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Microbiology Research is published by MDPI from Volume 11 Issue 2 (2020). Previous articles were published by another publisher in Open Access under a CC-BY (or CC-BY-NC-ND) licence, and they are hosted by MDPI on mdpi.com as a courtesy and upon agreement with PAGEPress.

Microbiol. Res., Volume 4, Issue 1 (April 2013) – 6 articles

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Article
Molecular Investigation of Zoonotic Genotypes of Giardia intestinalis Isolates in Humans, Dogs and Cats, Sheep, Goats and Cattle in Araçatuba (São Paulo State, Brazil) by the Analysis of β-Giardin Gene Fragments
Microbiol. Res. 2013, 4(1), e6; https://doi.org/10.4081/mr.2013.e6 - 09 Oct 2013
Cited by 2 | Viewed by 1
Abstract
In the period from July 2009 to October 2010, fecal samples from 61 animals and 154 humans from the municipality of Aracatuba (São Paulo State, Brazil) were studied. Fecal samples from animals were collected in the Municipal Animal Shelter and the Veterinary Hospital [...] Read more.
In the period from July 2009 to October 2010, fecal samples from 61 animals and 154 humans from the municipality of Aracatuba (São Paulo State, Brazil) were studied. Fecal samples from animals were collected in the Municipal Animal Shelter and the Veterinary Hospital of the Universidade Estadual Paulista. Human fecal specimens were collected in playschools in the outskirts of the city by the private network of clinical analysis laboratories of the municipal. Diagnosis was done by optical microscopy using the Faust and Hoffmann, Pons and Janer techniques. The genotypes of Giardia intestinalis were characterized by PCR-RFLP and confirmed by sequencing the β-giardin gene. Human specimens were positive in 25.3% (39/154) of the cases with 26.8% (36/134) of the specimens from children and 15% (3/20) from adults being positive. The frequency of G. intestinalis among the animals was 23.0% (14/61). A total of 32 isolates of G. intestinalis obtained from human feces and six from dogs and cats were characteristic of the A genotype (AI and AII/AIII). The results of this study in respect to frequency of giardiasis are similar to reported in most studies in Brazil. The prevalence observed in animal populations conforms to worldwide infection rates. G. intestinalis genotypes considered zoonotic were detected in both pets and humans from the city of Aractuba, suggesting a possible zoonotic transmission of the parasite in the northwestern region of São Paulo State. The absence of these genotypes in farm animals may imply that they are not involved in the chain of transmission to humans in this region. Full article
Article
Cloning, Expression of the Major Capsid Protein Gene from Marine Algae Emiliania huxleyi Virus and the Possible Use in Detection of Virus Infection
Microbiol. Res. 2013, 4(1), e5; https://doi.org/10.4081/mr.2013.e5 - 09 Sep 2013
Cited by 2 | Viewed by 1
Abstract
Here we described the cloning, bioinformatic characterization and expression in Escherichia coli of the major capsid protein (MCP) from marine unicellular algae Emiliania huxleyi virus EhV-99B1 isolate. The purified recombinant MCP was used to develop a polyclonal antibody for testing viral infection. The [...] Read more.
Here we described the cloning, bioinformatic characterization and expression in Escherichia coli of the major capsid protein (MCP) from marine unicellular algae Emiliania huxleyi virus EhV-99B1 isolate. The purified recombinant MCP was used to develop a polyclonal antibody for testing viral infection. The full length open-reading frame (ORF) of MCP encodes a protein of 496 amino acids with a calculated molecular mass of 55 kDa and Ip 6.34. Hydropathy analysis of MCP showed that there were 6 largely hydrophobic domains, which may be important for the interaction with the envelope protein. The conserved region of EhV strains MCP had high similarity in amino acid sequence and secondary structure which allow us to develop a specific biomarker for EhVs infection detection. The full length ORF was subcloned into expression vector pGEX-4T-3 for overexpression in E. coli as glutathione-Stransferase-L1 (GST-L1) fusion protein and the soluble recombinant protein was used to generate polyclonal antibodies in mice. The obtained antisera reacted in Western immunoblots with the same protein both in purified EhV-99B1 virions and infected host cells sample. These shows that the antiserum against recombinant EhV-MCP offers the potential to develop immunofluorescence techniques for the detection of EhVs infected cells. Full article
Article
Escherichia coli as An Indicator of Bacteriological Quality of Water: An Overview
Microbiol. Res. 2013, 4(1), e2; https://doi.org/10.4081/mr.2013.e2 - 11 Jun 2013
Cited by 160 | Viewed by 1
Abstract
Monitoring the microbiological quality of drinking water relies largely on examination of indicator bacteria such as coliforms, Escherichia coli, and Pseudomonas aeruginosa. E. coli is a member of the faecal coliform group and is a more specific indicator of faecal pollution [...] Read more.
Monitoring the microbiological quality of drinking water relies largely on examination of indicator bacteria such as coliforms, Escherichia coli, and Pseudomonas aeruginosa. E. coli is a member of the faecal coliform group and is a more specific indicator of faecal pollution than other faecal coliforms. Two key factors have led to the trend toward the use of E. coli as the preferred indicator for the detection of faecal contamination, not only in drinking water, but also in other matrices as well: first, the finding that some faecal coliforms were non faecal in origin, and second, the development of improved testing methods for E. coli. The faecal coliform definition has also been revised to coincide better with the genetic make-up of its members and now includes newly identified environmental species. As a result, faecal coliforms are increasingly being referred to as thermotolerant coliforms. This, combined with improved detection methods for E. coli, has started a trend toward the use of E. coli in place of thermotolerant coliforms as a more reliable indicator of faecal pollution in drinking water. At present, E. coli appears to provide the best bacterial indication of faecal contamination in drinking water. This is based on the prevalence of thermotolerant (faecal) coliforms in temperate environments as compared to the rare incidence of E. coli, the prevalence of E. coli in human and animal faeces as compared to other thermotolerant coliforms, and the availability of affordable, fast, sensitive, specific and easier to perform detection methods for E. coli. Full article
Article
Antibacterial Activities of Serum from the Komodo Dragon (Varanus komodoensis)
Microbiol. Res. 2013, 4(1), e4; https://doi.org/10.4081/mr.2013.e4 - 30 Apr 2013
Cited by 4 | Viewed by 1
Abstract
Komodo dragons (Varanus komodoensis) are able to feed on large prey items by injecting a dose of toxic bacteria with their bite that, over time, kills the prey by systemic infection. Dragons also suffer bites from other members of their own [...] Read more.
Komodo dragons (Varanus komodoensis) are able to feed on large prey items by injecting a dose of toxic bacteria with their bite that, over time, kills the prey by systemic infection. Dragons also suffer bites from other members of their own species during territorial disputes and feeding frenzies. However, they do not suffer the same fate as their prey, suggesting that they have developed a strong immunity to bacterial infections. This study was undertaken to determine the antibacterial activities of serum from the Komodo dragon. Bacterial cultures were treated with different volumes serum from Varanus komodoensis and the growth was monitored by optical density at 430 nm. In addition, the serum was treated with proteaseinfections. This study was undertaken to determine the antibacterial activities of serum from the Komodo dragon. Bacterial cultures were treated with different volumes serum from Varanus komodoensis and the growth was monitored by optical density at 430 nm. In addition, the serum was treated with protease, chelators of divalent metal ions, or with mild heat to determine the mechanism of antibacterial activities. Treatment of bacterial cultures with serum from Komodo dragons, chelators of divalent metal ions, or with mild heat to determine the mechanism of antibacterial activities. Treatment of bacterial cultures with serum from Komodo dragons (Varanus komodoensis) resulted in a volume-dependent decrease in bacterial growth. Cultures of Escherichia coli, Staphylococcus aureus, and Klebsiella oxytoca exhibited moderate-strong growth inhibition by V. komodoensis serum, while cultures of Streptococcus epidermitis, Salmonella typhimurium, Providencia stuartii, and Shigella flexneri were nearly completely obliterated for 24 h by only 10% (v/v) serum. The antibacterial activity of V. komodensis serum occurred very rapidly, as 18% of E. coli growth was inhibited by a five min exposure to serum. Furthermore, 10- and 20-min incubations of E. coli with serum from V. komodoensis resulted in 43 and 68% inhibition of bacterial growth, respectively. The bactericidal capacity of the serum against E. coli was 2,075,000 bacteria/μL serum, and was inhibited by mild heat treatment, pronase, EDTA, and phosphate, indicating that the anti-bacterial action is most probably due to the presence of a potent serum complement protein system. Full article
Article
Effect of Catanospermine, 1-Deoxynojirimycin or 1-Deoxymannojirimycin on Biological and Functional Activities of Japanese Encephalitis Virus in Porcine Stable Kidney Cells
Microbiol. Res. 2013, 4(1), e3; https://doi.org/10.4081/mr.2013.e3 - 15 Apr 2013
Cited by 1 | Viewed by 1
Abstract
In the present study, effect of catanospermine (CST), 1-deoxynojirimycin (DNJ) or 1-deoxymannojirimycin (DMJ) was studied on porcine stable kidney (PS) cells infected with Japanese encephalitis virus (JEV). As both CST and DNJ are potent inhibitors of ER alpha-glucosidases 1 and II, while DMJ [...] Read more.
In the present study, effect of catanospermine (CST), 1-deoxynojirimycin (DNJ) or 1-deoxymannojirimycin (DMJ) was studied on porcine stable kidney (PS) cells infected with Japanese encephalitis virus (JEV). As both CST and DNJ are potent inhibitors of ER alpha-glucosidases 1 and II, while DMJ is an inhibitor of Golgi mannosidase which removes alpha (1, 2) Man residues from the N-glycan precursor. Treatment of infected cells with CST (200 uM/mL), DNJ (100 uM/mL) or DMJ (200 uM/mL) did not produce much effect on viral gpE epitope presentation within the cells as well as on the cell surface as detected in the immunofluorescence employing monoclonal (MAbs) and polyclonal (PAbs) antibodies. As well the treated (infected) cells showed only a marginal decrease in infectious virus yield along with a slight decrease in haemagglutination activity of the virus that was recorded in comparison to the untreated infected (control) cells and the cells infected with Dengue virus. Immuno-blotting of the separated proteins from infected lysed cells and probed with anti-gpE MAbs also revealed a band corresponding to JEV gpE (MW 53 kDa) both with inhibitor treated and the untreated cells; the reactivity with the former however, was somewhat less intense and prominent in comparison to latter (control untreated) indicating some effect on JEV. The present results indicate that these inhibitors by in large, do not affect maturation and the release of infective JE virions in PS cells. Full article
Article
Molecular Epidemiology of GB Type C Virus among Individuals Exposed to Hepatitis C Virus in Cameroon
Microbiol. Res. 2013, 4(1), e1; https://doi.org/10.4081/mr.2013.e1 - 02 Apr 2013
Viewed by 1
Abstract
GB Virus Type C (GBV-C), a blood-borne flavivirus currently infects about one sixth of the world’s population. Its transmission has been reported through parenteral, sexual and vertical routes. Unusually for RNA viruses, it exhibits a high degree of conservation of the polyprotein sequence. [...] Read more.
GB Virus Type C (GBV-C), a blood-borne flavivirus currently infects about one sixth of the world’s population. Its transmission has been reported through parenteral, sexual and vertical routes. Unusually for RNA viruses, it exhibits a high degree of conservation of the polyprotein sequence. The geographical distribution of GBV-C suggests an African origin and a long-term co-evolution in the human population but without any known pathogenicity. The aim of this study was to describe the different sub-types of this virus in Southern Cameroon. We studied the genetic epidemiology of GBV-C among rural populations where many HIV-1 and HCV genotypes have been identified. Plasma samples of 345 subjects with evidence of HCV exposure were tested for GBV-C infection. To detect GBV-C RNA, reverse transcription followed by a nested PCR of 5’UTR were performed. Direct sequencing and phylogenetic studies using PHYLIP, PAUP* and SimPlot were carried out. In total, 31 GBV-C RNA-positive samples were detected giving a prevalence of 9.0% among HCV-exposed individuals. Phylogenetic analysis of the 5’UTR showed two distinct clusters: Genotype 1 and Genotype 2. Twenty-eight isolates (8.0%) clustered with Genotype 1 and 3 (1.0%) with Genotype 2. More than one genotype of GBV-C is prevalent in Cameroon of which GBV-C Genotype 1 is more common, confirming reports in the literature. Studying the near full-length genome sequences of GBV-C isolates from primates in this region may provide clues of viral recombination, evolution and origin. Full article
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