Short-Term Storage of Rooster Ejaculates: Sperm Quality and Bacterial Profile Differences in Selected Commercial Extenders.

Bacterial contamination of semen has become an important contributor to the reduced shelf life of insemination doses in the poultry industry, which is why antibiotics (ATBs) are an important component of semen extenders. Due to a global rise in antimicrobial resistance, the aim of this study was to assess the efficiency of selected commercially available semen extenders to prevent possible bacterial contamination of rooster ejaculates. Two selected extenders free from or containing 31.2 µg/mL kanamycin (KAN) were used to process semen samples from 63 healthy Lohmann Brown roosters. Phosphate-buffered saline without ATBs was used as a control. The extended samples were stored at 4 °C for 24 h. Sperm motility, viability, mitochondrial activity, DNA integrity and the oxidative profile of each extended sample were assessed following 2 h and 24 h of storage. Furthermore, selective media were used to quantify the bacterial load and specific bacterial species were identified with matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. The results indicate that semen extenders enriched with KAN ensured a significantly higher preservation of sperm quality in comparison to their KAN-free counterparts. Bacterial load was significantly decreased in diluents supplemented with ATBs (p ≤ 0.001); however, KAN alone was not effective enough to eradicate all bacteria since several Escherichia coli, Enterococcus faecalis, Enterococcus faecium and Micrococcus luteus were retrieved from samples extended in KAN-supplemented commercial extenders. As such, we may suggest that more focus should be devoted to the selection of an optimal combination and dose of antibiotics for poultry extenders, which should be accompanied by a more frequent bacteriological screening of native as well as extended poultry semen.

Salvia sclarea Essential Oil Chemical Composition and Biological Activities.

Salvia sclarea essential oil (SSEO) has a long tradition in the food, cosmetic, and perfume industries. The present study aimed to analyze the chemical composition of SSEO, its antioxidant activity, antimicrobial activity in vitro and in situ, antibiofilm, and insecticidal activity. Besides that, in this study, we have evaluated the antimicrobial activity of SSEO constituent (E)-caryophyllene and standard antibiotic meropenem. Identification of volatile constituents was performed by using gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS) techniques. Results obtained indicated that the main constituents of SSEO were linalool acetate (49.1%) and linalool (20.6%), followed by (E)-caryophyllene (5.1%), p-cimene (4.9%), a-terpineol (4.9%), and geranyl acetate (4.4%). Antioxidant activity was determined as low by the means of neutralization of the DDPH radical and ABTS radical cation. The SSEO was able to neutralize the DPPH radical to an extent of 11.76 ± 1.34%, while its ability to decolorize the ABTS radical cation was determined at 29.70 ± 1.45%. Preliminary results of antimicrobial activity were obtained with the disc diffusion method, while further results were obtained by broth microdilution and the vapor phase method. Overall, the results of antimicrobial testing of SSEO, (E)-caryophyllene, and meropenem, were moderate. However, the lowest MIC values, determined in the range of 0.22-0.75 µg/mL for MIC50 and 0.39-0.89 µg/mL for MIC90, were observed for (E)-caryophyllene. The antimicrobial activity of the vapor phase of SSEO (towards microorganisms growing on potato) was significantly stronger than that of the contact application. Biofilm analysis using the MALDI TOF MS Biotyper showed changes in the protein profile of Pseudomonas fluorescens that showed the efficiency of SSEO in inhibiting biofilm formation on stainless-steel and plastic surfaces. The insecticidal potential of SSEO against Oxycarenus lavatera was also demonstrated, and results show that the highest concentration was the most effective, showing insecticidal activity of 66.66%. The results obtained in this study indicate the potential application of SSEO as a biofilm control agent, in the shelf-life extension and storage of potatoes, and as an insecticidal agent.

Chemical Composition and Biological Activities of Eucalyptus globulus Essential Oil.

Eucalyptus globulus essential oil (EGEO) is considered as a potential source of bioactive compounds with significant biological activity. The aim of this study was to analyze the chemical composition of EGEO, in vitro and in situ antimicrobial activity, antibiofilm activity, antioxidant activity, and insecticidal activity. The chemical composition was identified using gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS). The main components of EGEO were 1,8-cineole (63.1%), p-cimene (7.7%), a-pinene (7.3%), and a-limonene (6.9%). Up to 99.2% of monoterpenes were present. The antioxidant potential of essential oil and results indicate that 10 µL of this sample can neutralize 55.44 ± 0.99% of ABTS•+, which is equivalent to 3.22 ± 0.01 TEAC. Antimicrobial activity was determined via two methods: disk diffusion and minimum inhibitory concentration. The best antimicrobial activity was shown against C. albicans (14.00 ± 1.00 mm) and microscopic fungi (11.00 ± 0.00 mm-12.33 ± 0.58 mm). The minimum inhibitory concentration showed the best results against C. tropicalis (MIC 50 2.93 µL/mL, MIC 90 3.17 µL/mL). The antibiofilm activity of EGEO against biofilm-forming P. flourescens was also confirmed in this study. The antimicrobial activity in situ, i.e., in the vapor phase, was significantly stronger than in the contact application. Insecticidal activity was also tested and at concentrations of 100%, 50%, and 25%; the EGEO killed 100% of O. lavaterae individuals. EGEO was comprehensively investigated in this study and information regarding the biological activities and chemical composition of the essential oil of Eucalyptus globulus was expanded.

Biological Activity of Cupressus sempervirens Essential Oil.

The aim of this study was to evaluate the antioxidant, antibiofilm, antimicrobial (in situ and in vitro), insecticidal, and antiproliferative activity of Cupressus sempervirens essential oil (CSEO) obtained from the plant leaf. The identification of the constituents contained in CSEO was also intended by using GC and GC/MS analysis. The chemical composition revealed that this sample was dominated by monoterpene hydrocarbons α-pinene, and δ-3-carene. Free radical scavenging ability, performed by using DPPH and ABTS assays, was evaluated as strong. Higher antibacterial efficacy was demonstrated for the agar diffusion method compared to the disk diffusion method. The antifungal activity of CSEO was moderate. When the minimum inhibitory concentrations of filamentous microscopic fungi were determined, we observed the efficacy depending on the concentration used, except for B. cinerea where the efficacy of lower concentration was more pronounced. The vapor phase effect was more pronounced at lower concentrations in most cases. Antibiofilm effect against Salmonella enterica was demonstrated. The relatively strong insecticidal activity was demonstrated with an LC50 value of 21.07% and an LC90 value of 78.21%, making CSEO potentially adequate in the control of agricultural insect pests. Results of cell viability testing showed no effects on the normal MRC-5 cell line, and antiproliferative effects towards MDA-MB-231, HCT-116, JEG-3, and K562 cells, whereas K562 cells were the most sensitive. Based on our results, CSEO could be a suitable alternative against different types of microorganisms as well as suitable for the control of biofilms. Due to its insecticidal properties, it could be used in the control of agricultural insect pests.

Seminal Bacterioflora of Two Rooster Lines: Characterization, Antibiotic Resistance Patterns and Possible Impact on Semen Quality.

This study aimed to characterize the bacterial profiles and their association with selected semen quality traits among two chicken breeds. Thirty Lohmann Brown and thirty ROSS 308 roosters were selected for semen quality estimation, including sperm motility, membrane and acrosome integrity, mitochondrial activity, and DNA fragmentation. The oxidative profile of the semen, including the production of reactive oxygen species (ROS), antioxidant capacity, protein, and lipid oxidation, were assessed as well. Moreover, the levels of pro-inflammatory cytokines, including tumor necrosis factor alpha (TNF-α), interleukins 1 and 6 (IL-1, IL-6) and C-reactive protein, as well as the concentrations of selected antibacterial proteins (cathelicidin, ß-defensin and lysozyme) in the seminal plasma were evaluated with the enzyme-linked immunosorbent assay. The prevailing bacterial genera identified by the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry were Citrobacter spp., Enterococcus spp., Escherichia spp. and Staphylococcus spp. While the bacterial load was significantly higher in the ROSS 308 line (p < 0.05), a higher number of potentially uropathogenic bacteria was found in the Lohmann Brown roosters. Antimicrobial susceptibility tests revealed a substantial resistance of randomly selected bacterial strains, particularly to ampicillin, tetracycline, chloramphenicol, and tobramycin. Furthermore, Lohmann Brown ejaculates containing an increased proportion of Escherichia coli presented with significantly (p < 0.05) elevated levels of TNF-α and IL-6, as well as ROS overproduction and lipid peroxidation. Inversely, significantly (p < 0.05) higher levels of ß-defensin and lysozyme were found in the semen collected from the ROSS 308 roosters, which was characterized by a higher quality in comparison to the Lohmann Brown roosters. In conclusion, we emphasize the criticality of bacteriospermia in the poultry industry and highlight the need to include a more complex microbiological screening of semen samples designated for artificial insemination.

Chemical Composition and Biological Activity of Tanacetum balsamita Essential Oils Obtained from Different Plant Organs.

The aim of this study is to evaluate the chemical composition of Tanacetum balsamita L. essential oils (EOs) obtained from different plant organs, flowers (FEO), leaves (LEO), and stems (SEO), as well as to assess their biological properties. The results obtained by using GC and GC/MS analysis indicate that this plant belongs to the carvone chemotype. Moreover, we examined the oil's antimicrobial and antitumor potential. Antimicrobial effects were determined using minimum inhibitory concentrations assay and the vapor phase method. Obtained results indicate better antimicrobial activity of investigated EO samples compared to the commercially available antibiotics. On the treatment with FEO, Y. enterocolitica and H. influenzae showed high sensitivity, while treatment with LEO and SEO showed the highest effects against S. aureus. The vapor phase method, as an in situ antibacterial analysis, was performed using LEO. Obtained results showed that this EO has significant activity toward S. pneumoniae in the apple and carrot models, L. monocytogenes in the pear model, and Y. enterocolitica in the white radish model. The potential antitumor mechanisms of FEO, LEO, and SEO were determined by the means of cell viability, redox potential, and migratory capacity in the MDA-MB-231 and MDA-MB-468 cell lines. The results show that these EOs exert antiviability potential in a time- and dose-dependent manner. Moreover, treatments with these EOs decreased the levels of superoxide anion radical and increased the levels of nitric oxide in both tested cell lines. The results regarding total and reduced glutathione revealed, overall, an increase in the levels of total glutathione and a decrease in the levels of reduced glutathione, indicating strong antioxidative potential in tested cancer cells in response to the prooxidative effects of the tested EOs. The tested EOs also exerted a drop in migratory capacity, which indicates that they can be potentially used as chemotherapeutic agents.

GC, GC/MS Analysis, and Biological Effects of Essential Oils from Thymus mastchina and Elettaria cardamomum.

Spanish marjoram (Thymus mastichina) and cardamom (Elettaria cardamomum) are traditional aromatic plants with which several pharmacological properties have been associated. In this study, the volatile composition, antioxidative and antimigratory effects on human breast cancer (MDA-MB-468 cell line), antimicrobial activity, and antibiofilm effect were evaluated. Results obtained via treatment of human breast cancer cells generally indicated an inhibitory effect of both essential oils (EOs) on cell viability (after long-term treatment) and antioxidative potential, as well as the reduction of nitric oxide levels. Antimigratory effects were revealed, suggesting that these EOs could possess significant antimetastatic properties and stop tumor progression and growth. The antimicrobial activities of both EOs were determined using the disc diffusion method and minimal inhibition concentration, while antibiofilm activity was evaluated by means of mass spectrometry. The best antimicrobial effects of T. mastichina EO were found against the yeast Candida glabrata and the G+ bacterium Listeria monocytogenes using the disc diffusion and minimal inhibitory concentration methods. E. cardamomum EO was found to be most effective against Pseudomas fluorescens biofilm using both methods. Similarly, better effects of this oil were observed on G- compared to G+ bacterial strains. Our study confirms that T. mastichina and E. cardamomum EOs act to change the protein structure of older P. fluorescens biofilms. The results underline the potential use of these EOs in manufactured products, such as foodstuffs, cosmetics, and pharmaceuticals.

Antimicrobial and Antioxidant Activity of Different Honey Samples from Beekeepers and Commercial Producers.

Honey contains compounds with antioxidant and antibacterial capacities, such as phenolic compounds and carotenoids. The current analysis evaluates the antioxidant and antibacterial activity of 100 honey samples from beekeepers from Slovakia and commercially purchased ones. Honey samples were diluted to 50%, 25%, 12.5%, and 6.25% concentrations. The antimicrobial activity of honey samples was evaluated against three Gram-positive, three Gram-negative bacteria, and four Candida spp. by well diffusion method. The highest antimicrobial effect of all honey concentrations was expressed as the size of the inhibition zone and was found against Pseudomonas aeruginosa among Gram-negative bacteria, Bacillus subtilis among Gram-positive bacteria, and Candida tropicalis among yeasts. Antibiotics used in the study showed the highest antimicrobial effect compared to all concentrations of honey samples. Slovakian honey from beekeepers and commercial honey samples from the Slovak market showed variable inhibitory effectiveness against microorganisms. The honey concentration of 50% was found the most effective. Lower concentrations of honey exhibited no effect against yeasts. The best antioxidant activity was found in a sample of buckwheat honey yielding 70.83% of DPPH inhibition and 2373.85 µg/g TEAC. Overall, better antioxidant activity was evaluated in honeydew honey.

Determination of Antioxidant, Antimicrobial Activity, Heavy Metals and Elements Content of Seaweed Extracts.

The aim of the research was to determine the antioxidant and antimicrobial activity, determination of chemical elements and heavy metals in seaweed extracts of wakame, arame, dulse, laminaria, kombu, and hijiki. Antioxidant activity was determined by DPPH method and the activity ranged from 0.00 to 2641.34 TEAC. The highest antioxidant activity was observed in kombu (2641.34 TEAC) and arame (2457.5 TEAC). Antimicrobial activity was analyzed by disk diffusion method and MIC method. Three G+ bacteria (Staphylococcus aureus, Enterococcus faecalis, Bacillus subtilis), three G- bacteria (Salmonella enterica, Pseudomonas aeruginosa, Yersinia enterocolitica), and four yeasts (Candida tropicalis, C. krusei, C. glabrata, C. albicans) were used as model organisms. The size of inhibition zones ranged from 0.00 to 8.67 mm. The minimum inhibitory concentrations of the selected seaweeds ranged from MIC50 98.46 (MIC90 100.25) to MIC50 3.43 µL/mL (MIC90 5.26 µL/mL). The content of selected elements was determined in seaweed samples by ICP-OES. The chemical composition of the algae showed differences between species and the presence of heavy metals. Arsenic, cadmium, and aluminum were confirmed. All seaweed samples contained arsenic ranging from 6.6546 to 76.48 mg/kg. Further investigation of seaweeds is needed to identify the active substances present in the algae that are responsible for antioxidant and antimicrobial activity. This study was carried out to evaluate the antimicrobial and antioxidant activity of extracts from five commonly consumed seaweeds for their ability to inhibit selected microorganisms and to determine the health risk due to heavy metals content. Our study contributes to the evidence that seaweeds have antimicrobial and antioxidant activity and seaweed extracts have for pharmacological applications.

Role of Litsea cubeba Essential Oil in Agricultural Products Safety: Antioxidant and Antimicrobial Applications.

The essential oil from Litsea cubeba (LCEO) has good antioxidant, antimicrobial, anti-insect properties, which gives it the potential for use as a natural additive to food resources and food products in order to prevent spoilage and extend shelf life. In this study the biological activity related to food preservation was observed. The main volatile organic compounds were geranial (39.4%), neral (29.5%), and limonene (14.3%). Antioxidant activity was 30.9%, which was equal to 167.94 µg of Trolox per mL of sample. Antimicrobial activity showed the strongest inhibition against Serratia marcescens by disk diffusion method and minimum inhibitory concentrations MIC 50 and MIC 90 were the lowest for Micrococcus luteus with values 1.46 and 3.52 µL/mL, respectively. Antimicrobial activity of the LCEO vapor phase showed strong inhibition of microorganisms on apples, pears, potatoes, and kohlrabies. Over 50% of gram-positive and gram-negative bacteria and yeasts were inhibited by a concentration of 500 µL/mL. The inhibition of microorganisms was concentration dependent. Anti-insect activity was also strong, with 100% lethality of Pyrrhocoris apterus at a concentration of 25%. These results suggest that LCEO could be potentially used as a food preservative.

Assessment of Ocimum basilicum Essential Oil Anti-Insect Activity and Antimicrobial Protection in Fruit and Vegetable Quality.

Basil (Ocimum basilicum) is a commonly used herb; it also contains essential oils and other valuable compounds. The basil oil obtained has a pleasant aroma, but also a broad spectrum of biological activity. This work reports on the chemical composition, antioxidant, antimicrobial and anti-insect activity in vitro and in situ of Ocimum basilicum essential oil (OBEO) obtained by steam distillation of fresh flowering plants. Gas chromatography-mass spectrometry, DPPH, agar and disc diffusion and vapor phase methods were used to analyze the OBEO properties. The analysis of the chemical composition of OBEO showed that its main components were methyl chavicol (88.6%), 1,8-cineole (4.2%) and α-trans-bergamotene (1.7%). A strong antioxidant effect was demonstrated at the level of 77.3%. The analysis of antimicrobial properties showed that OBEO exerts variable strength of inhibiting activity against various groups of microorganisms. The growth inhibition zones ranged from 9.67 to 15.33 mm in Gram-positive (G+) and Gram-negative (G-) bacteria and from 5.33 to 7.33 mm in yeast. The lowest measured minimal inhibition concentration (MIC) was 3.21 µL/mL against Gram-negative Azotobacter chrococcum and Gram-positive Micrococcus luteus. The antimicrobial activity of in situ vapor phase of OBEO was also confirmed on apples, pears, potatoes and kohlrabi. The highest insecticidal activity against Pyrrhocorisapterus, observed at the concentration of 100%, caused the death of 80% of individuals. Due to its broad spectrum of activity, OBEO seems an ideal candidate for preserving fruit and vegetables.

Chemical and Biological Characterization of Melaleuca alternifolia Essential Oil.

The essential oil of Melaleuca alternifolia, commonly known as tea tree oil, has many beneficial properties due to its bioactive compounds. The aim of this research was to characterize the tea tree essential oil (TTEO) from Slovakia and its biological properties, which are specific to the chemical composition of essential oil. Gas chromatography/mass spectroscopy revealed that terpinen-4-ol was dominant with a content of 40.3%. γ-Terpinene, 1,8-cineole, and p-cymene were identified in contents of 11.7%, 7.0%, and 6.2%, respectively. Antioxidant activity was determined at 41.6% radical inhibition, which was equivalent to 447 µg Trolox to 1 mL sample. Antimicrobial activity was observed by the disk diffusion method against Gram-positive (G+), Gram-negative (G-) bacteria and against yeasts, where the best antimicrobial activity was against Enterococcus faecalis and Candida albicans with an inhibition zone of 10.67 mm. The minimum inhibitory concentration showed better susceptibility by G+ and G- planktonic cells, while yeast species and biofilm-forming bacteria strains were more resistant. Antibiofilm activity was observed against Pseudomonas fluorescens and Salmonella enterica by MALDI-TOF, where degradation of the protein spectra after the addition of essential oil was obtained. Good biological properties of tea tree essential oil allow its use in the food industry or in medicine as an antioxidant and antimicrobial agent.

Chemical Composition, Antioxidant, In Vitro and In Situ Antimicrobial, Antibiofilm, and Anti-Insect Activity of Cedar atlantica Essential Oil.

The present study was designed to evaluate commercial cedar essential oil (CEO), obtained by hydrodistillation from cedar wood, in relationship to its chemical composition and antioxidant, in vitro and in situ antimicrobial, antibiofilm, and anti-insect activity. For these purposes, gas chromatography-mass spectrometry, DPPH radical-scavenging assay, agar and disc diffusion, and vapor phase methods were used. The results from the volatile profile determination showed that δ-cadinene (36.3%), (Z)-ß-farnesene (13.8%), viridiflorol (7.3%), and himachala-2,4-diene (5.4%) were the major components of the EO chemical constitution. Based on the obtained results, a strong antioxidant effect (81.1%) of the CEO was found. CEO is characterized by diversified antimicrobial activity, and the zones of inhibition ranged from 7.33 to 21.36 mm in gram-positive and gram-negative bacteria, and from 5.44 to 13.67 mm in yeasts and fungi. The lowest values of minimal inhibition concentration (MIC) were noted against gram-positive Micrococcus luteus (7.46 µL/mL) and against yeast Candida krusei (9.46 µL/mL). It seems that the vapor phase of CEO can inhibit the growth of the microscopic filamentous fungi of the genus Penicillium according to in situ antifungal analysis on bread, carrots, and celery. This finding confirms the impact of CEO on the change in the protein structure of older biofilms of Pseudomonas fluorescens and Salmonella enterica subsp. enterica. Insecticidal activity of a vapor phase has also been demonstrated against Pyrrhocoris apterus. CEO showed various advantages on antimicrobial activity, and it is an ideal substitute for food safety.

Curcumin Attenuates Damage to Rooster Spermatozoa Exposed to Selected Uropathogens.

Artificial insemination, as an essential pillar of the modern poultry industry, primarily depends on the quality of semen collected from stud roosters. Since the collection and storage of ejaculates is not a sterile process, antimicrobial agents have become essential supplements to semen extenders. While the use of traditional antibiotics has been challenged because of rising bacterial resistance, natural biomolecules represent an appealing alternative because of their antibacterial and antioxidant properties. As such, this study strived to compare the effects of 50 µmol/L curcumin (CUR) with 31.2 µg/mL kanamycin (KAN) as a conventional antibiotic on rooster sperm quality in the presence of Salmonella enterica, Escherichia coli and Pseudomonas aeruginosa. Changes in sperm structural integrity and functional activity were monitored at 2 and 24 h of culture. Computer-assisted semen analysis revealed significant sperm motility preservation following treatment with KAN, particularly in the case of Salmonella enterica and Pseudomonas aeruginosa (p < 0.001) after 24 h. On the other hand, CUR was more effective in opposing ROS overproduction by all bacteria (p < 0.05), as determined by luminol-based luminometry, and maintained sperm mitochondrial activity (p < 0.001 in the case of Salmonella enterica; p < 0.05 with respect to Escherichia coli and Pseudomonas aeruginosa), as assessed by the fluorometric JC-1 assay. The TUNEL assay revealed that CUR readily preserved the DNA integrity of rooster sperm exposed to Salmonella enterica (p < 0.01) and Escherichia coli (p < 0.001). The bacteriological analysis showed higher efficiency of KAN in preventing the growth of all selected bacterial species (p < 0.0001) as opposed to CUR. In conclusion, CUR provided protection to rooster spermatozoa against alterations caused by uropathogens, most likely through its antioxidant activity. Hence, CUR supplementation to poultry semen extenders in combination with properly selected antibacterial substances may become an interesting strategy in the management of bacterial contamination during semen storage.

Staphylococcus-Induced Bacteriospermia In Vitro: Consequences on the Bovine Spermatozoa Quality, Extracellular Calcium and Magnesium Content.

Bacterial contamination of bovine ejaculates intended for artificial insemination may be reflected in a significant economic loss due to unsuccessful fertilization as well as health issues of the recipients. The Staphylococcus genus represents a large part of bacteriocenosis of bovine ejaculates. Therefore, this study aims to get a closer look on the effects of Staphylococcus-induced bacteriospermia under in vitro conditions on bovine sperm quality. Prior to inducing bacteriospermia, spermatozoa were separated from each ejaculate using Percoll® Plus gradient medium in order to limit the effects only to the selected bacterial species. Seven Staphylococcus species previously isolated from bovine semen were used for our experiments at a turbidity of 0.5 McFarland (equivalent to 1.5 × 108 colony-forming units per mL). The contaminated semen samples were incubated at 37 °C and at times of 0, 2, and 4 h, motility, mitochondrial membrane potential, reactive oxygen species (ROS) generation, sperm DNA fragmentation, and magnesium (Mg) and calcium (Ca) extracellular concentration were analyzed and compared with the control group (uncontaminated). The results showed no significant changes at the initial measurement. However, significant adverse effects were observed after 2 h and 4 h of incubation. Most notably, the presence of S. aureus, S. warneri, S. kloosii, and S. cohnii caused a significantly increased ROS production, leading to sperm DNA fragmentation, changes in the mitochondrial membrane potential, and a decreased sperm motility. Furthermore, the presence of Staphylococcus species led to lower extracellular concentrations of Mg and Ca. In conclusion, the overgrowth of Staphylococcus bacteria in bovine semen may contribute to oxidative stress resulting in sperm DNA fragmentation, altered mitochondrial membrane potential, and diminished sperm motility.

Antioxidant and antimicrobial activities of fruit extracts from different fresh chili peppers.

BACKGROUND: The aim of the current study was to assess the antimicrobial and antioxidant potential of ethanol extracts obtained from the fruit of five species of fresh chili pepper, Capsicum (C.) baccatum L. (Aji Amarillo), C. chinense (Fidalgo Roxa), C. annuum (Cherry Chocolate), C. pubescens (Rocoto Orange) and C. frutescens (Peruvian Purple). METHODS: To obtain the ethanol extracts, accelerated solvent extraction (ASE) was applied. DPPH assay was used to determine the antioxidant activity of the extract samples. The disc diffusion method was used to measure antimicrobial activity against nine investigated microorganism species. RESULTS: The tested extract samples exhibited DPPH radical scavenging activities ranging from 0.24 ±0.01 (Peruvian Purple) to 0.72 ±0.02 (Aji Amarillo) mg TEAC∙g–1 dw. The differences between all the varieties were statistically significant (P < 0.05; except for the Cherry Chocolate and Rocoto Orange), and the potential of antioxidant capacity increased in the following manner: Peruvian Purple < Fidalgo Roxa < Rocoto Orange < Cherry Chocolate < Aji Amarillo. The results from the antimicrobial evaluation showed that the Capsicum extracts had no uniform inhibition activity against tested gram-negative, gram-positive bacteria, and yeast. Specifically, Aji Amarillo fruit extract revealed the strongest antimicrobial activity against S. pneumoniae (6.33 ±0.58 mm), followed by Cherry Chocolate against S. pneumoniae (5.33 ±0.58 mm), Rocoto Orange against S. enterica (5.27 ±0.58 mm), Fidalgo Roxa against C. albicans (4.67 ±0.58 mm), and Peruvian Purple against S. pneumoniae (4.57 ±0.58 mm). CONCLUSIONS: Considering these results, Capsicum spp. can be used as a source of novel antioxidant and antimicrobial compounds.

Chemical Composition, In Vitro and In Situ Antimicrobial and Antibiofilm Activities of Syzygium aromaticum (Clove) Essential Oil.

The essential oil of Syzygium (S.) aromaticum (CEO) is known for its good biological activity. The aim of the research was to evaluate in vitro and in situ antimicrobial and antibiofilm activity of the essential oil produced in Slovakia. The main components of CEO were eugenol 82.4% and (E)-caryophyllene 14.0%. The antimicrobial activity was either weak or very strong with inhibition zones ranging from 4.67 to 15.78 mm in gram-positive and gram-negative bacteria and from 8.22 to 18.56 mm in yeasts and fungi. Among the tested bacteria and fungi, the lowest values of MIC were determined for Staphylococcus (S.) aureus and Penicillium (P.) expansum, respectively. The vapor phase of CEO inhibited the growth of the microscopic filamentous fungi of the genus Penicillium when tested in situ on bread. The strongest effect of mycelia inhibition in a bread model was observed against P. expansum at concentrations of 250 and 500 µL/mL. The best antimicrobial activity of CEO in the carrot model was found against P. chrysosenum. Differences between the mass spectra of Bacillus (B.) subtilis biofilms on the tested surfaces (wood, glass) and the control sample were noted from the seventh day of culture. There were some changes in mass spectra of Stenotrophomonas (S.) maltophilia, which were observed in both experimental groups from the fifth day of culture. These findings confirmed the impact of CEO on the protein structure of older biofilms. The findings indicate that, besides being safe and sensorially attractive, S. aromaticum has antimicrobial activity, which makes it a potential substitute for chemical food preservatives.

Thymus vulgaris Essential Oil and Its Biological Activity.

Thymus vulgaris essential oil has potential good biological activity. The aim of the research was to evaluate the biological activity of the T. vulgaris essential oil from the Slovak company. The main components of T. vulgaris essential oil were thymol (48.1%), p-cymene (11.7%), 1,8-cineole (6.7), γ-terpinene (6.1%), and carvacrol (5.5%). The antioxidant activity was 85.2 ± 0.2%, which corresponds to 479.34 ± 1.1 TEAC. The antimicrobial activity was moderate or very strong with inhibition zones from 9.89 to 22.44 mm. The lowest values of MIC were determined against B. subtilis, E. faecalis, and S. aureus. In situ antifungal analysis on bread shows that the vapor phase of T. vulgaris essential oil can inhibit the growth of the microscopic filamentous fungi of the genus Penicillium. The antimicrobial activity against S. marcescens showed 46.78-87.80% inhibition at concentrations 62.5-500 µL/mL. The MALDI TOF MS analyses suggest changes in the protein profile of biofilm forming bacteria P. fluorescens and S. enteritidis after the fifth and the ninth day, respectively. Due to the properties of the T. vulgaris essential oil, it can be used in the food industry as a natural supplement to extend the shelf life of the foods.

Thymus serpyllum Essential Oil and Its Biological Activity as a Modern Food Preserver.

The aim of this study was to analyze the chemical composition and biological and antibiofilm activity of the essential oil (EO) of Thymus serpyllum with the use of a MALDI-TOF MS Biotyper. The main compounds of the EO were thymol, 18.8%; carvacrol, 17.4%; o-cymene, 15.4%; and geraniol, 10.7%. It was found that free-radical scavenging activity was high. The highest antimicrobial activity was observed against Pseudomonas aeruginosa, Salmonella enteritidis, and biofilm-forming bacteria. The changes in the biofilm structure after T. serpyllum EO application confirmed the inhibitory action and the most pronounced effect was observed on Bacillus subtilis biofilm. The antifungal activity of the vapor phase was the most effective against Penicillium crustosum. T. serpyllum should be a suitable alternative to synthetic antioxidants as well as antimicrobials. The EO of T. serpyllum can be used in the vapor phase in the storage of root vegetables as well as a growth inhibitor of Penicillium on bread.

In Vitro Antimicrobial Activity of Lavender, Mint, and Rosemary Essential Oils and the Effect of Their Vapours on Growth of Penicillium spp. in a Bread Model System.

The chemical composition, antioxidant activity, and antimicrobial properties of three commercially available essential oils: rosemary (REO), lavender (LEO), and mint (MEO), were determined in the current study. Our data revealed that the major components of REO, MEO, and LEO were 1,8-cineole (40.4%), menthol (40.1%), and linalool acetate (35.0%), respectively. The highest DPPH radical-scavenging activity was identified in MEO (36.85 ± 0.49%) among the investigated EOs. Regarding antimicrobial activities, we found that LEO had the strongest inhibitory efficiencies against the growth of Pseudomonas aeruginosa and Candida (C.) tropicalis, MEO against Salmonella (S.) enterica, and REO against Staphylococcus (S.) aureus. The strongest antifungal activity was displayed by mint EO, which totally inhibited the growth of Penicillium (P.) expansum and P. crustosum in all concentrations; the growth of P. citrinum was completely suppressed only by the lowest MEO concentration. The lowest minimal inhibitory concentrations (MICs) against S. enterica, S. aureus, and C. krusei were assessed for MEO. In situ analysis on the bread model showed that 125 µL/L of REO exhibited the lowest mycelial growth inhibition (MGI) of P. citrinum, and 500 µL/L of MEO caused the highest MGI of P. crustosum. Our results allow us to make conclusion that the analysed EOs have promising potential for use as innovative agents in the storage of bakery products in order to extend their shelf-life.