Sargassum siliquosum grown in Australian tropical waters was tested in a rat model of metabolic syndrome. Forty-eight male Wistar rats were divided into four groups of 12 rats and each group was fed a different diet for 16 weeks: corn starch diet (C); high-carbohydrate, high-fat diet (H) containing fructose, sucrose, saturated and trans fats; and C or H diets with 5% S. siliquosum mixed into the food from weeks 9 to 16 (CS and HS). Obesity, hypertension, dyslipidaemia, impaired glucose tolerance, fatty liver and left ventricular fibrosis developed in H rats. In HS rats, S. siliquosum decreased body weight , fat mass , abdominal fat deposition and liver fat vacuole size but did not reverse cardiovascular and liver effects. H rats showed marked changes in gut microbiota compared to C rats, while S. siliquosum supplementation increased gut microbiota belonging to the family Muribaculaceae. This selective increase in gut microbiota likely complements the prebiotic actions of the alginates. Thus, S. siliquosum may be a useful dietary additive to decrease abdominal and liver fat deposition.The therapeutic potential of  Saccharina japonica, accounts for more than one-third of total world production, with over 11 million tonnes in 2018 [Ascophyllum nodosum and Fucus vesiculosis [Seaweeds are now a major industry worldwide, including a rediscovery of regional seaweed cuisines , but the in 2018 . Brown s in 2018 , improvi in 2018 , as a su in 2018 , as bios in 2018  and as ciculosis .Sargassum genus that inhabits the Atlantic, Pacific and Indian oceans, including temperate, subtropical and tropical habitats; its biogeography suggests an origin at around 6.7 million years ago [Sargassum species, primarily S. natans and S. fluitans, produced more than 20 million metric tonnes of biomass extending from West Africa to the Gulf of Mexico in 2018 [S. horneri have occurred since 2015 off the southeastern coast of Korea [Sargassum (fusiforme) is already a traditional and highly prized food in Asia, with commercial aquaculture production of 54,624 tonnes of S. fusiforme in 2017 at a value of USD 28.7 million [Sargassum species produce many compounds with a wide range of biological actions, including protection against heat, pollution, stress, decreased oxygen concentration and ultraviolet radiation [Brown seaweeds also include the ears ago . Sargass in 2018 . Furtherof Korea . Howeverof Korea . Sargass million . FurtherSargassum species such as S. fusiforme contain alginates, laminarins and fucoidans as the major polysaccharides [S. siliquosum, is found predominantly in the Sargasso Sea (western North Atlantic Ocean) and the Coral Sea  [Commercial charides ; the xanstralia) .Sargassum as a functional food has not been defined, although Sargassum products have shown effectiveness against metabolic diseases [Sargassum species could exhibit antioxidant responses such as nitric oxide scavenging [Sphaerotrichia divaricata [Sargassum species on the range of pathophysiological changes that characterise metabolic syndrome.Functional foods are claimed to ameliorate metabolic syndrome , defineddiseases . Sargassavenging  and antiavenging ; both acavenging . Fucoxanvaricata  may be svaricata . HoweverSarconema filiforme [Caulerpa lentillifera [S. siliquosum, as a source of non-sulphated  and sulphated polysaccharides (fucoidans) and fucoxanthin, also reverses the signs of metabolic syndrome in this rat model [S. siliquosum, including the structure and function of heart and liver, plasma biochemistry, glucose and insulin responses and body composition. Further, we characterised the changes in the composition of gut microbiota after seaweed treatment, as functional foods have the potential to reverse the changes induced in obesity [S. siliquosum could include the prebiotic actions of polysaccharides including fucoidans and alginates in the colon by reducing the intestinal absorption of carbohydrates and fats and also changing the gut microbiota, in combination with the anti-inflammatory effects of fucoxanthin [We have reported that sulphated polysaccharides from the red seaweed iliforme  and the illifera  reversedat model . This seat model . To test obesity ,31. The oxanthin . The comS. siliquosum was collected by snorkel from a rocky reef at approximately 2 m depth in Nelly Bay, Magnetic Island, QLD, Australia , on 28 November 2017, and immediately transported to James Cook University, Townsville, QLD, Australia. Twelve specimens with intact holdfasts and fresh new growth were photographed (ographed  and seleSargassum specimens to a genetic species group. The forward primer GazF2 (5′CCAACCAYAAAGATATWGGTAC3′) and reverse primer GazR2 (5′GGATGACCAAARAACCAAAA3′) were used [One blade from each of the twelve specimens selected for molecular barcoding was excised and cleaned in autoclaved seawater, using a soft paintbrush to remove debris and epiphytes. Each blade was dried to remove excess water and subsequently kept in individually labelled zip-lock bags with 50 g of silica beads to desiccate. Dried samples were sent to Victoria University of Wellington, New Zealand, for DNA extraction using a modified CTAB protocol . The molere used . Maximumn = 5) of milled and homogenised material, except for amino acid and constituent sugar analyses (n = 3) and fibre analysis (n = 2). For sugar analysis, hydrolysis of the biomass (10 mg) was performed in a two-step process: step 1: hydrolysis in 300 µL of 72% sulphuric acid at 20 °C for 1 h; step 2: hydrolysis in 3.6 mL of 1M sulphuric acid at 100 °C for 2 h [v:v). The extract was filtered, washed with 0.9% NaCl (w:v), allowed to separate into two phases, and the lower (organic) phase was collected for evaporation under nitrogen and subsequently weighed to give the total lipid content [v/v)) with 300 µL nonadecanoic acid  as internal standard. After cooling, 1 mL of hexane and 2 mL of deionised water were added, and the upper (hexane) phase was collected for analysis by GC–MS [All analyses were performed on five subsamples  were sourced and housed as in previous studies [S. siliquosum for the final eight weeks . Detailed composition of diets has been described in our published study [Male Wistar rats  specimens and all sequences were identical. A phylogenetic tree was compiled from comparison with sequences downloaded from Genbank. A trimmed phylogenetic tree  including S. siliquosum. The COI sequences of this grouping were identical, except for our samples and S. siliquosum, which shared a single synapomorphy. Based on these data and distributional data, we identified this species as S. siliquosum. These samples of S. siliquosum contained (in % of dry weight) 57.8% carbohydrate , 1.7% lipid, 4.02% protein, 41.4% dietary fibre including 33.3% as soluble fibre with 7.5% K, 1.9% Ca, 1.2% Na and 0.97% S as the major elements  gene was used for identification of the tic tree  shows a elements . Hydrolyted salt . In S. sno acids  and potano acids . Total xPLC data .S. siliquosum. During weeks 9–16, the food consumption was unchanged in C and CS, and in H and HS  compared to groups supplemented with Sargassum  (Muribaculaceae in HS rats (24.34%) compared to H rats (10.35%) (p = 00063). Further information on the gut microbiota, including at genus level  . Within us level , and chaus level , are incS. siliquosum supplementation in rats with diet-induced metabolic syndrome decreased body weight and decreased retroperitoneal fat and liver fat but had no effect on systolic blood pressure, liver enzyme activities, lipid profile or glucose and insulin metabolism. Similar responses have been shown with other Sargassum species. As examples, S. thunbergii decreased obesity, serum insulin, triglycerides and cholesterol in high-fat-diet-fed mice [S. polycystum decreased damage to the liver in high-sugar, high-fat diet + streptozotocin-induced type 2 diabetic rats [Seaweeds containing complex mixtures of polysaccharides, peptides, pigments, minerals and omega-3 fatty acids have been shown to improve the signs of metabolic syndrome . This stfed mice  and S. ptic rats .Sargassum species contain polysaccharides, predominantly (in decreasing content) alginates, fucoidans and laminarans; these are the major constituents of the high fibre content of S. siliquosum [liquosum  as in otliquosum . Oral inliquosum . In healliquosum . In a huliquosum . This huliquosum .S. fusiforme changed the gut microbiota, particularly by increasing the Bacteroidetes to Firmicutes ratio, to decrease streptozotocin-induced hyperglycaemia in mice [S. fusiforme decreased high-fat-diet-induced insulin resistance in mice by activating the Nrf2 pathway, changing the gut microbiota and reducing intestinal inflammation [S. siliquosum.Fucoidan from  in mice . Furtherammation . Fucoidaammation . HoweverS. siliquosum and S. polycystum inhibited angiotensin-converting enzyme, α-amylase and α-glucosidase [Inhibiting digestive enzymes including α-amylase, α-glucosidase, pepsin and lipase by seaweed polysaccharides and polyphenols may modulate obesity and cardiovascular risk . In an icosidase . The rolS. siliquosum are primarily due to the actions of fucoxanthin. The metabolic actions of phenolic acids and other xanthophyll compounds present in this seaweed are unknown but they could be additive to responses produced by the alginates.The biological responses could be caused by the xanthophyll, fucoxanthin. In high-fat-diet-induced obese rats, fucoxanthin at doses of 0.083 and 0.167 mg/kg reduced white adipose tissue weight, accumulation of hepatic lipid droplets and perirenal adipocyte size . These dS. siliquosum are unlikely to be due to an increased intake of polyunsaturated fatty acids.Polyunsaturated fatty acids such as EPA and DHA decreased abdominal obesity and total body fat in the same rat model of metabolic syndrome as the current study . Howevertrans and saturated fats changed the gut microbiome, similar to results in our previous study [S. siliquosum caused a further change in the gut microbiota, including an increase in bacteria belonging to the family Muribaculaceae . Members of this family are understudied but have recently been recognised for their versatile metabolism of complex carbohydrates [S. siliquosum.Not unexpectedly, a diet with higher simple sugars and us study . Supplemhydrates ,56 and thydrates . Membershydrates  and, basSargassum act as prebiotics to alter the changed gut microbiota in obesity, as shown by other seaweeds [Muribaculaceae, thereby increasing concentrations of short-chain fatty acids, which could lead to human health benefits [S. thunbergia by colonic microbiota to short-chain fatty acids [One potential mechanism would be that the polysaccharides from seaweeds . Alginatbenefits . Unsaturbenefits . An imprty acids .The current study could be extended by including the measurement of obesity-related plasma and tissue biomarkers such as adiponectin and C-reactive protein, inflammatory mediators such as TNF and functional changes in the colon such as short-chain fatty acid production. These data could allow further interpretation of the mechanisms of action of this seaweed.S. siliquosum, reduced liver and abdominal fat accumulation in high-carbohydrate high-fat fed rats. As this seaweed contains alginates as the major polysaccharide, these alginates may act as prebiotics in the intestine to increase concentrations of short-chain fatty acids. These actions are possibly complemented by an increase in specific bacteria such as Muribaculaceae in the gut microbiome. These intestinal changes could then lead to systemic anti-inflammatory effects in the heart and liver. We conclude that the tropical Australian seaweed, S. siliquosum, should now be included with other local tropical seaweeds such as Sarconema filiforme [Caulerpa lentillifera [The tropical Australian brown seaweed, iliforme  and Caulillifera  as dieta


Mycoplasma genitalium, trichomoniasis) among infertile males; to analyze the effect of STIs on semen parameters and blood PSA. Case-control study. Study group (n = 2000): males with fertility problems or desire for fertility check. Control group (n = 248): male partners of pregnant women. Analyses: polymerase chain reaction for STI, seminal interleukin 6 (IL-6), semen and fractionated urine, blood analyses . The prevalence of M. genitalium and chlamydia in the study group was 1.1% and 1.2%, respectively. The prevalence of chlamydia in the control group was 1.6%, while there were no M. genitalium cases. No cases with gonorrhoeae or trichomoniasis or combined infections were observed in neither group. There was a higher seminal concentration of neutrophils and IL-6 among M. genitalium positives compared with STI negatives. There was a trend toward a lower total count of spermatozoa and progressive motility among STI positives. No impact of STIs on PSA was found. The prevalence of STIs among infertile males is low. M. genitalium is associated with seminal inflammation. The impact of STIs on semen parameters deserves further investigations.The impact of sexually transmitted infections (STI) on male fertility is controversial. Aims: To investigate the prevalence of urethritis-associated STIs (chlamydia, gonorrhoeae,  Infertility is defined as the inability of a couple to become pregnant despite unprotected intercourse for a period of more than twelve months . The preChlamydia trachomatis other than nucleic acid amplification tests (NAAT) including culture or serological methods is of significant concern. Culture for C. trachomatis detects only viable infectious chlamydial elementary bodies and the sensitivity is at most 70–85%. The major drawback of serological tests for C. trachomatis is that a positive antibody test will not distinguish a previous from a current infection ; M. genitalium DNA by Sacace Biotehnologies/Mycoplasma genitalium Real-TM ; T. vaginalis DNA by Sacace Biotehnologies/Trichomonas vaginalis Real-TM ) according to the manufacturer’s instructions, at the United Laboratory of Tartu University Hospital. All STI-positive patients were contacted for the appropriate treatment and follow-up.FVU was used for the detection of STIs. STIs were detected from first-voided urine using the polymerase chain reaction (PCR) method (3 and 0.4% (v/v) formaldehyde in distilled water. The spermatozoa concentration was assessed using the improved Neubauer hemocytometer. Motility was assessed in order to report the number of progressively motile spermatozoa. Smears for the morphology assessment were made. Following the fixation and Papanicolaou staining, the morphology was assessed according to strict criteria , CFTR mutations ) were performed at the United Laboratories of Tartu University Hospital. A detailed description of genetic analyses is provided elsewhere [Tests for the known genetic causes of male infertility  entered the collected epidemiological, laboratory, and clinical examination data into two separate, but identically structured, databases. Prior to statistical analysis, the two databases were merged and duplicate entries were eliminated. The entered laboratory data were counter–controlled from primary sources (lab databases) and, if needed, edited by a specially trained researcher (MP and KP). Clinical data relevant to defining the cause of male infertility were controlled retrospectively for all study subjects from their medical case histories one by one by one author of the study (MP).U-test was used for non-normally distributed data, while the parametric unpaired t-test was used for normally distributed data. For categorical nominal data, Fisher’s exact test and chi-squared test were used. Two-sided p-value ˂ 0.05 was accepted as statistically significant. Comparing C. trachomatis-positive vs. STI-negative and M. genitalium-positive vs. STI-negative patients, the Bonferroni correction of p-values for two tests was used. To find the best cut-off value for semen neutrophils’ concentration and semen interleukin-6 for predicting C. trachomatis and/or M. genitalium infection, the construction of receiver operating curves (ROC) was performed. Due to the relatively small number of STI-positive patients, the construction of multivariate logistic regression models was avoided.Statistical analyses were performed using Microsoft Excel software  and RStudio software (R version 3.6.1. (2019–07–05), RStudio Inc., Massachusetts, USA). For comparison of numerical continuous parameters between different STI groups, the non-parametric Mann–Whitney M. genitalium 1.1% and C. trachomatis 1.2%. Markers of inflammation displayed higher levels in the semen of M. genitalium-positive men while currently accepted seminal leucocytes’ threshold proposed by the WHO does not allow for reliable prediction of the presence of urethritis-associated STIs. Patients infected with M. genitalium and C. trachomatis had a lower total count of spermatozoa and total count of spermatozoa with progressive motility compared to STI-negative subjects, therefore, the impact of these atypical bacteria on male fertility cannot be ruled out. However, the more precise effect of these bacteria separately still requires further investigation.The prevalence of urethritis-associated STI among infertile patients in Estonia is low, only two out of four tested causative agents were found in them—


Camellia species are no exception. Several species and cultivars are widespread in Asia, namely in China, Japan, Vietnam and India, being also identified in western countries like Portugal. Tea and oil are the most valuable and appreciated Camellia subproducts extracted from Camellia sinensis and Camellia oleifera, respectively. The economic impact of these species has boosted the search for additional information about the Camellia genus. Many studies can be found in the literature reporting the health benefits of several Camellia species, namely C. sinensis, C. oleifera and Camellia japonica. These species have been highlighted as possessing antimicrobial  and antitumoral activity and as being a huge source of polyphenols such as the catechins. Particularly, epicatechin (EC), epigallocatechin (EGC), epicatechin-3-gallate (ECG), and specially epigallocatechin-3-gallate (EGCG), the major polyphenols of green tea. This paper presents a detailed review of Camellia species’ antioxidant properties and biological activity.Medicinal plants have been used since antiquity to cure illnesses and injuries. In the last few decades, natural compounds extracted from plants have garnered the attention of scientists and the  Camellia genus of the family Theaceae is among the most traditional and famous plants in Asia [Camellia genus have a high economic value due to their subproducts such as tea, oil seed, iconic flowering shrubs and ornamental plants [Camellia genus were focused in three species; namely, C. sinensis (the tea plant), C. oleifera (used to obtain edible oil) and C. japonica . Tea is the second most consumed beverage worldwide, just after water [C. sinensis or C. sinensis var assamica leaves, the different types of teas are obtained according to the fermentation process used. The edaphoclimatic conditions and the agricultural practices strongly impact the composition and properties these plants [Camellia oil is obtained from C. oleifera seeds and is one of the four edible tree oils . It is mainly composed by neutral lipids (88.2%), especially oleic acid . and is commonly named as “Asian olive oil” because of the high oleic acid concentration, very similar to that of olive oil [C. oleifera seeds still are of interest  and have been used over the years as a material for washing hair, for itching and pain relief and to produce cosmetics and food supplements [Camellia species other than C. sinensis, C. japonica and C. oleifera.The  in Asia , especial plants . The higer water  and is mer water ,5. Blacker water . Despitee plants . Tea leae plants , includie plants . Camellilive oil ,10. Afteplements . Over thplements ,12,13. TCamellia species are unanimously considered a natural source of antioxidant compounds. Most of the studies focus on C. sinensis, C. japonica and C. oleifera being the leaves the best characterized part of the plant through their extracts or purified compounds  under different experimental conditions of temperature and extraction time. Chromatographic fractions of the extracts are also frequently investigated. C. sinensis and C. sinensis var assamica seems to be the best characterized species due to the commercial relevance of their leaves. Indeed, the comparison between seeds and leaves extracts (aqueous and organic) as well as their eluted fractions revealed a higher TPC and DPPH scavenging activity in the leaves [2O2 scavenging activity at different pH values [C. sinensis as possessing high antioxidant potential when compared with other plant extracts [ompounds . Extracte leaves . DespiteH values . Beyond H values  reportedH values ,18,19 asH values  than blaH values . Among bH values  being thH values  as alreaH values . A 50% oH values . Other sextracts ,22.C. sinensis var assamica, is a quite different plant which possess wider and larger leaves and is frequently found in warmer regions with tropical climates. This variety is frequently pointed as possessing higher polyphenol and lower amino acids content. Several studies reported its antioxidant properties which seems also to be highly impacted by the extraction solvent [C. sinensis, the kind [C. sinensis var assamica. Despite the low consistency of the results (highly dependent on the solvent used), the TPC seems to be lower for mature leaves [C. sinensis var. assamica identifying eighteen phenolic compounds. Soluble phenolics clearly exhibited higher TPC and TFC and also higher radical scavenging activity. Regarding the work developed by Zhou et al. [Despite being the same species,  solvent ,23,24,25 solvent ,23 as wethe kind  of leavethe kind  and the the kind  also leae leaves  while the leaves  and withe leaves . The dece leaves  and flave leaves . Yang ane leaves  exploredu et al. , five noCamellia species due to its ornamental flowers, C. japonica, was the target of some studies aiming at characterizing its antioxidant potential. Leaves aqueous and alcoholic extracts [C. japonica were undertaken using organic solvents; however, the different or non-specified maturity of the leaves and cultivars diversity used prevents a direct comparison among the obtained results. Páscoa and colleagues [C. sinensis, that green leaves possess higher scavenging activity [Another popular extracts ,32,33,34extracts ,36,37. Aextracts  in FRAP lleagues  performeactivity ,36,37 weC. oleifera is a source of cooking oil which has already proven effects on skin diseases and as a burn treatment. Due to its economic relevance, this species was also quite studied regarding its antioxidant properties, particularly its seed oil and cake, the byproduct obtained after the oil refining process. Regarding the oil, several organic compound were used for the extraction steps and a higher radical scavenging activity (DPPH and ABTS assays) was obtained with methanol [methanol . Other amethanol . Yu and methanol . An uncomethanol . Defattemethanol . Severalmethanol , saponinmethanol , ad purimethanol ,46 and pmethanol . Zhu andmethanol . Other smethanol , the extmethanol  all impamethanol . On the methanol . A uniqumethanol . The autC. sinensis, C. japonica and C. oleifera, other Camellia species were investigated [Camellia taliensis leaves and concluded that they possess significant DPPH radical scavenging activity. Hydrolysable tannins and flavan-3-ols with catechol and/or pyrogallol groups in the molecule presented stronger DPPH radical scavenging activities. Also, other authors reported the DPPH and ABTS scavenging activity of a flavan-3-ol dimer isolated from C. talensis leaves [Camellia tenuifloria organic and aqueous fractions were reported by Chiou [Camellia species are also found in literature [Beyond these three relevant species, stigated ,53,54,55stigated  evaluates leaves . Fruit aby Chiou  as posseby Chiou . StudiesCamellia species was also widely reported; namely, antibacterial, antiviral and antifungal. Similarly, to the antioxidant activity, most of the studies were dedicated to the three species with higher economic impact being C. sinensis and its compounds of higher relevance [The antimicrobial activity of different elevance ,57.https://www.ecdc.europa.eu/en/healthcare-associated-infections-acute-care-hospitals/surveillance-disease-data/report, accessed on 12 March 2021). The antibacterial activity of natural products has been largely explored due to the growing antibiotic resistance noticed in several bacterial species [Camellia species have been highly explored over the years as a natural source of new compounds with antibiotic activity. Published studies evaluated the activity of several Camellia species  against Gram negative and Gram positive bacteria . In 2009, Erol et al. [S. aureus, Listeria monocytogenes, E. coli, Hafnia alvei, Salmonella enteritidis, E. coli Type 1 and Bacillus cereus. Water extracts did not show antibacterial activity for any of the bacteria while ethyl acetate fractions of extracts showed antibacterial activity on S. aureus and B. cereus. It should be noted that the absence of antimicrobial activity reported for the water extract was in accordance with the findings of Yildirim et al.’s [S. aureus and Gram-negative bacteria. Of note, the best pH value was an acidic one, namely pH = 4.8. Dhaouadi et al. [S. aureus and Staphylococcus epidermidis and the Gram negative P. aeruginosa, E. coli and Salmenolla wiam. The authors noticed that the inhibition zones were proportional to the phenolic content, suggesting the implication of these molecules in the observed activities. Kiran et al. [C. sinensis) on virulence traits of clinical isolates of Shigella dysenteriae and E. coli. The tea-treated cultures showed a decrease of the time necessary for bacterial killing. Indeed, crude tea polyphenols of black tea improved (p < 0.05) phagocytic uptake of tea-adapted S. dysenteriae cells in 30 min by 37.68% and phagocytic killing by ten-fold in 3 h. Chan et al. [C. sinensis against the Gram positive bacteria Micrococcus luteus, S. aureus, and B. cereus, and Gram negative bacteria E. coli, Salmonella typhi and P. aeruginosa. The results showed no inhibitory activity of teas against any of the Gram negative bacteria. However, M. luteus and B. cereus, were inhibited by black tea and herbal teas and all of the Gram positive were inhibited by green tea, being the best results reported for M. luteus. The teas were obtained with hot water which validated the inactivity of C. sinensis reported by previous studies against Gram negative bacteria; however, this study reported activity against Gram positive ones. Similar results were obtained by Orak et al. [Porphyromonas gingivalis and Prevotella intermedia). These somehow contradictory results obtained by several authors regarding the antimicrobial activity of aqueous extracts need further clarification. Abidi et al. [C. sinensis against S. epidermidis; however, in combination with current antibiotics such as vancomycin, erythromycin, tetracycline, chloramphenicol, ampicillin, ofloxacin, cephalexin and gentamicin. The authors noticed an appreciable synergistic activity between the plant extracts and the tested antibiotics. Granato et al. 2016 [Salmonella Typhimurium; however, the combination of green/white and green/black teas also showed a significant inhibitory effect depending on the considered serotype.One of the first attempts to explore the bacterial activity of m et al. . These ai et al.  reportedl et al.  studied et al.’s  studies et al.’s  proved ti et al.  reportedn et al.  also pron et al.  studied k et al.  regardink et al.  reportedi et al.  also stuB. subtilis, S. aureus, E. coli and Klebsiella pneumoniae). The best result was obtained for tea flower extract (maximum zone of inhibition at 20 mg/mL) against S. aureus and K. pneumonia.Most of the reported studies were performed with plant leaves; however other plant parts revealed to possess antimicrobial activity. Namely, Rana et al.  evaluateShigella flexneri and B. cereus and the effects of solvent  on the antimicrobial activity. Water and ethanolic extracts showed antimicrobial activity against all bacteria tested being the best results obtained for aqueous ethanol extracts which possess higher phenolic content.Üstündag˘ et al.  also disS. aureus with ~9.4 folds’ improvement compared to pure green tea oil and ~4.7 fold against E. coli. From this study arises that not only the compounds extraction procedure is a relevant step on the microbial activity but also the way they are used.In 2019, Shetta et al.  comparedC. sinensis var assamica. Kristanti and Punbusayakul [S. aureus, S. typhimurium, E. coli and L. monocytogenes. All samples possess higher activity against Gram-positive bacteria being the better results reported in samples with higher total polyphenol content which is in accordance with previously reported studies. Mehrotra et al. [C. sinensis variety against Vibrio cholerae, S. aureus and P. aeruginosa. Abdulbaqi et al. [C. sinensis var. assamica) and Salvadora persica L. against primary colonizers of dental plaque Streptococcus mitis, S. sanguinis and Actinomyces viscosus. Also, Kawarai et al. [S. mutans using extracts from Assam tea and green tea. The results revealed that Assam tea  possess stronger biofilm inhibition activity against than green tea . These studies suggest that Camellia extracts may also have potential applications in preventing dental caries, dental plaque control and other oral health problems associated with serious damage.Four additional studies were found in literature discussing the antimicrobial activity of usayakul  studied a et al.  also repi et al.  reportedi et al.  studied C. japonica was evaluated regarding its antimicrobial activity against several bacteria . Kim et al. [C. japonica L. petals . None of the microorganisms was totally inhibited and the basic fraction showed no antibacterial activity. Gram negative S. typhimurium and E. coli demonstrated slightly greater sensitivity than the Gram positive L. monocytogenes and S. aureus to the extracts. Moon and Kim [C. japonica L. leaf extracts (70% ethanol and 100% methanol) against S. epidermidis, B. subtilis, K. pneumonia and E. coli. The authors concluded that the ethanol extracts possess higher antimicrobial activities against all bacteria and that the antimicrobial activity was doses dependent for both extracts. Sharma et al. [C. japonica L. leaf extract. The activity seems to be similar for all the tested bacteria.Also, m et al.  describe and Kim  studied a et al.  also repC. oleifera [C. reticulata and C. sasanqua [C. semiserrata [Three additional studies were found in literature reporting some antimicrobial activity of other Camellia species as oleifera ,73; C. rsasanqua  and C. siserrata .Camellia species extracts were undertaken in vitro, Thakur et al. [C. sinensis against carbapenem resistant E. coli, in vivo, in a murine model (Sprague Dawley rats). The results showed that a single doses extract  was very effective against this pathogen. Also, Khan et al. [E. coli, S. aureus, and six serovars of Salmonella: S. typhimurium, S. enteritidis, Salmonella gallinarum, Salmonella choleraesuis, Salmonella pullorum and Salmonella dublin) Results showed that the green tea isolated saponins fractions possess antibacterial activity both in in vitro and in vivo experiments.Although most of the studies to evaluate the antimicrobial activity of r et al.  tested an et al.  undertooCamellia species extracts was mainly explored against human pathogens. A single study was found in literature that reports the activity of green tea extracts in Pseudomonas syringae pv. actinidiae (Psa), the agent that causes kiwifruit bacterial cancer [The antimicrobial activity of l cancer . The resCamellia spp was evaluated by several authors, being most of the studies focused on the popular plant tea, C. sinensis. Fungus and yeasts were studied and the reported results were not always consensual. The heterogeneity of the results seems not to be related with the use of crude extracts versus purified compounds once antifungal activity was reported using both. Nevertheless, the works showing no Camellia antifungal activity only used crude extracts.The antifungal activity of C. albicans, one of the most opportunistic pathogens, as well as against other microorganisms [C. sinensis) has no activity against two species of Aspergillus. Both studies included four distinct extracts concentrations which somehow reinforce their conclusions.Rize tea and young shoot tea, two Turkish black tea extracts, were reported as ineffective against rganisms . Also, Crganisms  concludeCamellia species have been pointed as a great source of bioactive compounds including antifungal ones. C. albicans seems to be susceptible to several catechins (pH dependent), which also enhance the antifungal effect of amphotericin B and fluconazole [C. sinensis seeds) against C. albicans. In this study, low pH values lead to lower minimum inhibitory concentrations while in the Hirasawa and Takada [C. albicans, Cryptococcus neoformans, Alternaria alternate and Rhizoctonia solani cultures. The results shown a positive correlation between the concentration and inhibitory activity being fractions F2 and F3 the most active.Despite these two works, conazole . Also, Lconazole  reportedd Takada  work, acd Takada  also stuFusarium, Aspergillus fumigatus, Aspergillus niger and C. albicans. The fresh green tea methanolic extract was found to have higher activity. Later, Orak et al. [Aspergillus parasiticus with a positive correlation with the extract concentration. The results obtained by Orak and colleagues were in agreement with those obtained by Archana and Abraham in 2011 [The previous studies reporting the antifungal activity were performed using somehow purified compounds; however, other works also report such activity using crude extracts. Archana and Abraham in 2011  did a cok et al.  also stu in 2011  being thC. sinensis due to their highly appreciated subproduct, tea, totally eclipses the remaining Camellia species and few studies were dedicated to them. Feás et al. 2013 [C. oleifera, C. reticulata and C. sasanqua against clinical strains of C. albicans. The three oils evidenced antifungal activity exhibiting different MICs. C. reticulata and C. oleifera oils showed the best and similar antifungal activity (20.833 ± 7.217 mg/mL) followed by (20.833 ± 7.217 mg/mL) C. sasanqua (29.167 ± 19.094 mg/mL). In 2015, Meng et al. [C. semiserrata Chi (Nanshancha) seed cake (NSC) against three postharvest pathogens  performing in vitro and in vivo (Fruit Test) tests. The in vitro tests showed a higher inhibition rate for C. musae, followed by C. gloeosporioides and P. italicum. Such values were quite similar to those obtained with tea saponin for C. musae and P. italicum being even higher for C. gloeosporioides (90.9 ± 1.7% versus 49.8 ± 3.2%). The in vivo antifungal activity was evaluated by means of the fruit lesion caused by the three fungi after exposition to NSC crude extractum. The extractum proved to be able to eliminate the lesion diameter in banana; to kept it constant and lower when compared with the untreated fruit for mango and to maintain it slightly lower for the Shatang mandarin when compared with the untreated fruit and tea saponin effect.The popularity of al. 2013  studied g et al.  studied Camellia species  or roots as well as in their sub products like wine or tea. Indeed, tea is pointed as an innate immunity modulator which can enhance immune response in order to mitigate COVID-19 (SARS-COV-2) . Its pheEpigallocatechin-gallate (EGCG) is the major flavonoid present in green tea and seems to possess a highly antiviral potential through several action mechanisms.  also demonstrated activity against Influenza H5N1, a highly pathogenic avian influenza virus, inoculated in MDCK cells [Influenza viruses were also the target of several studies using g et al.  studied g et al.  which cog et al.  and Imang et al. . The firg et al.  reportedCK cells . Indeed,CK cells  comparedCK cells  which de50 = 5.0 ± 1.0 µM and 1.6 ± 0.3 µM for XTT and plaque reduction assays, respectively). Prodelphinidin B-2 3′-O-gallate seems affected the late stage of HSV-2 infection and also shown to inhibit the virus from attaching and penetrating into the cell. Oliveira et al. [Tea also demonstrated activity against herpes simplex viruses type 1 and 2. Prodelphinidin B-2 3′-O-gallate) isolated from green tea leaves was reported by Cheng et al.  as possea et al.  demonstra et al. .In 1998, Clark et al.  studied Camellia spp. namely, C. oleifera and C. sinensis. Epigallocatechin gallate (EGCG), the main polyphenol of green tea, was referenced as a “cancer preventing agent”. Later, many other studies proved the antitumor activity of Camellia spp being most of them associated with C. sinensis. The antitumor potential of this species was highly explored whether using raw C. sinensis leaves extracts, their purified compounds or their commercially available polyphenols alone or synergistically with the most used antitumor drugs.Okuda et al.  was one In 1999, Suganuma et al.  studied C. oleifera were also reported as possessing cytotoxic activity on MCF-7 cells [Camellia extracts was also evaluated against distinct tumour cell lines. Extracts were obtained were obtained from different species  and different parts of the plant. Choi et al. [50 = 54 ± 10 for A-498 and 129 ± 28 µg/mL for 769-P cells) It should be noted that, along with the antitumor activity studies, the authors identified the extract phenolic and methylxanthine content allowing a deeper knowledge on the compounds responsible for the reported biological activity. In 2016, Rana et al. [Despite the synergistic antitumor effects of tea polyphenols, mostly EGCG, noticed by the previous reported studies, their potential was also evaluated alone. Mittal et al. , dedicat-7 cells . Saponini et al.  studied i et al.  studied a et al. , evaluatC. oleifera, in 2012, Jin reported the antitumor activity of the polysaccharide obtained from seed cake and fruit shell of C. oleifera Abel in two different studies [50 = 5.826 µg/mL), followed by saponin (IC50 = 26.754 µg/mL) and protein (IC50 = 36.794 µg/mL)The inhibition rate of the three compounds on Hep G2 cells was higher than 80% at the higher concentration tested (200 µg/mL) being on IAR20 cells dependent on the considered compound . Later, Chaikul et al. [C. oleifera seed oil with the ultimate goal of utilize it as a functional oil in health, cosmetic products and feed. Cytotoxicity was evaluated by the sulforhodamine B (SRB) assay in B16-F10 melanoma cells and 3T3-L1 cells demonstrating cell viabilities of 94.59 ± 3.41% and 97.57 ± 1.62%, respectively. The melanogenesis revealed to be via inhibition of tyrosinase and tyrosinase-related protein-2 activities. According to the authors, their results clearly indicated the potential of the seed oil as a functional oil.Regarding  studies ,110. An  studies  reportedl et al.  studied C. japonica is a Camellia species commonly used as an ornamental plant and is often overlooked regarding its potential as a bioactive compound source, due to the popularity of C. sinensis and C. oleifera. A single study, regarding its antitumor activity performed by Thao et al. in 2010 [C. japonica triterpenoids obtained from stem barks. Three newly identified compounds along with seven known ones were tested. From them, the newly identified 3β-O-acetyl-16β-hydroxyolean-12-en presented cytotoxicity against LLC and HL-60 cancer cell lines . The remaining compounds shown weak or moderate cytotoxic activity against some of the tested cell lines. An additional study was found reporting the antitumor activity of a very uncommon camellia species, Camellia osmantha [ in 2010  was founosmantha . The autLastly, an uncommon study was undertaken by Chitsazan in 2015  to proveCamellia genus have been pointed as possessing a set of benefits including anti-osteoporotic, anti-inflammatory and antidiabetic activity, as blood pressure and body fat reducing agents among others  was explored by Miura et al. [C. oleifera seed cake measuring glucose uptake in cultured HepG2 cells and concluded that the extracts can stimulate the consumption of the glucose in the medium by the cells. In 2007, Nagao et al. [Camellia kucha and its main chemical components also on the lipid accumulation in 3T3-L1 adipocytes but in rats. Kucha tea (KT) revealed anti-adipogenic activity (dose-dependent) demonstrating significant decrease in lipid droplet accumulation. KT also diminished the mRNA and protein levels of fatty acid synthase, fatty acid translocase, steroylcoenzyme A desaturase-1, lipoprotein lipase and acetyl-CoA carboxylase-1. Chan et al. 2010 [C. oleifera. The results showed anti-inflammatory and analgesic activities with production of pro-inflammatory cytokines and PGE2 inhibited. Levites et al. 2003 [C. japonica leaf extracts (ECJL). In vitro, ECJL inhibited xanthine oxidase activity by 41.3 ± 5.5% (dose-dependent) and in vivo, at the doses of 100 and 300 mg/kg, inhibited hepatic xanthine oxidase activity, highly attenuating hyperuricemia. Jeon et al. 2018 [C. japonica on induced pluripotency and wound healing in vivo mouse wound model and proved that the extracts enhanced mouse and human induced pluripotent stem cell (iPSC) generation and promoted effective wound healing. Wang et al. [Camellia brevistyla) against indomethacin-induced gastrointestinal mucosal damage in vitro and in vivo . The experiments revealed that a pre-treatment with Camellia oil can increased cell viability, wound healing and reduced reactive oxygen species production in the cells. In vivo results demonstrated that pre-administration of camellia oil prevented gastric wound damage by decreasing inflammatory mediators as interleukin-6, tumor necrosis factor-α and cyclooxygenase-2 levels. Kim et al. [C. japonica flower extract (CJFE) against urban air pollutants by urban-induced reactive oxygen species (ROS) production in vitro cultured normal human dermal fibroblasts (NHDFs) and in an ex vivo model. CJFE was able to stop the urban air pollutants-induced ROS generation and matrixmetalloproteinase-1 (MMP-1) production and demonstrated very good protective activity against pollutants-induced deteriorating effects in the ex vivo model by reducing the lipid peroxidation marker and the level of pollutants-induced malondialdehyde (MDA). Zhang et al. [C. oleifera Abel (WECO) and find no toxic effects or significant differences on clinical chemistry value, body weight, hematology value or organ/body weight ratio. The NOAEL for WECO was 2 g/kg/BW for subacute toxicity study. These studies included different parts of different Camellia species demonstrating the importance of the genus Camellia in research for its benefits on health and diseases. Wang et al. [camellia seed meal (C. oleifera), in piglets. The dietary supplementation of about 500 mg·kg−1 had beneficial effects on piglets improving liver glutathione peroxidase (GSH-Px) activity but higher supplementations (as 2500 or 5000 mg·kg−1) could lead to growth retardation, hematological abnormalities and organ injuries.Medicinal plants have been used since antiquity exhibiting high benefits on health and proved pharmacological activities. Several species among g others . Negishia et al.  in geneta et al.  also stuo et al.  studied o et al.  studied al. 2010  studied al. 2010  studied al. 2003  studied al. 2003  studied al. 2018  evaluateg et al.  reportedm et al.  studied g et al.  evaluateg et al.  evaluateCamellia species have been extensively studied over the years, being most of the works focused in the three species with higher economic value, namely, C. sinensis, C. oleifera and C. japonica. It should be stressed that few studies have reported differences among cultivars of the same species which is known to highly impact the biological properties of the plants. Also, no comparisons were found between distinct sample collecting seasons nor geographic regions, other known relevant features. Furthermore, the differences encountered in the presentation of the results prevent easy comparisons of the results of the diverse studies. These gaps may justify additional studies on distinct Camellia species that have been somehow neglected, as well as further works comparing the impact of cultivars, collecting seasons and geographic regions on the plant properties. Also, and of high relevance, the already proven biological activity among Camellia genus, clearly show that these species should be further considered in future developments of new natural based drugs.The antioxidant properties and biological activity of


Attenborolimulus superspinosus gen. et sp. nov. A geometric morphometric analysis of 23 horseshoe crab genera illustrates that the new taxon is distinct from limulid and paleolimulid morphologies, supporting the assignment within Austrolimulidae. In considering Triassic austrolimulids, we suggest that the hypertrophy or reduction in exoskeletal sections illustrate how species within the family evolved as opportunistic taxa after the end-Permian extinction.Horseshoe crabs are extant marine euchelicerates that have a fossil record extending well into the Palaeozoic. Extreme xiphosurid morphologies arose during this evolutionary history. These forms often reflected the occupation of freshwater or marginal conditions. This is particularly the case for Austrolimulidae—a xiphosurid family that has recently been subject to thorough taxonomic examination. Expanding the austrolimulid record, we present new material from the Olenekian-aged Petropavlovka Formation in European Russia and assign this material to  Konservat Lagerstätte within the Petropavlovka Formation, Cis-Urals of Russia to promote the study of Austrolimulidae and their role in the Triassic recovery of Xiphosurida. This material is also examined using geometric morphometrics to mathematically illustrate the austrolimulid position of these fossils within xiphosurid morphospace. This evidence, coupled with a thorough taxonomic consideration, prompted us to place the Petropavlovka Formation material within a novel genus and species: Attenborolimulus superspinosus gen. et sp. nov.Examining ecological recovery from the “mother of all extinctions” (the end-Permian extinction) during the Triassic is important for understanding how biological systems can redevelop after major devastating events . TriassiParotosuchus Otschev & Shishkin . Red beds exposed here yield tetrapods, lungfish, clam shrimps (conchostracans), and ostracods . Along oThe studied specimens were collected by the field parties of, and are housed in, the Borissiak Paleontological Institute (PIN), Russian Academy of Sciences, Moscow, Russia. The material was photographed with a Nikon D800 camera mounted with a Nikon AF-S ED Micro Nikkor 60 mm f/2.8G lens. Images were z-stacked with Helicon Focus Pro 6.7. Furthermore, a Leica DFC425 camera coupled to Leica M165C stereomicroscope was used. Finally, to examine possible evidence for finer structures, specimens were examined under a TESCAN VEGA scanning electron microscope (SEM) housed at the PIN. A backscattered electron detector was used as the specimens were not coated.When describing the material, we followed the systematic taxonomy of http://life.bio.sunysb.edu/morph/index.html). A TPS file was constructed using tpsUtil64 (v.1.7). The TPS file was imported into tpsDig2 (v.2.26). This software was used to place four landmarks on the right prosomal section, as well as 50 semi-landmarks along the right prosomal shield border  will represent a published work according to the International Commission on Zoological Nomenclature (ICZN), and hence the new names contained in the electronic version are effectively published under that Code from the electronic edition alone. This published work and the nomenclatural acts it contains have been registered in ZooBank, the online registration system for the ICZN. The ZooBank LSIDs (Life Science Identifiers) can be resolved and the associated information viewed through any standard web browser by appending the LSID to the prefix Family Austrolimulidae Riek, 1955Attenborolimulus gen. nov.Genus Etymology: The generic name is given in honour of Sir David Attenborough and his unparalleled contributions to natural history and conservation. His last name is combined with Limulus—the most-well documented extant xiphosurid genus.Type species:Attenborolimulus superspinosus, new species.Diagnosis. Austrolimulid with anteriorly effaced, ridge-less cardiac lobe, slightly splayed genal spines extending posteriorly to three-fourths of thoracetron length with occipital bands extending to spine terminus, tubercle structures along posterior prosomal and anterior thoracetronic border, medial thoracetronic lobe lacking a sagittal ridge, and long, strongly keeled telson.Attenborolimulus superspinosus sp. nov.8Etymology: Species name reflects the hypertrophied (super-) genal spine (-spinosus) morphology.Holotype: PIN 5640/220 (part and counterpart).Paratypes: PIN 5640/217, PIN 5640/200 (part and counterpart).Type locality and horizon. Petropavlovka III near the village of Petropavlovka, Orenburg region, Russia; Petropavlovka Formation, upper Olenekian, Lower Triassic.Diagnosis. Same as for genus.Preservation. Specimens are preserved as partly domed exoskeletons as part and counterpart on yellowish or grey siltstone.Description. PIN 5640/220 (part and counterpart): An articulated prosoma, thoracetron, and distally incomplete telson : Isolated prosoma preserved more completely in part  and 6. PPIN 5640/217: Central and left side of prosoma , 15.4 mmRemarks: The horseshoe crab material documented herein displays hypertrophied genal spines, a feature common in Belinurina and Austrolimulidae. The examined material lacks the expression of thoracetronic tergites extending from the medial lobe to the thoracetron edge and a rounded thoracetron common to Belinurina. This suggests the material likely belong within Austrolimulidae. Batracholimulus fuchsbergensis .ergensis , Boeotiagispinus , Dubboliakovlevi . Comparimaterial . Franconic lobes . These aingensis . HoweverLimulitellaLimulitella fell into Austrolimulidae, suggesting that the family consisted of limuloids with “apodemal pits present on thoracetron; thoracetron lacking tergopleural fixed spines; posteriormost thoracetron tergopleurae swept back and elongated to form ‘swallowtail’; axis of thoracetron bearing dorsal keel”  and a neutral PC2 space (PC2 = 0.002) (The PCA plot illustrates that PC1 (48.3% shape variation) describes how laterally the most distal genal spine point extends from the sagittal line . PC2 (27= 0.002)  and 10. = 0.002)  and with= 0.002) .Attenborolimulus gen. nov. is a rare occurrence of grey lithologies among the red beds of the Petropavlovka Formation. The red beds yield the lungfish Ceratodus, temnospondyl amphibians, and procolophonid and erythrosuchid reptiles autochthonous fossils of ceratodontid lungfishes, numerous schizophoroid beetle adults, clam shrimp, ostracods, horseshoe crabs, microdriles, and microconchids. The microdrile specimens represent the earliest fossil record of oligochaete annelids. This small worm is similar to modern tubificids, and its relatively well-developed body wall musculature suggests that sediment burrowing was originally another way to escape desiccation on the bottom of seasonally drying ponds  suggesting that the morphological stock required for Triassic diversification had arisen prior to the end-Permian (EuproopsBelinurusVacant ecological space is a key factor in allowing evolutionary innovation to develop . Triassi-Permian . The hig-Permian  and the -Permian . The hyp10.7717/peerj.11709/supp-1Supplemental Information 1Click here for additional data file.10.7717/peerj.11709/supp-2Supplemental Information 2Click here for additional data file.10.7717/peerj.11709/supp-3Supplemental Information 3Click here for additional data file.10.7717/peerj.11709/supp-4Supplemental Information 4Click here for additional data file.10.7717/peerj.11709/supp-5Supplemental Information 5Includes family, generic and temporal data.Click here for additional data file.


Ciliated protists are a widely distributed, morphologically diverse, and genetically heterogeneous group of unicellular organisms, usually known for containing two types of nuclei: a transcribed polyploid macronucleus involved in gene expression and a silent diploid micronucleus responsible for transmission of genetic material during sexual reproduction and generation of the macronucleus. Although studies in a few species of culturable ciliated protists have revealed the highly dynamic nature of replicative and recombination events relating the micronucleus to the macronucleus, the broader understanding of the genomic diversity of ciliated protists, as well as their phylogenetic relationships and metabolic potential, has been hampered by the inability to culture numerous other species under laboratory conditions, as well as the presence of symbiotic bacteria and microalgae which provide a challenge for current sequencing technologies. Here, we optimized single-cell sequencing methods and associated data analyses, to effectively remove contamination by commensal bacteria, and generated high-quality genomes for a number of Euplotia species.We obtained eight high-quality Euplotia genomes by using single-cell genome sequencing techniques. The genomes have high genomic completeness, with sizes between 68 and 125 M and gene numbers between 14K and 25K. Through comparative genomic analysis, we found that there are a large number of gene expansion events in Euplotia genomes, and these expansions are closely related to the phenotypic evolution and specific environmental adaptations of individual species. We further found four distinct subgroups in the genus Euplotes, which exhibited considerable genetic distance and relative lack of conserved genomic syntenies. Comparative genomic analyses of Uronychia and its relatives revealed significant gene expansion associated with the ciliary movement machinery, which may be related to the unique and strong swimming ability.We employed single-cell genomics to obtain eight ciliate genomes, characterized the underestimated genomic diversity of Euplotia, and determined the divergence time of representative species in this subclass for the first time. We also further investigated the extensive duplication events associated with speciation and environmental adaptation. This study provides a unique and valuable resource for understanding the evolutionary history and genetic diversity of ciliates.The online version contains supplementary material available at 10.1186/s12915-021-01202-1. Ciliates, the unicellular organisms with the most complex lifestyles, are found anywhere there is water . Unlike Tetrahymena [Paramecium [There have been a large number of gene duplication events in ciliate genomes, bringing their gene numbers close to those of many plants and metazoans . These grahymena  and Pararamecium . There aramecium , but genIn recent years, single-cell sequencing technology has been applied to the sequencing of unculturable ciliates, which has helped us to solve controversial problems in ciliate phylogenetic and genomic studies , 10. HowCertesia quadrinucleata, Diophrys sp., Euplotes cf. woodruffi, Euplotes parawoodruffi, Euplotes weissei, Euplotes woodruffi, Uronychia binucleate, and Uronychia setigera, respectively. These eight species represent four major genera  in the order Euplotida, whose morphological characteristics have been described previously   and multEuplotes (Table Euplotes cultured species: 95% for Euplotes focardii (accession number in NCBI: GCA_001880345.1), 95% for Moneuplotes crassus (accession number in NCBI: GCA_001880385.1), and 84% for Euplotes vannus [. We employed a comprehensive strategy to annotate the genome by combining homolog-based and ab initio approaches. We finally obtained 9382~25,327 protein-coding genes for each species. Most contigs contained 1~3 genes, which is consistent to previous studies  and GC content (32.0%~45.9%) were similar to those of other macronuclear genomes of 25 Mb ands vannus  . Clans correlated to motor protein genes, tubulin genes, and tubulin modification genes were found to be significantly expanded in Euplotia  contain only one homologous group, and 585 (9.8%) contain more than five homologous groups. Among the 39 clans with a significant discrepancy between Euplotia and Hypotrichia, the cytochrome P450 clan contains 57 homologous groups and ATP-binding cassette transporters contain 42 homologous groups. To find out the relationship between these genes and speciation, we performed phylogenetic analysis of ABC transporter genes and cytochrome P450 genes. As shown in Fig. Euplotes exhibits many genus-specific characteristics, such as unique genetic codons, a high frequency of programmed ribosomal frameshifts, well-developed AZMs, and a single undulating membrane [Euplotes is still unknown because of the lack of genomic data. Here, the successful assembly of four new genomes in this genus enables a comprehensive comparison of genome structure and gene content among the different species. We first performed a synteny analysis of the seven Euplotes genomes and found that they could be divided into four subgroups based on their syntenic similarities  and ion transporter (Ion_trans) were always adjacent to each other among the four subgroups despite their limited sequence identities Fig. B. Previot test, P < 0.01)  (01) Fig. C. Then, 60% Fig. D. We furUronychia is a cosmopolitan ciliated organism that inhabits marine environments or saltwater. Although there are numerous studies concerning its morphology, morphogenesis, and taxonomy, the genus Uronychia remains one of the most mysterious ciliates due to the lack of whole-genome or transcriptome data [Uronychia has more rigid and sculptured body extracellular matrices, which makes its cell very stable. Meanwhile, Uronychia swims faster than other species and has rarely seen sudden jump behavior [Uronychia may have unique genome features related to cell wall generation and ciliary movement.ome data , 31. Combehavior , 4, 32. Uronychia and its relatives in Euplotia and Hypotrichia, we first compared the genomic synteny of two Uronychia species to the seven Euplotes and 10 Hypotrichia species we described above. The genomic structure between U. setigera and U. binucleate was highly conserved but divergent from that of other species. We further analyzed the genes shared in these species and their protein similarity. The number of homologous proteins and their protein identity between the two Uronychia is higher than that of other species. For example, U. setigera has 172 syntenic regions with U. binucleate, but only has five syntenic regions with E. parawoodruffi and five syntenic regions with O. trifallax   Fig. B.Fig. 5Uronychia species. Nine thousand eight hundred seventy-four genes of U. binucleate were classified into 711 GO terms, and 9838 genes of U. setigera were classified into 777 terms. We compared the enriched GO terms between Uronychia species and other 22 Spirotricha species using Fisher’s exact test and found 109 terms were significantly different (P < 0.01), including terms related to chitin synthesis, microtubule, signal transduction, sulfur metabolism, and carboxypeptidase. Chitin is an important component of ciliates [Uronychia, which is consistent with the morphological structure of Uronychia. Genes related to carboxypeptidase in Uronychia are six times higher than those in other Spirotricha species, indicating Uronychia may have diverse functions in protein degradation. Both adenylate cyclase genes and phospholipase genes were significantly expanded, which may suggest that Uronychia has a strong regulation of cyclic adenosine monophosphate (cAMP). In addition, genes related to transsulfuration, vacuolar transport, lipid catabolic, and antioxidant activity underwent contraction and genes related to ion transport and signal transduction underwent expansion  enrichment analysis for the two ciliates , and theion Fig. C.Uronychia exhibited a significant expansion in adenylate cyclase, phosphodiesterase, protein kinase, and ion transporter, which may explain its unique and strong swimming ability.We further proposed a model of ciliary movement machinery based on comparative genomic analysis Fig. D. The axAs one of the most diverse ciliate subclass , the stuSingle-cell sequencing has been applied to explore ciliate genomes and transcriptomes. Due to insufficient DNA content and high amplification bias, it is impossible to obtain a complete genome from a single cell. In addition, amplification of too many cells may increase the error rate caused by single-cell amplification and genomic differences between cells. By combining the advantages of MDA and MALBAC, we successfully obtained both better genome coverage and less amplification bias when processing the DNA samples of these uncultured ciliate cells. The incorporation of whole-genome sequence from a few more cells (2~4) can improve the final genome assembly without sacrificing homogeneity. In addition, excluding bacterial contamination is an important challenge in nonmodel organism genomics research. In this study, we found that the percentage of bacteria-derived reads in the raw sequencing data was generally below 10%, which was significantly lower than that in bulk whole-genome sequencing. Using the iterative filtration method, we were able to remove most of the nonciliate contaminants and obtain eight high-quality Euplotia genomes.Euplotes contains more than 80 species, most of which have similar cell shapes, cirri counts, and well-developed AZMs [Through the combination of homology search approach and functional annotation approach, we found that genes related to environmental adaption exhibited huge expansion. When a gene duplication event occurs after speciation, the genes tend to vary greatly between species due to evolution. In contrast, when gene duplication events occur before speciation, the genes tend to divide into several lineages shared by all species Fig. C. In higped AZMs . Based oIn this study, we optimized the single-cell sequencing methods and corresponding data analyses to effectively remove contamination by commensal bacteria and to generate high-quality ciliate genomes. We obtained eight ciliate genomes using this approach, and through comparative genomics analysis, we characterized the underestimated genomic diversity of Euplotia and determined the divergence time of representative species in this subclass for the first time. We further investigated the extensive duplication events associated with speciation and environmental adaptation. This study provides a unique and valuable resource for understanding the evolutionary history and genetic diversity of ciliates.E. woodruffi was collected from a freshwater pond in Taipingjiao Park in Qingdao, China. E. cf. woodruffi was collected from 17‰ brackish water pond in Shenzhen Binhai Park, China. E. parawoodruffi was collected from15‰ brackish water from Qingdao Coast, China. The other samples were collected from coastal seawater in Qingdao, China  and merged. Fastp v0.20.1 [Sequencing data from the same species were evaluated using FastQC v0.11.9  as its codon, which is the same as Tetrahymena. The other four species, Euplotes cf. woodruffi, Euplotes parawoodruffi, Euplotes weissei, and Euplotes woodruffi, used “translation table 10” (TGA encodes cystine). For species using translation table 6, we used the Tetrahymena dataset to predict genes. For species using translation table 10, we modified the configuration file of the Tetrahymena dataset by choosing translation table 10, and then performed gene prediction. To ensure the quality of the predicted genes, we also trained AUGUSTUS by using nucleic acid and protein sequences of Oxytricha trifallax (http://oxy.ciliate.org/index.php/home/downloads) and Euplotes vannus (http://evan.ciliate.org/index.php/home/downloads). The NR database was used to annotate genes using DIAMOND v0.9.26 in protein search mode . Motifs and protein domains were annotated against the PANTHER, Pfam, Gene3D, and CDD databases using InterProScan v10.0.2 [AUGUSTUS v3.3.3  was used v10.0.2 . GO and OrthoFinder v2.3.12  was appl7. The convergence of parameters was evaluated using Tracer v1.7.1 [We used two different multiple alignment datasets for phylogenetic analysis. The first dataset contained single-copy genes of 24 Spirotricha species using protein by protein search process in OrthoFinder. The second dataset was obtained from GPSit v1.0  pipeliner v1.7.1 .JCVI utility libraries v1.0.9  were useAdditional file 1: Tables S1-S3, Figures S1-S6. Supplementary tables and figures. Table S1. Single-cell amplification of eight Ciliates. Table on MDA and MALBAC methods for amplification of single cells. "1" means single-cell amplification success, "0" means amplification failure. For each amplification experiment, only one cell was used. Table S2. Information of 24 ciliate genomes. Table summarizing the data category, data source, accession number and hyperlink of the species used in this research. Table S3. Commands for analyzing process. Software, versions and parameters used for each analysis step. Fig. S1. Genome completeness assessed by BUSCO and EukCC. The completeness of the eleven Euplotia genomes assessed using EukCC and BUSCO software. Bold font indicates the species newly sequenced in this research, and regular font indicates species already published. Fig. S2. Percentage of contigs containing the number of genes. The proportion of contigs containing different gene counts out of the total contigs in each genome. Contigs containing more than 5 genes were grouped together. Fig. S3. Phylogenetic tree of all the sequenced ciliate species. Phylogenetic tree based on 157 gene sequences. The tree was constructed by the maximum likelihood method (C60+LG+G+F model). The numbers on the nodes represent the bootstrap values. Fig. S4. Phylogenetic tree based on single-copy genes. Phylogenetic tree of 19 Euplotia species base on single-copy genes obtained from compared genetic analysis. Fig. S5. Gene ontology enrichment. Gene ontology enrichment of housekeeping genes in Euplotia. The tree was constructed by the maximum likelihood method (C60+LG+G+F model). The numbers on the nodes represent the bootstrap values. Fig. S6. Gene gain/loss in this study. Gene gain/loss in Euplotia and Hypotrichia. Red letter represents gene gain, and blue letter represents gene loss.


Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) to which were given the highest “priority status” since they represent the great threat to humans. Understanding the resistance mechanisms of these bacteria is a key step in the development of new antimicrobial drugs to tackle drug-resistant bacteria. In this review, both the mode of action and the mechanisms of resistance of commonly used antimicrobials will be examined. It also discusses the current state of AMR in the most critical resistant bacteria as determined by the WHO’s global priority pathogens list.Antibiotics have made it possible to treat bacterial infections such as meningitis and bacteraemia that, prior to their introduction, were untreatable and consequently fatal. Unfortunately, in recent decades overuse and misuse of antibiotics as well as social and economic factors have accelerated the spread of antibiotic-resistant bacteria, making drug treatment ineffective. Currently, at least 700,000 people worldwide die each year due to antimicrobial resistance (AMR). Without new and better treatments, the World Health Organization (WHO) predicts that this number could rise to 10 million by 2050, highlighting a health concern not of secondary importance. In February 2017, in light of increasing antibiotic resistance, the WHO published a list of pathogens that includes the pathogens designated by the acronym ESKAPE ( S. aureus developed resistance to penicillin that in the past had normally been used to treat them [Acinetobacter, Pseudomonas, some Enterobacteriaceae such as: K. pneumoniae, E. coli and Enterobacter spp. [Enterococcus faecium and Staphylococcus aureus that are resistant to various antibiotics, such as vancomycin and fluoroquinolones. The medium priority category includes bacteria such as Streptococcus pneumoniae and Shigella that, although they may have some resistance, effective antibiotics are still available that can kill them [Although antimicrobial resistance (AMR) is a natural process, the public health emergency due to the uncontrolled spread of this phenomenon depends primarily on the overuse of antibiotics . Howevereat them . Nevertheat them . Unfortueat them . In 2017eat them . The WHOeat them ,9. Crititer spp. . These pill them . In 2019, due to its impact on human health, the World Health Organization (WHO) included antimicrobial resistance (AMR) as one of the top ten threats to global health . Escherichia coli and ampicillin, 1st and 2nd generation cephalosporins resistance in Pseudomonas aeruginosa [According to the World Health Organization, antimicrobial resistance is a natural phenomenon that occurs when microorganisms no longer respond to antibiotics to which they were previously susceptible and that were previously active in treating infections caused by these microorganisms ,12. As aruginosa ,20. Naturuginosa . The acqruginosa ,22. The Antibiotic-resistant genetic material is then transferred from the antibiotic-resistant bacteria to the non-resistant bacteria that become resistant to antibiotics .The rapid spread of AMR through bacterial populations cannot be attributed to a single mechanism. It is often the result of complex processes. It is therefore necessary to subdivide antibiotics into groups based on the different mechanism of action before analyzing the factors that affect resistance to these molecules. Although there are many different classes of antibiotics, in this review, we have chosen to describe those most closely involved in the occurrence of antibiotic resistance. Since the mechanism of action of different antibiotics is largely dependent on both the nature of their structure and the affinity of these agents for different bacterial structures, it follows that knowledge of the mechanism of action of these agents is “the condition sine qua non” for understanding the emergence of resistance to these drugs ,26. A deIn this review, the resistance of bacterial pathogens is discussed according to their categories established by the WHO . A. baumannii is an opportunistic pathogen that causes hospital-acquired infections worldwide and can develop resistance to antibiotics by different mechanisms such as:(1)Acinetobacter spp. have been identified both the genes encoding for narrow-spectrum β-lactamases  and those encoding for ESBL (GES-11 and CTX-M) [A. baumannii also possesses Class D or OXAs β-lactamases that can hydrolyze extended-spectrum cephalosporins and carbapenems [A. baumannii has an intrinsic ampC cephalosporinase [the production of enzymes that degrade beta-lactam antibiotics. The production of all four classes of β-lactamases  through the incorporation of exogenous DNA into its genome would underlie the rapid evolution of this strain toward multi-resistance ,29. Mored CTX-M) . As statd CTX-M) . Class Cd CTX-M) ,33. A. bbapenems . Moreoveporinase ;(2)A. baumannii efflux pumps are involved in bacterial resistance to a number of antibiotics belonging to different chemical classes such as aminoglycosides, tetracyclines, erythromycin, chloramphenicol, trimethoprim, fluoroquinolones and different beta-lactams [A. baumannii antimicrobial resistance: the major facilitator superfamily (MFS), the resistance nodulation division (RND) superfamily, the multidrug and toxic compound extrusion (MATE) family and the small multidrug resistance (SMR) family transporters [A. baumannii [the expression of efflux pumps. In -lactams ,37. Diffsporters ,38. Moreaumannii ;(3)A. baumannii to aminoglycosides [the enzymatic modification of aminoglycosides. Enzymatic modification is the most common type of aminoglycoside resistance . Acetyltycosides . The genycosides .(4)A. baumannii the reduced expression of porins, proteins that allow the transport of molecules across the outer membrane, is associated with carbapenem resistance [A. baumannii may acquire resistance to colistin, a polypeptide antibacterial agent that targets LPS, as a result of mutation of the genes involved in LPS biosynthesis [the production of modified porins that decreases the permeability of the outer membrane ,43. In Asistance ,44. Moreynthesis ,46;(5)A. baumannii this mechanism of resistance is mediated by overexpression of penicillin-binding proteins that results in imipenem resistance or by mutations of DNA gyrase that prompts quinolone and tetracycline resistance [the modification of the antibiotic target . In A. bsistance ,30.Acinetobacter baumannii is an aerobic gram-negative bacillus that belongs to the group of pathogens grouped under the acronym “ESKAPE” , which refers to the ability of these bacteria to escape the effect of bactericidal activity of antibiotics ,27. A. bA. baumannii infections [A. baumannii bacteremia [A. baumannii infections are treated with a combination of minocycline and colistin while colistin/rifampin is the most effective treatment for colistin-resistant A. baumannii [A. baumannii [A. baumannii.Until a few years ago, carbapenems like imipenem and meropenem were the most effective agents to treat fections . These afections ,49. Ampicteremia . Minocyccteremia . Minocycaumannii . Moreoveaumannii ,53. HoweP. aeruginosa is an aerobic gram-negative bacterium commonly found in the environment and one of the most common pathogens responsible for a variety of acute and chronic nosocomial infections including severe respiratory infections in patients with compromised host defenses [P. aeruginosa is the third most common gram-negative bacteria causing nosocomial bloodstream infections [P. aeruginosa has shown intrinsic resistance to many antibiotics that is due to different mechanisms of resistance that are both intrinsic and acquired from other microorganisms [P. aeruginosa coverage [A. baumanni, also in P. aeruginosa all four major classes of β-lactamases  have been identified [P. aeruginosa can acquire resistance through a gene mutation which leads to overexpression of AmpC β-lactamases [Pseudomonas resistance to aminoglycosides is mediated by transferable aminoglycoside modifying enzymes (AMEs) that decrease the binding affinity to their target in the bacterial cell [P. aeruginosa involves colistin in combination with an anti-pseudomonas agent like imipenem, piperacillin, aztreonam, ceftazidime or ciprofloxacin [P. aeruginosa have been successfully treated with fosfomycin in combination with aminoglycosides, cephalosporins and penicillins [defenses ,55. In tfections . P. aerurganisms ,58. The rganisms ,60. Ceftcoverage . Like A.entified . Endogenentified . Moreovectamases . Pseudomial cell ,66. The floxacin . Drug reicillins ,66. S. aureus, a major human pathogen, is a gram-positive, facultative anaerobe, catalase- and coagulase-positive coccus that tends to form irregular grape-like clusters [S. aureus causes infections ranging from mild to life-threatening such as skin and soft tissue infections, bacterial endocarditis, pleuropulmonary and device-related infections [S. aureus carrying a plasmid-encoded beta-lactamases capable of hydrolyzing the β-lactam ring of penicillin [S. aureus strain was identified [S. aureus by binding to the penicillin-binding proteins (PBPs). S. aureus became resistant to methicillin (MRSA) by acquiring, via horizontal gene transfer, the genes mecA and mecC which inactivate methicillin by the synthesis of an alternative PBP, designated PBP2a, that has very low affinity for almost all β-lactam antibiotics [S. aureus (VRSA) [VanA operon carried on the mobile genetic element Tn1546 acquired from vancomycin-resistant Enterococcus [S. aureus (VISA) which is not inhibited in vitro at vancomycin concentration below 4–8 µg/mL. In contrast, vancomycin-resistant S. aureus (VRSA) is inhibited only at concentrations of 16 µg/mL or more [clusters . S. aurefections . This mifections . For exanicillin . This genicillin ,72. In 1entified . Methiciibiotics ,74. For ibiotics . Howevers (VRSA) . The resrococcus ,78. In 1 or more . VISA an or more .S. aureus poses a severe threat to human health worldwide [gyrA and gyrB genes of topoisomerse II); (2) target alterations; (3) overexpression of efflux pump (NorA pump) [S. aureus is uncommon, resistance to this drug during therapy is increasing due to mutations of different proteins that result in a reduced drug binding to its target site [S. aureus is well known for its ability to acquire resistance to other antibiotics such as trimethoprim-sulphamethoxazole and tetracyclines by aforementioned different mechanisms of resistance [S. aureus skin infections [S. aureus and MRSA. However, numerous reports indicate that clindamycin resistance is also increasing among health care-associated MRSA strains. MLS resistance is due to three main mechanisms: target modification, active efflux and enzymatic antibiotic inactivation [erm genes  is the main mechanism [According to the World Health Organization (WHO), the pathogenicity and antibiotic resistance pattern of orldwide . MRSA, Vorldwide . MRSA inorldwide ,80,81. IrA pump) . DaptomyrA pump) . Howeverget site ,29. Moresistance . Since hfections ,83. In rfections ,84. Givetivation . Among techanism . These gechanism . Inducibechanism . Howeverechanism ,87. Infeechanism .K. pneumoniae is a member of the family Enterobacterales, non-fastidious, commonly encapsulated gram-negative bacillus [K. pneumoniae can cause different types of nosocomial and community acquired infections, including urinary tract infections, pneumonia, liver abscess, surgical site infections and bloodstream infections especially in immunocompromised patients [Klebsiella infection person-to-person contact is required [Klebsiella has become highly resistant to antibiotics by the widespread acquisition of genes encoding enzymes, such as ESBLs and carbapenemases [K. pneumoniae strains are the most clinically prominent carbapenem-resistant Enterobacteriaceae (CRE) [K.pneumoniae (KPC) strains harboring the carbapenemase encoding blaKPC-3 gene, is a major threat to public health [bacillus . K. pneupatients ,89. Sincrequired . Klebsieenemases . Carbapeae (CRE) . Carbapec health ,94.Enterobacter species are motile aerobic gram-negative bacilli belonging to Enterobacteriaceae family. The Enterobacter cloacae complex (ECC) includes different pathogens, capable of producing a wide variety of infections, the most frequent of which are Enterobacter cloacae and Enterobacter aerogenes [E. aerogenes was re-classified as Klebsiella aerogenes owing to its higher genotypic similarity with the genus Klebsiella [Enterobacter species are non-fastidious gram-negative rods that are sometimes encapsulated [Enterobacter strains produce ESBLs and carbapenemases, including VIM, OXA, metallo-β-lactamase-1, and KPC [K. aerogenes has also emerged, displaying resistance to the last-resort antibiotic colistin [K. aerogenes is capable of harboring subpopulations of colistin-resistant bacteria which are undetectable using current diagnostic testing strategies [erogenes . In 2019ebsiella . Enterobpsulated . They capsulated . Many En and KPC . Further and KPC . These M and KPC ,99. Morecolistin . To furtrategies .E. faecalis and E. faecium [E. faecalis is the most pathogenic species although E. faecium is more resistant to many antimicrobial agents and especially in immunocompromised hosts the latter can cause severe morbidity and mortality [E. faecium by greatly increasing the normal gram-negative intestinal microbiota (mutated PBP and the overexpression of β-lactamase enzymes lead to high levels of resistance to β-lactam antibiotics) [E. faecium, at least three different pathways involved in cephalosporins resistance have been identified [E. faecium can acquire genes through mobile genetic elements such as plasmids and transposons (i.e vancomycin resistance can be transferred by the vanA gene cluster on the transposon Tn1546) [van gene clusters such as vanR, vanS, vanH, vanX and vanZ that are responsible for the replacement of D-Ala-D-Ala with D-alanyl-D-lactate termini. Vancomycin binds to d-Ala–d-Lac much more weakly than it does to the normal dipeptide product resulting in a low binding affinity of vancomycin [Van A gene cluster is the most common type and was located on transposon on a 10,581-bp transposon (Tn1546) of E. faecium [Enterococci are gram-positive cocci, facultative anaerobes gastrointestinal commensals capable of persisting in a range of stressful and hostile environments . Althoug faecium . E. faecortality ,102. In biotics) ; (2) thebiotics) ; (3) thebiotics) . In E. fentified ,104. In entified . Then, ientified . E. faec Tn1546) . Vancomy Tn1546) . Vancomyncomycin . Van A g faecium .E. faecium is considered a MDR bacteria since it is intrinsically resistant to aminoglycoside like tobramycin, kanamycin, gentamicin being capable of producing aminoglycoside-modifying enzymes (AMEs) including aminoglycoside nucleotidyltransferases (ANTs) aminoglycoside acetyltransferases (AACs) and aminoglycoside phosphotransferases (APHs) [rpsL gene, which encodes the ribosomal protein S12, can result in high level resistance to streptomycin [E. faecium is most frequently linked with point mutations in gyrA and parC genes that encode subunits A of DNA gyrase and topoisomerase IV or with efflux transporter NorA that pump out these drugs [s (APHs) . Moreoveptomycin ,109. Morse drugs . Antibiotic resistance is the ability of bacteria to resist exposure to antibiotics designed to kill them or inhibit their growth. Although antibiotic resistance is a natural process due to genetic changes in the bacteria following antibiotics exposure, however, this phenomenon is being accelerated through the overuse and misuse of antibiotics. Overuse of antibiotics causes susceptible bacteria to be killed and allows drug-resistant bacteria to proliferate. Poor sanitation, poor infection control and the use of antibiotics in farm animals are among the main reasons for the spread of antimicrobial resistance. In addition, there are novel and often underrecognized mechanisms of resistance that further contribute to drug resistance such as the heteroresistance (HR) and the mutant prevention concentration (MPC). The first of these two factors can be defined as resistance to certain antibiotics by a preexisting subpopulation of resistant cells, within a larger population of antimicrobial-susceptible microorganisms . This suThe ESKAPE pathogens represent deadly bacteria with rapidly growing multi-drug resistant properties. Although these bacteria are genetically different, the resistance strategies that underlie the emergence and persistence of these pathogens are widely shared among them including decreased drug uptake, drug target alteration, drug inactivation and drug efflux pumps activation. To limit the spread of ESKAPE pathogens and antibiotic resistance more generally, it has become imperative to be more careful in surveillance and implementation of antimicrobial stewardship in both human health and food animals. Implementation of these programs together with the development of new antibiotics or new approaches (e.g. inhibition of biofilm formation and bacteriophage therapy) are likely the only way to slow the spread of multi-drug resistant strains worldwide.


Listeria monocytogenes F2365 by utilizing the asymptotic deceleration point (PDA) in a logistic model (LM), namely MIC-PDA. The experimental data obtained from the inhibitory rate (IR) versus OLE concentration against L. monocytogenes were sufficiently fitted (R2 = 0.88957). Five significant critical points were derived by taking the multi-order derivatives of the LM function: the inflection point (PI), the maximum acceleration point (PAM), the maximum deceleration point (PDM), the absolute acceleration point (PAA), and the asymptotic deceleration point (PDA). The PDA ([OLE] = 37.055 mg/mL) was employed to approximate the MIC-PDA. This MIC value was decreased by over 42% compared to the experimental MIC of 64.0 mg/mL, obtained using the conventional 2-fold dilution method . The accuracy of MIC-PDA was evaluated by an in vitro L. monocytogenes growth inhibition assay. Finally, the logistic modeling method was independently validated using our previously published inhibition data of OLE against the growths of Escherichia coli O157:H7 and Salmonella enteritidis. The MIC-PDA (for [OLE]) values were estimated to be 41.083 and 35.313 mg/mL, respectively, compared to the experimental value of 62.5 mg/mL. Taken together, MIC-PDA, as estimated from the logistic modeling, holds the potential to shorten the time and reduce cost when OLE is used as an antimicrobial in the food industry.Olive leaf extract (OLE) has been increasingly recognized as a natural and effective antimicrobial against a host of foodborne pathogens. This study attempts to predict the minimum inhibitory concentration (MIC) of OLE against  The resulting parameters , contrasting our previously experimentally determined IR of 95% for E. coli O157:H7  = 0.0 mg/mL), i.e., the positive control. Once again, e.g., [OLE] > 0.0 mg/mL) against a bacterial strain (when IR > 0.0%).Consistent with the LM results of R2 = 0.88957) for the experimental data of L. monocytogenes F2364 inhibitory rate (%) as a function of OLE concentration. Taking multi-order derivatives of the logistic function  yielded four meaningful critical points, i.e., PI, PAM, PDM, and PAD, and five phases. The asymptotic deceleration point (PAD) proved to be of particular significance as it (MIC-PDA) approximated the minimum inhibitory concentration (MIC) of OLE and agreed reasonably well with the MIC value obtained from the experimental inhibition assays. This work demonstrated combined approaches to yield MIC, i.e., estimation by taking the derivatives of a logistic function fitted with the inhibition data and direct determination by inhibition essays. We further independently validated the modeling method by applying the logistic function , 41.083 and 35.313, respectively, were significantly lower than those experimental MIC values, 62.5 mg/mL, similar to the case of OLE inhibition of L. monocytogenes studied in this work. Moreover, the LM results supported the observation that OLE was less effective against E. coli O157:H7 than S. enteritidis and L. monocytogenes. This logistic modeling procedure may be of particular interest in those cases when the observed variables follow a logistic relationship and help understand the biological events involved.A logistic relationship was established Click here for additional data file.


Streptococcus pyogenes can cause severe illnesses such as toxic-shock syndrome and necrotizing fasciitis due to pyrogenic exotoxins. Clindamycin is added to penicillin for treatment of severe S. pyogenes infections as it is a bacterial protein synthesis inhibitor which reduces toxin production. However, clindamycin is associated with several adverse effects including C. difficile infection. Linezolid is a bacterial protein synthesis inhibitor that has been shown to provide excellent coverage of S. pyogenes including toxin inhibition in vitro, but clinical evidence is lacking. We compared outcomes of patients treated with linezolid versus clindamycin for serious S. pyogenes infections. S. pyogenes admitted to the Shock Trauma Center at University of Maryland Medical Center treated with at least 48 hours of either clindamycin or linezolid. Data collected included Sequential Organ Failure Assessment (SOFA) and Laboratory Risk Indicator for Necrotizing Fasciitis (LRINEC) severity scores, time to resolution of infection, number of surgeries, C. difficile infection, other antibiotic associated adverse effects, and mortality. Associations between patient characteristics, antibiotic groups, and outcomes were analyzed using the chi-square test, Fisher’s exact test and t-test or Wilcoxon rank-sum test as appropriate (SAS v 9.4). This was a retrospective study of patients with necrotizing fasciitis or toxic shock syndrome caused by C. difficile infection (3.9% vs. 0% p = 1.0) and thrombocytopenia .52 patients were included, 26 treated with clindamycin and 26 with linezolid. Most patients (85% clindamycin and 96.2% linezolid) were treated for necrotizing fasciitis. Baseline characteristics, including SOFA and LRINEC scores, were similar between the groups. There was no difference in mortality between patients treated with clindamycin versus linezolid , and resolution of infection was similar between the groups . There was no difference in adverse effects between the clindamycin and linezolid groups, including S. pyogenes infections. Further prospective studies are needed. Linezolid could be an alternate to clindamycin for the treatment of serious toxin producing Emily Heil, PharmD, MS, BCIDP, Nothing to disclose