scholarly journals Bacteriophage genotyping using BOXA repetitive-PCR

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Dragica Damnjanovic ◽  
Xabier Vázquez-Campos ◽  
Daniel L. Winter ◽  
Melissa Harvey ◽  
Wallace J. Bridge
Keyword(s):  
Similarity Index ◽  
Pcr Amplification ◽  
Dna Fingerprint ◽  
Bacterial Strains ◽  
Phage Types ◽  
Phage Dna ◽  
Different Temperatures ◽  
Phage Propagation ◽  
Selection For ◽  
Genetic Profiles

Abstract Background Repetitive-PCR (rep-PCR) using BOXA1R and BOXA2R as single primers was investigated for its potential to genotype bacteriophage. Previously, this technique has been primarily used for the discrimination of bacterial strains. Reproducible DNA fingerprint patterns for various phage types were generated using either of the two primers. Results The similarity index of replicates ranged from 89.4–100% for BOXA2R-PCR, and from 90 to 100% for BOXA1R-PCR. The method of DNA isolation (p = 0.08) and the phage propagation conditions at two different temperatures (p = 0.527) had no significant influence on generated patterns. Rep-PCR amplification products were generated from different templates including purified phage DNA, phage lysates and phage plaques. The use of this method enabled comparisons of phage genetic profiles to establish their similarity to related or unrelated phages and their bacterial hosts. Conclusion The findings suggest that repetitive-PCR could be used as a rapid and inexpensive method to preliminary screen phage isolates prior to their selection for more comprehensive studies. The adoption of this rapid, simple and reproducible technique could facilitate preliminary characterisation of a large number of phage isolates and the investigation of genetic relationship between phage genotypes.

Mode of action of bacteriophage &phi149 on cholera and E1 Tor vibrios

1978 ◽  
Vol 24 (12) ◽  
pp. 1583-1589 ◽  
Author(s):  
M. Maiti
Keyword(s):  
Vibrio Cholerae ◽  
Phage Particle ◽  
Small Degree ◽  
Cellular Respiration ◽  
Bacterial Strains ◽  
Killing Effect ◽  
Rate Dependent ◽  
Phage Dna ◽  
Rna And Protein Synthesis ◽  
Phage Propagation

Bacteriophage [Formula: see text] which was propagated in Vibrio cholerae (classical) OGAWA 154 strain, killed Vibrio cholerae (E1 Tor) strain MAK 757 without phage propagation. E1 Tor vibrios underwent a small degree of lysis only when infected by the phage [Formula: see text] at a high multiplicity of infection and lost their viability at a rate-dependent multiplicity of phage infection. Evidence was obtained with 32P-labelled bacteriophage [Formula: see text] for penetration of phage DNA into both bacterial strains. In host strain (OGAWA 154) phage particle synthesis occurred normally. In E1 Tor strain MAK 757 the phage DNA was not degraded but its expression was blocked. The killing effect of ([Formula: see text] on E1 Tor strain MAK 757 is supposed to be due to damage of the cytoplasmic membrane, which could not be repaired under the influence of phage information. This was indicated by a blockage of cellular respiration and RNA and protein synthesis.

scholarly journals Single and Double Infections with Wolbachia in the Parasitic Wasp Nasonia vitripennis Effects on Compatibility

Genetics ◽  
1996 ◽  
Vol 143 (2) ◽  
pp. 961-972 ◽  
Author(s):  
Marie-Jeanne Perrot-Minnot ◽  
Li Rong Guo ◽  
John H Werren
Keyword(s):  
Genomic Dna ◽  
Rapid Development ◽  
Parasitic Wasp ◽  
Pcr Amplification ◽  
Bacterial Strains ◽  
Nasonia Vitripennis ◽  
Larval Diapause ◽  
Wide Range ◽  
Reproductive Incompatibility ◽  
Infected Line

Abstract Wolbachia are cytoplasmically inherited bacteria responsible for reproductive incompatibility in a wide range of insects. There has been little exploration, however, of within species Wolbachia polymorphisms and their effects on compatibility. Here we show that some strains of the parasitic wasp Nasonia vitripennis are infected with two distinct bacterial strains (A and B) whereas others are singly infected (A or B). Double and single infections are confirmed by both PCR amplification and Southern analysis of genomic DNA. Furthermore, it is shown that prolonged larval diapause (the overwintering stage of the wasp) of a double-infected strain can lead to stochastic loss of one or both bacterial strains. After diapause of a double-infected line, sublines were produced with AB, A only, B only or no Wolbachia. A and B sublines are bidirectionally incompatible, whereas males from AB lines are unidirectionally incompatible with females of A and B sublines. Results therefore show rapid development of bidirectional incompatibility within a species due to segregation of associated symbiotic bacteria.

Bactericidal substance produced by Haemophilus influenzae b

1975 ◽  
Vol 21 (10) ◽  
pp. 1587-1594 ◽  
Author(s):  
R. A. Venezia ◽  
R. G. Robertson
Keyword(s):  
Haemophilus Influenzae ◽  
Bactericidal Activity ◽  
High Speed ◽  
Indicator Strain ◽  
Bacterial Strains ◽  
Small Molecular Weight ◽  
Plaque Test ◽  
Haemophilus Species ◽  
Phage Propagation ◽  
Thermostable Protein

During bacteriophage studies on Haemophilus influenzae, it was observed that encapsulated type b and unencapsulated Rb strains released a bactericidal substance active against types a, c, d, e, and f H. influenzae, non-typable H. influenzae strains, other Haemophilus species, and certain members of the Enterobacteriaceae. The bactericidal activity was assayed by a plaque test utilizing an Rd strain as an indicator lawn and was also demonstrated in mixed broth cultures of a producer strain and an indicator strain. Immediate lysis of sensitive bacteria by the factor was not evident. The factor is sensitive to trypsin but resistant to deoxyribonuclease, treatment with 2-mercaptoethanol, lipase, α-amylase, and heating in a 100 °C water bath for 20 min. The activity is not dependent upon increased Ca2+ or Mg2+ concentration as is necessary for HP1C1 and S2 phage propagation. The bactericidal factor is not pelleted by high-speed centrifugation at 150 000 × g for 6 h. Treatment with ultraviolet light or mitomycin C does not result in observable phage, phage-like particles, or increased bactericidal activity. The bactericidal factor is not a typical small molecular weight "colicin-like" bacteriocin in that it is not inducible, has a wider range of activity, and does not kill by "single-hit" kinetics. On preliminary characterization, it is a thermostable protein toxic to certain bacterial strains.

scholarly journals Kefiran Ekstraktının Bazı Bitki Patojeni Bakterilerine Karşı Antimikrobiyal Etkisinin Değerlendirilmesi

2020 ◽  
Vol 8 (4) ◽  
pp. 889-894
Author(s):  
Bilgin Taşkın
Keyword(s):  
Antibacterial Effect ◽  
Antimicrobial Effect ◽  
Acetic Acid Bacteria ◽  
Fermented Milk ◽  
Diffusion Method ◽  
Milk Product ◽  
Bacterial Strains ◽  
Private Households ◽  
Different Temperatures

Kefir; is a fermented milk product which is produced by granules containing a wide variety of microorganisms such as lactic acid bacteria, acetic acid bacteria and yeasts. It is traditionally consumed in many countries. It has been shown in many studies that the polysaccharide structure surrounding the granules which is composed mainly of kefiran molecule has antimicrobial effect against various pathogens as well as many health promoting effects. In this study, 24 h fermented kefir was used with two types of kefir granules for production of kefiran extract. One of them is being sold commercially and the other was collected from private households in a different region of Turkey. Kefiran extraction was carried out from matured kefir granules using three different temperatures, 80°C, 90°C and 100°C. Also, the protein contents of the extracted solutions were determined by Bradford method. Protein content of the extract solutions obtained were measured as 0.001 g/ml. The antibacterial effect of 0.05, 0.1, 1 and 2 mg of this extract against several plant pathogenic bacterial strains belonging to genus Pseudomonas, Xanthomonas, Erwinia ve Clavibacter was investigated in vitro for the first time. For this purpose, two methods, disc diffusion method and spreading method were used. The AN and SD kefir supernatants used as the positive controls in the experiments showed an average of 13-17 mm and 10-14 mm inhibition zones on the isolates, respectively, but the antibacterial effect of kefiran extracts was not observed.

Diversity of nodular bacteria ofScorpiurus muricatusin western Algeria and their impact on plant growth

2017 ◽  
Vol 63 (5) ◽  
pp. 450-463 ◽  
Author(s):  
Zoulikha Bouchiba ◽  
Zineb Faiza Boukhatem ◽  
Zohra Ighilhariz ◽  
Nouria Derkaoui ◽  
Benaissa Kerdouh ◽  
...  
Keyword(s):  
Plant Growth ◽  
Rhizobium Leguminosarum ◽  
Root Nodules ◽  
Pcr Amplification ◽  
Nucleotide Sequencing ◽  
Rrna Gene ◽  
Bacterial Strains ◽  
Single Strain ◽  
Western Algeria ◽  
First Time

A total of 51 bacterial strains were isolated from root nodules of Scorpiurus muricatus sampled from 6 regions of western Algeria. Strain diversity was assessed by rep-PCR amplification fingerprinting, which grouped the isolates into 28 different clusters. Partial nucleotide sequencing of the 16S rRNA gene and BLAST analysis revealed that root nodules of S. muricatus were colonized by different species close to Rhizobium vignae, Rhizobium radiobacter, Rhizobium leguminosarum, Phyllobacterium ifriqiyense, Phyllobacterium endophyticum, Starkeya sp., and Pseudomonas sp. However, none of these strains was able to form nodules on its host plant; even nodC was present in a single strain (SMT8a). The inoculation test showed a great improvement in the growth of inoculated plants compared with noninoculated control plants. A significant amount of indole acetic acid was produced by some strains, but only 2 strains could solubilize phosphate. In this report we described for the first time the diversity of bacteria isolated from root nodules of S. muricatus growing in different regions in western Algeria and demonstrated their potential use in promoting plant growth.

scholarly journals Removal of Microcystin-LR by a Novel Native Effective Bacterial Community Designated as YFMCD4 Isolated from Lake Taihu

Toxins ◽  
2018 ◽  
Vol 10 (9) ◽  
pp. 363 ◽  
Author(s):  
Fei Yang ◽  
Jian Guo ◽  
Feiyu Huang ◽  
Isaac Massey ◽  
Ruixue Huang ◽  
...  
Keyword(s):  
Bacterial Community ◽  
High Performance ◽  
Lake Taihu ◽  
Degradation Process ◽  
Alcaligenes Faecalis ◽  
Natural Ecosystem ◽  
The Novel ◽  
Bacterial Strains ◽  
Different Temperatures ◽  
Cost Efficient

Microcystin-LR (MC-LR) is the most toxic and frequently detected monocyclic heptapeptide hepatotoxin produced by cyanobacteria, which poses a great threat to the natural ecosystem and public health. It is very important to seek environment-friendly and cost-efficient methods to remove MC-LR in water. In this study, the MC-degrading capacities of a novel indigenous bacterial community designated as YFMCD4 and the influence of environmental factors including various temperatures, MC concentrations and pH on the MC-degrading activities were investigated utilizing high-performance liquid chromatography (HPLC). In addition, the MC-degrading mechanism of YFMCD4 was also studied using HPLC coupled with a mass spectrometry equipped with electrospray ionization interface (HPLC-ESI-MS). The data showed MC-LR was completely removed at the maximum rate of 0.5 µg/(mL·h) under the optimal condition by YFMCD4. Two pure bacterial strains Alcaligenes faecalis and Stenotrophomonas acidaminiohila were isolated from YFMCD4 degraded MC-LR at a slower rate. The MC-degrading rates of YFMCD4 were significantly affected by different temperatures, pH and MC-LR concentrations. Two intermediates of a tetrapeptide and Adda appeared in the degradation process. These results illustrate that the novel YFMCD4 is one of the highest effective MC-degrading bacterial community, which can completely remove MC-LR and possesses a significant potential to treat water bodies contaminated by MC-LR.

Determination of Histamine in Japanese Spanish Mackerel (Scomberomorus niphonius) Meat Implicated in a Foodborne Poisoning

2019 ◽  
Vol 82 (10) ◽  
pp. 1643-1649 ◽  
Author(s):  
CHIU-CHU HWANG ◽  
PEI-HUI TSENG ◽  
YI-CHEN LEE ◽  
HSIEN-FENG KUNG ◽  
CHUN-YUNG HUANG ◽  
...  
Keyword(s):  
Meat Products ◽  
Pcr Amplification ◽  
Enterobacter Aerogenes ◽  
Potential Hazard ◽  
Bacterial Strains ◽  
Spanish Mackerel ◽  
Rdna Sequencing ◽  
Fish Samples ◽  
Raw Fish ◽  
Scomberomorus Niphonius

ABSTRACT An incident of foodborne poisoning causing illness in seven victims due to ingestion of fried Japanese Spanish mackerel (JS mackerel; Scomberomorus niphonius) meat occurred in September 2014 in Hualien County, eastern Taiwan. Of the two suspected fish meats, one raw sample contained 3,318 ppm of histamine and one fried sample contained 1,906 ppm of histamine, levels which are greater than the potential hazard action level (500 ppm) in most illness cases. Given the allergy-like symptoms of the victims and the high histamine content in the suspected fish samples, this foodborne poisoning was strongly suspected to be caused by histamine intoxication. In addition, five histamine-producing bacterial strains isolated from suspected raw fish samples, capable of producing 152 to 1,020 ppm of histamine in Trypticase soy broth supplemented with 1.0% l-histidine, were identified as Hafnia alvei (one strain), Enterobacter aerogenes (two strains), Raoultella ornithinolytica (one strain), and Morganella morganii (one strain) by 16S rDNA sequencing with PCR amplification. Moreover, 12 raw fish samples and 39 fried fish samples from retail stores were collected and tested to determine the occurrence of histamine. Two of 12 commercial raw fish samples (16.7%) had histamine levels greater than the U.S. Food and Drug Administration guideline for decomposition of 50 ppm for scombroid fish or product or a combination of both. To our knowledge, this is the first report in Taiwan to demonstrate that the JS mackerel meat products could cause histamine intoxication.

scholarly journals Streptomyces sp. Strain MUSC 125 from Mangrove Soil in Malaysia with Anti-MRSA, Anti-Biofilm and Antioxidant Activities

Molecules ◽  
2020 ◽  
Vol 25 (15) ◽  
pp. 3545
Author(s):  
Hefa Mangzira Kemung ◽  
Loh Teng-Hern Tan ◽  
Kok-Gan Chan ◽  
Hooi-Leng Ser ◽  
Jodi Woan-Fei Law ◽  
...  
Keyword(s):  
Polyketide Synthase ◽  
Methanolic Extract ◽  
Antioxidant Activities ◽  
Pcr Amplification ◽  
Gene Clusters ◽  
Bacterial Strains ◽  
Phenotypic Analysis ◽  
Streptomyces Sp ◽  
Mangrove Soil ◽  
Promising Source

There is an urgent need to search for new antibiotics to counter the growing number of antibiotic-resistant bacterial strains, one of which is methicillin-resistant Staphylococcus aureus (MRSA). Herein, we report a Streptomyces sp. strain MUSC 125 from mangrove soil in Malaysia which was identified using 16S rRNA phylogenetic and phenotypic analysis. The methanolic extract of strain MUSC 125 showed anti-MRSA, anti-biofilm and antioxidant activities. Strain MUSC 125 was further screened for the presence of secondary metabolite biosynthetic genes. Our results indicated that both polyketide synthase (pks) gene clusters, pksI and pksII, were detected in strain MUSC 125 by PCR amplification. In addition, gas chromatography-mass spectroscopy (GC-MS) detected the presence of different chemicals in the methanolic extract. Based on the GC-MS analysis, eight known compounds were detected suggesting their contribution towards the anti-MRSA and anti-biofilm activities observed. Overall, the study bolsters the potential of strain MUSC 125 as a promising source of anti-MRSA and antibiofilm compounds and warrants further investigation.

Changes in the structure and diversity of bacterial communities during the process of adaptation to organic wastewater

2010 ◽  
Vol 56 (4) ◽  
pp. 352-355 ◽  
Author(s):  
Junmin Li ◽  
Zexin Jin ◽  
Binbin Yu
Keyword(s):  
Bacterial Communities ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Similarity Index ◽  
Pcr Amplification ◽  
Genotypic Diversity ◽  
Diversity Indices ◽  
Inhibitory Effect ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Gradient Gel Electrophoresis

To explore changes in the structure and diversity of activated sludge-derived microbial communities during adaptation to gradual increases in the concentration of wastewater, RAPD–PCR and the combination of PCR amplification of 16S rRNA genes with denaturing gradient gel electrophoresis (DGGE) analysis were used. In bacterial communities exposed to 0%, 5%, 10%, 20%, or 40% wastewater, there were 27, 25, 18, 17 and 16 bands, respectively, based on DGGE data, while there were 69, 83, 97, 86, and 88 bands, respectively, based on RAPD data. The community similarity index among bacterial communities during the process of adaptation to different concentrations of wastewater was different based on DGGE and RAPD data. Based on DGGE and RAPD profiles, the Shannon–Weiner and Simpson’s diversity indices decreased sharply upon exposure to 10% wastewater, indicating that 10% wastewater might be a critical point at which the growth of bacteria could be significantly inhibited and the genotypic diversity could change. This indicated that changes in structure and diversity might have an inhibitory effect on the toxicity of organic matter and that selection and adaptation could play important roles in the changes.

scholarly journals The Action of Bacteriophage Ω8 on Two Strains of Escherichia coli 08

Microbiology ◽  
2000 ◽  
Vol 81 (1) ◽  
pp. 131-144 ◽  
Author(s):  
Barbara Wallenfels ◽  
K. Jann
Keyword(s):  
Escherichia Coli ◽  
Protein Synthesis ◽  
Phage Particle ◽  
Cytoplasmic Membrane ◽  
Cellular Respiration ◽  
Bacterial Strains ◽  
Viral Dna ◽  
Killing Effect ◽  
E Coli ◽  
Phage Propagation

Bacteriophage Ω8 is propagated in Escherichia coli E56b (08: K27-:H-), a non-capsulated strain. Another non-capsulated strain, E. coli 2398 (08:K?-:H-), is killed by bacteriophage Ω8 without phage propagation. This strain was formerly believed to be E. coli 093:K?-:H-, cross-reacting with strain E56b. We have established chemical and serological identity of the 08-specific lipopolysaccharides of the two strains. The 08-specific lipopolysaccharides of both strains inhibited the infection of Escherichia coli E56b with bacteriophage Ω8 equally well. The adsorption rate constants of Ω8 were identical for the two strains of E. coli 08. Evidence was obtained with 32P-labelled bacteriophage Ω8 for penetration of viral DNA into both bacterial strains. In host strain E56b, phage particle synthesis occurred normally. In strain 2398 the viral DNA was not degraded but its expression was blocked. The killing effect of Ω8 on E. coli strain 2398 is supposed to be due to damage of the cytoplasmic membrane, which could not be reversed under the influence of viral information. This was indicated by a blockage of cellular respiration, β-galactoside transport and RNA as well as protein synthesis.

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