ABSTRACT
AIM: To investigate the impact of inoculating peanut seeds with the biocontrol agent Trichoderma harzianum ITEM 3636 on the structure of bacterial and fungal communities from agricultural soils. METHODS AND RESULTS: Polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (PCR-DGGE) and next-generation sequencing (NGS) of amplicons (or marker gene amplification metagenomics) were performed to investigate potential changes in the structure of microbial communities from fields located in a peanut-producing area in the province of Córdoba, Argentina. Fields had history of peanut smut (caused by Thecaphora frezii) incidence. The Shannon indexes (H'), which estimate diversity, obtained from the PCR-DGGE assays did not show significant differences neither for bacterial nor for fungal communities between control and inoculation treatments. On the other hand, the number of operational taxonomic units obtained after NGS was similar between all the analysed samples. Moreover, results of alpha and beta diversity showed that there were no significant variations between the relative abundances of the most representative bacterial and fungal phyla and genera, in both fields. CONCLUSIONS: Trichoderma harzianum ITEM 3636 decreases the incidence and severity of agriculturally relevant diseases without causing significant changes in the microbial communities of agricultural soils. SIGNIFICANCE AND IMPACT OF THE STUDY: Our investigations provide information on the structure of bacterial and fungal communities in peanut-producing fields after inoculation of seeds with a biocontrol agent.
Subject(s)
Biological Control Agents , Soil Microbiology , Trichoderma , Agriculture , Arachis , Argentina , Bacteria/genetics , Bacteria/isolation & purification , Denaturing Gradient Gel Electrophoresis , Fungi/genetics , Fungi/isolation & purification , High-Throughput Nucleotide Sequencing , Microbiota , Polymerase Chain Reaction , Seeds , Soil/chemistryABSTRACT
The in vitro antiviral activity of the essential oil from Minthostachys verticillata was investigated against herpes simplex virus type 1 (HSV-1) and pseudorabies virus (PrV). The viral inhibition was assayed employing viral plaque reduction assay. The antiviral activity of the essential oil specifically affects PrV and HSV-1 multiplication, since it was found that non toxic effects on cells were observed at the concentrations assayed. The therapeutic index values were 10.0 and 9.5 for HSV-1 and PrV, respectively. The antibacterial activity was studied using a diffusion assay and the broth tube dilution method. Gram-positive bacteria were more sensitive to inhibition by plant essential oil than the gram-negative bacteria. The essential oil of M. verticillata was analyzed by gas chromatography (GC) technique. Of the six components identified in the volatile oil, pulegone (44.56%) and menthone (39.51%) were the major constituents. The antimicrobial activity can be explained to some extent by the presence of pulegone. Results suggest that further investigations concerning the isolation of the substance responsible for the antimicrobial activity and an effort to define the mechanisms of action are warranted.
Subject(s)
Anti-Bacterial Agents/pharmacology , Antiviral Agents/pharmacology , Gram-Negative Bacteria/drug effects , Gram-Positive Bacteria/drug effects , Herpesvirus 1, Suid/drug effects , Monoterpenes , Plant Oils/pharmacology , Plants, Medicinal/chemistry , Simplexvirus/drug effects , Bacillus cereus/drug effects , Chromatography, Gas , Cyclohexane Monoterpenes , Escherichia coli/drug effects , Herpesvirus 1, Suid/physiology , Menthol/analogs & derivatives , Menthol/isolation & purification , Menthol/pharmacology , Microbial Sensitivity Tests , Plant Oils/chemistry , Proteus mirabilis/drug effects , Pseudomonas aeruginosa/drug effects , Simplexvirus/physiology , Staphylococcus aureus/drug effects , Terpenes/isolation & purification , Terpenes/pharmacology , Viral Plaque Assay , Virus Replication/drug effectsABSTRACT
The in vitro antiviral activity of the essential oil from Minthostachys verticillata was investigated against herpes simplex virus type 1 (HSV-1) and pseudorabies virus (PrV). The viral inhibition was assayed employing viral plaque reduction assay. The antiviral activity of the essential oil specifically affects PrV and HSV-1 multiplication, since it was found that non toxic effects on cells were observed at the concentrations assayed. The therapeutic index values were 10.0 and 9.5 for HSV-1 and PrV, respectively. The antibacterial activity was studied using a diffusion assay and the broth tube dilution method. Gram-positive bacteria were more sensitive to inhibition by plant essential oil than the gram-negative bacteria. The essential oil of M. verticillata was analyzed by gas chromatography (GC) technique. Of the six components identified in the volatile oil, pulegone (44.56) and menthone (39.51) were the major constituents. The antimicrobial activity can be explained to some extent by the presence of pulegone. Results suggest that further investigations concerning the isolation of the substance responsible for the antimicrobial activity and an effort to define the mechanisms of action are warranted.
Subject(s)
Anti-Bacterial Agents/pharmacology , Antiviral Agents , Gram-Negative Bacteria/drug effects , Gram-Positive Bacteria/drug effects , Herpesvirus 1, Suid , Plant Oils/pharmacology , Plants, Medicinal , Simplexvirus , Bacillus cereus , Chromatography, Gas , Escherichia coli , Herpesvirus 1, Suid , Menthol , Microbial Sensitivity Tests , Plant Oils/chemistry , Proteus mirabilis , Pseudomonas aeruginosa , Virus Replication/drug effects , Simplexvirus , Staphylococcus aureus , Terpenes , Viral Plaque AssayABSTRACT
The in vitro antiviral activity of the essential oil from Minthostachys verticillata was investigated against herpes simplex virus type 1 (HSV-1) and pseudorabies virus (PrV). The viral inhibition was assayed employing viral plaque reduction assay. The antiviral activity of the essential oil specifically affects PrV and HSV-1 multiplication, since it was found that non toxic effects on cells were observed at the concentrations assayed. The therapeutic index values were 10.0 and 9.5 for HSV-1 and PrV, respectively. The antibacterial activity was studied using a diffusion assay and the broth tube dilution method. Gram-positive bacteria were more sensitive to inhibition by plant essential oil than the gram-negative bacteria. The essential oil of M. verticillata was analyzed by gas chromatography (GC) technique. Of the six components identified in the volatile oil, pulegone (44.56) and menthone (39.51) were the major constituents. The antimicrobial activity can be explained to some extent by the presence of pulegone. Results suggest that further investigations concerning the isolation of the substance responsible for the antimicrobial activity and an effort to define the mechanisms of action are warranted.(AU)
Subject(s)
Anti-Bacterial Agents/pharmacology , Antiviral Agents/pharmacology , Gram-Negative Bacteria/drug effects , Gram-Positive Bacteria/drug effects , Herpesvirus 1, Suid/drug effects , Plant Oils/pharmacology , Plants, Medicinal/chemistry , Simplexvirus/drug effects , Bacillus cereus/drug effects , Chromatography, Gas , Escherichia coli/drug effects , Herpesvirus 1, Suid/physiology , Menthol/analogs & derivatives , Menthol/isolation & purification , Menthol/pharmacology , Microbial Sensitivity Tests , Plant Oils/chemistry , Viral Plaque Assay , Proteus mirabilis/drug effects , Pseudomonas aeruginosa/drug effects , Simplexvirus/physiology , Staphylococcus aureus/drug effects , Terpenes/isolation & purification , Terpenes/pharmacology , Virus Replication/drug effectsABSTRACT
Mitogenic and antigenic lymphocyte stimulation was examined in pigs that had been immunized with 2 inactivated vaccines which had been prepared with Aujesky's disease virus (ADV), strain RC/79. One vaccine was partially purified by ultra-centrifugation (Vaccine A) the other one was not (Vaccine B). A second dose of vaccine had no influence over the blastogenic response when the lymphocytes were stimulated with phytohemoagglutinin (PHA). Lymphocyte response to the ADV antigen in the immunized pigs was significantly higher at day 30 post inoculation than at day 0 indicating that it was highly specific. Cellular antigens contained in the viral cultures produced a slight non-specific response as shown by a low increase in the levels of lymphocyte blastic transformation (LBT) in the control group at day 30 p.i., this group only received a non infected Vero cell suspension. This was the case in pigs that received vaccine A as well as in those that were vaccinated with vaccine B. Vaccine B contains a greater quantity of contaminating cellular antigens, since it is an impure vaccine. Such antigens could act as non-specific immunomodulators, potentiating cell-mediated immunity (CMI). This assay demonstrated that inactivated vaccines produced with VPR-RC/79, partially purified and unpurified are capable of inducing a humoral immune response. The blastogenic reaction of the peripheral blood lymphocytes to antigens of ADV strain RC/79, indicated that the employed immunogens also induced the CMI. Results indicate that the analyzed immunogens could be considered for the possible implementation of epidemiological measures, which imply the use of vaccines to prevent pseudo-rabies in Argentina.
Subject(s)
Herpesvirus 1, Suid/immunology , Lymphocyte Activation , Pseudorabies/prevention & control , Sus scrofa/immunology , Swine Diseases/prevention & control , Viral Vaccines/immunology , Adjuvants, Immunologic , Animals , Antibody Formation , Antigens, Surface/immunology , Antigens, Viral/immunology , Chlorocebus aethiops , Female , Immunity, Cellular , Lymphocyte Activation/drug effects , Lymphocytes/drug effects , Lymphocytes/immunology , Male , Phytohemagglutinins/pharmacology , Vaccines, Inactivated/immunology , Vaccines, Inactivated/isolation & purification , Vero Cells/immunology , Viral Vaccines/isolation & purificationABSTRACT
The in vitro antiviral activity of the essential oil from Minthostachys verticillata was investigated against herpes simplex virus type 1 (HSV-1) and pseudorabies virus (PrV). The viral inhibition was assayed employing viral plaque reduction assay. The antiviral activity of the essential oil specifically affects PrV and HSV-1 multiplication, since it was found that non toxic effects on cells were observed at the concentrations assayed. The therapeutic index values were 10.0 and 9.5 for HSV-1 and PrV, respectively. The antibacterial activity was studied using a diffusion assay and the broth tube dilution method. Gram-positive bacteria were more sensitive to inhibition by plant essential oil than the gram-negative bacteria. The essential oil of M. verticillata was analyzed by gas chromatography (GC) technique. Of the six components identified in the volatile oil, pulegone (44.56
) and menthone (39.51
) were the major constituents. The antimicrobial activity can be explained to some extent by the presence of pulegone. Results suggest that further investigations concerning the isolation of the substance responsible for the antimicrobial activity and an effort to define the mechanisms of action are warranted.
ABSTRACT
The antiviral activity of alcoholic extracts of several species belonging to the Asteraceae, Labiatae, Plantaginaceae, Schizaceae, Umbelliferae, Usneaceae and Verbenaceae families has been studied. The tests were carried out in Vero celís-pseudorabies virus strain RC/79 (herpes suis virus) system. Eight plant extracts (Achyrocline satureioides, Ambrossia tenuifolia, Baccharis articulata, Eupatorium buniifolium, Mynthostachys verticillata, Plantago brasiliensis, Plantago mayor L and Verbascum thapsus) were able to inhibit at least 2 log, the viral infectivity.
Subject(s)
Antiviral Agents/isolation & purification , Herpesvirus 1, Suid/drug effects , Plant Extracts/pharmacology , Plants, Medicinal/chemistry , Animals , Antiviral Agents/pharmacology , Argentina , Chlorocebus aethiops , Herpesvirus 1, Suid/physiology , Vero Cells , Virus Replication/drug effectsABSTRACT
BACKGROUND: Most mucose-cutaneous infections can be diagnosed by clinical history with explorative technique. Nevertheless the definitive etiologic diagnostic can only be established with the help of the isolation and identification of the causal agent. PATIENT AND METHODS: We reported a case of generalized infection in a new born, with is clinical characteristics, virological diagnostic techniques and treatment of the disease. RESULTS: Herpes simplex virus was isolated in Vero cellular culture from CRL and vesiculose lesions of the child's thorax region and from the mothers endocervical scrapes. The agent was identified by seroneutralization test. This confirmed a perinatal transmission of Herpes simplex virus type 2. The tomographic studies revealed characteristic alterations of a viral encephalitis. An antiviral treatment was useful in this therapy. CONCLUSIONS: Our results demonstrate the importance of an early viral diagnostic which permits the applications of specific treatment and thus the prevention of severe complications.
Subject(s)
Herpes Simplex/congenital , Herpes Simplex/diagnosis , Herpes Simplex/drug therapy , Herpesvirus 2, Human/isolation & purification , Humans , Infant, Newborn , Neutralization TestsABSTRACT
Colorimetric determinations of proteolytic activity were performed to measure the effects on dye protean substrates including tissue powders. The substrates were assayed with 98 strains obtained from the milk of cows with mastitis. Trypsin was employed as positive control and it verified the susceptibility of the method. Enzymatic activity was estimated in trypsin units per milliliter of incubation mixture. The percentages of strains active on specific proteins were 47.8% for elastin, 61.6% for collagen and when hide powder and udder extract were used as dye substrates, the proteolytic staphylococci were 76.5 and 92.4% respectively. There was no significant difference in hydrolytic activity on proteins between coagulase positive and coagulase negative cocci.
