Terminalia ferdinandiana Exell. extracts reduce pro-inflammatory cytokine and PGE2 secretion, decrease COX-2 expression and down-regulate cytosolic NF-κB levels.

Based on their high antioxidant capacity and noteworthy phytochemistry, Terminalia ferdinandiana fruit and leaves have attracted considerable recent interest for their therapeutic potential. Whilst those studies have reported a variety of therapeutic properties for the fruit, the anti-inflammatory potential of T. ferdinandiana has been largely neglected and the leaves have been almost completely ignored. This study investigated the immune-modulatory and anti-inflammatory properties of T. ferdinandiana fruit and leaf extracts by evaluating their inhibition of multiple pro- and anti-inflammatory cytokines and chemokines secretion in lipopolysaccharide (LPS)-stimulated and unstimulated RAW 264.7 macrophages using multiplex bead immunoassays and ELISA assays. The methanolic extracts were particularly good immune-modulators, significantly inhibiting the secretion of all the cytokines and chemokines tested. Indeed, the methanolic extracts completely inhibited IL-10, IFN-γ, IL-1ß, IL-6, MCP-1, and MIP-2a secretion, and almost completely inhibited the secretion of TNF-α. In addition, the methanolic T. ferdinandiana extracts also significantly inhibited cytosolic COX-2 levels (by 87-95%) and the synthesis of the PGE2 (by ~ 98%). In contrast, the methanolic extracts stimulated LTB4 secretion by ~ 60-90%, whilst the aqueous extracts significantly inhibited LTB4 secretion (by ~ 27% each). Exposure of RAW 264.7 cells to the methanolic T. ferdinandiana extracts also significantly down-regulated the cytosolic levels of NF-κB by 33-44%, indicating that the immune-modulatory and anti-inflammatory properties of the extracts may be regulated via a decrease in NF-κB transcription pathways. Taken together, these results demonstrate potent anti-inflammatory properties for the extracts and provide insights into their anti-inflammatory mechanisms.

Leaf extracts of eight selected southern African medicinal plants modulate pro-inflammatory cytokine secretion in LPS-stimulated RAW 264.7 macrophages.

This study investigates the anti-inflammatory properties of extracts prepared from the leaves of eight southern African medicinal plants used traditionally to treat inflammation and pain. The inhibitory effect of aqueous and ethanol extracts on the release of pro-inflammatory cytokines was determined in lipopolysaccharide (LPS) stimulated and unstimulated RAW 264.7 murine macrophage cells. The levels of interleukin (IL)-1ß, IL-6, tumour necrosis factor-α (TNF-α), interferon-gamma (IFN-γ), monocyte chemoattractant protein 1 (MCP-1) and macrophage inflammatory protein (MIP)-2 release were determined using cytokine multiplex-bead assays. The ethanol extracts of Melianthus comosus Vahl (commonly known as honey flower), Tetradenia riparia (Hochst.) Codd (misty plume bush) and Warburgia salutaris (G. Bertol.) Chiov. (pepper-bark tree), demonstrated the most significant inhibitory activity, with over 50-fold inhibition of IL-1ß, IL-6 and TNF-α levels in LPS-stimulated RAW 264.7 macrophages. The aqueous extract of M. comosus also significantly inhibited the secretion of all the tested cytokines and chemokines. Phytochemical investigation of M. comosus ethanol leaf extract using ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-HRMS) led to the detection of crassolide, deoxylimonoic acid D-ring-lactone, 2-hydroxynonanoic acid and 5-noniloxytryptamine. To the best of our knowledge, the cytokine inhibition properties of most of the medicinal plants screened in this study are reported for the first time. Our results support the use of southern African medicinal plants as anti-inflammatory remedies and provide an insight into the immunomodulatory mechanisms of action.

A Review of the Antimicrobial Properties of Cyanobacterial Natural Products.

The development of multiple-drug-resistant pathogens has prompted medical research toward the development of new and effective antimicrobial therapies. Much research into novel antibiotics has focused on bacterial and fungal compounds, and on chemical modification of existing compounds to increase their efficacy or reactivate their antimicrobial properties. In contrast, cyanobacteria have been relatively overlooked for antibiotic discovery, and much more work is required. This may be because some cyanobacterial species produce environmental toxins, leading to concerns about the safety of cyanobacterial compounds in therapy. Despite this, several cyanobacterial-derived compounds have been identified with noteworthy inhibitory activity against bacterial, fungal and protozoal growth, as well as viral replication. Additionally, many of these compounds have relatively low toxicity and are therefore relevant targets for drug development. Of particular note, several linear and heterocyclic peptides and depsipeptides with potent activity and good safety indexes have been identified and are undergoing development as antimicrobial chemotherapies. However, substantial further studies are required to identify and screen the myriad other cyanobacterial-derived compounds to evaluate their therapeutic potential. This study reviews the known phytochemistry of cyanobacteria, and where relevant, the effects of those compounds against bacterial, fungal, protozoal and viral pathogens, with the aim of highlighting gaps in the literature and focusing future studies in this field.

Correction: Tran et al. Antimicrobial Properties of Bacillus Probiotics as Animal Growth Promoters. Antibiotics 2023, 12, 407.

In the original publication [...].

Hamamelis virginiana L. Leaf Extracts Inhibit the Growth of Antibiotic-Resistant Gram-Positive and Gram-Negative Bacteria.

Virginian witch hazel (WH; Hamamelis virginiana L.; family: Hamamelidaceae) is a North American plant that is used traditionally to treat a variety of ailments, including bacterial infections. Solvents of varying polarity (water, methanol, ethyl acetate, hexane and chloroform) were used to prepare extracts from this plant. Resuspensions of each extract in an aqueous solution were tested for growth-inhibitory activity against a panel of bacteria (including three antibiotic-resistant strains) using agar disc diffusion and broth microdilution assays. The ethyl acetate, hexane and chloroform extracts were completely ineffective. However, the water and methanolic extracts were good inhibitors of E. coli, ESBL E. coli, S. aureus, MRSA, K. pneumoniae and ESBL K. pneumoniae growth, with the methanolic extract generally displaying substantially greater potency than the other extracts. Combining the active extracts with selected conventional antibiotics potentiated the bacterial growth inhibition of some combinations, whilst other combinations remained non-interactive. No synergistic or antagonistic interactions were observed for any WH extracts/antibiotic combinations. Gas chromatography-mass spectrometry analysis of the extracts identified three molecules of interest that may contribute to the activities observed, including phthalane and two 1,3-dioxolane compounds. Putative modes of action of the active WH extracts and these molecules of interest are discussed herein.

Antimicrobial Properties of Bacillus Probiotics as Animal Growth Promoters.

Antibiotic growth promoters (AGPs) suppress the growth of infectious pathogens. These pathogens negatively impact agricultural production worldwide and often cause health problems if left untreated. Here, we evaluate six Bacillus strains (BPR-11, BPR-12, BPR-13, BPR-14, BPR-16 and BPR-17), which are known for their ability to survive harsh environmental conditions, as AGP replacements in animal feed. Four of these Bacillus strains (BPR-11, BPR-14, BPR-16 and BPR-17) showed antimicrobial activity against the pathogenic strains Clostridium perfringens, Escherichia coli and Staphylococcus aureus at 25 µg/mL, with BPR-16 and BPR-17 also able to inhibit Pseudomonas aeruginosa and Salmonella enterica at 100 µg/mL. Further chemical investigation of BPR-17 led to the identification of eight metabolites, namely C16, C15, C14 and C13 surfactin C (1-4), maculosin (5), maculosine 2 (6), genistein (7) and daidzein (8). Purified compounds (1-4) were able to inhibit all the tested pathogens with MIC values ranging from 6.25 to 50 µg/mL. Maculosin (5) and maculosine 2 (6) inhibited C. perfringens, E. coli and S. aureus with an MIC of 25 µg/mL while genistein (7) and daidzein (8) showed no activity. An animal trial involving feeding BPR-11, BPR-16 and BPR-17 to a laboratory poultry model led to an increase in animal growth, and a decrease in feed conversion ratio and mortality. The presence of surfactin C analogues (3-4) in the gut following feeding with probiotics was confirmed using an LC-MS analysis. The investigation of these Bacillus probiotics, their metabolites, their impacts on animal performance indicators and their presence in the gastrointestinal system illustrates that these probiotics are effective alternatives to AGPs.

Antimicrobial Bacillus: Metabolites and Their Mode of Action.

The agricultural industry utilizes antibiotic growth promoters to promote livestock growth and health. However, the World Health Organization has raised concerns over the ongoing spread of antibiotic resistance transmission in the populace, leading to its subsequent ban in several countries, especially in the European Union. These restrictions have translated into an increase in pathogenic outbreaks in the agricultural industry, highlighting the need for an economically viable, non-toxic, and renewable alternative to antibiotics in livestock. Probiotics inhibit pathogen growth, promote a beneficial microbiota, regulate the immune response of its host, enhance feed conversion to nutrients, and form biofilms that block further infection. Commonly used lactic acid bacteria probiotics are vulnerable to the harsh conditions of the upper gastrointestinal system, leading to novel research using spore-forming bacteria from the genus Bacillus. However, the exact mechanisms behind Bacillus probiotics remain unexplored. This review tackles this issue, by reporting antimicrobial compounds produced from Bacillus strains, their proposed mechanisms of action, and any gaps in the mechanism studies of these compounds. Lastly, this paper explores omics approaches to clarify the mechanisms behind Bacillus probiotics.

A review of the traditional use of southern African medicinal plants for the treatment of inflammation and inflammatory pain.

ETHNOPHARMACOLOGICAL RELEVANCE: Inflammation is a serious global concern due to its debilitating symptoms, resulting in considerable suffering and lost productivity. Chronic and auto-immune inflammatory diseases are of particular concern. Several pharmaceutical therapies are already available. However, the use of non-steroidal anti-inflammatory drugs (NSAID's) is accompanied by harmful and toxic side effects. Hence, the search for safer alternative therapeutics with limited side effects is imperative. The use of medicinal plants is common practice amongst the southern African population and may provide targets for drug development. AIM OF THE STUDY: This study aims to review and document the medicinal uses and pharmacological properties of southern African medicinal plants used for inflammation and pain-related ailments. MATERIAL AND METHODS: An extensive literature review was undertaken to identify southern African plants used traditionally to treat inflammation. A variety of ethnobotanical books and grey literature, as well as ScienceDirect, Google Scholar and Scopus search engines were used as sources of information. RESULTS: This review identified 555 medicinal plants from 118 families which were traditionally used in southern Africa to treat inflammation and pain. Fabaceae was the most prominent family with 63 species, followed by Asteraceae (54 species) and Apocynaceae (33 species). The top category of ailments indicated include non-specific inflammation with 150 species, followed by inflammatory pain (148 species), headache (114 species) and toothache (114 species). CONCLUSION: Despite a large number of southern African medicinal plants used to treat inflammation and pain, relatively few have been screened for their anti-inflammatory properties. Furthermore, biologically active plant extracts have been tested against relatively few inflammatory markers and considerable further work is required.

An assessment of the growth inhibition profiles of Hamamelis virginiana L. extracts against Streptococcus and Staphylococcus spp.

Staphylococcal and streptococcal species trigger a wide variety of infections involving epithelial tissues. Virginian witch hazel (WH; Hamamelis virginiana L.; family: Hamamelidaceae) is a plant that has been used traditionally by Native Americans to treat a variety of skin conditions. Extracts from the leaves were examined for their inhibitory effects on these bacterial species. Solvents of different polarity (water, methanol, ethyl acetate, hexane and chloroform) were used to prepare extracts from WH leaves, and the aqueous resuspensions were screened for antibacterial activities using disc diffusion and liquid dilution assays. Extract phytochemical profiles and toxicities were also examined, and combinations of extracts with conventional antibiotics were tested against each bacterial strain. The methanolic and aqueous extracts inhibited the growth of S. oralis, S. pyogenes, S. epidermidis and S. aureus, but not S. mutans. The extracts were especially active against staphylococcal species, with MIC values between 200 and 500 µg/ml. Combinations of active extracts with conventional antibiotics failed to yield beneficial interactions, except for two cases where additive interactions were observed (aqueous WH extract combined with chloramphenicol against S. oralis, and methanolic WH extract combined with ciprofloxacin against S. aureus). Phytochemical assays indicated an abundance of tannins, triterpenoids and phenolics in the water and methanol extracts, with trace amounts of these components in the ethyl acetate extract. Phytochemicals were not detected in hexane and chloroform extracts. Thus, phytochemical abundance in extracts was concordant with antibacterial activities. All extracts were found to be non-toxic in Artemia nauplii assays. These findings indicate the potential for WH leaf extracts for clinical use in treating staphylococcal and streptococcal infections, while substantiating their traditional Native American uses.

The medicinal plant Tabebuia impetiginosa potently reduces pro-inflammatory cytokine responses in primary human lymphocytes.

Bark from the Handroanthus impetiginosus (Mart. ex DC.) Mattos (Bignoniaceae) tree has long been used in traditional South American healing practises to treat inflammation. However, its anti-inflammatory activity has not been closely examined. Here we use chemical extraction, qualitative phytochemical examination, toxicity testing and quantitative examination of anti-inflammatory activity on human cells ex vivo. All extracts were found to be nontoxic. We found different extracts exhibited unique cytokine profiles with some extracts outperforming a positive control used in the clinic. These results verify the immunomodulatory activity of Handroanthus impetiginosus (Mart. ex DC.) Mattos (Bignoniaceae) tree bark-derived compounds. Collectively, combining a lack of toxicity and potency in human immune cells supports further fractionation and research.

Batch Effect Adjustment to Lower the Drug Attrition Rate of MCF-7 Breast Cancer Cells Exposed to Silica Nanomaterial-Derived Scaffolds.

Drug attrition rate is the calculation or measure of the clinical efficacy of a candidate drug on a screen platform for a specific period. Determining the attrition rate of a prospective cancer drug is a reliable way of testing the clinical efficacy. A low attrition rate in the last phase of a preclinical trial increases the success of a drug discovery process. It has been reported that the attrition rates of antineoplastic drugs are much higher than for other therapeutic drugs. Among the factors identified for the high attrition rates in antineoplastic drugs are the nature of the screen-based platforms involving human-derived xenografts, extracellular matrix-derived scaffold systems, and the synthetic scaffolds, which all have propensity to proliferate tumor cells at faster rates than in vivo primary tumors. Other factors that affect the high attrition rates are induced scaffold toxicity and the use of assays that are irrelevant, yet affect data processing. These factors contribute to the wide variation in data and systematic errors. As a result, it becomes imperative to filter batch variations and to standardize the data. Importantly, understanding the interplay between the biological milieu and scaffold connections is also crucial. Here the cell viability of MCF-7 (breast cancer cell line) cells exposed to different scaffolds were screened before cisplatin dosing using the calculated p-values. The statistical significance (p-value) of data was calculated using the one-way analysis of variance, with the p-value set as: 0 < p < 0.06. In addition, the half-maximal inhibitory concentration (IC50) of the different scaffolds exposed to MCF-7 cells were calculated with the probit extension model and cumulative distribution (%) of the extension data. The chemotherapeutic dose (cisplatin, 56 mg/m2) reduced the cell viability of MCF-7 cells to 5% within 24 h on the scaffold developed from silica nanoparticles (SNPs) and polyethylene glycol (PEG) formulation (SNP:PEG) mixtures with a ratio of 1:10, respectively.

Profiling the Neoplasm Microenvironment of Silica Nanomaterial-Derived Scaffolds of Single, 2-, and 3-Composite Systems.

The challenges with scaffold profiling of cell-based assay includes accelerated cancer cell proliferation, induced scaffold toxicity, and identifying irrelevant cancer cell-based assays in batch assessments. This study investigates profiling carcinoma of breast cancer cells of MCF-7 model systems using silica nanoparticles scaffold sourced from synthetic materials and plant extracts. Herein, the engineered tissue scaffolds were used to create temporary structures for cancer cell attachments, differentiation, and subsequently to assess the metabolic activity of the cancer cell colonies. The cell viability of the cancer cells was assessed using the tetrazolium compound (MTS reagent), which was reduced to colored formazan, to indicate metabolically active cancer cells in a proliferating assay. We aimed to develop cancer cell-based scaffolds that not only mimic the neoplastic activity, but that also allowed synergistic interaction with cisplatin for in vitro assay screening.

Combating Antibiotic-Resistant Gram-Negative Bacteria Strains with Tetracycline-Conjugated Carbon Nanoparticles.

Nontoxic carbon nanoparticle samples prepared by both bottom-up and top-down approaches do not inhibit Gram-negative bacterial growth, indicating excellent biocompatibilities. However, cell growth inhibitory efficacies increase considerably when the carbon nanoparticles are conjugated with the antibiotic tetracycline. In tetracycline-resistant bacteria, these efficacies can approach tenfold higher activities when compared to tetracycline alone. No structural abnormality such as membrane disruptions is evident in the tested bacterial strains; this is in contrast with other nanocarbon systems such as graphene oxides, carbon nanotubes, and amine-functionalized carbon nanoparticles which do exhibit membrane disruptions. In comparison, the tetracycline-conjugated carbon nanoparticles induce membrane perturbations (but not membrane disruptions), inhibiting bacterial efflux mechanisms. It is proposed that when tetracycline is conjugated to the surface of carbon nanoparticles, it functions to direct the nanoparticles to membrane-associated tetracycline efflux pumps, thereby blocking and subsequently inhibiting their function. The conjugation between biocompatible carbon nanoparticles and subtherapeutic but well-established antibiotic molecules may provide hybrid antibiotic assembly strategies resulting in effective multidrug efflux inhibition for combating antibiotic resistance.

The traditional use of southern African medicinal plants in the treatment of viral respiratory diseases: A review of the ethnobotany and scientific evaluations.

ETHNOPHARMACOLOGICAL RELEVANCE: Viral respiratory infections are amongst the most common infections globally, with most of the world's population contracting at least one infection annually. Numerous plant species are used in traditional southern African healing systems to treat these diseases and to alleviate the symptoms. Despite this, the therapeutic potential of these plants against viral respiratory diseases remains poorly explored. AIM OF THE STUDY: The aim of this study was to document the southern African plant species used in traditional medicine to treat viral respiratory infections. We also examined the extent of scientific evaluations of southern African plant species against the respiratory-infective viruses, with the aim of stimulating interest in this area and focusing on future studies. MATERIALS AND METHODS: We undertook an extensive review of ethnobotanical books, reviews and primary scientific studies to identify southern African plants which are used in traditional southern African medicine to treat viral respiratory diseases. This information was used to identify gaps in the current research that require further study. RESULTS: Two hundred and fifty-seven southern African plant species were identified as traditional therapies for viral respiratory diseases. Surprisingly, only one of those species (as well as twenty-one other species not recorded for these purposes) has been evaluated for the ability to block respiratory virus production. Furthermore, most of these studies screened against a single viral strain and none of those studies examined the mechanism of action of the plant preparations. CONCLUSIONS: Despite well documented records of the use of southern African plants to treat respiratory viral diseases, the field is poorly explored. Nearly all of the plant species used in traditional healing systems to treat these diseases are yet to be tested. Substantial further work is required to verify the efficacy of these traditional medicines.

The traditional use of southern African medicinal plants for the treatment of bacterial respiratory diseases: A review of the ethnobotany and scientific evaluations.

ETHNOPHARMACOLOGICAL RELEVANCE: Multiple plant species were used traditionally in southern Africa to treat bacterial respiratory diseases. This review summarises this usage and highlights plant species that are yet to be verified for these activities. AIM OF THE STUDY: This manuscript reviews the traditional usage of southern African plant species to treat bacterial respiratory diseases with the aim of highlighting gaps in the literature and focusing future studies. MATERIALS AND METHODS: An extensive review of ethnobotanical books, reviews and primary scientific studies was undertaken to identify southern African plants which are used in traditional southern African medicine to treat bacterial respiratory diseases. We also searched for southern African plants whose inhibitory activity against bacterial respiratory pathogens has been conmfirmed, to highlight gaps in the literature and focus future studies. RESULTS: One hundred and eighty-seven southern African plant species are recorded as traditional therapies for bacterial respiratory infections. Scientific evaluations of 178 plant species were recorded, although only 42 of these were selected for screening on the basis of their ethnobotanical uses. Therefore, the potential of 146 species used teraditionally to treat bacterial respiratory diseases are yet to be verified. CONCLUSIONS: The inhibitory properties of southern African medicinal plants against bacterial respiratory pathogens is relatively poorly explored and the antibacterial activity of most plant species remains to be verified.

Cancer biomarker profiling using nanozyme containing iron oxide loaded with gold particles.

Nanozymes are nanomaterials with intrinsic magnetism and superparamagnetic properties. In the presence of an external magnet, nanozyme particles aggregate and redisperse without a foreign attraction. We evaluated the performances of nanozyme by changing the biosensing platforms and substituting other biological variants for a complete cancer assay detection. We investigated the expression of morphological variants in the transmission of signals using an electrochemical method. The signal responses, including signal enhancement with the nanozyme (Au-Fe2O3), showed a wide capturing range (greater than 80%, from 102 to 105 cells ml-1 in phosphate-buffered saline buffer, pH 7.4). The platform showed a fast response time within a dynamic range of 10-105 cells ml-1 for the investigated T47D cancer cell line. We also obtained higher responses for anti-HER2 (human epidermal receptor 2)/streptavidin interface as the biosensing electrode in the presence of T47D cancer cells. The positive assay produced a sixfold increase in current output compared to the negative target or negative biological variant. We calculated the limit of detection at 0.4 U ml-1, and of quantitation at 4 U ml-1 (units per millilitre). However, blood volume amounts in clinical settings may constrain diagnosis and increase detection limit value significantly.

Use of specific combinations of the triphala plant component extracts to potentiate the inhibition of gastrointestinal bacterial growth.

ETHNOPHARMACOLOGICAL RELEVANCE: Triphala is used in Ayurveda to treat a wide variety of diseases, including numerous bacterial infections. Interestingly, the plant components of triphala (Terminalia bellirica, Terminalia chebula and Emblica officinalis) are also good inhibitors of bacterial growth when used individually, yet plant preparations are generally used in combination in traditional medicine. Surprisingly, no previous studies have addressed the reason why the combination is preferred over the individual components to treat bacterial infections. AIM OF THE STUDY: To test and compare the antibacterial efficacy of triphala and its component parts to quantify their relative efficacies. The individual plant components will also be tested as combinations, thereby determining whether combining the individual components potentiates the antibacterial activity of the components used alone. MATERIALS AND METHODS: Triphala and the three individual plant components were extracted using solvents of varying polarity (methanol, water, ethyl acetate) and the antibacterial activity of the aqueous resuspensions was quantified by disc diffusion and broth microdilution MIC assays. Combinations of extracts produced from the individual components were also tested against each bacterial species and the ΣFICs was calculated to determine the class of interaction. Where synergy was detected, isobologram analysis was used to determine the optimal component ratios. The Artemia nauplii bioassay was used to test for toxicity and GC-MS headspace profiling analysis was used to highlight terpenoid components that may contribute to the antibacterial activity of triphala. RESULTS: The aqueous and methanolic triphala, T. bellirica, T. chebula and E. officinalis extracts displayed good inhibitory activity against all bacterial strains, with MICs often in the 250-750 µg/mL range. The methanolic extracts were generally more potent than the aqueous extracts and T. chebula was the most potent of the individual plant components. Combining the extracts of the different plant species resulted in potentiation of the growth inhibitory activity of most combinations compared to that of the individual components. Indeed, with the exception of S. flexneri, all bacterial species were potentiated by at least one combination of methanolic plant extracts, with a substantial proportion of these displaying synergistic interactions. All extracts were found to be either non-toxic, or of low to moderate toxicity in Artemia nauplii assays. CONCLUSION: Whilst the individual plant components of triphala all inhibit the growth of multiple pathogenic bacteria, the activity is potentiated for multiple combinations. Therefore, the traditional usage of the combination of the three plant materials in triphala not only extends the activity profile of the mixture over that of the individual components, but it also substantially potentiates the inhibitory activity towards multiple bacteria, partially explaining the preference of triphala compared to the individual components.

Terminalia ferdinandiana Fruit and Leaf Extracts Inhibit Methicillin-Resistant Staphylococcus aureus Growth.

The development of multiple antibiotic-resistant bacteria has vastly depleted our repertoire of effective antibiotic chemotherapies. The development of multi-ß-lactam-resistant strains are particularly concerning due to our previous reliance on this class of antibiotics because of their initial efficacy and broad-spectrum activity. With increases in extended-spectrum ß-lactam-resistance and an expanded resistance to other classes of antibiotics, there is an urgent need for the development of effective new antibiotic therapies. Terminalia ferdinandiana is an endemic Australian plant known for its high antioxidant and tannin contents. T. ferdinandiana fruit and leaf extracts have strong antibacterial activity against a wide variety of bacterial pathogens. However, T. ferdinandiana extracts have not been tested against ESBL and MRSA antibiotic-resistant pathogens. An objective of this study was to screen T. ferdinandiana fruit and leaf extracts for bacterial growth inhibitory activity by disc diffusion assay against ß-lactam-sensitive and -resistant E. coli strains and against methicillin-sensitive and -resistant S. aureus. The minimum inhibitory concentration (MIC) was quantified by liquid dilution techniques. The fruit methanolic extract, as well as the methanolic, aqueous, and ethyl acetate leaf extracts strongly inhibited the growth of the MRSA, with MICs as low as 223 µg/mL. In contrast, the extracts were ineffective inhibitors of ESBL growth. Metabolomic fingerprint analysis identified a diversity and relative abundance of tannins, flavonoids, and terpenoids, several of which have been reported to inhibit MRSA growth in isolation. All extracts were nontoxic in the Artemia nauplii and HDF toxicity assays, further indicating their potential for medicinal use.

Interactive antimicrobial and toxicity profiles of Pittosporum angustifolium Lodd. extracts with conventional antimicrobials.

OBJECTIVE: Pittosporum angustifolium Lodd. is used to treat a variety of pathogenic diseases and inflammation by Australian aborigines. Practitioners of complementary medicine frequently use herbal medicines concurrently with conventional antibiotics. There is a need to evaluate their effects in combination. METHODS: The bacterial growth inhibitory activity of P. angustifolium leaf extracts was assessed against a panel of pathogenic triggers of some autoimmune diseases by standard disc diffusion and liquid dilution minimum inhibitory concentration (MIC) methods. Combinational effects between the extracts and conventional antimicrobials were classified using the sum of the fractional inhibitory concentration. Synergistic interactions were further assessed across a range of ratios by isobologram analysis. The toxicity of the individual samples and combinations was evaluated by Artemia lethality and 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) human dermal fibroblast cell viability assays. RESULTS: P. angustifolium leaf extracts strongly inhibited the growth of several bacterial triggers of autoimmune diseases. The methanolic, aqueous and ethyl acetate extracts were particularly good inhibitors of Proteus mirabilis and Klebsiella pneumonia growth (MIC = 26 and 57 µg/mL respectively). Some combinations of the extracts and conventional antibiotics significantly potentiated the combined inhibitory activity compared to the individual components. Of the 250 combinations studied, approximately 0.02% showed synergistic interactions, 49.6% were additive, 46.8% showed indifferent interactions and antagonism occurred in only 0.02% of the combinations. Interestingly, all of the synergistic and antagonistic combinations contained tetracycline as their antibiotic component. CONCLUSION: P. angustifolium leaf extracts inhibit the growth of pathogenic triggers of some autoimmune diseases. Some extracts also potentiated the activity of conventional antibiotics, without significantly affecting the toxicity of the combination.

Plantago squarrosa Murray extracts inhibit the growth of some bacterial triggers of autoimmune diseases: GC-MS analysis of an inhibitory extract.

Ankylosing spondylitis, multiple sclerosis, rheumatoid arthritis and rheumatic fever are autoimmune inflammatory diseases that may be triggered in genetically susceptible individuals by specific bacterial pathogens. Inhibiting the growth of these bacteria with high antioxidant plant extracts may inhibit the aetiology of these diseases, as well as inhibiting the later phase symptoms. P. squarrosa extracts were analysed for antioxidant activity using a DPPH free radical scavenging assay. Bacterial growth inhibitory activity was evaluated using disc diffusion assays and the activity was quantified by MIC determination. The extracts were screened for toxicity by A. franciscana nauplii assays. The most potent antibacterial extract (ethyl acetate) was analysed by GC-MS headspace profile analysis and compounds were identified with reference to a phytochemical database. All extracts displayed strong DPPH radical scavenging activity. The ethyl acetate extract was particularly potent (IC50 1.4 µg/mL), whilst the other extracts also had significant radical scavenging activity (IC50 values between 11 and 22 µg/mL). Notably, the bacterial growth inhibitory activity of the extracts correlated with their DPPH radical scavenging activity. The ethyl acetate extract, which had the greatest DPPH scavenging activity, generally displayed the most potent bacterial growth inhibitory activity. This extract was particularly potent against P. mirabilis, P. vulgaris and A. baylyi (MIC values of 484, 575 and 880 µg/mL, respectively). It also inhibited P. aeruginosa and S. pyogenes growth, albeit with higher MICs (1600-3700 µg/mL). All other extract-bacteria combinations were either inactive or resulted in mid-low potency inhibition. All extracts were non-toxic in the A. franciscana bioassay (LC50 substantially > 1000 µg/mL). In total, 89 unique mass signals were identified in the P. squarrosa ethyl acetate extract by non-biased GC-MS headspace analysis. A number of compounds which may contribute to the antibacterial activity of this extract have been highlighted.