The synthesized titanium dioxide nanotubes (TiO2-NTs) were emerged as wound healing enhancer as well as exhibited significant wound healing activity on Sprague Dawley rats. In our present study, the blends of GG and TiO2-NTs bio-nanocomposite film was characterised by fourier transform infrared (FTIR), x-ray diffraction (XRD), scanning electron microscopy (SEM), thermogravimetric analysis, atomic force microscopy (AFM). The morphology of TiO2-NTs was investigated using transmission electron microscopy (TEM). The mechanical properties study shows that the GG + TiO2-NTs (20 w/w %) bio-nanocomposite film possessed the highest tensile strength and young modulus which are (4.56 ± 0.15) MPa and (68 ± 1.63) MPa, respectively. GG + TiO2-NTs (20 w/w %) also displays the highest antibacterial activity with (16 ± 0.06) mm, (16 ± 0.06) mm, (14 ± 0.06) mm, and (12 ± 0.25) mm inhibition zone were recorded against Staphylococcus aureus, Streptococcus, Escherichia coli, and Pseudomonas aeruginosa. The prepared bio-nanocomposite films have good biocompatibility against 3T3 mouse fibroblast cells and caused accelerated healing of open excision type wounds on Sprague Dawley rat model. The synergistic effects of bio-nanocomposite film like good swelling and WVTR properties, excellent hydrophilic nature, biocompatibility, wound appearance and wound closure rate through in vivo test makes it a suitable candidate for wound healing applications.
Introduction: Silver nanoparticles has been proven to be an effective agent for antimicrobial efficacy against bacte-ria, viruses and other eukaryotic microorganisms. Green synthesis is one of the methods that has been developed to synthesize silver nanoparticles in environmentally-friendly conditions. It uses plant extracts as reducing and capping agents. Besides act as reducing and capping agents, bioactives such as phenolic compounds may bind to silver nanoparticles and enhance its medicinal properties. Strobilanthes crispus is a Malaysian native plant. Previous stud-ies had shown that S. crispus contains polyphenols, catechins, alkaloids, caffeine, tannins and vitamins. Therefore, the aim of this study is to determine antibacterial activities of silver nanoparticles-Strobilanthes crispus (AgNP-SC) against clinically important pathogens such as Escherichia coli, Pseudomonas aeruginosa and Streptococcus mutans. Methods: The disc diffusion assay (DDA) was performed to investigate the inhibition zone of AgNps-Sc towards E. coli, P. aeruginosa and S. mutans. Minimum inhibition concentration (MIC) and minimum bactericidal concentration (MBC) was used to determine bactericidal/bacteriostatic profile of AgNP- SC against E. coli, P. aeruginosa and S. mu-tans. Results: AgNP-SC (40mg/mL) shows the greatest inhibition properties (12.67±0.6mm) against S. mutans when compared to Strobilanthes crispus leaves extract (6.0±0.001mm) and blank silver nanoparticles (6.0±0.001mm). MIC values for AgNP-SC against S. mutans and E. coli were at 0.625 mg/mL and 1.25 mg/mL, respectively. Whereas the MIC value of AgNP- SC against P. aeruginosa was at 2.5 mg/mL. MBC values of AgNP-SC against E. coli, P. aerugino-sa and S. mutans were at 1.25, 2.5 mg/mL respectively. Results are concentration-dependent, with higher concentra-tion demonstrating better inhibition property. Conclusion: It can be concluded that AgNP-SC possesses bactericidal properties against S. mutans, E. coli and P. aeruginosa.
Microorganisms have begun to develop resistance because of inappropriate and extensive use of antibiotics in the hospital setting. Therefore, it seems to be necessary to find a way to tackle these pathogens by developing new and effective antimicrobial agents. Carbon nanotubes (CNTs) have attracted growing attention because of their remarkable mechanical strength, electrical properties, and chemical and thermal stability for their potential applications in the field of biomedical as therapeutic and diagnostic nanotools. However, the impact of carbon nanotubes on microbial growth has not been fully investigated. The primary purpose of this research study is to investigate the antimicrobial activity of CNTs, particularly double-walled and multi-walled nanotubes on representative pathogenic strains such as Gram-positive bacteria Staphylococcus aureus, Gram-negative bacteria Pseudomonas aeruginosa, Klebsiella pneumoniae, and fungal strain Candida albicans. The dispersion ability of CNT types (double-walled and multi-walled) treated with a surfactant such as sodium dodecyl-benzenesulfonate (SDBS) and their impact on the microbial growth inhibition were also examined. A stock concentration 0.2 mg/mL of both double-walled and multi-walled CNTs was prepared homogenized by dispersing in surfactant solution by using probe sonication. UV-vis absorbance, Fourier transform infrared spectroscopy (FTIR), and transmission electron microscopy (TEM) were used for the characterization of CNTs dispersed in the surfactant solution to study the interaction between molecules of surfactant and CNTs. Later, scanning electron microscopy (SEM) was used to investigate how CNTs interact with the microbial cells. The antimicrobial activity was determined by analyzing optical density growth curves and viable cell count. This study revealed that microbial growth inhibited by non-covalently dispersed CNTs was both depend on the concentration and treatment time. In conclusion, the binding of surfactant molecules to the surface of CNTs increases its ability to disperse in aqueous solution. Non-covalent method of CNTs dispersion preserved their structure and increased microbial growth inhibition as a result. Multi-walled CNTs exhibited higher antimicrobial activity compared to double-walled CNTs against selected pathogens.
Despite intensive research, cancer incidence and mortality continue to rise. Consequently, the necessity to develop effective anti-cancer therapy is apparent. We have recently shown that the gut bacteria of animals living in polluted environments, such as crocodiles, are a potential source of novel anti-tumor molecules. To extend this work to other resilient species, we investigated the anti-tumor effects of gut bacteria of Heterometrus spinifer (a scorpion). Bacteria from the feces and gut were isolated, identified and evaluated for their anti-tumor effects. Bacterial-conditioned media was prepared in Roswell Park Memorial Institute (RPMI) 1640 media, and cytotoxicity and growth inhibitory properties were examined against cervical (HeLa) cancer cells. Liquid chromatography-mass spectrometry (LC-MS) was conducted to establish the identity of the molecules. Eighteen bacteria species from the gut (HSG01-18) and ten bacteria species from feces (HSF01-10) were tested for anti-tumor effects. Bacterial-conditioned media from scorpion gut and feces exhibited significant growth inhibitory effects against HeLa cells of 66.9% and 83.8%, respectively. Microscopic analysis of cancer cells treated with conditioned media HSG12 and HSG16 revealed apoptosis-like effects. HSG12 was identified as Pseudomonas aeruginosa and HSG16 was identified as Bacillus subtilis. Both conditioned media exhibited 100% growth inhibitory effects versus a selection of cancer cells, comprising cervical, breast and prostate cancer cells. LC-MS indicated the presence of 72 and 38 compounds, detected from HSG12 and HSG16, respectively. Out of these compounds, 47 were successfully identified while the remainder were unidentified and are possibly novel. This study suggests that the fecal and gut microbiota of scorpions might possess molecules with anti-cancer properties, however, further intensive research is needed to assess these expectations.
A new 2-arylbenzofuran, sesbagrandiflorain C (1), together with four known compounds, 2-(3,4-dihydroxy-2-methoxyphenyl)-4-hydroxy-6-methoxybenzofuran-3-carbaldehyde (2), 2-(4-hydroxy-2-methoxyphenyl)-5,6-dimethoxybenzofuran-3-carboxaldehyde (3), sesbagrandiflorain A (4) and sesbagrandiflorain B (5), have been isolated from the stem bark of an Indonesian plant, Sesbania grandiflora (L.) Pers. The chemical structure of compound 1 was elucidated by UV, IR, MS, and NMR spectroscopic techniques. The proton and carbon NMR resonances of 1 were also compared with the predicted chemical shifts obtained from DFT quantum mechanical calculations with Gaussian. None of the compounds showed antibacterial activity against Bacillus subtilis, Escherichia coli, Mycobacterium smegmatis, Pseudomonas aeruginosa, and Staphylococcus aureus in an agar diffusion assay. However, sesbagrandiflorains A (4) and B (5) exhibited moderate activity against Mycobacterium tuberculosis H37Rv. In addition, compounds 1 - 5 have moderate cytotoxicity against HeLa, HepG2, and MCF-7 cancer cell lines.
We present a case study of a 26-year-old morbidly obese man with a three-day history of right leg pain and swelling. The swelling was associated with low grade fever. He was alert and conscious upon presentation to the hospital. His physical examination showed gross swelling of the entire right lower limb with no systemic manifestations. There was no discharge and bullae from the swelling area of the leg. He had high blood sugar and was newly diagnosed with type 2 diabetes mellitus. He was diagnosed with necrotizing fasciitis. An intravenous imipenem-cilastatin 500 mg every 6 h together with clindamycin 900 mg every 8 h was started empirically. Extensive wound debridement was performed. The swab culture obtained intraoperatively grew Pseudomonas aeruginosa. He required an above knee amputation due to worsening infection despite wound debridement. Post-operatively, he developed acute kidney injury with severe metabolic acidosis, which required daily hemodialysis. However, the patient deteriorated due to septic shock with multi-organ failure, resulting in his death.
Ginger essential oils (GEO) are natural products with antibacteria properties consisting of many different volatile
compounds have high potential to be used in many applications. In this study, the ginger GEO was successfully
encapsulated in chitosan as a carrier agent using a spray drying technique. The extraction of Zingiber officinale
(ginger) essential oil is performed by steam distillation method. The GEO was encapsulated in chitosan as a carrier
agents at 1:3, GEO:chitosan ratio by using spray drying technique. GEO together with encapsulated GEO were
further assayed for antimicrobial activity by disc-diffusion method. For characterization of encapsulated GEO,
Fourier transform infrared spectroscopy (FTIR) and Field emission scanning electron microscopy (FESEM) were
used. FTIR analysis revealed that there was no existence of a new functional group in the encapsulated GEO
showing that there is only physical interaction between GEO and chitosan. Besides, FESEM analysis showed the
encapsulated GEO were in micro in sizes and possessed spherical shape with smooth and porous surface.
Furthermore, Both GEO and encapsulated GEO showed in vitro antimicrobial activity against Escheriachia coli,
Pseudomonas aeruginosa, Staphylococcus aureus and Salmonella typhi with encapsulated GEO possessed higher in
the activities for all studied bacteria compared to GEO. The encapsulated GEO demonstrated a superior performance
against Salmonella typhi with the inhibition zone of 22.5 mm compared to GEO only 13.5 mm. The results obtained
indicated that due to the volatility and instability of the GEO when exposed to environmental factors, its encapsulation considerably improve and enhanced its performance.
Diopatra claparedii which is colloquially known as Ruat Sarung can be found along the west coast of Peninsular Malaysia. The species has a unique ability to regenerate anterior and posterior segments upon self-amputation or injury, thus having potential as a wound healing promoter. In this study, the wound healing potential of D. claparedii aqueous extract on acute wound model in rats was revealed for the first time. Various concentrations (0.1%, 0.5%, and 1.0% w/w) of D. claparedii ointment were formulated and tested on Sprague Dawley rats through topical application on full-thickness skin wounds for 14 days. The wound healing effects were investigated via behaviour observation, wound contraction, and histopathological analysis. Quality assessment was performed via skin irritation test, microbial contamination test (MCT), and heavy metal detection. The study also included test for antibacterial activities and detection of bioactive compounds in D. claparedii. One percent of D. claparedii ointment showed rapid wound healing potential with good soothing effects and more collagen deposition in comparison to the commercial wound healing ointments such as acriflavine (0.1% w/v) and traditional ointment gamat (sea cucumber extract) (15.0% w/v). No local skin irritation, microbial contamination, and insignificant concentration of heavy metals were observed, which indicate its safe application. Moreover, the aqueous extract of D. claparedii exhibited antibacterial activities against Escherichia coli and Pseudomonas aeruginosa with minimum inhibitory concentration (MIC) value at 0.4 g/ml. 1H NMR analysis of the aqueous extract of D. claparedii revealed some metabolites that might be responsible for its wound healing properties such as amino acids, halogenated aromatics, organic acids, vitamins, and others. Altogether, these results suggested that the aqueous extract of D. claparedii could be utilised as an alternative natural wound healing promoter.
Two cocoa bean fermentation methods (spontaneous fermentation and the use of starter culture) for 7 days fermentation were compared in terms of safety and quality fermented beans. Candida sp. was used as a starter culture in this study. The safety of the fermented cocoa beans were measured by the growth colonies of pathogenic microorganisms namely Bacillus cereus, Escherichia coli, Salmonella sp., Staphylococcus aureus, and Pseudomonas sp., on Bacillus cereus agar, eosin-methylene blue (EMB) agar, xylose lysine deoxycholate (XLD) agar, Baird-Parker agar (BPA), and Pseudomonas agar, respectively. B. cereus, E. coli and Salmonella sp. were early present in both fermentations. Candida sp.-fermentation showed detection of B. cereus at 5.34 log10 CFU/g and absence after 24 hours of fermentation while in spontaneous-fermentation B. cereus was too few to count. Moreover, the log10 E. coli number in Candida sp.-fermentation and spontaneous-fermentation were reduced from 5.72 to 3.66 and from 7.15 to 4.46 on day 1 to day 3, respectively. There were no presences of pathogenic microorganisms on day 5 and day 7 for both fermentations. In term of quality, proximate analysis of spontaneous-fermentation resulted that the content of moisture, ash, fat, crude protein, crude fibre and carbohydrate was 56.47%, 2.32%, 3.17%, 7.02%, 28.14% and 2.88%, meanwhile for the Candida sp.-fermentation was 53.96%, 2.19%, 3.44%, 8.25%, 25.46% and 6.70%, respectively. This study showed that both fermentations are considered to be safe and there is no significant difference in proximate value in fermented cocoa beans from spontaneous-fermentation and Candida sp.-fermentation.
There are relatively little data on bacteria with antimicrobial activities from Antarctic, especially from the South Shetland Islands when compared to the other parts of the world. Hence, this project was set to isolate and characterize bacteria that produce anti-microbial compounds from Greenwich Island (one of the South Shetland Islands), Antarctica. A total of 356 strains of bacteria were isolated from Greenwich Island. They were screened for antimicrobial activities against 13 Gram-negative and one Gram-positive indicator food-borne pathogens. Two out of the 356 Antarctic bacterial strains exhibited an antagonistic effect on the indicator strains, Escherichia coli, Salmonella spp., Klebsiella pneumoniae, Enterobacter cloacae, Vibrio parahaemolyticus and Bacillus cereus. The two Antarctic bacterial strains were designated as SS157 and SR13. Biochemical and 16S rDNA analysis indicated that the strain SS157 was closely related to Pseudomonas congelans while the strain SR13 was closely related to Pseudomonas tremae. The anti-microbial compounds produced by the two Antarctic bacteria were not sensitive to temperature and were not degraded by trypsin or pronase indicating that they were likely to be chemical compounds or antibiotics. Antimicrobial compounds from strains SS157 and SR13 were broad spectrum, and targeted both Gram-positive and negative pathogens.
A cross-sectional study of 109 patients with Pseudomonas aeruginosa bacteremia from the University of Malaya Medical Centre (UMMC) in the years 2000 and 2001 was conducted to describe epidemiological features, underlying diseases, possible source of infection, early mortality among patients as well as the antibiotic susceptibility pattern of patients' isolates. Further analysis of the 87 patient records that were available revealed that the mean age was 48.5 years (SD ± 25.1). Fifty-two per cent of cases were male and 48% female. Seventy-nine per cent of infections were nosocomially acquired, 33% of bacteremias were polymicrobial, 47% of patients had a continuous bladder drainage catheter (CBD) in situ, 33% had a central venous catheter (CVL) present at the time of bacteremia and 30% were ventilated. Sixty-eight per cent of patients had an underlying immunosuppressed state and 26% had undergone surgery involving general anesthesia in the week prior to isolating P. aeruginosa. Among the 23 patients with early mortality, 61% were on inappropriate antimicrobials. Most of the patients' isolates were sensitive to imipenem (86%), ciprofloxacin (81%), ceftazidime (79%), gentamicin (78%) and cefoperazone (77%). Among the community acquired strains, however, there was 100% sensitivity to imipenem, ceftazidime, cefoperazone and ciprofloxacin.
Edible medicinal plants are often used in the treatment of various ailments and spice in traditional food preparation. In this study, 45 of tropical edible medicinal plants extracts from Indonesia, Malaysia, and Thailand were screened for their antimicrobial activity against five standard microorganisms for food preservative namely Aspergillus niger, Candida albicans, Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus. The methanol extracts of Piper nigrum L. seed, Piper cubeba L. seed, and the root of Ligusticum acutilobum Siebold and Zucc. showed antimicrobial activity against five species of standard microorganisms. Among them, P. cubeba L. extract demonstrated the most susceptible against all tested microorganisms. Minimal inhibitory concentration (MIC) and minimal bactericidal or fungicidal concentration (MBC or MFC) were performed by the broth microdilution techniques as described by the Clinical and Laboratory Standard Institute. MIC values of P. cubeba L. extract to A. niger, C. albicans, E. coli, P. aeruginosa and S. aureus were 12.8, 1.6, 3.2, 6.4, and 1.6 mg/ml, respectively. P. cubeba extract killed A. niger, C. albicans, E. coli, P. aeruginosa and S. aureus with MBC values of 25.6, 3.2, 6.4, 12.8, and 3.2 mg/ml, respectively. The potent antimicrobial activity of P. cubeba L. extract may support its use for natural food preservative.
Abstract: Ampicillin-sulbactam combination is the most frequently prescribed antibiotic in diabetic foot ulcers. We conducted a retrospective study to evaluate the antibiotic sensitivity of bacteria isolated to this antibiotic. In 33 patients with diabetic foot ulcer (September 2008-March 2009), 67% were culture positive in which Citrobacter spp accounted for 36% of these isolates. The rest isolated included Pseudomonas aeruginosa (22%), Proteus spp (18%), Acinetobacter spp (9%), Klebsiella pneumoniae (5%), Escherichia coli (5%) and Staphylococcus aureus (5%). These isolates were more likely to be ampicillin-resistant (n=18) than were ampicillin-sensitive isolates (n=4). Ampicillin resistance has raised our concern about current practice of prescribing ampicillin/ sulbactam as monotherapy for majority of our patients with such ulcers.
Synthesis of a natural proline-rich cyclopolypeptide - rolloamide A was carried out by coupling of tri- and tetrapeptide units Boc-Phe-Pro-Val-OMe and Boc-Pro-Leu-Pro-Ile-OMe after proper deprotection at carboxyl and amino terminals using carbodiimide chemistry in alkaline environment followed by cyclization of linear heptapeptide segment in the presence of base. The structure of synthesized peptide was confirmed by spectral techniques including FTIR, 1H NMR, 13C NMR, MS analyses. Newly synthesized peptide was subjected to biological screening against pathogenic microbes and earthworms. Cyclopeptide 8 possessed promising activity against pathogenic fungi Candida albicans (ZOI: 24 mm, MIC: 6 μg/mL) and Gram-negative bacteria Pseudomonas aeruginosa (ZOI: 27 mm, MIC: 6 μg/mL) and Klebsiella pneumoniae (ZOI: 23 mm, MIC: 12.5 μg/mL), in comparison to reference drugs - griseofulvin (ZOI: 20 mm, MIC: 6 μg/mL) and ciprofloxacin (ZOI: 25 mm, MIC: 6 μg/mL/ZOI: 20 mm, MIC: 12.5 μg/mL). Also, newly synthesized heptacyclopeptide exhibited potent anthelmintic activity against earthworms Megascoplex konkanensis,Pontoscotex corethruses, and Eudrilus species (MPT/MDT ratio - 8.22-16.02/10.06-17.59 min), in comparison to standard drugs - mebendazole (MPT/MDT ratio - 10.52-18.02/12.57-19.49 min) and piperazine citrate (MPT/MDT ratio - 12.38-19.17/13.44-22.17 min).
Chitin-encapsulated cadmium sulfide quantum dots (CdS@CTN QDs) were successfully synthesized from chitin and Cd(NO3)2 precursor using the colloidal chemistry method, toward the development of biocompatible and biodegradable QDs for biomedical applications. CdS@CTN QDs exhibited the nanocrystalline cubic CdS encapsulated by α-chitin. The average particle size of CdS@CTN QDs was estimated using empirical Henglein model to be 3.9 nm, while their crystallite size was predicted using Scherrer equation to be 4.3 nm, slightly larger compared to 3-mercaptopropionic acid-capped CdS QDs (3.2 and 3.6 nm, respectively). The mechanism of formation was interpreted based on the spectroscopic data and X-ray crystal structures of CdS@CTN QDs fabricated at different pH values and mass ratios of chitin to Cd(NO3)2 precursor. As an important step to explore potential biomolecular and biological applications of CdS@CTN QDs, their antibacterial activities were tested against four different bacterial strains; i.e. Escherichia coli, Bacillus subtillus, Staphylococcus aureus and Pseudomonas aeruginosa.
Biodiesel side stream waste glycerol was identified as a cheap carbon source for rhamnolipids (RLs) production which at the same time could improve the management of waste. The present study aimed to produce RLs by using Pseudomonas aeruginosa RS6 utilizing waste glycerol as a substrate and to evaluate their physico-chemicals properties. Fermentation conditions such as temperature, initial medium pH, waste glycerol concentration, nitrogen sources and concentrations resulted in different compositions of the mono- and di-RLs produced. The maximum RLs production of 2.73 g/L was obtained when P. aeruginosa RS6 was grown in a basal salt medium supplemented with 1% waste glycerol and 0.2 M sodium nitrate at 35 °C and pH 6.5. At optimal fermentation conditions, the emulsification index (E24) values of cooking oil, diesel oil, benzene, olive oil, petroleum, and kerosene were all above E24=50%. The surface tension reduction obtained from 72.13 mN/m to 29.4-30.4 mN/m was better than the surface activity of some chemical-based surfactants. The RLs produced possessed antimicrobial activities against Gram-negative and Gram-positive bacteria with values ranging from 37% to 77% of growth inhibition when 1 mg/mL of RLs was used. Concentrations of RLs below 1500 μg/mL did not induce phytotoxicity effects on the tested seeds (Vigna radiata) compared to the chemical-based- surfactant, SDS. Furthermore, RLs tested on zebrafish (Danio rerio) embryos only exhibited low acute toxicity with an LC50 value of 72.97 μg/mL at 48 h of exposure suggesting a green and eco-biochemical worthy of future applications to replace chemical-based surfactants.
The epidermal mucus of fish contains antimicrobial agents that act as biological defence against disease. This study aims to identify antibacterial activity and protein concentration of epidermal mucus of Barbodes everetti, a Bornean endemic freshwater fish. The epidermal mucus was extracted with 3% acetic acid, 0.85% sodium chloride and crude solvents. The mucus activity against eight strains of human pathogenic bacteria, including Bacillus cereus ATCC 33019, Escherichia coli O157:H7, Listeria monocytogenes ATCC 7644, Pseudomonas aeruginosa ATCC 27853, Salmonella braenderup ATCC BAA 664, Salmonella typhimurium, Staphylococcus aureus ATCC 25933, and Vibrio cholerae, were tested. The acetic acid mucus extract of B. everetti was able to inhibit five strains of bacteria and show no activity toward E. coli O157:H7, B. cereus ATCC 33019 and L. monocytogenes ATCC 7644. Moreover, the highest protein concentration was quantified in crude extract, followed by aqueous and acetic acid extracts. This study provides a preliminary knowledge on the activity of epidermal mucus of B. everetti towards five out of the eight human pathogens tested, therefore it may contain potential sources of novel antibacterial components which could be further extracted for the production of natural antibiotics towards human-related pathogenic bacteria.
Curcuma species (family: Zingiberaceae) are widely utilized in traditional medicine to treat diverse immune-related disorders. There have been many scientific studies on their immunomodulating effects to support their ethnopharmacological uses. In this review, the efficacy of six Curcuma species, namely, C. longa L., C. zanthorrhiza Roxb., C. mangga Valeton & Zijp, C. aeruginosa Roxb. C. zedoaria (Christm.) Roscoe, and C. amada Roxb., and their bioactive metabolites to modulate the immune system, their mechanistic effects, and their potential to be developed into effective and safe immunomodulatory agents are highlighted. Literature search has been carried out extensively to gather significant findings on immunomodulating activities of these plants. The immunomodulatory effects of Curcuma species were critically analyzed, and future research strategies and appropriate perspectives on the plants as source of new immunomodulators were discussed. Most of the pharmacological investigations to evaluate their immunomodulatory effects were in vivo and in vitro experiments on the crude extracts of the plants. The extracts were not chemically characterized or standardized. Of all the Curcuma species investigated, the immunomodulatory effects of C. longa were the most studied. Most of the bioactive metabolites responsible for the immunomodulating activities were not determined, and mechanistic studies to understand the underlying mechanisms were scanty. There are limited clinical studies to confirm their efficacy in human. Of all the bioactive metabolites, only curcumin is undergoing extensive clinical trials based on its anti-inflammatory properties and main use as an adjuvant for the treatment of cancer. More in-depth studies to understand the underlying mechanisms using experimental in vivo animal models of immune-related disorders and elaborate bioavailability, preclinical pharmacokinetics, and toxicity studies are required before clinical trials can be pursued for development into immunomodulatory agents.
Among several animals, Rattus rattus (rat) lives in polluted environments and feeds on organic waste/small invertebrates, suggesting the presence of inherent mechanisms to thwart infections. In this study, we isolated gut bacteria of rats for their antibacterial activities. Using antibacterial assays, the findings showed that the conditioned media from selected bacteria exhibited bactericidal activities against Gram-negative (Escherichia coli K1, Klebsiella pneumoniae, Pseudomonas aeruginosa, Serratia marcescens, and Salmonella enterica) and Gram-positive (Bacillus cereus, methicillin-resistant Staphylococcus aureus, and Streptococcus pyogenes) pathogenic bacteria. The conditioned media retained their antibacterial properties upon heat treatment at boiling temperature for 10 min. Using MTT assays, the conditioned media showed minimal cytotoxic effects against human keratinocyte cells. Active conditioned media were subjected to tandem mass spectrometry, and the results showed that conditioned media from Bacillus subtilis produced a large repertoire of surfactin and iturin A (lipopeptides) molecules. To our knowledge, this is the first report of isolation of lipopeptides from bacteria isolated from the rat gut. In short, these findings are important and provide a platform to develop effective antibacterial drugs.
A thermostabilized, multiplex polymerase chain reaction (mPCR) assay was developed in this study for the detection of six respiratory bacterial pathogens. Specific primers were designed for an internal amplification control (IAC) and six target sequences from Klebsiella pneumoniae, Staphylococcus aureus, Streptococcus pneumoniae, Pseudomonas aeruginosa, Mycobacterium tuberculosis, and Haemophilus influenzae. The resultant seven-band positive amplification control (PAC) of this heptaplex PCR assay corresponded to 105 base pairs (bp) of IAC, 202 bp of K. pneumoniae, 293 bp of S. aureus, 349 bp of S. pneumoniae, 444 bp of P. aeruginosa, 505 bp of M. tuberculosis, and 582 bp of H. influenzae. Results found that 6% (w/v) of the stabilizer was optimum to preserve the functional conformation of Taq DNA polymerase enzyme. This assay was stable at ambient temperature for at least 6 months. The sensitivity and specificity of this assay were both 100% when testing on the intended target organisms (n = 119) and non-intended species (n = 57). The mPCR assay developed in this study enabled accurate, rapid, and simple detection of six respiratory bacteria.