scholarly journals Isolation and Characterization of Phages InfectingBacillus subtilis

2015 ◽  
Vol 2015 ◽  
pp. 1-10 ◽  
Author(s):  
Anna Krasowska ◽  
Anna Biegalska ◽  
Daria Augustyniak ◽  
Marcin Łoś ◽  
Malwina Richert ◽  
...  
Keyword(s):  
Food Production ◽  
Burst Size ◽  
Industrial Applications ◽  
Isolation And Characterization ◽  
Latent Time ◽  
Food Borne ◽  
Alternative Approach ◽  
Sds Page

Bacteriophages have been suggested as an alternative approach to reduce the amount of pathogens in various applications. Bacteriophages of various specificity and virulence were isolated as a means of controlling food-borne pathogens. We studied the interaction of bacteriophages withBacillusspecies, which are very often persistent in industrial applications such as food production due to their antibiotic resistance and spore formation. A comparative study using electron microscopy, PFGE, and SDS-PAGE as well as determination of host range, pH and temperature resistance, adsorption rate, latent time, and phage burst size was performed on three phages of theMyoviridaefamily and one phage of theSiphoviridaefamily which infectedBacillus subtilisstrains. The phages are morphologically different and characterized by icosahedral heads and contractile (SIOΦ, SUBω, and SPOσphages) or noncontractile (ARπphage) tails. The genomes of SIOΦ and SUBωare composed of 154 kb. The capsid of SIOΦ is composed of four proteins. Bacteriophages SPOσand ARπhave genome sizes of 25 kbp and 40 kbp, respectively. Both phages as well as SUBωphage have 14 proteins in their capsids. Phages SIOΦ and SPOσare resistant to high temperatures and to the acid (4.0) and alkaline (9.0 and 10.0) pH.

scholarly journals Isolation and characterization of staphylococcus aureus from two large-scale food poisoning outbreaks in Vietnam

2020 ◽  
pp. 139-147
Author(s):  
Lam Quoc Hung ◽  
◽  
Huong Minh Nguyen ◽  
Ta Thi Yen ◽  
Le Vinh Hoa ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Food Production ◽  
Large Scale ◽  
Virulence Genes ◽  
Food Poisoning ◽  
Isolation And Characterization ◽  
Food Borne ◽  
Different Strains ◽  
Spa Type

In Vietnam and around the world, Staphylococcus aureus remains a major hazard of food safety and food poisoning. S. aureus is present in many places and easily contaminates food production during processing chains. In this study, we successfully isolated S. aureus strains from suspected samples of two food borne poisoning outbreaks in Ha Giang and Vinh Phuc in 2017 and 2018, respectively. The collected samples were examined for presence of staphylococcal enterotoxins (SEs) by using 3MTMTECRATM Staph Enterotoxin kit, from there all the samples were positive with SEs. Different strains of S. aureus were isolated and then confirmed by MALDI-TOF technique. Those strains then were stored in Brain heart solution with 15% glycerol until further analysis. Our results identified three STs, ST96, ST88 (spa type t7558), and ST72 (spa type t3092), were responsible for two outbreaks. Two virulence genes detected from the above strains were sea and sec. Furthermore, these strains are test for antibiotic resistance susceptibility with commonly antibiotics. Penicillin are found to be resisted by all three STs, in particularly, ST96 and ST88 are both resistant to erythromycin while ST72 is resistant to gentamicin. Taken together, our study highlights the usefulness of molecular characterization to study and monitor bacterial pathogens associated with food poisoning outbreaks in Vietnam.

Isolation and characterization of a rare waterborne lytic phage of Salmonella enterica serovar Paratyphi B

2013 ◽  
Vol 59 (5) ◽  
pp. 318-323 ◽  
Author(s):  
Sangeeta S. Ahiwale ◽  
Ashok V. Bankar ◽  
Sujata N. Tagunde ◽  
Smita Zinjarde ◽  
Hans W. Ackermann ◽  
...  
Keyword(s):  
Salmonella Enterica ◽  
Burst Size ◽  
Lytic Phage ◽  
Isolation And Characterization ◽  
Bacterial Contaminants ◽  
Sds Page ◽  
Wide Range ◽  
Paratyphi B ◽  
Drug Resistant Strains

A lytic phage of Salmonella serovar Paratyphi B, named φSPB, was isolated from surface waters of the Pavana River in India. Phage φSPB is a member of the Podoviridae family and is morphologically similar to the 7-11 phages of the C3 morphotype of tailed phages, characterized by a very long, cigar-shaped head. The head measured approximately 153 × 57 nm, and the tail size was 12 × 7 nm. The phage was stable over a wide range of pH (4–9) and temperature (4–40 °C). The adsorption rate constant was 4.7 × 10−10. Latent and eclipse periods were 10 and 15 min, respectively, and the burst size was 100 plaque-forming units/infected cell after 25 min at 37 °C. The phage DNA was 59 kb in size. Ten major proteins were observed on SDS–PAGE, although some of these proteins could be bacterial contaminants. This is the first report of Salmonella enterica subsp. enterica serovar Paratyphi B phage of C3 morphotype from India that has many unique features, such as high replication potential, short replication time, and stability over a wide range of pH and temperature, making it a promising biocontrol agent against the drug-resistant strains of Salmonella Paratyphi B.

scholarly journals Isolation and characterization of staphylococcus aureus from two large-scale food poisoning outbreaks in Vietnam

2020 ◽  
pp. 139-147
Author(s):  
Lam Quoc Hung ◽  
◽  
Huong Minh Nguyen ◽  
Ta Thi Yen ◽  
Le Vinh Hoa ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Food Production ◽  
Large Scale ◽  
Virulence Genes ◽  
Food Poisoning ◽  
Isolation And Characterization ◽  
Food Borne ◽  
Different Strains ◽  
Spa Type

In Vietnam and around the world, Staphylococcus aureus remains a major hazard of food safety and food poisoning. S. aureus is present in many places and easily contaminates food production during processing chains. In this study, we successfully isolated S. aureus strains from suspected samples of two food borne poisoning outbreaks in Ha Giang and Vinh Phuc in 2017 and 2018, respectively. The collected samples were examined for presence of staphylococcal enterotoxins (SEs) by using 3MTMTECRATM Staph Enterotoxin kit, from there all the samples were positive with SEs. Different strains of S. aureus were isolated and then confirmed by MALDI-TOF technique. Those strains then were stored in Brain heart solution with 15% glycerol until further analysis. Our results identified three STs, ST96, ST88 (spa type t7558), and ST72 (spa type t3092), were responsible for two outbreaks. Two virulence genes detected from the above strains were sea and sec. Furthermore, these strains are test for antibiotic resistance susceptibility with commonly antibiotics. Penicillin are found to be resisted by all three STs, in particularly, ST96 and ST88 are both resistant to erythromycin while ST72 is resistant to gentamicin. Taken together, our study highlights the usefulness of molecular characterization to study and monitor bacterial pathogens associated with food poisoning outbreaks in Vietnam.

Characterization of Glycoprotein Fraction from Carp Pituitaries Isolated Using Concanavalin A as the Affinity Ligand

1998 ◽  
Vol 63 (3) ◽  
pp. 434-440 ◽  
Author(s):  
Irena Hulová ◽  
Jana Barthová ◽  
Helena Ryšlavá ◽  
Václav Kašička
Keyword(s):  
Concanavalin A ◽  
Reversed Phase ◽  
Amino Acid Sequences ◽  
Gel Chromatography ◽  
Affinity Ligand ◽  
Sds Page ◽  
Terminal Amino ◽  
Α And Β Subunits

Glycoproteins that have affinity to Concanavalin A were isolated from the acetone-dried pituitaries of common carp (Cyprinus carpio L.). Two fractions of glycoproteins were separated using gel chromatography on Superdex 75HR. The fraction with lower molecular weight (30 000) corresponding to the carp gonadotropin cGtH II was composed of two subunits as determined using SDS-PAGE. This protein fraction was further divided into four components using reversed-phase HPLC. Two fractions were pure α and β subunits of cGtH II as follows from immunodetection and from determination of N-terminal amino acid sequences. The other two were a mixture of α and β subunits as was also revealed by N-terminal analysis. Capillary electrophoresis was also used for characterization of isolated glycoproteins.

scholarly journals Isolation and characterization of pathogenic Escherichia coli bacteriophages from chicken and beef offal

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Celosia Lukman ◽  
Christopher Yonathan ◽  
Stella Magdalena ◽  
Diana Elizabeth Waturangi
Keyword(s):  
Escherichia Coli ◽  
Foodborne Pathogens ◽  
High Efficiency ◽  
Multiplicity Of Infection ◽  
Isolation And Characterization ◽  
Transmission Electron ◽  
Pathogenic Escherichia Coli ◽  
Family Based

Abstract Objective This study was conducted to isolate and characterize lytic bacteriophages for pathogenic Escherichia coli from chicken and beef offal, and analyze their capability as biocontrol for several foodborne pathogens. Methods done in this research are bacteriophage isolation, purification, titer determination, application, determination of host range and minimum multiplicity of infection (miMOI), and bacteriophage morphology. Results Six bacteriophages successfully isolated from chicken and beef offal using EPEC and EHEC as host strain. Bacteriophage titers observed between 109 and 1010 PFU mL−1. CS EPEC and BL EHEC bacteriophage showed high efficiency in reduction of EPEC or EHEC contamination in meat about 99.20% and 99.04%. The lowest miMOI was 0.01 showed by CS EPEC bacteriophage. CI EPEC and BL EPEC bacteriophage suspected as Myoviridae family based on its micrograph from Transmission Electron Microscopy (TEM). Refers to their activity, bacteriophages isolated in this study have a great potential to be used as biocontrol against several foodborne pathogens.

scholarly journals Isolation and Characterization of Bacteriophage ZCSE6 against Salmonella spp.: Phage Application in Milk

Biologics ◽  
2021 ◽  
Vol 1 (2) ◽  
pp. 164-176
Author(s):  
Abdallah S. Abdelsattar ◽  
Anan Safwat ◽  
Rana Nofal ◽  
Amera Elsayed ◽  
Salsabil Makky ◽  
...  
Keyword(s):  
Pathogenic Bacteria ◽  
Raw Milk ◽  
Salmonella Spp ◽  
Isolation And Characterization ◽  
Food Borne ◽  
Wide Range ◽  
Almost All ◽  
And Control ◽  
Milk And Dairy Products

Food safety is very important in the food industry as most pathogenic bacteria can cause food-borne diseases and negatively affect public health. In the milk industry, contamination with Salmonella has always been a challenge, but the risks have dramatically increased as almost all bacteria now show resistance to a wide range of commercial antibiotics. This study aimed to isolate a bacteriophage to be used as a bactericidal agent against Salmonella in milk and dairy products. Here, phage ZCSE6 has been isolated from raw milk sample sand molecularly and chemically characterized. At different multiplicities of infection (MOIs) of 0.1, 0.01, and 0.001, the phage–Salmonella interaction was studied for 6 h at 37 °C and 24 h at 8 °C. In addition, ZCSE6 was tested against Salmonella contamination in milk to examine its lytic activity for 3 h at 37 °C. The results showed that ZCSE6 has a small genome size (<48.5 kbp) and belongs to the Siphovirus family. Phage ZCSE6 revealed a high thermal and pH stability at various conditions that mimic milk manufacturing and supply chain conditions. It also demonstrated a significant reduction in Salmonella concentration in media at various MOIs, with higher bacterial eradication at higher MOI. Moreover, it significantly reduced Salmonella growth (MOI 1) in milk, manifesting a 1000-fold decrease in bacteria concentration following 3 h incubation at 37 °C. The results highlighted the strong ability of ZCSE6 to kill Salmonella and control its growth in milk. Thus, ZCSE6 is recommended as a biocontrol agent in milk to limit bacterial growth and increase the milk shelf-life.

scholarly journals Isolation and characterization of escherichia coli in ready-to-eat foods vended in Islamic University, Kushtia

1970 ◽  
Vol 18 ◽  
pp. 99-103 ◽  
Author(s):  
S Biswas ◽  
MAK Parvez ◽  
M Shafiquzzaman ◽  
S Nahar ◽  
MN Rahman
Keyword(s):  
Escherichia Coli ◽  
Pattern Analysis ◽  
University Campus ◽  
Rflp Pattern ◽  
E Coli ◽  
Fish Meat ◽  
Isolation And Characterization ◽  
Food Borne ◽  
Identification And Characterization

Context: Escherichia coli is shed in the feces of warm blooded animals and humans and thus potential for public health. Detection and characterization of E. coli in the ready-to-eat (RTE) foods concerns due to their presence indicates fecal contamination of the food.   Objective: To identify, characterize and RFLP pattern analysis of E. coli isolated from RTE foods vended in Islamic University campus, Kushtia.   Materials and Methods: Fifty samples from four types of consumed foods in six student halls of residence, some temporary restaurants of Islamic University, Kushtia were assessed for bacterial contamination by standard methods. Identification and characterization of E. coli isolates were performed using IMViC tests. Genomic DNA was used to perform RFLP pattern analysis.   Results: Thirty seven out of 50 (74%) examined samples of RTE foods had E. coli contamination. The highest number of E. coli was isolated from vegetable oriented RTE foods (90.90%) and fish, meat and cereals samples were also significantly E. coli positive. RFLP profiling of two E. coli isolates were observed.   Conclusion: The results of this study provide evidence that some RTE foods had unsatisfactory levels of contamination with E. coli. Thus street vended RTE food could be important potential vehicles for food-borne diseases. Molecular characterization may be exploited to identify food borne pathogen among different species.  Keywords: Ready-to-eat foods; Escherichia coli; RFLP pattern DOI: http://dx.doi.org/10.3329/jbs.v18i0.8783 JBS 2010; 18(0): 99-103

scholarly journals PENDEKATAN PENINGKATAN PRODUKSI LELE PADA KELOMPOK PEMBUDIDAYA IKAN SURYA PERKASA DESA GENDONGKULON, BABAT, LAMONGAN MELALUI IMPLEMENTASI PROBIOTIK DAN TEKNOLOGI PASCAPANEN

2020 ◽  
Vol 2 (1) ◽  
pp. 45
Author(s):  
Heru Pramono ◽  
Dwi Yuli Pujiastuti ◽  
Sofi Maulana ◽  
Abdul Azis
Keyword(s):  
Digestive Tract ◽  
Community Service ◽  
Food Production ◽  
Bacterial Resistance ◽  
Fish Processing ◽  
Isolation And Characterization ◽  
Water Conditions ◽  
Nutritional Needs ◽  
And Training

Food production through the cultivation industry is one of the promising options because it is able to support the nutritional needs of the community, especially in terms of the adequacy of animal protein. One promising aquaculture product is catfish culture (Clarias batracus) because catfish are known to be able to grow in relatively unfavorable water conditions, have relatively short growth times (2–3 months). Nonetheless, the emergence of diseases that cause mass death and decreased cultivator income have become the main problems lately. Therefore, the purpose of community service is to overcome mass death in order to increase catfish production and diversify post-harvest fish processing to increase community income. The approach includes isolation and characterization of bacteria that cause mass death, bacterial resistance to antibiotics, indigenous probiotic production, and training in making catfish. The results of the dedication showed that there were bacteria that were resistant to antibiotics from isolation from catfish that experienced disease during July to October. In addition, efforts have been made to produce indigenous probiotics from the catfish digestive tract that survive mass death, and training in processing catfish snacks on site. A good response is shown in processing training and it is hoped that in the future these two efforts will be able to increase the economic independenceof the community.AbstrakProduksi pangan melalui industri budidaya merupakan salah satu opsi menjanjikan karena mampu mendukung kebutuhan gizi masyarakat, terutama dari segi kecukupan protein hewani. Salah satu produk budidaya yang menjanjikan adalah budidaya ikan lele (Clarias batracus) karena ikan lele dikenal mampu tumbuh dalam kondisi air yang relatif kurang baik, memiliki waktu pertumbuhan relatif pendek (2-3 bulan). Meskipun demikian munculnya penyakit yang menyebabkan kematian massal serta penghasilan pembudidaya yang menurunmenjadi permasalahan utama belakangan ini. Oleh karena itu, tujuan pengabdian kepada masyarakat ini adalah upaya mengatasi kematian massal guna meningkatkan produksi lele serta diversifi kasi olahan ikan pasca panen untuk meningkatkan penghasilan masyarakat. Pendekatan yang dilakukan meliputi isolasi dan karakteristik bakteri penyebab kematian massal, resistansi bakteri terhadap antibiotik, produksi probiotik indigenous, serta pelatihan pembuatan olahan lele. Hasil pengabdian menunjukkan bahwa terdapat bakteri yang tahan terhadap antibiotik hasil isolasi dari lele yang mengalami penyakit selama bulan Juli sampai Oktober. Selain itu juga telah dilakukan upaya produksi probiotik indigenous dari saluran pencernaan lele yang bertahan hidup dari kematian massal, dan pelatihan pengolahan snack berbahan ikan lele di lokasi. Respons yang baik ditunjukkan pada pelatihan pengolahan serta diharapkan ke depan kedua upaya ini mampu meningkatkan kemandirian ekonomi masyarakat.

Isolation and Characterization of a Chlorogenic Acid Esterase from Aspergillus niger

1980 ◽  
Vol 35 (3-4) ◽  
pp. 209-212 ◽  
Author(s):  
B. Schöbel ◽  
W. Pollmann
Keyword(s):  
Enzyme Activity ◽  
Chlorogenic Acid ◽  
Divalent Cations ◽  
Hydrolysis Product ◽  
The Other ◽  
Temperature Optimum ◽  
Pig Liver ◽  
Isolation And Characterization

Abstract The isolation and characterization of a specific chlorogenic acid esterase is described. The enzyme activity is measured by determination of the hydrolysis product caffeic acid. The enzyme had been concentrated by means of ultrafiltration and column-chromatography. The pH- and temperature optimum were 6.5 and 45 °C respectively. Divalent cations were not required for the enzyme activity. As other esterases, this enzyme is inhibited by di-isopropyl-phosphorofluoridate. The Km-value is 0.70 mᴍ chlorogenic acid, the molecular weight 240000. The described enzyme is specific for chlorogenic acid. On the other hand a typical unspecific esterase like the pig liver esterases does not split chloro­genic acid. The isoelectric focusing reveals several isoenzymes of chlorogenase within a pI-range of 4.0-4.5.

Cloning and Characterization of a Novel N-acetylglucosaminidase (AtlD) from Enterococcus faecalis

2018 ◽  
Vol 28 (1) ◽  
pp. 14-27 ◽  
Author(s):  
Carlos Eduardo Serrano-Maldonado ◽  
Israel García-Cano ◽  
Augusto González-Canto ◽  
Eliel Ruiz-May ◽  
Jose Miguel Elizalde-Contreras ◽  
...  
Keyword(s):  
Enterococcus Faecalis ◽  
Food Industry ◽  
Pathogenic Bacteria ◽  
Heterologous Protein ◽  
Biochemical Characterization ◽  
Ph Values ◽  
Food Borne ◽  
Sds Page ◽  
Exclusion Chromatography

The <i>atlD</i> gene from<i></i> an <i>Enterococcus faecalis</i> strain isolated from a Mexican artisanal cheese was cloned, sequenced and expressed in <i>Escherichia coli</i> in order to perform a biochemical characterization<i>.</i> A partial amino acid sequence of the heterologous protein was obtained by LC-MS/MS, and it corresponded to a novel peptidoglycan hydrolase designated AtlD. Its molecular mass was 62–75 kDa, as determined by SDS-PAGE, zymography, Western blot, and exclusion chromatography. Electrofocusing rendered an isoelectric point (pI) of 4.8. It exhibited N-acetylglucosaminidase activity, with an optimal pH and temperature between 6–7 and 50°C, respectively. It retained 85% activity with NaCl at 1,000 mM, but it was susceptible to divalent ions, particularly Zn<sup>2+</sup>. It showed antibacterial activity against <i>Listeria monocytogenes</i>,<i> Staphylococcus aureus</i>, and enterococcal<i></i> strains of clinical origin. Due to the fact that it showed activity versus pathogenic bacteria, and because of its capabilities under ionic strength, temperature, and pH values present in food matrices, it could be applied as an additive in the food industry. This study will aid in the design of new antibacterial agents of natural origin to combat food-borne diseases, and it could be used as an industrial or hospital hygiene agent as well.

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