Thermosonication as an effective substitution for fusion in Brazilian cheese spread (Requeijão Cremoso) manufacturing: The effect of ultrasonic power on technological properties.

In this initial study, the impact of thermosonication as an alternative to the traditional fusion in Brazilian cheese spread (Requeijão Cremoso) manufacture was investigated. The effect of ultrasound (US) power was evaluated considering various aspects such as gross composition, microstructure, texture, rheology, color, fatty acid composition, and volatile compounds. A 13 mm US probe operating at 20 kHz was used. The experiment involved different US power levels (200, 400, and 600 W) at 85 °C for 1 min, and results were compared to the conventional process in the same conditions (85 °C for 1 min, control treatment). The texture became softer as ultrasound power increased from 200 to 600 W, which was attributed to structural changes within the protein and lipid matrix. The color of the cheese spread also underwent noticeable changes for all US treatments, and treatment at 600 W resulted in increased lightness but reduced color intensity. Moreover, the fatty acid composition of the cheese spread showed variations with different US power, with samples treated at 600 W showing lower concentrations of saturated and unsaturated fatty acids, as well as lower atherogenicity and thrombogenicity indexes, indicating a potentially healthier product. Volatile compounds were also influenced by US, with less compounds being identified at higher powers, especially at 600 W. This could indicate possible degradation, which should be evaluated in further studies regarding US treatment effects on consumer perception. Hence, this initial work demonstrated that thermosonication might be interesting in the manufacture of Brazilian cheese spread, since it can be used to manipulate the texture, color and aroma of the product in order to improve its quality parameters.

Impact of adding xylooligosaccharides encapsulated in butter: Microstructural, optical, rheological and sensory aspects.

This study investigated the microstructure, rheological properties, and sensory characteristics of butters produced with free and encapsulated xylooligosaccharides (XOS). Four formulations of butter were processed: BCONT: 0 % w/w XOS (control); BXOS: 20% w/w free XOS; BXOS-ALG: 20% w/w XOS microencapsulated with alginate (XOS-alginate ratio of 3:1 w/w); and BXOS-GEL: 20% w/w XOS microencapsulated with alginate-gelatin (XOS-alginate-gelatin ratio of 3:1:1.5 w/w). The microparticles showed a bimodal distribution, low size and low span values, demonstrating physical stability to be included in emulsions. The XOS-ALG presented surface weighted mean diameter (D3.2) of 90.24 µm, volume-weighted mean diameter (D4.3) of 131.8 µm, and Span of 2.14. In contrast, the XOS-GEL presented D3.2 of 82.80 µm, D4.3 of 141.0 µm, and a Span of 2.46. Products with XOS were characterized by higher creaminess, sweet taste, and lower salty taste than the control. However, the addition form significantly impacted the other evaluated parameters. The utilization of XOS in a free form (BXOS) resulted in smaller droplet sizes (1.26 µm) than encapsulated XOS and control (XOS-ALG = 1.32 µm / XOS-GEL = 1.58 µm, / BCONT = 1.59 µm), and changes in the rheological parameters (higher values of shear stress, viscosity, consistency index, rigidity (J0), and Newtonian viscosity (ηN) and lower elasticity (τ)). Furthermore, it changed the color parameters (more yellow and dark color, lower L* and higher b* values). On the other hand, the utilization of micropaticles of XOS (BXOS-ALG and BXOS-GEL) kept shear stress, viscosity, consistency index, rigidity (J0), and elasticity (τ) more similar to control. The products had a less intense yellow color (lower b* values) and was perceived with more consistency and butter taste. However, the presence of particles was perceived by consumers. The results suggest that consumers were more attentive to reporting flavor-related attributes than texture. In conclusion, adding microparticles of XOS could improve butter's rheological and sensory properties. In conclusion, adding microparticles of XOS could improve butter's rheological and sensory properties.

Applications of Silver Nanoparticles in Patent Research.

BACKGROUND: Silver nanoparticles (AgNPs) have been widely applied in research and industrial fields, finding applications in nanomedicine, drug delivery, biomedical devices, electronics, the energy sector, and environmental protection. Patents provide information on the industrial viability of product technologies, and the number of patent documents provides an estimate of the evolution of a specific technological field. AIMS: The present work aims to describe the current trends in AgNPs patent applications. In addition, a retrospective study of published patents in Brazil is presented. METHODS: Analyses of AgNPs-related patents were conducted using the free platform for patent search Lens® in 2010-2019 and articles published in same period using the Scholar® base. The patent applications and their evolution over time, major depositors and holders, and the main technological areas associated with AgNP applications have been described. RESULTS: China and United States are the major patent applicants for nanotechnologies. The worldwide distribution of publications of journal articles shows that China, India, and the United States are the leading countries in the total number of articles published, in that order. CONCLUSION: Our study of patent applications and published articles confirmed the growing global increase in new technologies involving NPs and AgNPs, particularly in the biotechnology area, in the fields of medicine and agriculture.

Dual action of pyroligneous acid in the eco-friendly synthesis of bactericidal silver nanoparticles.

In the present study, we demonstrate that pyroligneous acid (PA), also known as wood vinegar, functions efficiently as both reducing and stabilizing agent in the synthesis of silver nanoparticles (AgNPs). The synthesis and stabilization of AgNPs take place in the following fashion: 1) in alkaline environment, oxygenated species (phenols in the present case) contained in PA reduce silver ions to metallic silver; 2) acetic acid, abundantly present in PA, adsorb onto the AgNPs conferring electrostatic stabilization. This mechanism is supported by GC-MS and RAMAN analysis, with the former revealing the compounds lacking in PA after nanoparticle synthesis and the latter demonstrating acetic acid adsorbed on the nanoparticles. The AgNPs produced via this method were quite stable up to 150 days (zeta potential = -56 mV). The AgNPs were then found to inhibit the growth of Escherichia coli and Staphylococcus aureus. Concerning PA, we showed that it displays bactericidal properties only under acidic conditions. This study contributes to the development of more environmentally benign routes to produce nanomaterials.

Assessing the antiproliferative effect of biogenic silver chloride nanoparticles on glioblastoma cell lines by quantitative image-based analysis.

Glioblastoma is the most life-threatening tumour of the central nervous system. Temozolomide (TMZ) is the first-choice oral drug for the treatment of glioblastoma, although it shows low efficacy. Silver nanoparticles (AgNPs) have been shown to exhibit biocidal activity in a variety of microorganisms, including some pathogenic microorganisms. Herein, the antiproliferative effect of AgCl-NPs on glioblastoma cell lines (GBM02 and GBM11) and on astrocytes was evaluated through automated quantitative image-based analysis (HCA) of the cells. The cells were treated with 0.1-5.0 µg/ml AgCl-NPs or with 9.7-48.5 µg/ml TMZ. Cells that received combined treatment were also analysed. At a maximum tested concentration of AgCl-NPs, GBM02 and GBM11, the growth decreased by 93% and 40%, respectively, following 72 h of treatment. TMZ treatment decreased the proliferation of GBM02 and GBM11 cells by 58% and 34%, respectively. Combinations of AgCl-NPs and TMZ showed intermediate antiproliferative effects; the lowest concentrations caused an inhibition similar to that obtained with TMZ, and the highest concentrations caused inhibition similar to that obtained with AgCl-NPs alone. No significant changes in astrocyte proliferation were observed. The authors' findings showed that HCA is a fast and reliable approach that can be used to evaluate the antiproliferative effect of the nanoparticles at the single-cell level and that AgCl-NPs are promising agents for glioblastoma treatment.

Biomedical application of graphitic carbon nitrides: tissue depositionin vivo, induction of reactive oxygen species (ROS) and cell viability in tumor cells.

The urgency for new materials in oncology is immediate. In this study we have developed the g-C3N4, a graphitic-like structure formed by periodically linked tris-s-triazine units. The g-C3N4has been synthesized by a simple and fast thermal process. XRD has shown the formation of the crystalline sheet with a compacted structure. The graphite-like structure and the functional groups have been shown by Raman and FTIR spectroscopy. TEM image and AFM revealed the porous composed of five or six C-N layers stacked. DRS and Photoluminescence analyses confirmed the structure with band gap of 2.87 eV and emission band at 448 nm in different wavelengths excitation conditions. The biological results showed inhibitory effect on cancer cell lines and non-toxic effect in normal cell lines. To the best of our knowledge, this is the first work demonstrating the cytotoxic effects of 2D g-C3N4in a cancer cell line, without any external or synergistic influence. The biodistribution/tissue accumulation showed that g-C3N4present a tendency to accumulation on the lung in the first 2 h, but after 24 h the profile of the biodistribution change and it is found mainly in the liver. Thus, 2D-g-C3N4showed great potential for the treatment of several cancer types.

Ohmic heating as a method of obtaining paraprobiotics: Impacts on cell structure and viability by flow cytometry.

This study aimed to evaluate the effects of ohmic heating (OH) on probiotic inactivation, cell viability and morphology of the probiotic strains Lactobacillus acidophilus LA 05 (LA), Lacticaseibacillus casei 01 (LC), and Bifidobacterium animalis Bb 12 (BA) to develop paraprobiotics. OH at different electric field magnitudes (4, 8, and 12 V/cm at 60 Hz) and conventional heat treatment (CONV) were performed to determine the most adequate condition for the obtainment of paraprobiotics. Analysis of culturability, flow cytometry (FC), and Scanning electron microscope (SEM) was carried out. The complete inactivation by CONV was achieved only in the following conditions: LA - 95 °C/5 min, LC and BA - 95 °C/7 min. The same temperature profile was used in OH treatments to study the OH electrical effects. The OH treatment (8 V/cm) caused lower damage to the cell membrane integrity compared to the CONV treatment (p < 0.05). The OH showed to be adequate technology for the efficient production of paraprobiotics.

Ohmic heating increases inactivation and morphological changes of Salmonella sp. and the formation of bioactive compounds in infant formula.

The effect of ohmic heating (OH) (50, 55, and 60 °C, 6 V/cm) on the inactivation kinetics (Weibull model) and morphological changes (scanning electron microscopy and flow cytometry) of Salmonella spp. in infant formula (IF) was evaluated. In addition, thermal load indicators (hydroxymethylfurfural and whey protein nitrogen index, HMF, and WPNI) and bioactive compounds (DPPH, total phenolics, ACE, α-amylase, and α-glucosidase inhibitory activities) were also studied. OH presented a more intense inactivation rate than conventional heating, resulting in a reduction of about 5 log CFU per mL at 60 °C in only 2.91 min, being also noted a greater cell membrane deformation, higher formation of bioactive compounds, and lower values for the thermal load parameters. Overall, OH contributed to retaining the nutritional value and improve food safety in IF processing.

Can sucrose-substitutes increase the antagonistic activity against foodborne pathogens, and improve the technological and functional properties of sheep milk kefir?

The effect of different types of sugar (sucrose, demerara, brown, fructose, coconut sugar, and honey) on sheep milk kefir was evaluated. Microbial counts (Lactobacillus, Lactococcus, Leuconostoc, yeast), antagonistic activity against foodborne pathogens, microstructure (scanning electron microscopy), and antiproliferative activity of cancer cells were evaluated. Furthermore, the antioxidant activity (DPPH), inhibitory activity of angiotensin-converting enzyme (ACE), α-amylase, and α-glucosidase, lactose content, lactic and acetic acids and ethanol, fatty acid profile and volatile organic compounds were determined. The addition of sugars increased the Lactobacillus population (up to 2.24 log CFU/mL), metabolites concentration, antagonistic activity against pathogens, antioxidant activity (11.1 to 24.1%), ACE inhibitory activity (27.5 to 37.6%), α-amylase inhibition (18 to 37.4%), and anti-proliferative activity. Furthermore, it improved the fatty acid profile and volatile compounds. The results suggest that the replacement of sucrose with different types of sugar constitute an interesting option to be used in sheep milk kefir formulations.

Cellular toxicology and mechanism of the response to silver-based nanoparticle exposure in Ewing's sarcoma cells.

Ewing's sarcoma is the most aggressive connective tissue tumor, mainly affecting children and adolescents; the 5 year survival rate is only 50%. Current treatments have poor effectiveness, and more efficient treatments are being sought. Silver-based nanoparticles, such as silver chloride nanoparticles (AgCl-NPs) and silver/silver chloride (Ag/AgCl-NPs) nanoparticles, can be biologically produced and can release Ag+ ions into solution; however, their antitumor activity has been minimally investigated. The aim of this study was to evaluate the antitumor potential of AgCl-NPs and Ag/AgCl-NPs against Ewing's sarcoma cells. A673 cells (Ewing's sarcoma) were treated for 72 h with 0-12.5 µg ml-1 of Ag/AgCl-NPs or 0-40 µg ml-1 of AgCl-NPs. Human cells from the RPE-1 cell line (pigmented retinal epithelium) were used as a model of nontumor cells. The RPE-1 cells were less affected by the administration of AgCl-NPs or Ag/AgCl-NPs, with small reductions in the number of cells and viability and a small increase in apoptosis rates, while lysosomal damage, changes in reactive oxygen species (ROS) production, loss of mitochondrial membrane potential and alterations in microfilaments or cell areas were not observed. A673 tumor cells had significantly reduced number and viability levels when treated with AgCl-NPs, with reductions of 65.05% and 99.17%, respectively, whereas with Ag/AgCl-NP treatment, reductions of 65.53% and 92.51% were observed, respectively. When treated with silver-based nanoparticles, A673 cells also showed a significant increase in ROS production and loss of mitochondrial membrane potential, which culminated in an increase in the percentage of apoptosis among the population. Lysosomal damage was also observed when A673 cells were treated with the highest concentration of AgCl-NPs. In conclusion, the results showed that both AgCl-NPs and Ag/AgCl-NPs had some antitumor activity with minimal effects against healthy cells, which demonstrated the possibility of their use in cancer therapy.

Corrigendum: Quantifying the Responses of Three Bacillus cereus Strains in Isothermal Conditions and During Spray Drying of Different Carrier Agents.

[This corrects the article on p. 1113 in vol. 9, PMID: 29904375.].

Biologically produced silver chloride nanoparticles from B. megaterium modulate interleukin secretion by human adipose stem cell spheroids.

Stem cell tissue constructs are likely to come into contact with silver-based nanoparticles-such as silver chloride nanoparticles (AgCl-NPs)-used as microbicidals at the implant site or in cosmetics. However, the effect of silver-based nanoparticles on 3D cell cultures with potential for tissue engineering has received little attention. Here, we examined the effect of sub-lethal doses (5, 10 and 25 µg/mL, for 1, 7 and 21 days) of AgCl-NPs produced by 'green' bacterial-based synthesis on spheroid 3D cultures of human adipose tissue stem cells (ASCs). Light microscopy analysis revealed that the shape and diameter of ASC spheroids remained largely unchanged after AgCl-NP treatment. Flow cytometry analysis with 7-AAD and 2',7'-dichlorofluorescein diacetate revealed no statistically significant differences in cell death but showed an increase of ROS levels for the untreated group and significant differences for the groups treated with 5 and 10 µg/mL at day 7 (p = 0.0395, p = 0.0266, respectively). Electron microscopy analysis showed limited cell damage in the periphery of AgCl-NP-treated spheroids. However, treatment with AgCl-NP had statistically significant effects on the secretion of IL-6, IL-8, IL-1ß and IL-10 by spheroids, at specific treatment periods and concentrations, and particularly for IL-6, IL-8 and IL-1ß. TGF-ß1 and -ß2 secretion also changed significantly throughout the treatment period. Our results indicate that, despite having little effect on cell viability and morphology, sub-lethal AgCL-NP doses modulate ROS production at day 7 for the groups treated with 5 and 10 µg/mL and also modulate the secretory profile of ASC spheroids. Thus, the use of skin implants or products containing Ag-NPs may promote long-term disturbances in subcutaneous adipose tissue homeostasis.

Silver/silver chloride nanoparticles inhibit the proliferation of human glioblastoma cells.

Glioblastomas (GBM) are aggressive brain tumors with very poor prognosis. While silver nanoparticles represent a potential new strategy for anticancer therapy, the silver/silver chloride nanoparticles (Ag/AgCl-NPs) have microbicidal activity, but had not been tested against tumor cells. Here, we analyzed the effect of biogenically produced Ag/AgCl-NPs (from yeast cultures) on the proliferation of GBM02 glioblastoma cells (and of human astrocytes) by automated, image-based high-content analysis (HCA). We compared the effect of 0.1-5.0 µg mL-1 Ag/AgCl-NPs with that of 9.7-48.5 µg mL-1 temozolomide (TMZ, chemotherapy drug currently used to treat glioblastomas), alone or in combination. At higher concentrations, Ag/AgCl-NPs inhibited GBM02 proliferation more effectively than TMZ (up to 82 and 62% inhibition, respectively), while the opposite occurred at lower concentrations (up to 23 and 53% inhibition, for Ag/AgCl-NPs and TMZ, respectively). The combined treatment (Ag/AgCl-NPs + TMZ) inhibited GBM02 proliferation by 54-83%. Ag/AgCl-NPs had a reduced effect on astrocyte proliferation compared with TMZ, and Ag/AgCl-NPs + TMZ inhibited astrocyte proliferation by 5-42%. The growth rate and population doubling time analyses confirmed that treatment with Ag/AgCl-NPs was more effective against GBM02 cells than TMZ (~ 67-fold), and less aggressive to astrocytes, while Ag/AgCl-NP + TMZ treatment was no more effective against GBM02 cells than Ag/AgCl-NPs monotherapy. Taken together, our data indicate that 2.5 µg mL-1 Ag/AgCl-NPs represents the safest dose tested here, which affects GBM02 proliferation, with limited effect on astrocytes. Our findings show that HCA is a useful approach to evaluate the antiproliferative effect of nanoparticles against tumor cells.

Quantifying the Responses of Three Bacillus cereus Strains in Isothermal Conditions and During Spray Drying of Different Carrier Agents.

Spray drying is a widely used method for producing milk powder. This process is not aimed to cause microbial inactivation, thus sporeforming bacteria may be abundant in the microbiota of milk powder. The first aim of this study was to determine the inactivation kinetics parameters in capillary tubes of three Bacillus cereus strains (436, B63, 540) in three menstrua (whole milk, phosphate buffer, and talc suspension) at 90, 100, and 110°C. D-values for B. cereus in the three menstrua were not significantly different at the highest tested temperature (p > 0.05). Thus, talc was chosen as a carrier agent to allow the recovery of B. cereus from spray dried materials given its low interference on inactivation kinetics. B. cereus spores were also inoculated in whole milk and skim milk following spray drying at 95, 105, and 110°C (outlet temperature). After the spray drying runs, B. cereus spores were counted and the number of decimal reductions (γ) calculated. A correlation between the small diameter of the particles with the survival of spores of three B. cereus strains was found, and B. cereus 436 presented consistently the lowest γ no matter temperature and a carrier agent. The highest γ was found when talc powder was used, which suggest that this carrier agent does not protect B. cereus spores during spray drying. Spray drying of milk can lead to up to 4 γ (strain 540) of B. cereus spores but depending on the strain less than one γ (strain 436) could be observed. This study contributes to the knowledge on the microbiology of low water activity foods by providing novel findings regarding the fate of three B. cereus strains to different spray drying conditions. Acknowledging the variability of inactivation of B. cereus during spray drying is key in the current context of food safety in which the quantification of effects of unit operations must be known for the validation of processes and development of more robust formulations.

The addition of inulin and Lactobacillus casei 01 in sheep milk ice cream.

The effect of the Lactobacillus casei 01 and inulin addition on sheep milk ice cream during storage (-18 °C, 150 days) was investigated. Control, probiotic and synbiotic ice cream (10% w/w sheep milk cream; 10% w/w sheep milk cream, L. casei 01, 6 log CFU/mL; 10% w/w inulin, L. casei 01, 6 log CFU/mL, respectively) were manufactured. Microbiological counts (probiotic count, survival after in vitro gastrointestinal resistance, Caco-2 cell adhesion), bioactivity and microstructure were analysed. Physical and textural characteristics, colour parameters, thermal analysis and organic acids/volatile compounds were also evaluated. All formulations supported L. casei 01 viability and maintained above the minimum therapeutic level (>6 log CFU/mL) during storage. Inulin did not affect L. casei 01 survival after the passage through simulated gastrointestinal tract and adhesion to Caco-2 cells while improved the ACE-inhibitory and antioxidant activity. L. casei 01 addition produced several volatile compounds, such as carboxylic acids, alcohols, aldehydes and ketones. Also, scanning electron microscopy showed an interaction between probiotic bacteria and inulin fibre on synbiotic ice cream and the adhesion of L. casei to Caco-2 cells was observed.

On the synergy between silver nanoparticles and doxycycline towards the inhibition of Staphylococcus aureus growth.

In a previous paper (RSC Adv., 2015, 5, 66886-66893), we showed that the combination of silver nanoparticles (NanoAg) with doxycycline (DO) culminated in an increased bactericidal activity towards E. coli. Herein we further investigated the metabolic changes that occurred on Staphylococcus aureus upon exposure to NanoAg with the help of attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) coupled with multivariate data analysis. It has been discovered that the combination of DO with NanoAg produced metabolic changes in S. aureus that were not simply the overlap of the treatments with DO and NanoAg separately. Our results suggest that DO and NanoAg act synergistically to impede protein synthesis by the bacteria.

Anti-inflammatory, analgesic and anti-tumor properties of gold nanoparticles.

BACKGROUND: Gold nanoparticles (GNPs) are regarded as potential platforms for drug delivery. However, their interaction with live organisms must be understood prior to their utilization as drug carriers. The present study reports the anti-inflammatory, analgesic and anti-tumor effects of GNPs. The biodistribution of GNPs and their effect on various tissues have also been studied. METHODS: GNPs were synthesized through an environmentally friendly route and characterized with TEM and UV-vis. After HT-29 cells had been exposed to GNPs, apoptosis was assessed with Annexin V and propidium iodide staining and caspase-3 activity determined with a confocal laser scanning microscope. GNPs were administrated to male and female Swiss mice for posterior assessment of their anti-inflammatory and analgesic properties. The biodistribution of GNPs and their impact on tissues were studied with UV-vis and histopathological analysis, respectively. RESULTS: Cell apoptosis was observed in a dose-dependent manner for GNPs concentrations ranging from 40µg/mL to 80µg/mL (p<0.05). The best anti-inflammatory activity was observed at the dose of 1500µg/kg, which caused a reduction of 49.3% in leukocyte migration. GNPs showed peripheral analgesia at the dose of 1500µg/kg and have been found in liver, spleen, kidney and lungs. Histopathological examination revealed extravasation of red blood cells in lungs. CONCLUSION: The study draws attention to gold nanoparticles as a resource for technological innovation in the anti-inflammatory, analgesic and anti-tumor fields. GNPs have biological effects that deserve investigation to assess their full interaction with organic systems.

Impact of culture conditions on the chlorophyll content of microalgae for biotechnological applications.

Chlorophyll is a commercially important natural green pigment responsible for the absorption of light energy and its conversion into chemical energy via photosynthesis in plants and algae. This bioactive compound is widely used in the food, cosmetic, and pharmaceutical industries. Chlorophyll has been consumed for health benefits as a nutraceutical agent with antioxidant, anti-inflammatory, antimutagenic, and antimicrobial properties. Microalgae are photosynthesizing microorganisms which can be extracted for several high-value bioproducts in the biotechnology industry. These microorganisms are highly efficient at adapting to physicochemical variations in the local environment. This allows optimization of culture conditions for inducing microalgal growth and biomass production as well as for changing their biochemical composition. The modulation of microalgal culture under controlled conditions has been proposed to maximize chlorophyll accumulation. Strategies reported in the literature to promote the chlorophyll content in microalgae include variation in light intensity, culture agitation, and changes in temperature and nutrient availability. These factors affect chlorophyll concentration in a species-specific manner; therefore, optimization of culture conditions has become an essential requirement. This paper provides an overview of the current knowledge on the effects of key environmental factors on microalgal chlorophyll accumulation, focusing on small-scale laboratory experiments.

Green production of microalgae-based silver chloride nanoparticles with antimicrobial activity against pathogenic bacteria.

Silver nanoparticles are powerful antimicrobial agents. Here, the synthesis of silver chloride nanoparticles (AgCl-NPs) was consistently evidenced from a commercially valuable microalgae species, Chlorella vulgaris. Incubation of C. vulgaris conditioned medium with AgNO3 resulted in a medium color change to yellow/brown (with UV-vis absorbance at 415nm), indicative of silver nanoparticle formation. Energy-dispersive X-ray spectroscopy (EDS) of purified nanoparticles confirmed the presence of both silver and chlorine atoms, and X-ray diffraction (XRD) showed the typical pattern of cubic crystalline AgCl-NPs. Transmission electron microscopy (TEM) showed that most particles (65%) were spherical, with average diameter of 9.8±5.7nm. Fourier transform infrared spectroscopy (FTIR) of purified nanoparticle fractions suggested that proteins are the main molecular entities involved in AgCl-NP formation and stabilization. AgCl-NPs (from 10µg/mL) decreased by 98% the growth of Gram-positive Staphylococcus aureus and Gram-negative Klebsiella pneumoniae bacterial pathogens, and had a dose-dependent effect on cell viability, which was measured by automated image-based high content screening (HCS). Ultrastructural analysis of treated bacteria by TEM revealed the abnormal arrangement of the chromosomal DNA. Our findings strongly indicated that the AgCl-NPs from C. vulgaris conditioned medium is a promising 'green' alternative for biomedical application as antimicrobials.