 ppm with an inhibition zone area of 11.43 + 0.02. The absorbance value of black fruit seed extract in ethyl acetate solvent at 3430 cm-1 revealed the absorption band with the strongest vibration of the hydroxylate (-OH) functional group, according to FTIR research. Final thoughts with an average value of 13.65 + 0.06, black fruit seed extract using ethyl acetate can inhibit A. Hydrophila bacteria.

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Uji In Vitro Gabungan Ekstrak Boesenbergia pandurata, Solanum ferox, Zingimber zerumbet terhadap Bakteri Patogen pada Ikan Nila (IN VITRO TEST OF CONCOCTION PLANT EXTRACTS OF BOESENBERGIA PANDURATA, SOLANUM FEROX, ZINGIMBER ZERUMBET

jurnal veteriner ◽

10.19087/jveteriner.2018.19.1.35 ◽

2018 ◽

Vol 19 (1) ◽

pp. 35

Author(s):

Esti Handayani Hardi ◽

Gina Saptiani ◽

Nurkadina Nurkadina ◽

Irawan Wijaya Kusuma ◽

Wiwin Suwinarti

Keyword(s):

Antibacterial Activity ◽

Plant Extracts ◽

Aeromonas Hydrophila ◽

Inhibition Zone ◽

The Other ◽

In Vitro Test ◽

Pseudomonas Sp ◽

Vitro Test

The aim of this study was to evaluate the antibacterial activity of ethanol concoction (mixed extract) of three herbs plants, such as: Boesenbergia pandurata, Solanum ferox and Zingimber zerumbet against Aeromonas hydrophila, Pseudomonas sp. and both bacteria. The Plant extracts were obtained by using ethanol and the concentration of the extract was 600 mgL-1 of B. pandurata, 900 mgL-1 of S. ferox and 200 mgL-1 of Z. zerumbet. The inhibition zone was measured after 6, 12, 18, 24 h incubation at 30o C. The ratios of concoction S. ferox and B. pandurata (for 100 mL solution) were 90:10; 80:20; 70:30; 60:40; 50:50; 40:60; 30:70; 20:80; 10:90. The same ratios were made on the concoction of S. ferox and Z. zerumbet. The concoctions of S. ferox and B. pandurata in the ratio of 50:50 and 60:40, and the concoction of S. ferox and Z. Zerumbet with ratio 60:40 had higher antibacterial activity against A. hydrophila single isolate compared to the other concoctions. Furthermore, the concoction extract of S. ferox and B. pandurate 50:50 and the combined of S. ferox and Z. zerumbet ratio 50:50 and 90:10 were the best combination to inhibit the growth of a single bacterium Pseudomonas sp. The combined S. ferox and B. pandurata ratio 50:50; 10:90 and S. ferox mixed with Z. zerumbet ratio 50:50 and 40:60 were the best combination against the combined bacteria between A. hydrophila and Pseudomonas sp. The conclusion of this research was the combined extract of S. ferox and B. pandurata and S. ferox with Z. zerumbet are effective to suppress the growth of single or combination of A. hydropila and Pseudomonas sp.

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Metabolomics Guides Rational Development of a Simplified Cell Culture Medium for Drug Screening against Trypanosoma brucei

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00044-13 ◽

2013 ◽

Vol 57 (6) ◽

pp. 2768-2779 ◽

Cited By ~ 62

Author(s):

Darren J. Creek ◽

Brunda Nijagal ◽

Dong-Hyun Kim ◽

Federico Rojas ◽

Keith R. Matthews ◽

...

Keyword(s):

Cell Culture ◽

Trypanosoma Brucei ◽

Culture Medium ◽

Culture Media ◽

Culture Methods ◽

Content Type ◽

Drug Activity ◽

Rational Development ◽

High Concentrations

ABSTRACTIn vitroculture methods underpin many experimental approaches to biology and drug discovery. The modification of established cell culture methods to make them more biologically relevant or to optimize growth is traditionally a laborious task. Emerging metabolomic technology enables the rapid evaluation of intra- and extracellular metabolites and can be applied to the rational development of cell culture media. In this study, untargeted semiquantitative and targeted quantitative metabolomic analyses of fresh and spent media revealed the major nutritional requirements for the growth of bloodstream formTrypanosoma brucei. The standard culture medium (HMI11) contained unnecessarily high concentrations of 32 nutrients that were subsequently removed to make the concentrations more closely resemble those normally found in blood. Our new medium, Creek's minimal medium (CMM), supportsin vitrogrowth equivalent to that in HMI11 and causes no significant perturbation of metabolite levels for 94% of the detected metabolome (<3-fold change; α = 0.05). Importantly, improved sensitivity was observed for drug activity studies in whole-cell phenotypic screenings and in the metabolomic mode of action assays. Four-hundred-fold 50% inhibitory concentration decreases were observed for pentamidine and methotrexate, suggesting inhibition of activity by nutrients present in HMI11. CMM is suitable for routine cell culture and offers important advantages for metabolomic studies and drug activity screening.

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Effect of bovine colostrum, cheese whey, and spray-dried porcine plasma on the in vitro growth of probiotic bacteria and Escherichia coli

Canadian Journal of Microbiology ◽

10.1139/cjm-2014-0130 ◽

2014 ◽

Vol 60 (5) ◽

pp. 287-295 ◽

Cited By ~ 7

Author(s):

Claude P. Champagne ◽

Yves Raymond ◽

Yves Pouliot ◽

Sylvie F. Gauthier ◽

Martin Lessard

Keyword(s):

Escherichia Coli ◽

Lactic Acid ◽

Lactic Acid Bacteria ◽

Pathogenic Bacteria ◽

Cheese Whey ◽

Probiotic Bacteria ◽

E Coli ◽

Spray Dried ◽

Mrs Broth

The aim of this study is to evaluate the effects of defatted colostrum (Col), defatted decaseinated colostrum whey, cheese whey, and spray-dried porcine plasma (SDPP) as supplements of a growth medium (de Man – Rogosa – Sharpe (MRS) broth) on the multiplication of lactic acid bacteria, probiotic bacteria, and potentially pathogenic Escherichia coli. Using automated spectrophotometry (in vitro system), we evaluated the effect of the 4 supplements on maximum growth rate (μmax), lag time (LagT), and biomass (ODmax) of 12 lactic acid bacteria and probiotic bacteria and of an E. coli culture. Enrichment of MRS broth with a Col concentration of 10 g/L increased the μmax of 5 of the 12 strains by up to 55%. Negative effects of Col or SDPP on growth rates were also observed with 3 probiotic strains; in one instance μmax was reduced by 40%. The most effective inhibitor of E. coli growth was SDPP, and this effect was not linked to its lysozyme content. The positive effect of enrichment with the dairy-based ingredient might be linked to enrichment in sugars and increased buffering power of the medium. These in vitro data suggest that both Col and SDPP could be considered as supplements to animal feeds to improve intestinal health because of their potential to promote growth of probiotic bacteria and to inhibit growth of pathogenic bacteria such as E. coli.

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