CBD Oil Bacterial Infection

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Cannabis sativa CBD Extract Shows Promising Antibacterial Activity against Salmonella typhimurium and S. newington 1 The Microbiology Program, College of Science, Technology, Engineering, The researchers tested the CBD against some strains of staphylococcus, which cause skin infections, and streptococcus, which cause strep throat. Find out if CBD oil could be as effective at fighting bacterial infections as traditional antibiotics. Learn about CBD oil antibacterial dosage, research, and benefits.

Cannabis sativa CBD Extract Shows Promising Antibacterial Activity against Salmonella typhimurium and S. newington

1 The Microbiology Program, College of Science, Technology, Engineering, and Mathematics (C-STEM), Alabama State University, Montgomery, AL 36104, USA; [email protected] (L.G.); [email protected] (R.V.)

Joseph Atia Ayariga

2 The Biomedical Engineering Program, College of Science, Technology, Engineering, and Mathematics (C-STEM), Alabama State University, Montgomery, AL 36104, USA

Olufemi S. Ajayi

3 Department of Biological Sciences, Alabama State University, Montgomery, AL 36104, USA; [email protected] (J.X.); [email protected] (M.S.-F.)

Junhuan Xu

3 Department of Biological Sciences, Alabama State University, Montgomery, AL 36104, USA; [email protected] (J.X.); [email protected] (M.S.-F.)

Robert Villafane

1 The Microbiology Program, College of Science, Technology, Engineering, and Mathematics (C-STEM), Alabama State University, Montgomery, AL 36104, USA; [email protected] (L.G.); [email protected] (R.V.)

Michelle Samuel-Foo

3 Department of Biological Sciences, Alabama State University, Montgomery, AL 36104, USA; [email protected] (J.X.); [email protected] (M.S.-F.)

1 The Microbiology Program, College of Science, Technology, Engineering, and Mathematics (C-STEM), Alabama State University, Montgomery, AL 36104, USA; [email protected] (L.G.); [email protected] (R.V.)

2 The Biomedical Engineering Program, College of Science, Technology, Engineering, and Mathematics (C-STEM), Alabama State University, Montgomery, AL 36104, USA

3 Department of Biological Sciences, Alabama State University, Montgomery, AL 36104, USA; [email protected] (J.X.); [email protected] (M.S.-F.)

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Abstract

Products derived from Cannabis sativa L. have gained increased interest and popularity. As these products become common amongst the public, the health and potential therapeutic values associated with hemp have become a premier focus of research. While the psychoactive and medicinal properties of Cannabis products have been extensively highlighted in the literature, the antibacterial properties of cannabidiol (CBD) have not been explored in depth. This research serves to examine the antibacterial potential of CBD against Salmonella newington and S. typhimurium. In this study, we observed bacterial response to CBD exposure through biological assays, bacterial kinetics, and fluorescence microscopy. Additionally, comparative studies between CBD and ampicillin were conducted against S. typhimurium and S. newington to determine comparative efficacy. Furthermore, we observed potential resistance development of our Salmonella spp. against CBD treatment.

1. Introduction

Cannabis sativa L., a member of the genus Cannabis and a species in the family Cannabaceae, originated from Central Asia and is one of the oldest psychoactive plants known to humans [1]. Since its discovery, C. sativa has been utilized recreationally, medicinally, and industrially for numerous applications. Early uses of C. sativa focused on its use as an industrial material to produce textiles, ropes, and paper products [2,3,4]. Throughout early history, the use of C. sativa as an industrial product spread from Asia throughout Europe and Africa [5]. As industrial use of C. sativa became common, its use as a therapeutic against rheumatic pain, intestinal constipation, disorders of the female reproductive system, malaria, and other common health problems can be traced back to ancient China where it was one of the oldest pharmaceuticals recorded [6]. Over time, the use of C. sativa as a therapeutic option was introduced across the western world and by the 19th century had gained attention from medical science. The first clinical conference on C. sativa in 1860 led to an increased interest in the research and development of medicinal products derived from C. sativa [2]. This interest in C. sativa drastically declined in 1942 when it was removed from the United States Pharmacopoeia and lost its status due to research suggesting a correlation between C. sativa and ‘insanity’ [7]. This decision eventually led to the United Nations designating Cannabis as an illegal psychotropic substance in 1971 at the Convention on Psychotropic Substances [8,9]. These designations and findings drastically reduced research progression on Cannabis and produced a negative stigma on the substance from the perspective of the public [10,11].

As we move towards the mid-21st century, the acceptance of Cannabis and the potential of its byproducts has increased [12]. Continued research demonstrated the efficiency of Cannabis in the treatment of medical conditions including epilepsy, multiple sclerosis, Tourette’s syndrome, and other neurological diseases [13,14,15]. Additionally, research has further characterized the active compounds—tetrahydrocannabinol (THC) and cannabidiol (CBD)—of Cannabis and their psychological and physiological effects on humans [16,17,18,19]. While clinical research has primarily focused on the efficacy of Cannabis against neurological disorders, a current gap in knowledge is the efficacy of Cannabis and its byproducts as antibacterial agents. The current literature does suggest that Cannabis and more specifically its active compound, CBD, shows antibacterial function [20,21]. However, further research and characterization of this antibacterial function is crucial to the development of novel therapeutics against clinically relevant bacteria [22,23].

The increased prevalence and occurrence of antimicrobial resistance is a major health concern worldwide, making the development of alternative therapies a necessity [24,25,26,27]. The steady decrease in antibiotic efficacy alongside the decreased development of new antibiotics present a major obstacle in the treatment of multidrug-resistant bacteria [28,29]. Antibiotic-resistant and multidrug-resistant bacterial infections account for roughly 2.8 million infections and 35,000 deaths annually in the United States alone [30]. Antibiotic-resistant bacterial infections, while a major threat to public health, also present a major economic burden to the United States, resulting in an annual cost of USD 7.7 billion [31]. As resistant bacterial species persist, the development of novel antibacterial treatment methods is essential to the preservation of public health. The purpose of this research is to determine if CBD extracted from C. sativa demonstrates antibacterial activity against a common Gram-negative bacterial pathogen, Salmonella. CBD, initially not recognized as an active compound of cannabinoids, has been shown to possess potential therapeutic benefits [32,33,34]. Cannabis is commonly associated with psychotic effects. We now know that these effects can be attributed to THC, whereas CBD “exhibits no effects indicative of any abuse or dependence-potential and to date, there is no evidence of public health related problems associated with the use of pure CBD” according to the World Health Organization [35]. The literature suggests that CBD’s antibacterial function is carried out through the disruption of the cell membrane of both Gram-positive and Gram-negative bacteria [36]. The antibacterial activity of CBD against several bacterial pathogens including Staphylococcus aureus, Streptococcus pneumoniae, Clostridiosis difficile, Neisseria spp., Moraxella catarrhalis, and Legionella pneumophila has been observed [20,21]. Study of CBD as an antibacterial agent requires further determination of the range of bacteria that CBD exhibits antibacterial activity against. Examining the potential of CBD against a clinically relevant pathogen such as Salmonella is essential to further expand our knowledge on CBD as a potential novel therapeutic.

Salmonella species are one of the most common and prevalent foodborne pathogens worldwide, found in several food products including poultry, seafood, and other fresh or processed meats [26,27,37,38,39]. Salmonella accounts for 1.35 million infections, 26,500 hospitalizations, and 420 deaths annually in the United States alone [30]. Salmonella is a significant spoilage hazard. In addition, the increased prevalence of multidrug-resistant strains makes Salmonella a major threat to public health [25,30]. The CDC reported in 2019 that Salmonella typhimurium, one of the species examined in this study, accounted for 59% of ampicillin-resistant Salmonella infections in the United States [30]. The continued use of broad-spectrum antibiotics could drastically increase the prevalence of antibiotic and multidrug-resistant Salmonella strains, making the development of novel antibiotic alternatives a necessity [31,40].

In this study, we performed plate assays, fluorescence microscopy, and growth kinetic studies to determine the antibacterial activity of CBD extracted from C. sativa against S. typhimurium and S. newington. Additionally, we conducted comparative kinetic studies of the two Salmonella strains in the presence of CBD or a common broad-spectrum antibiotic, ampicillin. Finally, we examined resistance development of S. typhimurium and S. newington against CBD treatment over an extended time. We hypothesized that CBD would exhibit antibacterial activity against S. typhimurium and S. newington. The results of these studies suggest that CBD does exhibit antibacterial activity against these Salmonella species, further encouraging research and development of CBD as a potential antibacterial agent.

2. Results

2.1. Gas Chromatography

For confirmation of pure CBD in our C. sativa extract, gas chromatography (GC) was conducted. Figure 1 A represents the internal standard for CBD and THC, while Figure 1 B represents our sample. In these readings, we observe a strong single peak at the 8.23 min retention time (RT) in our sample. This is consistent with the expected RT of CBD at 8.22 min as shown in the internal control sample. The absence of other peaks represents the absence of other compounds or contaminants. Overall, we conclude through the GC analysis that our sample is pure CBD.

GC peaks confirm CBD as the primary compound present within our C. sativa extract. (A) represents CBD (in grey color) internal standard for GC with an 8.22000 expected RT (min). Peak detected at 10.09 min (A) represents the expected RT (min) of THC (in white color), which was not detected within our sample (B). (B) represents GC analysis of our CBD extract featuring an 8.2300 RT (min), confirming the sample is composed of CBD. Secondary readings around the base of the sample are consistent to those seen within our CBD internal standard (A).

2.2. Plate Assays

To examine the potential antibacterial activity of our CBD against S. typhimurium and S. newington, the Kirby–Bauer and spot assays were conducted. These assays allow us to visualize and quantify the inhibitory effects of CBD against our Salmonella strains. Both assays confirmed inhibitory activity of CBD against S. typhimurium and S. newington. In the Kirby–Bauer assay, we observed zones of inhibition (ZOI) around the CBD-treated disks suggesting inhibition of bacterial growth due to exposure to CBD ( Figure 2 C). These results also suggest a dose-dependent inhibition due to CBD treatment with ZOIs decreasing as CBD concentration decreased ( Figure 2 A,B). The results of the spot assay further confirmed the dose-dependent nature of CBD’s inhibitory activity. As CBD concentration decreased, we saw an increase in the density of bacterial colonies in both S. typhimurium and S. newington ( Figure 2 D). These data also suggested that S. newington was more susceptible to CBD treatment than S. typhimurium due to the larger ZOIs ( Figure 2 B) and lower colony density ( Figure 2 D).

Plate assays confirm inhibitory activity of CBD. Quantitative analysis of Kirby–Bauer assay results of S. typhimurium (A) and S. newington (B). Images from Kirby–Bauer assay and CBD-produced ZOIs against S. typhimurium and S. newington (C). Images from spot assay featuring S. typhimurium and S. newington treated with decreasing concentrations (left to right) of CBD (D). (A) * p-value = 0.000961, ** p-value = 0.0117, *** p-value = 0.7179. (B) * p-value = 0.00000343, ** p-value = 0.000480, *** p-value = 0.00832.

2.3. CBD Extract Reduces Bacterial Growth of Salmonella typhimurium and Salmonella newington

To determine the effect of CBD on S. typhimurium and S. newington, cultures were treated with CBD at concentrations of 1.25, 0.125, 0.0125, or 0.00125 μg/mL. The optical density (OD600) was recorded using a spectrometer (Molecular Devices SpectraMax ® ABS Plus) hourly for 6 h. Both S. typhimurium and S. newington cultures treated with CBD showed a significant reduction in OD600 within 6 h of treatment ( Figure 3 A,B). It was even observed that in S. newington, CBD at a concentration as low as 0.0125 μg/mL reduced bacterial growth over the 6 h period. These results suggest that S. newington might have a greater susceptibility to CBD than S. typhimurium, whose MIC was 0.125 μg/mL. It was observed that treatment of S. typhimurium with CBD at a concentration of 0.00125 μg/mL resulted in a higher OD600 than that of the control treatment. We infer that these results suggest a rapid development of resistance to low concentrations of CBD treatment resulting in an increase in bacterial fitness. The reduction of OD600 signifies that CBD extract does exhibit antibacterial characteristics. The antibacterial effect on S. typhimurium and S. newington was observed to be dose-dependent with the OD600 increasing in correlation with decreased concentrations of CBD.

Treatment of lag-phase S. typhimurium (A) and S. newington (B) with CBD. The effect of several CBD dilutions on OD600 was recorded in triplicate.

2.4. Assessment of Bacterial Membrane Integrity via Fluorescent Staining of CBD-Treated Salmonella Cells

To examine the effect of CBD treatment on Salmonella cells, S. typhimurium and S. newington samples were treated with CBD at concentrations of 1.25, 0.125, or 0.0125 μg/mL. These samples were visualized using immunofluorescent 4′,6-diamidino-2-phenylindole (DAPI) staining. This staining technique emits fluorescence when the stain binds to AT-rich DNA. This stain is membrane impermeable, and thus, fluorescence represents compromised membrane integrity. DAPI staining confirmed that CBD treatment resulted in degradation of membrane integrity after 5 min and 30 min of treatment ( Figure 4 A,B). Quantitative analysis using densitometry of these images showed increased DAPI fluorescence from the 5 min to the 30 min time points ( Figure 4 C). These images all indicate that CBD can successfully damage Salmonella cells outer lipopolysaccharide membranes.

(A). Fluorescence microscopy of CBD-treated Salmonella cells stained with DAPI. Images were acquired at the time points of 5 min. DAPI staining was utilized to assess membrane integrity following treatment with CBD. (B). Fluorescence microscopy of CBD-treated Salmonella cells stained with DAPI. Images were acquired at the time points of 30 min. DAPI staining was utilized to assess membrane integrity following treatment with CBD. (C). Quantitative analysis of membrane integrity via densitometry analysis of fluorescence after DAPI staining.

2.5. Comparative Study of CBD and Antibiotic Treatment against Salmonella

Salmonella infections are typically treated with broad-spectrum antibiotics such as ampicillin; however, the development of resistance to these treatments has become more prevalent, thus increasing the need for alternative treatments. To compare conventional antibiotics to CBD, we conducted comparative kinetic studies to observe antibacterial activity. In this study, Salmonella strains S. typhimurium and S. newington at an OD600 of 0.5 were treated with the minimum inhibitory concentration (MIC) of ampicillin (0.5 µg/mL) or CBD (0.125 µg/mL). Results suggested that CBD and ampicillin both successfully inhibited Salmonella growth in both strains ( Figure 5 ). In comparison, both treatments resulted in a similar OD600 after 6 h of treatment suggesting that CBD was able to inhibit bacterial growth to an extent similar to ampicillin.

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Comparative efficacy of CBD and antibiotic treatment of Salmonella typhimurium (A) and Salmonella newington (B). Salmonella cells were cultured overnight and then treated with either CBD or ampicillin. OD600 was measured hourly over a 6 h time period to observe bacterial growth following treatment.

2.6. Developed Resistance of Salmonella to CBD Treatment

A major concern with conventional antibiotic treatments is the development of resistance. To observe the development of resistance in Salmonella against CBD treatment, extended kinetic studies were conducted over a span of 48 h. Salmonella strains S. typhimurium and S. newington were treated with ampicillin (MIC 0.5 µg/mL) or CBD (1.25, 0.125, 0.0125, or 0.00125 μg/mL). Results confirmed a developed resistance to CBD in both Salmonella species ( Figure 6 ). At concentrations of 1.25 and 0.125 μg/mL of CBD, resistance development was not observed until the 48 h time point. These results suggest that CBD was effective in killing Salmonella and decreasing the rate of resistance development in both Salmonella strains over the initial 24 h time period. At 0.0125, 0.00125, and 0.000125 μg/mL concentrations of CBD, there was no significant reduction in bacterial growth and resistance was developed after only 12 h of exposure in both Salmonella strains. These results suggest that both Salmonella strains were able to develop resistance quickly to low concentrations of CBD.

Development of resistance to CBD and ampicillin treatments by S. typhimurium (A) and S. newington (B) examined through extended kinetics. Bacterial cultures were treated with either ampicillin, CBD dilutions, or dH2O. OD600 was recorded over a period of 48 h.

2.7. CBD Effectiveness against Salmonella Biofilm

The literature has determined that most hospital-acquired infections are the result of bacterial biofilm formation [41]. Typically, a result of bacteria reaching a non-replicatory state, biofilms are a major health threat that are characterized by antibiotic resistance, reoccurring infection, and sepsis [42]. Understanding CBD activity against bacterial biofilm is an important aspect of determining CBD viability against Salmonella spp. To determine the efficacy of CBD treatment against biofilm formation, S. typhimurium biofilms were grown in vitro and treated with CBD at a concentration of 0.125 µg/mL. S. typhimurium biofilms were then visualized through scanning electron microscopy (SEM) ( Figure 7 ).

SEM images depicting the comparison between untreated (control) and treated (0.125 µg/mL CBD) S. typhimurium biofilms. Untreated S. typhimurium biofilm resulted in continued persistence (A). CBD treatment at 1.25 µg/mL resulted in a reduction of S. typhimurium characterized by fragmented cells suggesting S. typhimurium cell death (B).

3. Discussion

The rapid increase of antibiotic- and multidrug-resistant bacteria over the early 21st century is a major threat to public health [30]. Knowing this, the study of potentially efficacious alternative therapeutics has been a topic of growing interest. C. sativa products, while typically associated with the treatment of neurological disorders, have shown promise as antibacterial agents against several notable pathogens [20,21]. Of the multiple metabolites of C. sativa, research suggests that CBD is the most promising [32,33,34]. This compound, unlike THC, holds no psychoactive properties but does possess antioxidant, anti-inflammatory, and antibacterial properties [22,43,44]. CBD’s antibacterial potential has shown some promise; however, research is relatively limited. Further investigation and confirmation of the specificity of this antibacterial activity is crucial as the field of medicine searches for new viable therapeutics for resistant bacterial infections.

C. sativa products have exhibited a wide variety of applications in numerous fields. This study helps expand our knowledge of C. sativa-derived CBD as an antibacterial agent against two common Gram-negative pathogenic Salmonella strains. Salmonella spp. have significance in terms of resistance to antibiotics and standard treatment protocols making these infections a major threat to the preservation of public health [25,28,30]. Discovery and development of novel antibacterial agents such as CBD are a major step towards the future of therapeutics in a world where antibiotics are no longer efficacious and cost effective [20,21]. This study serves primarily to determine the antibacterial potential of CBD against S. typhimurium and S. newington through plate assays, fluorescence microscopy, and kinetic assays. Additionally, this study examines the comparative antibacterial activity of CBD and ampicillin as well as resistance development of S. typhimurium and S. newington against CBD treatments.

In this study, we confirm that CBD does exhibit antibacterial activity against S. typhimurium and S. newington. This inference was derived from a combination of plate assays, fluorescence microscopy, and bacterial growth kinetic assays and thus allowed us to propose a potential mechanism of this antibacterial activity. Plate assays consisted of the Kirby–Bauer assay and the spot assay, both of which served as initial confirmation of bacterial inhibition of S. typhimurium and S. newington. Once this inhibitory activity was observed, a fluorescence microscopy assay for membrane integrity was utilized. This assay utilized DAPI stain, a membrane-impermeable DNA binding stain. Fluorescence of the DAPI stain confirmed that CBD treatment of S. typhimurium and S. newington resulted in a loss of membrane integrity and cell death, and thus proving our hypothesis that the CBD treatment leads to membrane disruption. Other researchers have reported this mechanism as the antibacterial mechanism of action of CBD [36]. Finally, kinetic assays were conducted to examine the effect of CBD treatment on lag-phase S. typhimurium and S. newington over a span of 6 h. Once again, it was observed that CBD was effective in growth inhibition of both species in a 6 h period with a MIC of 0.0125 μg/mL in S. typhimurium and 0.125 μg/mL in S. newington. These findings confirm that CBD does possess antibacterial activity through mechanisms similarly described in other Gram-negative bacterial species [20,21,36].

Within this study, we were able to exhibit the antibacterial effectiveness of CBD at a concentration of 0.125 μg/mL against a S. typhimurium biofilm ( Figure 7 ). This study is relevant in terms of clinical application of CBD where most hospital-acquired infections are in the form of biofilms [41]. Biofilms typically associated with antibiotic resistance result in infections that are difficult to treat with an increased chance of sepsis and mortality in some cases [42]. Considering the threat that biofilms pose, it is essential that novel therapeutics are developed for the prevention and treatment of these harmful infections. Our results suggest that CBD features antibacterial activity that is not only effective against lag- and log-phase Salmonella spp. but also against Salmonella biofilms.

An important facet of this study was the comparison between CBD and broad-spectrum antibiotic treatment. In this study, we examined the comparative kinetics between CBD treatment and ampicillin treatment of S. typhimurium and S. newington. We concluded from the results of these kinetic studies that both CBD and ampicillin at their MIC concentrations are successful at inhibition of S. typhimurium and S. newington growth. It is important to note that the MIC concentration of ampicillin (0.5 μg/mL) is significantly higher than the MIC of CBD (0.125 μg/mL) ( Figure 5 ). To further investigate the comparative efficacy of a novel therapeutic agent (i.e., CBD in this study) and a standard treatment (such as ampicillin), it was important to examine the resistance development of the pathogen to the therapeutic agents. To examine the resistance of S. typhimurium and S. newington against CBD and ampicillin treatments, extended growth kinetics under different dosages of CBD were carried out over a 48 h period (see Figure 6 ). These results varied across the two strains of Salmonella that were used. In S. typhimurium, CBD concentrations of 1.25 and 0.125 μg/mL were able to inhibit bacterial growth over the span of 24 h after a single treatment. These results are similar to the growth kinetics we observed with ampicillin treatment after 24 h, whereas lower concentrations of CBD showed a rapid rise in growth following 12 h of treatment, suggesting a rapid development of resistance. After 48 h of exposure to CBD treatment, S. typhimurium had developed resistance, suggested by the rapid rise in OD600 at the 48 h time point, to all CBD treatments and the ampicillin treatment. This result is consistent with the literature on S. typhimurium, which accounts for 59% of all ampicillin-resistant Salmonella infections in the United States [30]. These results raise the question of what mechanism S. typhimurium develops to resist the antibacterial activity of CBD. The literature has suggested several mechanisms for S. typhimurium resistance to antibiotics including the multidrug efflux pump AcrAB, OXA-1 β -lactamase, and other beta-lactamase genes (bla). Ampicillin, a beta-lactam class antibiotic, kills bacteria through binding to penicillin-binding proteins in the cytoplasmic membrane [45,46,47]. This mechanism of resistance relies on the inner mechanisms of the bacterial cell, whereas we hypothesize that CBD’s antibacterial activity is a result of membrane disruption. In this work, we demonstrated that the antibacterial activity of CBD was generated through membrane integrity disruption. Therefore, we suggest that S. typhimurium resistance to CBD might be conferred through a resistance mechanism different from that of the ampicillin-resistance mechanism of this bacteria. This could be significant in the application of CBD as a therapeutic agent against ampicillin-resistant S. typhimurium or in a potential co-therapy designed with CBD and ampicillin. Co-therapies typically reduce the ability of bacteria to persist due to bacteria having to rapidly develop multiple resistance mechanisms [27,48,49,50,51]. Additionally, co-therapy strategies have been shown to ‘revert’ resistant bacteria back to a state that is susceptible to antibiotic treatment [52]. These factors further buttress the need to employ CBD in a potential CBD–antibiotic co-therapy once the resistance mechanism of CBD is fully understood. The extended kinetics of S. newington revealed that this strain was more susceptible to CBD treatments of 1.25, 0.125, and 0.0125 μg/mL 24 h after treatment. However, at the 48 h time point, there was a strong development of resistance in all CBD treatments but no resistance development to ampicillin. These results suggest that the mechanism of resistance is different between ampicillin and CBD ( Figure 6 ). Further studies to characterize the resistance developed by S. typhimurium and S. newington against CBD are important to understand how to responsibly develop this potential therapeutic agent.

The previous literature has outlined the potential of CBD against several, mostly Gram-positive, bacterial pathogens including Staphylococcus aureus, Streptococcus pneumoniae, and Clostridioides difficile [20,21]. In these studies, there was a reported MIC of 1–5 μg/mL against Staphylococci spp., Clostridioides spp., and Streptococci spp. [20]. More recent studies have reported a MIC of 0.5–2 μg/mL against several clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA). This study focused on a relevant Gram-negative pathogen, Salmonella, whose tolerance of CBD has not previously been studied. Within our study, S. typhimurium and S. newington were shown to have a MIC of 0.125–1.25 μg/mL. These MIC results are consistent with the previous literature exploring the antibacterial capabilities of CBD [20,21]. Results from this study and the studies mentioned earlier [20,21] seem to suggest bacterial susceptibility to CBD within a MIC range of 0.1–5 μg/mL. As we continue to explore potential antibacterial agents, it is essential that we explore the effectiveness of these agents against a multitude of diverse bacterial pathogens. Considering this, our study confirms that CBD does have antibacterial activity against two Gram-negative Salmonella strains, filling a valuable gap in our knowledge of CBD as an antibacterial agent. While this study illuminates the potential of CBD as a therapeutic and fills a void in the current literature, future work is necessary for further development of this bioactive compound as a therapeutic agent.

While our study was successful in determining the presence of CBD’s antibacterial activity against S. typhimurium and S. newington, there were limitations. One limitation was that the specific resistance mechanism of S. typhimurium and S. newington against CBD was not investigated in this study. This study simply determined the ability of our Salmonella strains to develop resistance against the CBD treatments administered and did not define the mechanism of resistance. The development of CBD as a novel therapeutic option will require further studies and characterization. Some relevant studies for the progression of CBD as a potential therapeutic include resistance mechanisms to CBD, cytotoxicity of CBD especially to co-therapy situations, immunological response to CBD treatment, CBD function in pathophysiological conditions, and in vivo models. These future studies will serve to further expand our knowledge of CBD as an antibacterial agent with potential therapeutic benefits.

4. Materials and Methods

4.1. CBD Extraction

CBD extraction was carried out by Sustainable CBD LLC. C. sativa biomass was weighed, tagged, and recorded in a receiving trailer for processing. Following storage, the biomass was reduced to between 200 and 500 microns and underwent CO2 extraction in an Apeks Transformer for subcritical extraction (Gibraltar Industries Inc., Buffalo, NY, USA). Subcritical extraction was carried out at a target pressure of 1200 psi, chiller temperature of 20–25 °C, propylene glycol percentage of 10%, orifice size of 22, resultant separator pressure of 350–400 psi, resultant separator temperature of −6–4 °C, for an extraction time of approximately 2–3 h. Following subcritical extraction, samples were prepared for decarboxylation prior to supercritical extraction. Hemp biomass was placed in the oven for approximately 100 min at 265 °C to decarboxylate. Once decarboxylation was carried out, an Apeks Transformer was utilized for supercritical extraction (Gibraltar Industries Inc., Buffalo, NY, USA). Supercritical extraction was carried out at a target pressure of 1800 psi, chiller temperature of 37–42 °C, propylene glycol percentage of 10%, orifice size of 18, resultant separator pressure of 350–400 psi, resultant separator temperature of 0–10 °C, for an extraction time of approximately 1–2 h per pound of material. The resulting material then underwent winterization through addition of ethanol to the crude extract. This sample was frozen and then filtered through Buchner funnels and the remaining ethanol was evaporated using a Heidolph rotary evaporator (Heidolph Instruments GmbH & Co.KG, Kelheim, Germany). Distillation was carried out using the Lab Society 5 L short path distillation unit (Lab Society ® , Boulder, CO, USA) for further refinement. The resulting product of this procedure was winterized cannabinoid [53,54,55].

4.2. Gas Chromatography Analysis of CBD

CBD extracts were analyzed by gas chromatograph (GC)/mass spectrometry (MS) using Agilent 6890N GC and Agilent 5975 MS (Agilent Technologies Inc., Santa Clara, CA, USA) with a Restek Rxi-5Sil MS with integra guard column (15 m, 0.250 mmID, 0.25 µm df) (Restek Corporation, Bellefonte, PA, USA). A solution of Restek Qualitative Retention Time Index Standard (Restek Corporation, Bellefonte, PA, USA) was used to create the retention time index. The temperature of the injection port was set at 280 °C and the helium gas flow was constant at 1.1 mL/min. The samples were injected in split mode (2:1) with a volume of 1 µL of sample. The GC oven temperature was programmed as follows: initial temperature of 70 °C for 4 min, ramp to 200 °C at 20 °C/min, ramp to 300 °C at 8 °C/min, ramp to 325 °C at 50 °C/min with a 5 min hold, thus requiring a total run time of 30.5 min. The MS transfer line was set to 250 °C, source to 230 °C, and quads to 150 °C. The raw data were processed and analyzed using Agilent Enhanced ChemStation software (Agilent Technologies Inc., Santa Clara, CA, USA). The NIST 2.0 library was used with AMDIS for compound identification.

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4.3. Plate Assays for Antibacterial Screening of CBD

To qualitatively determine the antibacterial potential of CBD against S. typhimurium LT2 strain MS1868 (a kind gift from Dr. Anthony R. Poteete, University of Massachusetts) and S. newington (also known as S. enterica serovar Anatum var 15+ strain UC1698, a kind gift from Dr. Sherwood R. Casjens, University of Utah), plate assays were utilized. Prior to the assays, CBD was serially diluted to the concentrations of 1.25, 0.125, 0.0125, or 0.00125 μg/mL. Bacterial strains S. typhimurium and S. newington were incubated until the late log-phase (OD > 1) in sterile Luria broth (BD Difco, Franklin Lakes, NJ, USA).

To conduct the Kirby–Bauer assay, plates were inoculated with overnight cultures of either S. typhimurium or S. newington and top agar (BD Difco, Franklin Lakes, NJ, USA) to create a bacterial lawn. These plates were then divided into quarters representing the four CBD dilutions (1.25, 0.125, 0.0125, or 0.00125 μg/mL). Sterile paper discs were soaked in a designated dilution of CBD or treated with dH2O (control) and then applied to the agar plates in triplicate. The results were three discs per CBD dilution on the agar plate. Plates were then incubated at 37 °C for 24 h, and bacterial growth was observed to determine zones of inhibition (ZOI) around the CBD-treated discs [56,57,58]. ZOIs were quantitatively assessed using ImageJ (NIH Image, Bethesda, Maryland). This assay was completed in triplicate.

For further confirmation of antibacterial activity, spot assays were conducted using lag-phase S. typhimurium and S. newington cultures (OD > 1). Sterile agar plates were inoculated with four 10 μL aliquots of S. typhimurium or S. newington. Each ‘dot’ was then inoculated with 10 μL of one of the four dilutions (1.25, 0.125, 0.0125, or 0.00125 μg/mL) of CBD. Each culture was also spotted and treated with 10 μL of dH2O as the control. The plate was then incubated for 24 h at 37 °C and observed for inhibition of bacterial growth [59]. This assay was completed in triplicate.

4.4. Fluorescence Microscopy of CBD-Treated Salmonella Cells

To visualize the effects of CBD treatment on Salmonella cells, fluorescence microscopy was utilized. Salmonella cells were fixed and stained using DAPI (4′,6-diamidino-2-phenylindole) to determine cell membrane integrity. DAPI stain binds exclusively to dsDNA, which is only accessible as a result of compromised membrane integrity and expulsion of the cytoplasmic material including the nucleus out of the cell. Visualization of DAPI fluorescence under the fluorescence microscope (Biotek Cytation™ 3 Automated Fluorescence Microscope) (Agilent Technologies Inc., Santa Clara, CA, USA) confirmed cell membrane damage and hence cell death. Salmonella cells were treated with CBD dilutions (1.25, 0.125, or 0.0125 μg/mL) or left untreated (control). Following treatment, cells were fixed at either 5 min or 30 min and assessed using fluorescence microscopy [27].

4.5. Bacterial Growth Kinetics

To study bacterial growth kinetics, a 96-well plate (Fisherbrand™, Fisher Scientific, Fair Lawn, NJ, USA) was inoculated with 180μL of overnight bacterial cultures of either S. typhimurium or S. newington at an OD600 ≈ 0.5. The cultures were then incubated at 37 °C with rotary shaking at 121 rpm. Measurements of bacterial density (OD600) were taken every hour for 6 h and again at the 12, 24, and 48 h time points using a spectrometer (Molecular Devices SpectraMax ® ABS Plus) (Molecular Devices LLC., San Jose, CA, USA). This experiment was completed in triplicate [27,60].

4.6. Bacterial Growth Kinetics in the Prescence of CBD

To study how CBD affects bacterial growth kinetics, three 96-well plates were inoculated with overnight bacterial cultures of either S. typhimurium or S. newington at an OD600 ≈ 0.5. These cultures were then treated with varying concentrations of CBD (1.25, 0.125, 0.0125, or 0.00125 μg/mL). The cultures were then incubated at 37 °C with rotary shaking at 121 rpm. Measurements of bacterial density (OD600) were taken hourly for 6 h and at the 12, 24, and 48 h time points using a spectrometer (Molecular Devices SpectraMax ® ABS Plus). This experiment was completed in triplicate [27,60].

4.7. Comparative Bacterial Growth Kinetics in the Prescence of Ampicillin or CBD

To study how ampicillin affects bacterial growth kinetics, three 96-well plates were inoculated with overnight bacterial cultures of either S. typhimurium or S. newington at an OD600 ≈ 0.5. These cultures were then treated with either ampicillin (0.5 μg/mL) or CBD (0.125 μg/mL). The cultures were then incubated at 37 °C with rotary shaking at 121 rpm. Measurements of bacterial density (OD600) were taken hourly for 6 h and at 24 and 48 h time points using a spectrometer (Molecular Devices SpectraMax ® ABS Plus). This experiment was completed in triplicate [27,60].

4.8. SEM Imaging of CBD-Treated S. typhimurium Biofilm

S. typhimurium was grown in 6-well plates (Fisherbrand™, Fisher Scientific, Fair Lawn, NJ, USA) supplemented with LB broth until a biofilm was developed. S. typhimurium biofilm samples were left untreated or treated with CBD at a concentration of 0.125 μg/mL, and the morphology and surface microstructure of the bacteria were observed using scanning electron microscopy. Untreated S. typhimurium served as control. Prior to microscopy, the specimens were dried in a vacuum, and sprayed with gold using an EMS Quorum (EMS 150R ES) ion-sputtering instrument and observed through an Analytical Scanning Electron Microscope (SEM) (JEOL JSM-6010LA, Tokyo, Japan) installed with IntouchScope software (JSM-IT200 InTouchScope™ SEM Series, JEOL Solutions, Tokyo, Japan). Sample preparation for SEM was carried out as described by Li et al., 2021 [41] with few modifications. In short, the specimens for SEM were fixed with 10% formaldehyde solution at room temperature for 10 min, washed with PBS solution thrice, and dehydrated serially in 50%, 70%, and 95% absolute ethanol solutions for 10 min each. Finally, the specimens were dried in a vacuum, and sprayed with gold using an EMS Quorum (EMS 150R ES) ion-sputtering instrument and observed through an Analytical Scanning Electron Microscope (SEM) (JEOL JSM-6010LA, Tokyo, Japan) installed with IntouchScope software (JSM-IT200 InTouchScope™ SEM Series, JEOL Solutions, Tokyo, Japan) [61].

4.9. Statistical Analysis

All experiments were performed on independent biological replicates. Statistical significance was determined for control and experimental groups using a paired sample t-test. Data points were excluded if contamination was identified. Statistical analyses were preformed using OriginPro Plus version 2021b (OriginLab Corporation, North Hampton, MA, USA).

5. Conclusions

As this facultative anaerobe continues to cause serious public health problems, the need to investigate Salmonella has received much scientific scrutiny [61,62,63,64]. In this study, we demonstrated the antibacterial activity of CBD against two relevant pathogenic bacteria, S. typhimurium and S. newington. Despite the scarce knowledge of the molecular mechanisms of CBD’s mode of action, we propose that the antibacterial activity might be due to membrane integrity disruption, and this was verified through the utilization of plate assays, fluorescence microscopy, and kinetic studies. These experiments confirmed that CBD has antibacterial activity against our target bacteria. Additionally, our comparative studies showed that CBD has antibacterial activity similar to ampicillin with a MIC roughly one-fifth of ampicillin. We observed the resistance development of S. typhimurium and S. newington against CBD treatment. Resistance development was observed after 48 h. These results suggest that Salmonella resistance to CBD might be conferred through a different mechanism than antibiotic resistance. These results posed the question of CBD–antibiotic co-therapy as a potential novel application. Finally, we observed CBD to be effective against S. typhimurium biofilms. This study further progresses our current knowledge on the effectiveness of CBD as an antibacterial agent and demonstrates the effectiveness of CBD against Gram-negative bacteria, S. typhimurium and S. newington.

Acknowledgments

The authors acknowledge Benjamin Bramlett of Sustainable CBD LLC., Salem, AL for help extracting the CBD. We acknowledge Melissa Boersma of Auburn University for help with gas chromatography. In addition, we acknowledge Derrick Dean and Vida Dennis for their HBCU-RISE grant support. We also acknowledge Alabama State University, C-STEM for supplies and laboratory space. The authors acknowledge receiving funding from the United States Department of Education, Title III- HBGI-RES.

Author Contributions

Conceptualization, J.A.A.; methodology, L.G. and J.A.A.; software, L.G. and J.A.A.; validation, O.S.A. and M.S.-F.; formal analysis, L.G., J.A.A., O.S.A., J.X. and M.S.-F.; investigation, L.G., J.A.A., O.S.A., J.X. and M.S.-F.; resources, J.A.A., O.S.A., R.V. and M.S.-F.; data curation, L.G., J.A.A., O.S.A., J.X. and M.S.-F.; writing—original draft preparation, L.G.; writing—review and editing, L.G., J.A.A., J.X. and O.S.A.; visualization, L.G. and J.A.A.; supervision, J.A.A. and M.S.-F.; project administration, O.S.A. and M.S.-F.; funding acquisition, O.S.A. and M.S.-F. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by United States Department of Education, Title III- HBGI-RES, and National Science Foundation; and HBCU-RISE; grant number: 1646729.

CBD as a Superbug Antibiotic?

June 24, 2019 — Cannabidiol, or CBD, already being researched and used for anxiety, insomnia, epilepsy and pain, may be the next superbug fighter for resistant infections, a new study suggests.

The researchers tested CBD against a wide variety of bacteria, ”including bacteria that have become resistant to the most commonly used antibiotics,” says Mark Blaskovich, PhD, senior research officer at the Centre for Superbug Solutions at the Institute for Molecular Bioscience at the University of Queensland in Australia.

The development is important, as antibiotic resistance is reaching dangerously high levels, according to the World Health Organization.

What the Research Shows

CBD is a non-psychoactive compound taken from cannabis and hemp; it does not produce the high that regular marijuana does. To date, the FDA has only approved CBD for treating rare and severe forms of seizure, although it is promoted for many other health benefits.

Blaskovich presented the research Sunday at the American Society for Microbiology annual meeting. The research includes work in test tubes and animal models. Research presented at meetings should be viewed as preliminary until published in a peer-reviewed medical journal.

“The first thing we looked at is CBD’s ability to kill bacteria,” he says. “In every case, CBD had a very similar potency to that of common antibiotics.”

The researchers tested the CBD against some strains of staphylococcus, which cause skin infections, and streptococcus, which cause strep throat.

They compared how effective CBD was compared to common antibiotics, such as vancomycin and daptomycin. “We looked at how quickly the CBD killed the bacteria. It’s quite fast, within 3 hours, which is pretty good. Vancomycin (Vancocin) kills over 6 to 8 hours.”

The CBD also disrupted the biofilm, the layer of ”goop” around bacteria that makes it more difficult for the antibiotic to penetrate and kill.

Finally, the lab studies showed that “CBD is much less likely to cause resistance than the existing antibiotics,” Blaskovich says.

The CBD ”is selective for the type of bacteria,” he says.

He found it effective against gram-positive bacteria but not gram-negative. Gram-positive bacteria cause serious skin infections and pneumonia, among other conditions. Gram-negative bacteria include salmonella (found in undercooked foods) and E. coli (the cause of urinary tract infections, diarrhea, and other ailments), among other bacteria.

In another study, also presented at the meeting, the researchers tested topical CBD to treat a skin infection on mice. It cut the number of bacteria after 48 hours, Blaskovich says, although it did not clear the infection. That research is ongoing.

How It Might Work, Caveats

The researchers can’t say exactly how the CBD may prove to be a superbug infection fighter. “We thought it might work by damaging the outer membrane of the bacteria, to make it leaky,” Blaskovich says. “It doesn’t seem to do that. It might be a completely new mechanism of action.”

He says the research results are promising but in early stages. He also warns people that it’s much too early to self-treat infections with CBD.

The study was funded by Botanix Pharmaceuticals Ltd., which is researching uses of CBD for skin conditions, and the Australian government. Blaskovich is a consultant for Botanix.

Perspective

Brandon Novy, a microbiology researcher at Reed College in Portland, OR, calls the study findings ”very promising,” since the results show the bacteria were not able to form resistance to the CBD, and since the bacteria were not able to form a biofilm.

Both findings are important. “The biofilm is an important part of the whole infection process,” he says. “It helps the bacteria attach [to whatever surface or host] and survive.”

At the same meeting, Novy presented a preliminary study, finding that CBD also looks promising to fight some gram-negative infections.

“It is an important study that deserves to be followed up on,” says Amesh Adalja, MD, an infectious disease doctor and senior scholar at the Johns Hopkins Center for Health Security.

He was not involved in the new study. But he cautions that ”it is important to keep it all in context. I think it is a good thing that people are looking at the use of CBD for infectious uses in a systematic way.”

But the work so far is only in test tubes and animals. Many question remain, such as looking at whether it is toxic, doses, and the best way to deliver the CBD, Adalja says. He, too, cautions against self-treating with CBD for infections.

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Show Sources

Amesh Adalja, MD, infectious disease specialist and senior scholar, Johns Hopkins Center for Health Security.

American Society for Microbiology annual meeting, June 23, 2019, San Francisco.

Mark Blaskovich, PhD, senior research officer, Centre for Superbug Solutions, Institute for Molecular Bioscience, University of Queensland, Australia.

Bacterial Infections and CBD Oil: An Alternative to Antibiotics?

CBD oil is gaining ground as an effective treatment for a number of common ailments. Research continues into better understanding all the ways CBD oil can potentially benefit human health. One of the most significant developments in the use of CBD oil as a medicinal supplement has been in the area of bacterial infections.

According to the World Health Organization (WHO) , “ antibiotic resistance is one of the biggest threats to global health, world security, and development… ” Antibiotic resistance is far from being a modern-day issue as bacteria have been building resistance to treatment methods even before the discovery of penicillin in 1928.

Some argue the widespread use of antibiotics has sped up bacterias’ ability to resist manmade medications, with the Centers for Disease Control (CDC) reporting that approximately two million Americans per year are now being infected with antibiotic-resistant bacteria.

Luckily, CBD oil may be able to help.

Benefits of Taking CBD Oil for Bacterial Infections

Antibiotics indiscriminately attack both good and bad bacteria in your body, and can essentially wipe the slate clean. The good bacteria reside mostly in your gut, aiding in digestion—and possibly more. Recently, studies have been finding links between the presence of good gut bacteria and a person’s mood , suggesting the balance of these good bacteria could affect other parts and functions of your body.

Because good bacteria contribute to overall health, and because harmful, antibiotic-resistant bacteria are steadily on the rise, it’s important to consider whether you really need to take antibiotics when you’re feeling sick, or if you should allow your immune system to fight off the invaders on its own—or with the help of CBD oil.

If you have a mild to moderate bacterial infection that doesn’t warrant a course of antibiotics, or if you have a resistant strain of bacteria in your system, it may be worth considering CBD oil.

CBD oil shows promise as an effective bacteria fighter and is covered in ‘Effectiveness of CBD Oil for Bacterial Infections’ in the section below. It also won’t wipe out your good bacteria or produce some of the side effects (nausea, vomiting, stomach pain) associated with common antibiotics.

Effectiveness of CBD Oil for Bacterial Infections

CBD has been found to possess antimicrobial and anti-inflammatory properties . While the impact of CBD oil on inflammation has been an area of intense research, the last significant study on its use against bacteria was conducted in 2008. Though a bit older, this groundbreaking study helps point to the legitimacy of using CBD oil as an alternative to antibiotics.

CBD Oil as an Antimicrobial

A joint study by Italian and English researchers investigated the effect of various cannabinoids , including cannabidiol (CBD) and tetrahydrocannabinol (THC), on pathogenic bacteria. The cannabinoids were tested against 6 strains of the antibiotic-resistant “superbug” MRSA (methicillin-resistant Staphylococcus aureus). Amazingly, all 5 of the major cannabinoids showed “potent activity” against the bacterium. In fact, the results showed that “activity was exceptional against some of these strains, in particular, the multidrug-resistant (MDR) SA-1199B” and also “against EMRSA-15 and EMRSA-16, the major epidemic methicillin-resistant S. aureus strains occurring in U.K. hospitals.”

The study concluded that there were incredibly promising benefits of the use of cannabinoids to treat resistant bacteria, though it could not be determined exactly how the cannabinoids carried out this function.

A more recent 2014 study from the National Yang-Ming University School of Medicine in Taiwan investigated the use of CB2 receptor agonists (like CBD) in rats artificially infected with bacterial peritonitis. It found that using CBD decreased bacterial overgrowth and adhesion.

Because these findings are so promising, particularly with respect to cannabinoids’ effects on MRSA, it’s likely that more research will be done on CBD oil’s antibiotic qualities. This, as CBD and cannabis in general become more widely accepted on a legal level, which may mean that it could be just a short matter of time before CBD oil joins the ranks of revolutionary bacteria busters like penicillin.

CBD Oil as an Anti-Inflammatory

Inflammation plays a key role in helping the body fight a bacterial infection, but if left unchecked, it can also be problematic. Chronic inflammation can cause pain and stiffness in many areas of the body, and is also responsible for a host of internal conditions. Uncontrolled bacterial inflammation promotes apoptosis (cell death) and the generation of harmful free radicals, which has been linked to conditions such as Alzheimer’s Disease.

CBD may reduce inflammation that accompanies a bacterial infection by working both directly and indirectly on endocannabinoid receptors in the endocannabinoid system . These receptors are located throughout the body and central nervous system, with the CB2 cannabinoid receptor thought to play a significant role in the body’s regulation of inflammation. Cannabidiol (CBD) acts on CB2 receptors in a way that’s thought to be a major driving factor behind CBD oil’s anti-inflammatory qualities.

CBD has also been shown to increase levels of the brain neurotransmitter adenosine, which stimulates adenosine receptors. These receptors play a significant role in the reduction of cell inflammation , and when activated, could potentially help reduce effects or symptoms of an inflammatory response to a bacterial infection.

How to Take CBD Oil for Bacterial Infections

Before you start taking CBD oil, it is best to talk to your doctor. While generally considered safe with no significant side effects, CBD oil can interact with some medications, including certain antibiotics . The combination of CBD oil and antibiotics is not necessarily a dangerous one, but it is still important to understand how they function together and how it may impact your body.

By speaking with your doctor first, you can avoid any unwanted effects and establish a safe course of action going forward.

Once you have your physician’s approval to try using CBD oil to treat a bacterial infection, you’ll need to decide on a CBD delivery format. CBD oil is available to purchase in a large range of product types, and what you choose will ultimately come down to your personal preference and the types of symptoms you have.

For example, if a bacterial infection is affecting your skin, a CBD topical (lotion, cream, salve) will be the most direct way of addressing the issue. You can directly target the site of infection and let the CBD go to work for hours.

If the infection is wider spread, a more general delivery method like CBD oil drops or capsules may be a better choice. There is no shortage of options on the market, so do not be afraid to experiment in order to find what works best for you.

Dosage of CBD Oil for Bacterial Infections

Unfortunately, there is no universal dose of CBD oil for bacterial infections. Because CBD’s effects can depend on a number of factors like age, weight, and metabolism speed, it can be hard to come up with a single amount that works for everyone.

As a starting point, it is best to begin with the lowest recommended dosage on the information sheet included with a CBD oil product. You can also speak to a naturopathic doctor for a dosage recommendation that is specific to your condition and needs.

If you’re looking for further, more general guidance, we at CBD Oil Review have analyzed hundreds of products and have come up with a standard serving suggestion:

The CBD Oil Review Serving Standard is 25 mg of CBD, taken twice daily.

If you are not getting results from this amount, we recommend increasing the serving amount by 25 mg every 3 to 4 weeks until you attain relief.

For more information on dosages and some useful tips for getting it right, check out our guide here .

The Best CBD Oil for Bacterial Infections

The best CBD oil for bacterial infections will be the one that works best for you. While that may seem like a frustrating non-answer, there are some things to consider that will help narrow down your options.

First, do not be afraid to try a few different brands and products. CBD oil will not cause you to become “high” or “intoxicated,” nor cause serious side effects , so experimenting with different options presents very little risk to your mental and physical health.

You should also understand the laws in your state around cannabis and CBD oil . While the vast majority of CBD oil products on the market will follow the federal guidelines laid out in the 2018 Farm Bill , namely that CBD oil should contain 0.3% THC or less, some products may contain more. If you live in a state with legal access to recreational and medicinal cannabis, this will be less of a concern. But for states with more restrictive access, you’ll want to check and verify the THC content before making a purchase.

Understanding the law will help you narrow things down between a full-spectrum CBD oil, broad-spectrum CBD oil, or CBD oil isolate. A full-spectrum CBD oil is one that contains all of the cannabinoids, terpenes, flavonoids, and fatty acids found in the whole hemp plant, including the mind-altering compound THC. A broad-spectrum CBD oil contains everything a full-spectrum product contains except THC. A CBD oil isolate is just CBD without any of the other compounds of the hemp plant.

Most full-spectrum products will be in line with federal laws, meaning they should not contain THC in amounts that will make you feel “high.” However, there will always be outliers, which is why doing your research is important.

While researching CBD products to buy, look for the following to ensure you’re getting the best, high-quality CBD oil for bacterial infections:

  • Ingredients . Typically, the best CBD oils will have the fewest ingredients. You will want a CBD oil that contains CBD, a carrier oil like MCT oil or hemp seed oil, and other natural ingredients. Avoid anything that contains chemicals or artificial additives.
  • Product sourcing . Look for brands that source products from organically grown hemp, which is more agriculturally sustainable and free of harmful pesticides and herbicides.
  • Certificates of Analysis (CoA) . Most reputable brands will send out their CBD products for independent lab testing to ensure quality and potency. The lab results from these tests are then posted in a Certificate of Analysis. This analysis will tell you what hemp compounds (cannabinoids, terpenes, etc.) are in the CBD product as well as any harmful heavy metals or chemicals that may remain after the extraction process . This is also where you can verify the accuracy of the CBD amount claimed on the packaging, as well as check the THC content.

While research on the uses of CBD oil for bacterial infections is ongoing, it appears that it could present an effective and beneficial alternative or supplement to traditional antibiotics.

If you found this topic interesting, you may also enjoy:

Sources:

  1. (2020) Antibiotic resistance – World Health Organization https://www.who.int/news-room/fact-sheets/detail/antibiotic-resistance
  2. (2021) Biggest Threats and Data – Centers for Disease Control and Prevention https://www.cdc.gov/drugresistance/biggest-threats.html?CDC_AA_refVal=https%3A%2F%2Fwww.cdc.gov%2Fdrugresistance%2Fbiggest_threats.html
  3. Elizabeth Pennisi (2019) Evidence mounts that gut bacteria can influence mood, prevent depression – American Association for the Advancement of Science https://www.sciencemag.org/news/2019/02/evidence-mounts-gut-bacteria-can-influence-mood-prevent-depression
  4. Giovanni Appendino et al. (2008) Antibacterial Cannabinoids from Cannabis sativa: A Structure−Activity Study – The American Chemical Society and American Society of Pharmacognosy https://pubs.acs.org/doi/10.1021/np8002673
  5. Ying-Ying Yang et al. (2014) Long-term cannabinoid type 2 receptor agonist therapy decreases bacterial translocation in rats with cirrhosis and ascites – National Library of Medicine https://pubmed.ncbi.nlm.nih.gov/24953022/
  6. Fehmida Bibi et al. Link between chronic bacterial inflammation and Alzheimer disease – National Library of Medicine https://pubmed.ncbi.nlm.nih.gov/25230225/
  7. A Thomas et al. (2007) Cannabidiol displays unexpectedly high potency as an antagonist of CB1 and CB2 receptor agonists in vitro – National Library of Medicine https://pubmed.ncbi.nlm.nih.gov/17245363/
  8. Clementino Ibeas Bih et al. (2015) Molecular Targets of Cannabidiol in Neurological Disorders – National Center for Biotechnology Information https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4604182/
  9. 2018 Farm Bill – U.S. Department of Agriculture https://www.farmers.gov/working-with-us/farm-bill

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