Synergistic antimicrobial activity of <i>Camellia sinensis</i> and <i>Juglans regia</i> against multidrug-resistant bacteria

Synergistic combinations of antimicrobial agents with different mechanisms of action have been introduced as more successful strategies to combat infections involving multidrug resistant (MDR) bacteria. In this study, we investigated synergistic antimicrobial activity of Camellia sinensis and Juglans regia which are commonly used plants with different antimicrobial agents. Antimicrobial susceptibility of 350 Gram-positive and Gram-negative strains belonging to 10 different bacterial species, was tested against Camellia sinensis and Juglans regia extracts. Minimum inhibitory concentrations (MICs) were determined by agar dilution and microbroth dilution assays. Plant extracts were tested for synergistic antimicrobial activity with different antimicrobial agents by checkerboard titration, Etest/agar incorporation assays, and time kill kinetics. Extract treated and untreated bacteria were subjected to transmission electron microscopy to see the effect on bacterial cell morphology. Camellia sinensis extract showed higher antibacterial activity against MDR S. Typhi, alone and in combination with nalidixic acid, than to susceptible isolates.” We further explore anti-staphylococcal activity of Juglans regia that lead to the changes in bacterial cell morphology indicating the cell wall of Gram-positive bacteria as possible target of action. The synergistic combination of Juglans regia and oxacillin reverted oxacillin resistance of methicillin resistant Staphylococcus aureus (MRSA) strains in vitro. This study provides novel information about antimicrobial and synergistic activity of Camellia sinensis and Juglans regia against MDR pathogen

influenza monitoring in sardinia italy identifies h3 subtype in mediterranean wild migratory birds

Introduction: Wild migratory birds are global distributors of pathogens. Sardinia, Italy, is the second largest Island in the Mediterranean and is a land bridge between Europe and Africa. Methodology: We designed a surveillance protocol to investigate wild migratory birds for presence, frequency, and type of avian influenza viruses. We collected over 4,000 avian samples and compared three sampling methods, fecal, cloacal, and tracheal, to determine the most productive for virus identification. To determine frequency of infection, RNA was extracted and RT-PCRs for avian influenza virus genes were run. Positive samples were cultivated for live virus, sub typed and sequenced. Results: Forty-four samples were positive for influenza nucleoprotein gene. We identified two previously unidentified H3 subtype strains and found cloacae to have the highest rate of virus identification and fecal sampling to provide quality RNA and repeatable results for determination of virus presence. Conclusion: Our investigation provides information on the frequency of Mediterranean avian influenza viruses, and validates the initiation of an avian influenza surveillance protocol. Taken together with global avian influenza findings, these results give insight into infectious disease distributions which is important for viral pandemic monitoring and design of preventative measures

Inflammatory Cytokine Expression Is Associated with Chikungunya Virus Resolution and Symptom Severity

The Chikungunya virus infection zones have now quickly spread from Africa to parts of Asia, North America and Europe. Originally thought to trigger a disease of only mild symptoms, recently Chikungunya virus caused large-scale fatalities and widespread economic loss that was linked to recent virus genetic mutation and evolution. Due to the paucity of information on Chikungunya immunological progression, we investigated the serum levels of 13 cytokines/chemokines during the acute phase of Chikungunya disease and 6- and 12-month post-infection follow-up from patients of the Italian outbreak. We found that CXCL9/MIG, CCL2/MCP-1, IL-6 and CXCL10/IP-10 were significantly raised in the acute phase compared to follow-up samples. Furthermore, IL-1β, TNF-α, Il-12, IL-10, IFN-γ and IL-5 had low initial acute phase levels that significantly increased at later time points. Analysis of symptom severity showed association with CXCL9/MIG, CXCL10/IP-10 and IgG levels. These data give insight into Chikungunya disease establishment and subsequent convalescence, which is imperative to the treatment and containment of this quickly evolving and frequently re-emerging disease

Antimicrobial activity of <i>Juglans regia</i>.

<p>S_ pan susceptible; MRSA_ Methicillin resistance <i>Staphyococcus aureus</i>; MDR_ Multidrug resistant; MIC_ minimum inhibitory concentration; MBC_ minimum bactericidal concentration; Env._ environmental; R A C_ Rifampicin Ampicillin Chloramphenicol; R-type_ Resistance phenotype; n_number of isolates; zone_diameter of zone of inhibition</p><p>Antimicrobial activity of <i>Juglans regia</i>.</p

Effect of synergistic antimicrobial combinations on growth kinetics of MDR pathogens.

<p>Synergistic antimicrobial combinations of (a) <i>Camellia sinensis</i> and nalidixic acid and (b) <i>Juglans regia</i> and oxacillin were tested for antimicrobial effect on growth kinetics of MDR <i>Salmonella enterica</i> serovar Typhi and Methicillin Resistant <i>Staphylococcus aureus</i> respectively. Organisms were grown in MH broth in the presence of plants extracts alone and in combination with antibiotics and samples were taken for colony counts at different time intervals. Control organisms were grown in MH broth only. A 2Log₁₀ decrease at any time point from original CFU was considered as significant. Results are given as average for three experiments.</p

Effect of <i>Juglans regia</i> on bacterial growth kinetics.

<p>(a) Methicillin resistamt <i>Staphylococcus</i> aureus (MRSA) and (b) <i>S</i>. <i>aureus</i> ATCC 25923 (pan—susceptible) were treated with graded concentrations of aqueous extract of <i>Juglans regia</i> (Dandasa). Growth cycle of untreated organisms served as growth control. A 2Log₁₀ decrease at any time point from original CFU was considered as significant. Results are presented as an average for three experiments.</p

Effect of <i>Camellia sinensis</i> on bacterial growth kinetics.

<p>Representative bacterial strains of (a) <i>Salmonella enterica</i> serovar Typhi, (b) Methicillin resistant <i>Staphylococcus</i> aureus (MRSA), and (c) <i>S</i>. <i>aureus</i> ATCC 25923 (pan—susceptible) were treated with graded concentrations of methanolic extract of <i>Camellia sinensis</i> (green tea). Growth cycle of untreated organisms served as growth control. A 2 Log₁₀ decrease at any time point from original CFU was considered as significant. Results are presented as an average for three experiments.</p

Effect of <i>Juglans regia</i> on cell morphology of MRSA.

<p>Change in cellular morphology of MRSA was observed by Transmission Electron Microscopy, A: Control cell, B-D shows bacterial cells treated with <i>Juglans regia</i>. B: Intracellular thick material. C: covering on unknown material on cell surface. D: Hollow and de-shaped bacteria.</p

Influenza monitoring in Sardinia, Italy identifies H3 subtype in Mediterranean wild migratory birds

Introduction: Wild migratory birds are global distributors of pathogens. Sardinia, Italy, is the second largest Island in the Mediterranean and is a land bridge between Europe and Africa. Methodology: We designed a surveillance protocol to investigate wild migratory birds for presence, frequency, and type of avian influenza viruses. We collected over 4,000 avian samples and compared three sampling methods, fecal, cloacal, and tracheal, to determine the most productive for virus identification. To determine frequency of infection, RNA was extracted and RT-PCRs for avian influenza virus genes were run. Positive samples were cultivated for live virus, sub typed and sequenced. Results: Forty-four samples were positive for influenza nucleoprotein gene. We identified two previously unidentified H3 subtype strains and found cloacae to have the highest rate of virus identification and fecal sampling to provide quality RNA and repeatable results for determination of virus presence. Conclusion: Our investigation provides information on the frequency of Mediterranean avian influenza viruses, and validates the initiation of an avian influenza surveillance protocol. Taken together with global avian influenza findings, these results give insight into infectious disease distributions which is important for viral pandemic monitoring and design of preventative measures

CHIKV disease severity is associated with high CXCL10, CXCL9 and IgG levels at the 6-month time point.

<p>CHIKV patients were determined to be nonsymptomatic (N), to have mild symptoms (M) or to have severe symptoms (S). The cytokine and IgG levels were then grouped by symptom level and a Mann-Whitney U test was used to determine significance among the severity groups. CXCL10, CXCL9 and IgG were found to be significantly increased in the patients with mild and severe symptoms at 6 months following initial infection. The cross symbol indicates a p-value less than 0.05.</p