Synergistic interactions of essential oil components with antibiotics against multidrug-resistant Corynebacterium striatum.

AIM: In this study, it was aimed to examine the antibacterial activity of the essential oil components (EOCs), carvacrol (CAR), cinnamaldehyde (CIN), thymol (TH), alpha pinene (α-PN), eucalyptol (EU), limonene (LIM), and the antibiotics, linezolid (LZD), vancomycin (VAN), gentamicin (GEN), ciprofloxacin (CIP), clindamycin (CLN), and penicillin (PEN) against 50 multidrug resistant Corynebacterium striatum strains, and the synergistic interactions of CAR and CIN with the antibiotics against 10 randomly selected Coryne. striatum strains to explore synergistic interactions to determine if their combined use could enhance antibiotic activity and potentially reduce resistance. METHODS AND RESULTS: The activity of the EOCs and the antibiotics against Coryne. striatum strains isolated from clinical specimens, was examined by broth microdilution method. The synergistic interactions of the EOCs with the antibiotics against 10 randomly selected Coryne. striatum strains were determined by checkerboard method. EOCs, CIN, and CAR and antibiotics, LZD, VAN, GEN, CIP, and CLN were detected to have antibacterial activity against Coryne. striatum strains alone and either synergistic interactions were observed in combinations of the antibiotics with EOCs. CONCLUSIONS: All Coryne. striatum strains were determined to be susceptible to VAN and LZD and resistant to GEN, PEN, CIP, and CLN. Synergistic interactions were observed in all combinations of antibiotics tested with CAR and CIN.

[Investigation of Cytotoxic and Antileishmanial Activity of Hybrid Silver Nanoparticle Complexes: Potential Drug Candidates against Leishmania Species]. / Hibrit Gümüs Nanoparçacik Komplekslerinin Sitotoksik ve Antilaysmanyal Aktivitesinin Arastirilmasi: Leishmania Türlerine Karsi Potansiyel Ilaç Adaylari.

In recent years, isolation of resistant Leishmania species to drugs in use has made it necessary to search alternative molecules that may be drug candidates. In this study, it was aimed to investigate the cytotoxic and in vitro antileishmanial activity of hybrid silver nanoparticle (AgNP) complexes. In this study, three types of nanoparticles (NPs), oxidized amylose-silver (OA-Ag) NPs, oxidized amylose-curcumin (OA-Cur) NPs and oxidized amylose-curcumin-silver (OA-CurAgNP) nanoparticles were synthesized. The cytotoxic activity of the synthesized nanoparticles was determined against L929 mouse fibroblasts and the in vitro antileishmanial activity was determined against Leishmania tropica, Leishmania infantum and Leishmania donovani isolates by the broth microdilution method. It was observed that the hybrid OA-CurAgNP complex obtained by combining curcumin and silver nanoparticles showed cytotoxic effects against L929 mouse fibroblasts at concentrations of 1074 µg/mL and above. IC50 values expressing the antileishmanial activity of the hybrid OA-CurAgNP complex against L.tropica, L.infantum and L.donovani isolates, were found to vary between 95-121 µg/mL, 202-330 µg/mL and 210-254 µg/mL, respectively. Resistance development has emerged as a major challenge in the treatment of leishmaniasis in recent times. Metallic nanoparticles are considered excellent candidates for medical applications due to their chemical and physical properties, as well as their prolonged circulation in the body. The current drugs used for leishmaniasis treatment are highly toxic, while nanoparticles offer advantages such as low toxicity and easy cellular uptake due to their nanoscale dimensions. The identification of strong efficacy in these particles may contribute scientific evidence for their potential use in leishmaniasis treatment. Therefore, the therapeutical value of OA-CurAgNP complex alone in combination with existing drugs should be examined.

[Comparison of Conventional Methods with Molecular Methods in the Diagnosis of Trichomonas vaginalis and Investigation of Metronidazole Resistance]. / Trichomonas vaginalis'in Tanisinda Konvansiyonel Yöntemlerin Moleküler Yöntemlerle Karsilastirilmasi ve Metronidazol Direncinin Arastirilmasi.

Trichomoniasis is a sexually transmitted parasitic infection caused by Trichomonas vaginalis. In the diagnosis of trichomoniasis, direct microscopy (DM) is preferred, which is a cheap and fast method, although it has low sensitivity. Culture methods, which are accepted as the gold standard, can only be applied in certain centers due to the need for experienced personnel and the ability to get results within 2-7 days, despite their high sensitivity. In this study, it was aimed to compare conventional microscopic and culture methods used in the routine diagnosis of T.vaginalis with polymerase chain reaction (PCR) method and to investigate ntr4 and/or ntr6 gene polymorphism in the nitroreductase gene region, which are thought to be associated with metronidazole resistance in T.vaginalis strains isolated from clinical specimens. Vaginal swab specimens were collected from the posterior fornix of the vagina with two sterile ecuvion sticks during the gynecological examinations of 200 patients who applied to the Balikesir University Health Practice and Research Hospital, Obstetrics and Gynecology Polyclinic between March 2019 and August 2021. The first swab sample was used for direct microscopic examination, Giemsa staining and conventional PCR analysis, while the second swab specimen was taken into trypticase-yeast-extract-maltose (TYM) medium for T.vaginalis culture and followed for eight days at 37 °C. All specimens were screened for the presence of T.vaginalis using primers specific to the ß-tubulin (btub1) gene region and clinical isolates grown in TYM medium were examined for metronidazole resistance using primers specific for the nitroreductase gene region by using conventional PCR. Drug resistance test was also performed for the isolates in which polymorphism associated with metronidazole resistance was detected. Eight (4%) of 200 patient specimens were found positive by both culture/staining and PCR methods. The mean age of the patients included in the study was 39.9, while the mean age of the patients with positive T.vaginalis was 41.8. The most common clinical findings in the patients were foul-smelling vaginal discharge (36%), groin pain (21%), vaginal itching (19%), and burning sensation during urination (18%). In three out of eight T.vaginalis strains isolated from clinical samples, the presence of polymorphism in the ntr6 gene, which is thought to be associated with metronidazole resistance, was demonstrated by PCR. It was observed that three isolates with ntr6 gene polymorphism were phenotypically resistant to metronidazole (MLK= 390 µM). In this study, the fact that three of eight clinical isolates that were resistant to metronidazole by the broth microdilution method and as well as showing ntr6 gene polymorphism supported the thesis that there might be a close relationship between metronidazole resistance and ntr6 gene polymorphism. As a result, the use of culture and molecular methods in the diagnosis of T.vaginalis, in addition to the microscopy method, may contribute to a more accurate laboratory diagnosis of the agent, to detect metronidazole resistance molecularly and phenotypically, to determine metronidazole resistance rates in our country and to update treatment protocols within the framework of these data.

Integration of Meta-Multi-Omics Data Using Probabilistic Graphs and External Knowledge.

Multi-omics has the promise to provide a detailed molecular picture of biological systems. Although obtaining multi-omics data is relatively easy, methods that analyze such data have been lagging. In this paper, we present an algorithm that uses probabilistic graph representations and external knowledge to perform optimal structure learning and deduce a multifarious interaction network for multi-omics data from a bacterial community. Kefir grain, a microbial community that ferments milk and creates kefir, represents a self-renewing, stable, natural microbial community. Kefir has been shown to have a wide range of health benefits. We obtained a controlled bacterial community using the two most abundant and well-studied species in kefir grains: Lentilactobacillus kefiri and Lactobacillus kefiranofaciens. We applied growth temperatures of 30 °C and 37 °C and obtained transcriptomic, metabolomic, and proteomic data for the same 20 samples (10 samples per temperature). We obtained a multi-omics interaction network, which generated insights that would not have been possible with single-omics analysis. We identified interactions among transcripts, proteins, and metabolites, suggesting active toxin/antitoxin systems. We also observed multifarious interactions that involved the shikimate pathway. These observations helped explain bacterial adaptation to different stress conditions, co-aggregation, and increased activation of L. kefiranofaciens at 37 °C.

Listeria monocytogenes cross-contamination during apple waxing and subsequent survival under different storage conditions.

This study evaluated Listeria monocytogenes cross-contamination between inoculated fruits, waxing brush, and uninoculated fruits during apple wax coating and investigated the fate of L. monocytogenes on wax-coated apples introduced via different wax coating schemes. There were 1.8-1.9 log10 CFU/apple reductions of L. monocytogenes on PrimaFresh 360, PrimaFresh 606, or Shield-Brite AP-450 coated apples introduced before wax coating after 6 weeks of ambient storage (22 °C and ambient relative humidity). L. monocytogenes showed a similar trend (P > 0.05) on waxed apples under cold storage (1 °C and ∼ 90% relative humidity); there were 1.8-2.0 log10 CFU/apple reductions of L. monocytogenes during the 12 weeks of cold storage regardless of wax coating type. For cross-contamination study, a waxing brush was used to wax one inoculated apple (6.2 log10 CFU/apple); then, this brush was used to wax five uninoculated apples in a sequence. There were 3.7, 3.5, 3.3, 2.9, and 2.7 log10 CFU/apple and 3.6 log10 CFU/brush of L. monocytogenes transferred from the inoculated apple to uninoculated apple 1 to apple 5, and the waxing brush, respectively. The die-off rate of L. monocytogenes on wax-coated apples contaminated during wax coating was not significantly different from that contaminated on apples before wax coating, and 1.8-1.9 log10 CFU/apple reductions were observed during the 12 weeks of cold storage. The application of wax coatings, regardless of wax coating type, did not impact the survival of endogenous yeasts and molds on apples during ambient or cold storage. L. monocytogenes transferred onto waxing brushes during wax coating remained relatively stable during the 2-week ambient holding. Fungicide application during wax coating reduced (P < 0.05) yeast and mold counts but had a minor impact (P > 0.05) on the survival of L. monocytogenes on apples after 12 weeks of cold storage. Collectively, this study indicated that a high cross-contamination risk of L. monocytogenes during apple waxing, and L. monocytogenes on wax-coated apples introduced via different scenarios is stable during subsequent cold storage, highlighting the need for potential intervention strategies to control L. monocytogenes on wax-coated apples.

The effect of organic acids and storage temperature on lite salad dressing rheology and Zygosaccharomyces parabailii growth.

Zygosaccharomyces parabailii (Z. parabailii) causes spoilage in salad dressings due to its tolerance to osmotic pressure. The objective of this study was to determine the effect of organic acids and storage temperatures (4, 10, and 25 °C) on Z. parabailii growth and salad dressing mechanical properties. Acetic, lactic, and gluconic acids were used alone and in combination to acidify salad dressing. Z. parabailii-challenged formulations containing acetic acid alone tended to have lower counts of Z. parabailii when compared to Z. parabailii-challenged formulations containing other acid combinations. Overall, storage temperature had the most impact on Z. parabailii growth over a 45-day storage. Acidulant type and combination impacted salad dressing mechanical properties. During the 45-day storage period, all formulations showed increased viscosity, a Herschel-Bulkley viscosity profile, and elastic-dominant viscoelastic behavior. The degree of change in rheological behaviors over time was dependent on the type of acid used in the formulation. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-022-05459-4.

Growth of Trichomonas vaginalis in Basic Media Available in Routine Microbiology Laboratories

Objective: Trichomonas vaginalis is a protozoan parasite with unicellular, flagellate, and anaerobic metabolism. It is the second most prevalent pathogen among sexually transmitted agents after viruses. Microscopic examination, culture, and molecular methods are used in the laboratory diagnosis of T. vaginalis. However, in most routine microbiology laboratories, microscopy is preferred instead of culture and molecular methods for T. vaginalis diagnosis because microscopy is cheaper than other methods. This study aimed to produce T. vaginalis in media that can be detected frequently in microbiology laboratories. Methods: In this study, four media, namely, thioglucholate medium (THIO), brain heart infusion medium (BHI), tryptic soy broth medium (TSB), and Brucella broth medium (BRB) were modified and tested. Trypticase-yeast extract-maltose (TYM) medium was used as a reference medium. Each medium tested was enriched with three different serum additives. T. vaginalis trophozoite at a density of 104 parasites/mL was inoculated into each medium and incubated at 37 °C for 10 days. We determined the number of trophozoites using a hemocytometer, and the viability rates were determined using trypan blue. Results: Trichomonas vaginalis grew extremely well on THIO, BHI, and TSB media but not on BRB media. The time and number of parasites peaked were determined as 100×104 parasites/mL on THIO-ATS and THIO-FCS media on days five and four, respectively, 100×104 parasites/mL on BHI-ATS on day three, 98×104 parasite/mL on BHI-FCS media on day five, 100×104 parasites/mL on TSB-ATS on day four, and 82×104 parasite/mL on TSB-FCS media on day seven. Compared with the reference medium, TYM, T. vaginalis trophozoites survived significantly longer in THIO, BHI, and TSB media. Conclusion: The rich content of THIO, TSB, and BHI media, which are widely available in routine microbiology laboratories, may allow T. vaginalis growth.

[Antibacterial Activity and Synergistic Interaction of Various Essential Oil Components and Antibiotics]. / Çesitli Uçucu Yag Bilesenleri ile Antibiyotiklerin Antibakteriyel Etkinligi ve Sinerjik Etkilesimi.

Inappropriate and uncontrolled use of antibiotics in humans and animals leads to the emergence of multi-resistant bacteria. Before the discovery of antibiotics, plant extracts and essential oils were used for therapeutic purposes. Today, due to increasing antibiotic resistance, many studies are frequently carried out on the antimicrobial activities of natural active substances that can be a source for new drug candidates. The aim of this study was to investigate the antibacterial activity of components such as α-pinene (α-PN), p-cymene (p-CYM), carvacrol (CAR), thymol (TY) and eugenol (EG) found in the essential oils of many plants and their synergistic interaction with antibiotics. In this study, the antibacterial activity of these essential oil components and antibiotics in clinical use such as gentamicin (GEN), tetracycline (TET), tigecycline (TGC) and linezolid (LZD), against Staphylococcus aureus [methicillin resistant S.aureus (MRSA), and methicillin sensitive S.aureus (MSSA)], Escherichia coli and Pseudomonas aeruginosa strains were determined by disc diffusion and microdilution method. In addition, the interaction between the essential oil components and antibiotics was also determined by the checkerboard method. While CAR, TY and EG components showed strong antibacterial activity, the antibacterial activity of αPN and p-CYM was found to be weak. Combinations of α-pinene, carvacrol, thymol and eugenol with gentamicin and tetracycline mostly showed synergistic interactions against all bacteria. In αPN, CAR, TY and EG with GEN and TET, synergistic/partial synergistic interaction was observed against S.aureus strains, while indifferent interaction was detected in E.coli and P.aeruginosa strains. The combination of αPN and p-CYM with TGC showed synergistic interaction against E.coli and P.aeruginosa strains, and additive and indifferent interaction against S.aureus strains. On the other hand, synergistic interaction was observed against all bacterial strains in combinations of TGC and CAR, TY and EG components. Antagonistic interaction was not detected in any of the tested component-antibiotic combinations against the bacteria used in our study. A synergistic interaction between natural bioactive components and commonly used antibiotics may contribute to the effectiveness of antibiotics and components at lower doses, minimizing their potential toxic side effects and reducing treatment costs. However, more research is needed in terms of their pharmacokinetic and toxic properties to evaluate the therapeutic application potential of phytochemicals.

Bacterial Populations in International Artisanal Kefirs.

Artisanal kefir is a traditional fermented dairy product made using kefir grains. Kefir has documented natural antimicrobial activity and health benefits. A typical kefir microbial community includes lactic acid bacteria (LAB), acetic acid bacteria, and yeast among other species in a symbiotic matrix. In the presented work, the 16S rRNA gene sequencing was used to reveal bacterial populations and elucidate the diversity and abundance of LAB species in international artisanal kefirs from Fusion Tea, Britain, the Caucuses region, Ireland, Lithuania, and South Korea. Bacterial species found in high abundance in most artisanal kefirs included Lactobacillus kefiranofaciens, Lentilactobacillus kefiri,Lactobacillus ultunensis, Lactobacillus apis, Lactobacillus gigeriorum, Gluconobacter morbifer, Acetobacter orleanensis, Acetobacter pasteurianus, Acidocella aluminiidurans, and Lactobacillus helveticus. Some of these bacterial species are LAB that have been reported for their bacteriocin production capabilities and/or health promoting properties.

Antimicrobial Activity of Six International Artisanal Kefirs Against Bacillus cereus, Listeria monocytogenes, Salmonella enterica serovar Enteritidis, and Staphylococcus aureus.

Kefir, a fermented dairy beverage, exhibits antimicrobial activity due to many metabolic products, including bacteriocins, generated by lactic acid bacteria. In this study, the antimicrobial activities of artisanal kefir products from Fusion Tea (A), Britain (B), Ireland (I), Lithuania (L), the Caucuses region (C), and South Korea (K) were investigated against select foodborne pathogens. Listeria monocytogenes CWD 1198, Salmonella enterica serovar Enteritidis ATCC 13076, Staphylococcus aureus ATCC 25923, and Bacillus cereus ATCC 14579 were inhibited by artisanal kefirs made with kefir grains from diverse origins. Kefirs A, B, and I inhibited all bacterial indicator strains examined at varying levels, except Escherichia coli ATCC 12435 (non-pathogenic, negative control). Kefirs K, L, and C inhibited all indicator strains, except S. aureus ATCC 25923 and E. coli ATCC 12435. Bacteriocins present in artisanal kefirs were determined to be the main antimicrobials in all kefirs examined. Kefir-based antimicrobials are being proposed as promising natural biopreservatives as per the results of the study.

Dexmedetomidine as an Alternative Anesthetic Agent for Flap Surgery: An Intravital Evaluation in the Cremaster Muscle Flap.

INTRODUCTION: Flap surgery is one of the most commonly used techniques of reconstructive surgery for effective repair of damaged tissue. Optimal anesthetic technique and anesthetic agent plays an important role in flap perfusion. This study aimed to evaluate the effects of dexmedetomidine infusion on the microcirculation in the cremaster muscle flap by direct in vivo monitoring. MATERIALS AND METHODS: We randomly divided 9 Wistar albino rats into 3 groups. The rats in the control group underwent the surgical procedure (isolation of the cremaster muscle) alone; the rats in the experimental groups 1 and 2 received an infusion of dexmedetomidine (10 and 30 min) after the surgical procedure. RESULTS: The means of vessel diameters, number of functional capillaries, and movements of leukocytes in all groups were evaluated using intravital microscopic examination. The diameters of the arterioles and venules of the cremaster muscle significantly increased in the dexmedetomidine groups. The number of functional capillaries was higher in the dexmedetomidine groups than in the control group. No difference was observed in the movements of leukocytes between the control and experimental groups. Dexmedetomidine significantly increased the diameters of the arterioles and venules of the cremaster flap and the number of functional capillaries. CONCLUSION: On the basis of the effects of dexmedetomidine on microcirculation, we suggest that dexmedetomidine continue to be used as an anesthetic agent, and may be considered also for reconstructive procedures, particularly flap surgery.

In-vitro GIT Tolerance of Microencapsulated Bifidobacterium bifidum ATCC 35914 Using Polysaccharide-Protein Matrix.

Longevity of probiotic is the main concern for getting maximum benefits when added in food product. Bifidobacterium, a probiotic, tends to lose its viability during gastrointestinal track (GIT) transit and storage of food. Their viability can be enhanced through microencapsulation technology. In this study, Bifidobacterium bifidum (B. bifidum) ATCC 35914 was encapsulated by using two experimental plans. In the first plan, chitosan (CH) at 0.6, 0.8, and 1.0% and sodium alginate (SA) at 4, 5, and 6% were used. Based on encapsulation efficiency, 6% sodium alginate and 0.8% chitosan were selected for single coating of the bacteria, and the resulting micro beads were double coated with different concentrations (5, 7.5, and 10%) of whey protein concentrate (WPC) in the second plan. Encapsulation efficiency and GIT tolerance were determined by incubating the micro beads in simulated gastrointestinal juices (SIJ) at variable pH and exposure times, and their release (liberation of bacterial cells) profile was also observed in SIJ. The microencapsulated bacterial cells showed significantly (P < 0.01) higher viability as compared to the unencapsulated (free) cells during GIT assay. The double-coated micro beads SA 6%-WPC 5% and CH 0.8%-WPC 5% were proven to have the higher survival at pH 3.0 after 90 min of incubation time and at pH 7.0 after 3-h exposure in comparison to free cells in simulated conditions of the stomach and intestine, respectively. Moreover, double coating with whey protein concentrate played a significant role in the targeted (106-9 CFU/mL) delivery under simulated intestinal conditions.

Inhibition of Listeria monocytogenes in Hot Dogs by Surface Application of Freeze-Dried Bacteriocin-Containing Powders from Lactic Acid Bacteria.

Six lactic acid bacteria (LAB) strains, Lactococcus lactis BFE 920, L. lactis subsp. lactis ATCC 11454, L. lactis subsp. cremoris ATCC 14365, Lactobacillus curvatus L442, Lact. curvatus LTH 1174, and Lact. bavaricus MN, were grown in cheddar cheese whey supplemented with complex nutrient sources. Cell-free culture supernatants were freeze-dried, and the resulting bacteriocin-containing powders were applied on the surface of hot dogs that were inoculated (~4 log cfu/hot dog) with a five-strain Listeria monocytogenes cocktail. Hot dogs were vacuum-sealed and stored at 4 °C for 4 weeks. L. monocytogenes was enumerated, using both tryptic soy agar (TSA) and oxford listeria agar (OXA), on day 0 and at 1, 2, 3, and 4 weeks of the refrigerated storage. In hot dogs containing only the L. monocytogenes inoculum, L. monocytogenes counts increased from 4 up to 7 log cfu/hot dog. All samples containing freeze-dried bacteriocin-containing powders exhibited significantly lowered (P < 0.05) L. monocytogenes populations on the surface of hot dogs throughout the 4-week study except for bavaricin MN powder. Bacterial counts on hot dogs packed without any powder were statistically equal on day 0 when enumerated on OXA. Freeze-dried bacteriocin-containing powders from Lact. curvatus L442 and L. lactis subsp. cremoris ATCC 14365 decreased L. monocytogenes populations on the surface of hot dogs by greater than 2 log cfu/hot dog throughout the 4-week study. For the powdered bacteriocin preparations from L. lactis BFE 920, L. lactis subsp. lactis ATCC 11454, and Lact. curvatus LTH 1174, L. monocytogenes populations were determined to be approximately 3-log cfu/hot dog after 4 weeks of storage.

Thermal and Starvation Stress Response of Escherichia coli O157:H7 Isolates Selected from Agricultural Environments.

Pathogens exposed to agricultural production environments are subject to multiple stresses that may alter their survival under subsequent stress conditions. The objective of this study was to examine heat and starvation stress response of Escherichia coli O157:H7 strains isolated from agricultural matrices. Seven E. coli O157:H7 isolates from different agricultural matrices-soil, compost, irrigation water, and sheep manure-were selected, and two ATCC strains were used as controls. The E. coli O157:H7 isolates were exposed to heat stress (56°C in 0.1% peptone water for up to 1 h) and starvation (in phosphate-buffered saline at 37°C for 15 days), and their survival was examined. GInaFiT freeware tool was used to perform regression analyses of the surviving populations. The Weibull model was identified as the most appropriate model for response of the isolates to heat stress, whereas the biphasic survival curves during starvation were fitted using the double Weibull model, indicating the adaptation to starvation or a resistant subpopulation. The inactivation time during heating to achieve the first decimal reduction time (δ) calculated with the Weibull parameters was the highest (45 min) for a compost isolate (Comp60A) and the lowest (28 min) for ATCC strain 43895. Two of the nine isolates (ATCC 43895 and a manure isolate) had ß < 1, indicating that surviving populations adapted to heat stress, and six strains demonstrated downward concavity (ß > 1), indicating decreasing heat resistance over time. The ATCC strains displayed the longest δ2 (>1,250 h) in response to starvation stress, compared with from 328 to 812 h for the environmental strains. The considerable variation in inactivation kinetics of E. coli O157:H7 highlights the importance of evaluating response to stress conditions among individual strains of a specific pathogen. Environmental isolates did not exhibit more robust response to stress conditions in this study compared with ATCC strains.

Production of Antilisterial Bacteriocins from Lactic Acid Bacteria in Dairy-Based Media: A Comparative Study.

One hundred and eight strains of lactic acid bacteria (LAB) were screened for bacteriocin production by the modified deferred antagonism and agar well diffusion methods. When the modified deferred antagonism method was employed, 82 LAB strains showed inhibitory action against Listeria monocytogenes v7 ½a, whereas 26 LAB strains expressed no inhibition. Only 12 LAB strains exhibited inhibitory activity when the agar well diffusion method was used, 11 of which had been previously recognized as bacteriocin production positive (Bac(+)). Lactobacillus viridescens NRRL B-1951 was determined, for the first time, to produce an inhibitory compound with a proteinaceous nature. The inhibitory activity was observed in the presence of lipase, α-chymotrypsin, and trypsin, but no inhibition zone could be detected in the presence of proteinase K, indicating the proteinaceous nature of the inhibitory compound. The inhibitory compound was active against Lact. sake ATCC 15521 and Lact. plantarum NCDO 995. Bacteriocin production by the Bac(+) LAB strains was assessed in Lactobacillus MRS Broth as well as in dairy-based media such as nonfat milk, demineralized whey powder, and cheddar cheese whey supplemented with complex nutrient sources that are rich in nitrogen. Lact. sake ATCC 15521 and L. monocytogenes CWD 1002, CWD 1092, CWD 1157, CWD 1198, and v7 ½a were used as indicators. The inhibitory activities of the bacteriocins varied depending on the indicator strains and the growth media used. The LAB indicator strains were found to be more sensitive to inhibition by bacteriocins when compared to the listerial indicator strains. Among the listerial indicators, L. monocytogenes CWD 1002 and CWD 1198 were the most sensitive strains to the bacteriocins investigated in this study. Media composition had a significant influence on bacteriocin production and activity. When compared to demineralized whey powder medium and cheddar cheese whey medium supplemented with whey protein concentrate, cheddar cheese whey medium supplemented with complex nutrient sources such as yeast extract, polypeptone, proteose peptone nr. 3, or soytone appeared to be more supportive of bacteriocin production.

Kefir: a multifaceted fermented dairy product.

Kefir is a fermented dairy beverage produced by the actions of the microflora encased in the "kefir grain" on the carbohydrates in the milk. Containing many bacterial species already known for their probiotic properties, it has long been popular in Eastern Europe for its purported health benefits, where it is routinely administered to patients in hospitals and recommended for infants and the infirm. It is beginning to gain a foothold in the USA as a healthy probiotic beverage, mostly as an artisanal beverage, home fermented from shared grains, but also recently as a commercial product commanding shelf space in retail establishments. This is similar to the status of yogurts in the 1970s when yogurt was the new healthy product. Scientific studies into these reported benefits are being conducted into these health benefits, many with promising results, though not all of the studies have been conclusive. Our review provides an overview of kefir's structure, microbial profile, production, and probiotic properties. Our review also discusses alternative uses of kefir, kefir grains, and kefiran (the soluble polysaccharide produced by the organisms in kefir grains). Their utility in wound therapy, food additives, leavening agents, and other non-beverage uses is being studied with promising results.

Intravital microscopic evaluation of cremasteric microcirculation in experimental testicular torsion.

AIM: Although absent cremasteric reflex is a significant clinical finding for testicular torsion (TT), there is limited information about microcirculation of the cremasteric muscle (CM) after TT. This experimental study was performed to evaluate CM microcirculation by intravital microscopy after TT. MATERIALS AND METHODS: Twelve Wistar rats were allocated into two equal groups: control (CG) and torsion (TG). After anesthetization of the CG rats, the CM flap was dissected through a left ventral inguinal incision with its vascular pedicle. In TG rats, TT was performed by rotating left testicles 720(°) in clockwise direction for 1 h. Then, the CM flap was dissected as in CG, and was placed under an intravital microscope. Vessel diameters, functional capillary perfusion and leukocyte activation in post-capillary venules were measured and evaluated statistically. RESULTS: There was a significant decrease in vessel diameter in TG compared to CG (p < 0.05). The median of perfused capillaries in CG and TG was 13 (11.75-14.30) and 5.5 (4.75-7.25), respectively (p < 0.05). Number of granulocytes (rolling, sticking, transmigrated) was greater in TG than CG (p < 0.05). CONCLUSION: Intravital microscopic evaluation of CM after TT showed decrease in vessel diameter and number of perfused capillaries, and increase in granulocyte activation. Clinical, electrophysiological alterations in CM after TT can be explained by deterioration of microcirculation of CM.

Trout-skin gelatin-based edible films containing phenolic antioxidants: effect on physical properties and oxidative stability of cod-liver oil model food.

UNLABELLED: Trout-skin (Oncorhynchus mykiss) gelatin-based films containing antioxidants (epigallocatechin gallate (EGCG), 50 and 250 ppm w/w) and green tea powder (1% and 20% w/w of gelatin) were tested for tensile strength, elastic modulus, and elongation, and oxygen and water vapor transmission rates, in vitro antioxidant activity using the DPPH (2,2-diphenyl-1-picrylhydrazyl) assay and effect on stabilizing cod-liver oil held under mild thermal abuse conditions. Cod-liver oil overlaid with films was stored at 40 °C for 20 d and analyzed for peroxide value (PV) and thiobarbituric acid reactive substances (TBARS). Antioxidant activity was retained in films containing green tea powder, but was reduced (P < 0.05) in EGCG films (20 d, 23 °C). Water vapor transmission rate of the films incorporated with antioxidants did not change significantly (P > 0.05), but the oxygen transmission rate for films with 50 ppm EGCG and 20% green tea powder was significant (P < 0.05). Other physical properties varied with antioxidant incorporation. The TBARS and PV of control oil increased from 0.05 ± 0.01 to 4.71 ± 0.30 g MDA/kg oil and from 3.6 ± 0.2 to 178.3 ± 24.5 millieq peroxides/kg oil, respectively, after 20 d. For cod-liver oil covered with control or antioxidant-containing films, TBARS remained below 0.37 g MDA/kg oil and PV below 7 millieq peroxides/kg oil. Incorporation of antioxidants to the films did not reduce oil oxidation (P > 0.05) at the levels tested and this was confirmed by activation energy calculations. The rate of oil oxidation was more dependent upon the inherent oxygen barrier property of the films than the presence of antioxidants. PRACTICAL APPLICATION: This research has the potential to enhance the utilization of fish skins, a valuable food processing by-product, as edible films with natural antioxidants to extend the shelf life of foods. The film physical properties and barrier to oxygen and water are investigated.

Development of Freeze-Dried Bacteriocin-Containing Preparations from Lactic Acid Bacteria to Inhibit Listeria monocytogenes and Staphylococcus aureus.

There has been a recent movement to produce and consume "minimally processed" and more "natural" foods through the use of fewer chemical preservatives. The shift to more "natural" foods has resulted in a great interest in the use of bacteriocins from lactic acid bacteria as natural biopreservatives. The objective of this comparative study was to identify bacteriocins that can be produced in low-cost or no-cost dairy-based media (DBM), concentrated using freeze-drying, and applied to Cheddar cheese samples to concurrently inhibit Listeria monocytogenes and Staphylococcus aureus. Select bacteriocin producers were grown in DBM, their cell-free supernatants (CFS) were frozen, and the frozen CFS samples were freeze-dried to produce bacteriocin-containing powders. Cheddar cheese samples were challenged with L. monocytogenes or Staph. aureus cells. The challenged samples were exposed to buffered solutions of freeze-dried powders containing bacteriocins, incubated at 4 °C for 24-72 h, and plated onto appropriate selective media. All freeze-dried bacteriocin-containing powders tested were active against L. monocytogenes and Staph. aureus. Our research findings indicated that low-cost or no-cost DBM could successfully be used for production of bacteriocin-containing preparations. In addition, freeze-drying was determined to be a feasible approach to prepare concentrated and stable bacteriocin-containing powders for prospective food applications. The prevention of even a very small percentage of foodborne illnesses via the use of bacteriocins as natural biopreservatives would help reduce the number of foodborne illness-related hospitalizations, deaths, and financial loss due to medical expenses, lost income/productivity, cost of litigation/penalties, and loss of trade.