Lactobacillus-dominant in most Caucasian women. Previous studies suggest that the VMB of women with African ancestry is more likely to be non-Lactobacillus dominant (dysbiotic) compared to other populations, and possibly relate to the high incidence of pregnancy complications, such as preterm birth. This work reviewed the literature on VMB composition in pregnant women from sub-Saharan Africa.The vaginal microbiota (VMB) are the set of microorganisms residing in the human vagina. During pregnancy, their composition is A search was conducted in PubMed and Embase databases following PRISMA guidelines. Observational and intervention studies analysing VMB communities from sub-Saharan African pregnant women using molecular techniques were included.Lactobacillus-dominant VMB (particularly L. iners or L. crispatus) or VMB containing Lactobacilli are the most prevalent, followed by a more diverse anaerobe-dominant VMB, in the studied populations. The majority of pregnant women with a sexually-transmitted infection had a Lactobacillus-dominant VMB, but with a significantly higher presence of anaerobic species.Ten studies performed in seven sub-Saharan African countries were identified. They independently showed that Lactobacillus species are the most prevalent VMB species during pregnancy in sub-Saharan African women. The frequency of diverse anaerobe-dominant VMB is high in these populations. In Africa, studies on VMB in pregnancy are scant, heterogeneous in methodology, and knowledge remains limited. More insights on VMB composition and their possible sequalae among these populations is needed.In agreement with studies performed in other populations, The online version contains supplementary material available at 10.1186/s12884-021-04072-1. Lactobacillus species: Lactobacillus iners, Lactobacillus crispatus, Lactobacillus gasseri, or Lactobacillus jensenii [The number of studies on the microbial communities residing in the human vagina, the vaginal microbiota (VMB), and their role in female reproductive health has increased over the past two decades. Recent developments in molecular biology offer innovative opportunities for VMB profiling, but a full understanding of its role in reproductive health is still missing . Usuallyjensenii , 3. LactL. crispatus, L. gasseri, or L. jensenii relate to a healthy vaginal state, and that an overgrowth of (facultative) anaerobes, such as Gardnerella, Atopobium, and Prevotella species contributes to a dysbiotic vaginal state. A dysbiotic vaginal state is often defined as a prolonged deviation from a low-diversity, Lactobacilli-dominated VMB [Lactobacillus species are associated with a healthy vaginal state, low proinflammatory cytokine production, and desirable birth outcomes [L. iners-dominated VMB [L. iners is generally considered a vaginal symbiont, but it can also be a potential opportunist pathogen [L. crispatus has been shown to hinder colonization by anaerobe bacteria such as G. vaginalis, whereas L. iners co-exists with such aerobic microorganisms [There is increasing evidence that VMB dominated by ated VMB . This vaated VMB . While moutcomes , 7, it rated VMB –10. L. ipathogen . Furtherrganisms . Howeverrganisms .Gardnerella vaginalis, A. vaginae, and other anaerobic species instead of by Lactobacillus species [Lactobacillus-dominant VMB did not have any clinical symptoms associated with bacterial vaginosis. This suggests the possibility of a healthy non-Lactobacilli-dominant VMB state [Recent studies show that women from African countries or African ancestry more often have a VMB composition that contains BV-related bacteria compared to Caucasian women , 12–16.  species , 12–16. MB state , 18. It Lactobacillus species are increased and G. vaginalis and other (facultative) anaerobic bacteria progressively decrease from the first to the third trimester [Lactobacillus-dominant VMB among women of African ancestry is lower compared to women of European ancestry. Furthermore, women of African ancestry are more likely to switch from Lactobacillus dominated VMB to a VMB commonly associated with vaginal dysbiosis [Understanding the role of VMB in health and disease becomes complex when comparing women with different ethnic backgrounds , 17, 19.rimester . Followirimester , 21. Howysbiosis . Vaginalysbiosis , 23–25.Neisseria gonorrhoeae (NG), 6.5% for syphilis, 7.2% for C. trachomatis (CT), 25% for Trichomonas vaginalis (TV), and mother-to-child transmission of the human immunodeficiency virus (HIV) ranged between 5 and 30% in sub-Saharan Africa [An exhaustive understanding of what constitutes a healthy/normal VMB is lacking. Such knowledge, including appreciation of how VMB composition might impact pregnancy outcomes, is of great importance in areas where the overall burden of pregnancy complications and infections is high , 27. Subn Africa , 28–31.There is increasing research interest in understanding how the VMB composition might contribute to, and offer a predictive diagnostic potential for reproductive health outcomes. To date, most VMB pregnancy-related studies and reviews are based on data from North-American and European cohorts of women. Factors influencing the VMB, such as the host (including host-genetics), behavioural , sociodemographic, nutritional, and environmental ones, are nonetheless different across geographical regions and cultures , 32, 33.This review aims to compile available data from original research that characterized the VMB composition among pregnant women living in sub-Saharan Africa. Insights from this review are expected to offer background evidence to assist in designing future observational studies to investigate the VMB’s role on pregnancy outcomes and STIs. These are also expected to assist the design of future VMB-related intervention studies that aim to decrease vaginal dysbiosis and its related health problems in sub-Saharan African countries.Searches according to the Preferred Reporting Items for Systematic Review and Meta-Analysis Statement (PRISMA) guidelines were conducted in PubMed/Medline and Embase (Ovid) among studies published up to July 15th, 2020  capsules, and their control group placebo or no intervention , 41. TheLactobacillus species abundance [Pregnancy is characterized by high circulating estrogen levels produced by the ovaries and later by the placenta . High lebundance , 45. Botbundance , 46, 47.bundance –50.L. iners-dominant with a high abundance of G. vaginalis, A. vaginae, and Prevotella species (60%); this was followed by an L. crispatus-dominant VMB cluster (20%) [Lactobacillus genus was present in all 30 Kenyan pregnant women at 14 weeks of gestation, followed by P. bivia (97%), L. iners (80%), and G. vaginalis (50%) [P. bivia in their VMB [Two studies collected specimens in the first trimester )  . Jesper is (50%) . These mheir VMB .Fig. 1MLactobacillus-dominant VMB [Lactobacillaceae were again observed as the dominant VMB bacterial family among 478 Burkinabe pregnant, with a Lactobacillus-dominant VMB present in 78% of participants, L. crispatus in 44.9,% and L. iners in 33.1% [Two studies reported the VMB of pregnant women in the first and the second trimester (13–27 weeks GA)  or L. crispatus (17%) during the second trimester of the pregnancy [G. vaginalis, and 42% of women had a G. vaginalis-dominant VMB [Moreover 96 HIV-infected and 158 HIV-uninfected Zambian pregnant women had a regnancy  Lactobacillaceae as the dominant VMB bacterial family among 53 pregnant Kenyan women [Masha et al. included samples collected in the second and third trimester (≥14 weeks GA) and identified an women  Lactobacillus-dominant VMB. In Burkina Faso, 47% of women had a Lactobacillus-dominant VMB, while 37.5% of them had a VMB dominated by both Gardnerella species and L. iners [Lactobacillus species were further identified. The majority of pregnant women in that cohort had a VMB dominated by L. crispatus (31%), followed by a diverse VMB with various mixed anaerobes bacteria, including G. vaginalis (41%) [Lactobacillaceae as the dominant VMB bacterial family among 38 Rwandan pregnant women across all trimesters (20 women who received Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 and 18 women who received placebo) [The two studies that reported the VMB composition at the third pregnancy trimester (> 27 weeks GA) also reported that most Burkinabe and Zimbabwean women had is 41%) . Another% . AnothLactobacillus species were most prevalent among 21 Rwandese pregnant women, in particular L. iners [Gardnerella and Prevotella species [Lastly, Borgdorff et al. did not report sampling time  Fig. 1)1). But lL. iners . The sec species , 43.Lactobacillus-dominant VMB composition or high concentrations of Lactobacillus species in their VMB. However, based on the higher frequency of a diverse anaerobic VMB in the sub-Saharan African population and women with African ancestry, two hypotheses were drawn [These sub-Saharan African studies, irrespective of sampling time and population reported that most women had a VMB re drawn , 15, 51.re drawn , 12–16, Lactobacillus-dominant VMB is the most prevalent vaginal microbiota in tested pregnant women despite differences in methodology and definitions [L. iners was the most pervasive Lactobacillus species detected, followed by L. crispatus [G. vaginalis [Lactobacillus dominant vaginal milieu, such as genetic variation, like polymorphism in the immune and hormone-response genes, for instance single nucleotide polymorphisms in toll-like receptor (TLR)-4 (rs1554973 and rs7856729 or anti-inflammatory interleukin-10 (IL10–819 T/T and IL10–1082 A/A) [-Lactobacillus dominant VMB may mediate the host immunological reaction to microbiota species and nurture immunological tolerance [To date, there is no consensus on how best to classify VMB communities  Tables  and 3. Minitions –43, 52. initions , 53. Indrispatus , 39–42  . Several082 A/A) –63. Inte082 A/A) , 64. The082 A/A) , 33, 55.olerance , 64.FigL. crispatus-dominant VMB was slightly higher than the prevalence of L. iners-dominant VMB or VMB with a high abundance of G. vaginalis, Bacterial vaginosis-associated bacterium 1 (BVAB1), and mixed diverse anaerobic bacteria [L. crispatus dominant VMB [L. iners and diverse anaerobic VMB were the most dominant VMB [Lactobacillus genus was the highest in their cohort from Burkina Faso and Tanzania, respectively [Lactobacillus-dominant VMB clusters (L. crispatus and L. iners) in Burkinabe women [Lactobacillus genus in their VMB, P. bivia (70%), and G. vaginalis (50%) were also detected in the majority of women in South Africa [P. bivia present in their VMB [L. crispatus, L. gasseri, or L. jensenii relates to a healthy vaginal state and an overgrowth of anaerobes as Gardnerella, Atopobium, and Prevotella species most likely contributes to a dysbiotic-BV state. However, it is unclear whether the VMB clusters consistent with the presence of modest Lactobacillus species with a higher relative abundance of anaerobic bacteria, such as BVAB1, G. vaginalis, A. vaginae, but without an overgrowth of anaerobes bacteria, associate with a healthy vaginal state or with a transitional state of the vaginal milieu to a BV state [In pregnant Zimbabwean women, the prevalence of bacteria . Interesnant VMB . These onant VMB , 51. Unfnant VMB , 25, 39.be women . Jesper h Africa . In Kenyheir VMB . PetrovaBV state .L. iners-dominated VMB [Lactobacillus species, L. iners does secrete some amounts of hydrogen peroxide (H2O2), but not D-lactic acid, both common secretory products of most Lactobacilli [L. iners also been shown to promote the growth of E. coli and G. vaginalis biofilm and can produce pore-forming toxin similar to G. vaginalis can induce lysing of erythrocytes [L. iners are more likely to transition to a BV-associated VMB and offer limited protection against vaginal dysbiosis, clearance of urogenital pathogens, and might be a risk factor to adverse pregnancy outcomes [Lactobacillus iners dominance at 16 weeks of gestation was significantly associated with both a short cervix< 25 mm and preterm birth < 34 + 0 weeks [L. iners-dominant VMB and a diverse VMB are on pregnancy outcomes, especially in the sub-Saharan population where the prevalence of these VMB compositions are high.Moreover, it is unclear what type of vaginal state is associated with a ated VMB . Unlike obacilli . L. inerhrocytes –69. Seveoutcomes , 70, 71. 0 weeks . HoweverL. crispatus associates with suppression of viral replication and virus inactivation [Lactobacillus species, other than L. iners, are important to vaginal health as they produce antimicrobial molecules. For instance, H2O2 and other bacteriocins can destroy urogenital pathogens, and lactic acid can inhibit pathogenic bacteria’s growth and disrupt the bacterial cell membranes, thereby contributing to the host immune response to bacterial liposaccharides [Lactobacillus species, since the vaginal tracts of asymptomatic women with a diverse VMB are also dominated by taxa that produce lactic acid [Globally, 70% of all people living with HIV live in sub-Saharan Africa , 55, 73.tivation –77. As mcharides . Lactic tic acid . Women wtic acid , 78.Luckily, antiretroviral therapy (ART) and associated approaches for the prevention of Mother-To-Child Transmission of HIV infection (MTCT) are becoming more standard practice in most sub-Saharan African settings –82. PricP < 0.01) with a more diverse and anaerobic VMB composition in Burkinabe pregnant women [Besides HIV, other STIs are highly endemic in sub-Saharan Africa, including TV and CT, and have been associated multiple times with vaginal dysbiosis, lower Lactobacilli levels, and alkaline vaginal milieu , 83–85. nt women .Anaerococcus and Corynebacterium were most abundant compared to TV-infected women or controls. In line with other studies’ observations, in TV-infected women, Parvimonas and Prevotella were most abundant and significantly more present compared to controls [Prevotella, Anaerococcus, Parvimonas (formerly Peptostreptococcus), and Dialister have been observed frequently in women with vaginal dysbiosis (with CST IV) or BV; however, it is still unclear what the role of Parvimonas, Anaerococcus, Dialister, and Corynebacterium is in the pathology of TV and CT infections and its effect on the VMB state [The included study by Masha et al. characterized the VMB profiles in TV- and CT-infected, and non-infected pregnant women in Kenya . Lactobacontrols . The anaMB state , 25. TheBesides the high burden of STIs, malnutrition, or undernourishment is also prevalent in low-income areas in sub-Saharan Africa, especially among pregnant women . CurrentMcMillan et al. and Bisanz et al. examined whether probiotics containing specific Lactobacilli would alter the VMB composition during pregnancy in and Tanzanian women, respectively , 41. UnfLactobacillus-based probiotics, made with mixtures of Lactobacillus species, Bifidobacterium, and Streptococcus strains have been studied [Lactobacillus strains, mostly Lactobacillus rhamnosus) have been claimed to restore Lactobacillus-dominated VMB in women with BV [Lactobacillus species could restore VMB eubiosis and simultaneously nourish malnourished (pregnant) women in resources-constrained settings.Different  studied , 88. Lat with BV –91. The  with BV , 41. IdeVMB characteristics, or presence of pathobionts, have been linked to various maternal, fetal health issues and adverse pregnancy outcomes, primarily preterm birth, but also chorioamnionitis, premature rupture of membranes, stillbirth, preeclampsia , 92–96. The heterogeneity of results included in this review is high since different study designs were included, different study populations , and studies that used diverse molecular approaches. Pyrosequencing and Illumina MiSeq sequencing of the 16S ribosomal (r) RNA gene allow for an unprecedented high-resolution detection of the microbiota . HoweverL. iners and G. vaginalis among pregnant sub-Saharan women might indicate eubiosis or healthy VMB, rather than a transitional or abnormal VMB as seen in other ethnic populations [The present review identified many research gaps. It is still unclear what the mechanism behind a healthy and balanced VMB is , as it iulations , 103. A ulations . DecipheFurthermore, the role of pathogens on VMB composition, especially STIs that have a high incidence and prevalence, the latest estimate was made in 2012, where in Africa there were 357 million new episodes of four curable STIs  . These SL. iners or more diverse anaerobic communities, mostly with a high abundance of G. vaginalis. Future research should investigate the pathogenesis and host-immunological role of the VMB bacteria on various health conditions and outcomes in more detail. To allow for analysis across studies, consensus on how to test and report VMB composition in sub-Saharan African women should be reached for a better comparison of data sets. Furthermore, a large shared multi-county/international database could also help to minimize these problems. A systematic comparison of evidence across countries will help provide substantial qualitative evidence for public health strategies to improve reproductive and maternal health in sub-Saharan Africa.This study provides an overview of the current knowledge of VMB composition in pregnant women living in seven sub-Saharan Africa. It remains challenging to compare VMB characteristics across studies performed in this region since populations and experimental methods vary considerably . NeverthAdditional file 1: Figure S1. PRISMA based Flow diagram displaying the study selection [election .Additional file 2: Table S1. Search strategies and hits based on searches last conducted on July 15, 2020 [15, 2020 .


Glaciozyma antarctica, was isolated from various terrestrial materials collected from the Sôya coast, East Antarctica, and formed frost-columnar colonies on agar plates frozen at −1 °C. Thawed colonies were highly viscous, indicating that the yeast produced a large number of extracellular polysaccharides (EPS). G. antarctica was then cultured on frozen media containing red food coloring to observe the dynamics of solutes in unfrozen water; pigments accumulated in frozen yeast colonies, indicating that solutes were concentrated in unfrozen water of yeast colonies. Moreover, the yeast produced a small quantity of ice-binding proteins (IBPs) which inhibited ice crystal growth. Solutes in unfrozen water were considered to accumulate in the pore of frozen colonies. The extracellular IBPs may have held an unfrozen state of medium water after accumulation in the frost-columnar colony.The basidiomycetous yeast,  Pucciniomycotina, Ustilaginomycotina, and Tremellomycetes in Agricomycotina. Tremellomycetes and Agaricomycetes (known species were filamentous form) were detected in soils near glacial lakes in the McMurdo Dry Valley [Coprinellus and Pleurotus (both in Agaricomycetes) were included in dominant fungal taxa on rocks in the polar desert of the Ellsworth Mountains [Even in the continental Antarctic region, which represents one of the coldest places in the world, various eukaryotic microorganisms are known to exist. Isolated basidiomycetous fungi in this region occurred in yeast-form except for alien species , and basy Valley  and Victy Valley  by DNA mountains .Typhula ishikariensis secreted abundant ice-binding proteins (IBPs)  [Previously, we described that ecophysiological strategies for freezing tolerance of filamentous fungi differed among taxa in in the Northern Hemisphere . Filamenroteins)  to keep roteins) . HoweverMrakia spp. and Leucosporidium spp. to reduce the freezing point of the cytoplasm [M. blollopis were accelerated under freezing temperatures [M. psychrophila [Several cold-adapted yeasts, including basidiomycetes, could grow under freezing conditions ,13 becauytoplasm ,17. Bioseratures . Polyamihrophila .Glaciozyma antarctica  [Rhodotorula glacialis [Goffeauzyma gastrica (syn. Cryptococcus gastricus) [Mrakia spp. in Tremellomycetes could not produce IBPs [M. blollopis suggested that strain SK-4 did not have IBP genes [Mrakia spp. were less frost tolerant to freeze-thaw cycles among basidiomycetous yeasts isolated from Antarctica [IBPs or their activities were recorded in basidiomycetous yeasts in Pucciniomycotina, such as arcticum ,21,22, Ldium sp. , L. creainivorum , Psychrodensis’) , Rhodotolacialis ; Tremellstricus) . Howeveruce IBPs , and draBP genes . Therefotarctica .G. antarctica, which formed irregular frozen colonies like frost columns at subzero temperatures and elucidated their growth mechanism on frozen media. Our results indicated that G. antarctica had unique physiologic characteristics against freezing stress to adapt to continental Antarctica.In this article, we isolated the basidiomycetous yeast, Various terrestrial samples such as soil, mosses, and algal mats were collected from eight ice-free areas in the Coasts of Sôya and Prince Harald, and Lützow-Holm Bay, East Antarctica from December 2006 to February 2007 . SamplesG. antarctica formed frost-columnar colonies).DNA was extracted from 2–5 fungal isolates, each with similar morphological characteristics and grown on frozen PDA, according to the protocol of Isoplants II Kit . The internal transcribed spacer (ITS) region of genomic rDNA was amplified with the primer pairs ITS1F  and ITS4http://clustalw.ddbj.nig.ac.jp/) (access on 26 July 2021) and manually adjusted. A phylogenic tree was constructed by the Maximum parsimony method and using the program MEGA X [Multiple alignments of ITS region and D1/D2 domain sequences were performed using CLUSTAL W (m MEGA X  with booG. antarctica S4B was inoculated on a frozen PDA plate including 0.05% (w/v) red food color  and was incubated at −1 °C for two months. Three plates were used for this experiment in triplicate. Surface features of frost-columnar colonies were observed by scanning electron microscopy (SEM). Frost-columnar colonies were lyophilized and coated with platinum-palladium using JFC-1100 Ion Sputter . They were examined using JSM-T330A SEM  operating at 15 kV.One loopful of cells of g, 20 min at 2 Strain S4B was inoculated on frozen or unfrozen PDA plates . Three plates were used for each experimental condition in triplicate. Frost-columnar colonies on PDA were thawed and suspended in water. Yeast cells were removed by centrifugation , cut from the margin of an actively growing colony on a PDA plate, into potato dextrose broth and then incubating for 3 months at −1 °C without shaking. Yeast cells were removed by centrifugation  equipped with a Linkam LK600 temperature controller  . Each exp < 0.05).Relationships between frost-columnar colonies and sampling sites or sample types were analyzed using ANOVA and Tukey’s HSD test  composed of ca. 69% D-glucose, 30% D-galactose, and 1% D-mannose.During the growth of the yeast on frozen PDA containing red food coloring, the pigment was concentrated in the frozen colony C. This rG.antarctica on frozen PDA looked like frost columns, while on unfrozen PDA, the colonies were typical of yeast colonies  to adapt to freezing environments; however, ironment ,27. Possg damage . Both baG. antarctica, produced a large amount of EPS and developed frost-columnar colonies on frozen PDA. These frozen colonies were porous, and yeast cells were found aligned along the walls of the pores. We considered that unfrozen water containing nutrients was accumulated within the pores of frozen colonies through the capillary phenomenon and the Marangoni effect by EPS, and that yeast cells absorbed nutrients in liquid water for growth. IBP was detected in frozen colonies. IBPs in frozen colonies of G. antarctica may keep accumulated water in the liquid state.We found that the basidiomycetous yeast, Glaciozyma antarctica in continental Antarctica and T. ishikariensis in the Boreal zone were well adapted to subzero temperatures to utilize EPS and IBPs. However, while T. ishikariensis applied these substances to control the micro-environment around their mycelia under supercooling conditions, G. antarctica applied these substances to absorb nutrients in liquid water under freezing conditions, indicating that both fungi used the same substances for different purposes.


Lactobacillus reuteri delivered in a biofilm formulation increases probiotic survival in acidic gastric conditions, increases probiotic adherence to gastrointestinal epithelial cells, and reduces the incidence, severity, and neurocognitive sequelae of NEC in experimental models.Necrotizing enterocolitis (NEC) is a devastating disease predominately found in premature infants that is associated with significant morbidity and mortality. Despite decades of research, medical management with broad spectrum antibiotics and bowel rest has remained relatively unchanged, with no significant improvement in patient outcomes. The etiology of NEC is multi-factorial; however, gastrointestinal dysbiosis plays a prominent role in a neonate’s vulnerability to and development of NEC. Probiotics have recently emerged as a new avenue for NEC therapy. However, current delivery methods are associated with potential limitations, including the need for at least daily administration in order to obtain any improvement in outcomes. We present a novel formulation of enterally delivered probiotics that addresses the current limitations. A single enteral dose of  Necrotizing enterocolitis (NEC) is a disease that has been a major source of morbidity and mortality for premature neonates for decades. Affecting 10% of infants with birth weight < 1500 g, NEC is a neonatal intestinal disease that is manifested by excessive inflammation that may progress to tissue destruction, bacterial translocation, and sepsis. The disease carries a mortality rate as high as 20–30% .Lactobacillus and Bifidobacteria species, are present in healthy full-term breast-fed infants [Lactobacillus and Bifidobacterium species, and increased levels of bacteria that can become pathogenic including Gammaproteobacteria , which is evident in infants that develop NEC [The etiology of NEC is multi-factorial with prematurity, low birth weight, administration of enteral feeds, and antibiotic exposure associated with development of the disease ,5. Bacte infants ,12,13. A infants ,12. Convelop NEC ,18,19,20Lactobacillus and Bifidobacterium was shown to prevent NEC in very low birth weight infants [To counter the altered intestinal microbiome and to reduce the pathogenic bacterial colonization frequently seen in premature infants, administration of probiotics, or live microorganisms that confer a health benefit on the host, emerged as a means of NEC prevention in the late 1990s ,22. Sinc infants ,26, and  infants . Further infants ,30,31,32However, there are significant concerns and limitations regarding the current method of probiotic administration. An acidic gastric environment, interactions with bile acids, pressure from the host immune system, and competition with commensal and pathogenic bacteria can rapidly render probiotic bacteria ineffective, with a crippled capacity to adhere to and colonize the gut ,34. Due Lactobacillus reuteri (ATCC 23272) in a biofilm state rather than in a free-living planktonic state [To overcome these concerns and limitations, we introduced a novel probiotic delivery system that delivers beneficial health-promoting ic state ,41. Probic state ,43. The ic state ,45,46,47L. reuteri is a Gram-positive bacterium that is naturally found in a variety of hosts and environments, including the healthy human intestine [L. reuteri strains commonly found in humans are divided into two clades, i.e., clade II and clade IV, that are genetically and functionally distinct [L. reuteri ATCC 23272  is a clade II strain originally isolated from human breast milk that is a frequently used probiotic given its anti-inflammatory and antimicrobial properties. Anti-inflammatory abilities are in part attributable to its ability to produce histamine and diacylglycerol kinase that together lead to activation of histamine H2, but inhibition of histamine H1 receptors (respectively) [L. reuteri folate metabolism has been linked to its anti-inflammatory properties [L. reuteri has also been shown to induce anti-inflammatory T regulatory cells, suppress T helper (Th) 1 and Th2 cytokine responses, and to alter dendritic cell activity, but the mechanisms by which this occurs are not widely understood [L. reuteri has extracellular glucosyltransferase (GTF) proteins that catalyze the formation of exopolysaccharides of glucose (glucans) from disaccharide sugars  and possess glucan binding domains, that further allows for strong binding to other glucans.ntestine ,49,50. Ldistinct . L. reutctively) ,53,54. Tctively) . In addioperties , includioperties . L. reutderstood ,59,60,61derstood ,63. Reutderstood ,65. AddiL. reuteri attachment and biofilm formation [L. reuteri are introduced to DMs and undergo a brief incubation period to allow for adherence and biofilm formation  enhanced binding of L. reuteri to intestinal epithelial cells, (2) protection against low gastric pH, and (3) access to high concentrations of beneficial luminal substances to L. reuteri in order to augment its probiotic effects.We use dextranomer microspheres (DMs) as a surface for ormation . DMs areormation ,67,68. Iormation . Importaand grow . AdditioL. reuteri to DM is heavily GTF-dependent and is enhanced in the presence of certain disaccharides, e.g., maltose and sucrose  and DMs with Congo Red, confocal laser scanning microscopy revealed significantly increased L. reuteri adherence to DMs containing sucrose or maltose compared to DMs containing water only [Adherence of  sucrose . Bacteriin vitro . After dter only .L. reuteri to reach the distal gastrointestinal tract to be effective in preventing NEC. We investigated whether L. reuteri bound to DM would have increased survival under acidic conditions, and whether the addition of beneficial luminal cargo would further enhance survival in a GTF-dependent manner.A major hindrance to the efficacy of orally consumed probiotics is the acidic nature of the stomach . This isL. reuteri was evaluated after placement in synthetic gastric acid with a pH of 2 for 4 h. L. reuteri in its planktonic state had a survival of <0.1%, with no significant increase in survival in the presence of sucrose or maltose, or when adhered to water-filled DMs. However, L. reuteri in its biofilm state adhered to DMs loaded with sucrose or maltose demonstrated significantly improved survival in the acidic environment. Notably, no significant improvement in survival was seen with a mutated L. reuteri strain lacking GTF, even in the presence of DM loaded with beneficial cargo [L. reuteri to DMs loaded with beneficial luminal cargo.The viability of al cargo . These fL. reuteri in its biofilm state adhered to DM would improve adherence of the probiotic to human DLD-1 intestinal epithelial cells  and to FHs 74 Int cells  in vitro. We also examined whether enhanced adherence was promoted by the addition of beneficial luminal cargo within DM.We next investigated whether L. reuteri to the GTF mutant strain, we found significantly increased binding to DLD-1 cells with wild-type L. reuteri, regardless of whether L. reuteri was delivered in its planktonic or biofilm state. This signifies the importance of GTF in adherence of the probiotic to intestinal epithelial cells. When L. reuteri was bound to DMs containing sucrose or maltose, adherence to DLD-1 cells increased by 4.7-fold and 5.2-fold, respectively. Although there was lower probiotic adherence to FHs 74 Int small intestine epithelial cells compared to colonic cells, L. reuteri in its biofilm state bound to DMs loaded with sucrose or maltose demonstrated a 1.8-fold and 2.7-fold increase in adherence, respectively, compared to L. reuteri in its planktonic state [When comparing wild-type ic state .L. reuteri (2 × 108 CFU), either alone or with 0.5 mg of DM, or sterile water control treatment via gastric gavage [Sprague Dawley rat pups are delivered prematurely from timed-pregnant dams on gestational day 21 via terminal cesarean delivery. After delivery, pups are randomized into experimental groups that received a single 100 μL enteral dose of c gavage . Pups arc gavage ,41,70. Pc gavage .L. reuteri administered as a single dose shortly after delivery in preventing NEC in our experimental NEC model [L. reuteri in its planktonic state , DM alone, or L. reuteri in its biofilm state (L. reuteri +DM). The only significant decrease in the incidence of NEC was seen with pups receiving L. reuteri in biofilm state. In addition, differences in intestinal mucosal permeability were evaluated by quantifying systemic absorption of enterally administered fluorescein isothiocyanate (FITC) labeled dextran [L. reuteri in its biofilm state demonstrated significant reduction in intestinal permeability, indicative of improved gut barrier function [L. reuteri in its biofilm state significantly decreased the incidence of NEC and improved gut barrier function, in a murine model of experimental NEC.We initially evaluated the efficacy of EC model  . This improvement translated to enhanced survival, with nearly 60% of rat pups treated with L. reuteri in its biofilm state bound to DMs containing luminal sucrose or maltose surviving until the end of the 96-h protocol, whereas only 20% of untreated stressed pups survived [L. reuteri in its planktonic form, or in pups receiving DM loaded with sucrose or maltose alone. Furthermore, any significant protective effect seen with L. reuteri was absent when a GtfW mutated strain was administered, even in the presence of DMs loaded with beneficial cargo.Our in vitro studies showed that al cells . Based omulation . Approxisurvived . No signp = 0.001) [L. reuteri in its biofilm state bound to DMs containing sucrose or maltose had significantly reduced intestinal permeability compared to untreated pups , indicating improved gut barrier function. Pups treated with a single dose L. reuteri in its planktonic state did not demonstrate improved gut barrier function.To investigate gut barrier function, 48 h after initiating the experimental NEC protocol, rat pups received enteral FITC-dextran. Four hours after administration, pups were euthanized and serum FD70 levels analyzed. Untreated pups experienced significantly increased intestinal permeability compared to breast-fed control pups (= 0.001) . Pups trp < 0.002) [L. reuteri in its biofilm state bound to DMs containing sucrose or maltose had significantly lower expression of IL-6, IL-1B, CCL-2, CXCL-1, and IL-10 compared to untreated pups, or to pups treated with L. reuteri in its planktonic form (p < 0.05). The decrease in production of pro-inflammatory compounds is likely secondary to the intrinsic ability of L. reuteri to produce histamine, which suppresses pro-inflammatory cytokine production, in synergy with a biofilm state allowing for an enhanced anti-inflammatory effect. Further investigation is required as discussed in We next examined the expression of inflammatory cytokines interleukin (IL)-6, IL-1B, chemokine ligand (CCL)-2, CXCL-1, and IL-10 in intestinal specimens from the different treatment groups. Pups exposed to NEC receiving no treatment demonstrated significant elevation in the expression of IL-6, IL-1B, CCL-2, CXCL-1, and IL-10 in the small intestine compared to unstressed breast-fed rat pups (< 0.002) . Pups reL. reuteri in its biofilm compared to its planktonic state were investigated using 16S rRNA gene sequencing analysis [L. reuteri in its biofilm state bound to DMs containing maltose, and stressed pups treated with L. reuteri in its planktonic state. Notably, the microbiota community arrangement of pups treated with L. reuteri in its biofilm state clustered more closely with unstressed, vaginally delivered, breast-fed pups. Furthermore, taxa-level analysis demonstrated that Lactobacillus species abundance was more effectively maintained and closely related to unstressed, breast-fed controls in pups treated with L. reuteri in its biofilm state compared to its planktonic state (p < 0.05). In addition, Lactobacillus species abundance was inversely associated with NEC injury . Both the biofilm and planktonic formulations of L. reuteri effectively limited the abundance of Enterobacter species, a potential enteric pathogen, compared to unstressed pups and untreated stressed pups (p < 0.05).The gut microbiota community structure and taxa composition of pups treated with analysis . UnweighL. reuteri to adhere and form a biofilm is GTF-dependent and essential to provide intestinal protection from NEC. An experiment comparing the ability of wild type L. reuteri versus GTF-deficient L. reuteri in preventing NEC was conducted. Compared to untreated stressed pups, pups receiving L. reuteri +DM loaded with maltose had a significantly lower incidence of NEC  [L. reuteri deficient in GTF (blunted biofilm forming capacity), even in combination with DMs, failed to demonstrate any significant decrease in NEC incidence compared to untreated pups (p > 0.05), signifying the importance of GTF-dependent biofilm formation in the prevention of NEC.The ability of  < 0.05) . HoweverL. reuteri possesses anti-inflammatory and antimicrobial properties which we suspect play an integral role in intestinal protection from NEC. L. reuteri’s ability to produce histamine from L-histidine is believed to play a crucial role in anti-inflammation, as histamine suppresses tissue necrosis factor (TNF) production [L. reuteri with DMs loaded with L-histidine (to augment anti-inflammatory production) and (2) utilizing a histamine-deficient mutant of L. reuteri (to attenuate anti-inflammatory production). Both experiments will be investigated in an animal model to evaluate the effect that increased or decreased histamine production has on intestinal protection from NEC.oduction ,73,74,75L. reuteri also possesses antimicrobial properties due to its ability to produce reuterin [L. reuteri to evaluate whether intestinal protection from NEC is attenuated in the absence of reuterin.reuterin ,64. ReutOver 40% of NEC survivors are left with debilitating life-long neurocognitive and developmental impairments ,77. ThesL. reuteri in its biofilm state to down-regulate the expression of proinflammatory cytokines and up-regulate the expression of anti-inflammatory compounds such as histamine [L. reuteri in its biofilm state can attenuate microglial activation, preserve neuronal myelination, and protect against neurocognitive injury in rat pups surviving our experimental NEC model.There is increasing evidence suggesting that decreased myelination is largely responsible for the neurodevelopmental sequelae and white matter abnormalities in survivors of NEC ,84,85,86istamine . Future Lactobacillus reuteri in a biofilm formulation alleviates most of these concerns. Investigations using our experimental animal model have demonstrated the ability of our L. reuteri biofilm formulation to significantly reduce the incidence and severity of NEC, decrease NEC-related mortality, stabilize the intestinal mucosal barrier, and down-regulate the production of proinflammatory cytokines. Given the effects of NEC and of gut microbes on infant neurodevelopment, future studies will determine whether our enhanced probiotic formulation will help prevent the deleterious effects of NEC on neurocognitive development.Necrotizing enterocolitis continues to be a major source of morbidity and mortality for premature infants. Despite years of research and advancements in critical care, improvement in the outcomes of infants suffering from NEC are subtle at best. Probiotics have shown promise as a potential treatment to reduce the incidence and severity of NEC; however, current delivery methods present legitimate concerns. Our proposed method of delivering a single enteral dose of the probiotic Clostridioides difficile colitis [Since probiotics offer a potential benefit in other infectious or inflammatory conditions, additional investigation is underway for the use of our enhanced probiotic formulation in the treatment and management of several other gastrointestinal diseases, including  colitis  and infl


Mangifera species, mango (Mangifera indica) is an important commercial fruit crop. However, very few studies have been conducted on the Mangifera mitochondrial genome. This study reports and compares the newly sequenced mitochondrial genomes of three Mangifera species.Among the Mangifera mitochondrial genomes showed partial similarities in the overall size, genomic structure, and gene content. Specifically, the genomes are circular and contain about 63–69 predicted functional genes, including five ribosomal RNA (rRNA) genes and 24–27 transfer RNA (tRNA) genes. The GC contents of the Mangifera mitochondrial genomes are similar, ranging from 44.42–44.66%. Leucine (Leu) and serine (Ser) are the most frequently used, while tryptophan (Trp) and cysteine (Cys) are the least used amino acids among the protein-coding genes in Mangifera mitochondrial genomes. We also identified 7–10 large chloroplast genomic fragments in the mitochondrial genome, ranging from 1407 to 6142 bp. Additionally, four intact mitochondrial tRNAs genes  and intergenic spacer regions were identified. Phylogenetic analysis based on the common protein-coding genes of most branches provided a high support value.Mangifera species. The results showed that the gene content and the codon usage pattern of Mangifera mitochondrial genomes is similar across various species. Gene transfer from the chloroplast genome to the mitochondrial genome were identified. This study provides valuable information for evolutionary and molecular studies of Mangifera and a basis for further studies on genomic breeding of mango.We sequenced and compared the mitochondrial genomes of three The online version contains supplementary material available at 10.1186/s12864-022-08383-1. Plant mitochondrial genomes are large and variable in size [Mitochondria are semi-autonomous organelles found in virtually all eukaryotic cells, and their main function is to generate adenosine triphosphate (ATP) . With th in size , 3, and  in size , 5. The  in size  and nume in size , 7. The  in size , 9. An i in size , 11. TheMangifera, mango (M. indica) is an important tropical fruit [Mangifera mitochondrial genome. To date, the complete mitochondrial genome of only one Mangifera species has been deposited in GenBank [Among the genus al fruit , 13, natal fruit , 15. Manal fruit , high nual fruit . Thus, ial fruit . However GenBank .Mangifera species were sequenced and compared with M. indica [Mangifera species; (2) assess gene transfers between the chloroplast and mitochondrial genomes; (3) explore the evolutionary relationships among the Mangifera species based on the protein-coding genes of the mitochondrial genome.In this study, the mitochondrial genomes of three . indica . This stMangifera species  were collected from Xishuangbanna Tropical Flowers and Plants Garden. Total genomic DNA was extracted from all samples using cetyltrimethylammonium bromide (CTAB) method [Fresh leaves of the three ) method . DNA samAbout 5–10 μg of total DNA from each sample was used to construct a shotgun library with an average insertion size of 350 bp. Illumina Novaseq6000  was used to sequence the DNA samples in the paired-end sequencing mode. The generated mitochondrial genome raw reads were approximately 0.7–1.2 Gb per sample. Meanwhile, the mitochondrial genome depth of coverage was more than 900 × .The raw sequencing data were filtered using Trimmomatic v0.38 , and thehttps://www.ncbi.nlm.nih.gov/) using Dicotyledoneae species mitochondrial genomes as the reference sequences. The tRNAs were predicted using the tRNAscan-SE 2.0 [Mangifera was drawn using OGDRAW v1.3.1 [Mitochondrial genome annotation was performed using BLASTN and BLASTX alignment tools of the National Center for Biotechnology Information (NCBI) database (n-SE 2.0  softwaren-SE 2.0 . A circuW v1.3.1 .http://pgrc.ipk-gatersleben.de/misa/misa.html) [MEGA v7.0.26 was used to calculate the amino acid composition of the protein-coding genes and their relative synonymous codon usage (RSCU) values [sa.html) . Analysisa.html) .Mangifera plants.We mapped the mitochondrial genome to the plastid genome using the BLASTN tool with the default settings. Circos v0.69  softwareCarica papaya as outgroups. Protein-coding genes common to the mitochondrial genomes of all species were extracted to construct phylogenetic trees. MUSCLE v.3.8.31 [Phylogenetic analyses were performed for the 15 Dicotyledoneae species using v.3.8.31  was usedM. longipes (MZ751075), M. persiciformis (MZ751076), and M. sylvatica (MZ751077). The three newly sequenced Mangifera mitochondrial genomes have been deposited in the GenBank database.Raw sequence data were obtained from M. longipes, M. persiciformis, and M. sylvatica were 728,635 bp, 750,898 bp, and 714,426 bp, respectively  genes and 24–27 transfer RNA (tRNA) genes  and many partial genes and intergenic spacers. Four mitochondrial tRNAs genes  and intergenic spacer regions were also identified , such as TGC (Cys), CAC (Arg) and TAC (Tyr). This may be a kind of AT mutation bias that has an obvious influence on codon. Notably, the codon usage pattern was highly consistent across the Mangifera mitochondrial genomes. The protein-coding genes of the majority strands showed positive AT and negative GC skews, while those of the minority strands showed positive GC and negative AT skews. The AT skews of the protein-coding genes was highly consistent across the Mangifera mitochondrial genomes  analysis of the s Table S, S4. Themes Fig. .Fig. 2ReMangifera mitochondrial genomes. Mononucleotide repeats of A/T were more prevalent than the other repeat types, dinucleotide repeats (AT/TA) were the second most numerous, while tri- and hexa-nucleotide repeats were less numerous and occurred only in intergenic or intronic regions  .The ML analyses revealed that most branches had very high support. Within the Mangifera genus, M. sylvatica was evolutionarily closer to M. persiciformis . The results indicate a strong A or T bias in the third position of the codon in the protein-coding genes of the  genomes , 44. CodMangifera mitochondrial genomes were investigated in this study. SSRs are important molecular markers for species identification, evolutionary analysis and studying genetic diversity [Mangifera are 8348–13,655 bp and may be one of the reasons for isomerization. Previous studies have uncovered the genetic relationships among the Mangifera species through morphological, nuclear, internal ribosomal transcribed spacer (ITS), amplified fragment length polymorphism (AFLP), and chloroplast gene analyses [Mangifera and Dicotyledoneae species. There were several inconsistencies between the chloroplast and mitochondrial phylogenetic trees regarding the phylogenetic topology [Mangifera. Sequencing and assembling the three new complete mitochondrial genomes of Mangifera is the first step towards understanding the mitochondrial genome variation in this genus.Plant mitochondrial genomes are rich in repeat sequences , the vasiversity . Almost iversity . The lonanalyses , 47–50. analyses . Size ananalyses , 52. Hertopology . Differetopology , 54, andMangifera species. The results showed that the gene content and the codon usage pattern was highly consistent across the Mangifera mitochondrial genomes. We also identified 7–10 large fragments transferred from the chloroplast genome to the mitochondrial genome. The findings of this study provide valuable genetic resources for further studies on Mangifera species.Here, we sequenced and compared the mitochondrial genomes of three Additional file 1.


Cindy S Chu and co-authors review options for diagnosis, safe and radical cure, and relapse prevention of Plasmodium Vivax. Plasmodium vivax malaria has more than halved from an estimated 17.3 to 6.5 million cases between 2010 and 2019. This resulted from increased deployment of conventional malaria control measures  and significant global investment in malaria elimination. There is no generally available P. vivax vaccine, nor is there likely to be one in the near future.The worldwide burden of P. vivax diagnosis have sensitivities comparable to microscopy. Ultrasensitive PCR methods which can detect parasite densities as low as 28/ml have revealed a much higher prevalence of asymptomatic P. vivax infection in malaria endemic regions than previously estimated.The latest-generation RDTs used for P. vivax infections. Prevention of relapse requires “radical cure” with an 8-aminoquinoline . These drugs cause oxidant haemolysis in G6PD deficiency, so safe use requires G6PD testing. Overall, the risks of primaquine haemolysis have been overemphasised and the benefits of relapse prevention underappreciated.Chloroquine remains an effective schizonticide for vivax malaria, except in Indonesia, Sabah and Papua New Guinea where there is high-level chloroquine resistance. Artemisinin combination therapies are effective alternative schizonticides that unify the treatment of all malarias. Relapses contribute significantly to the burden of Qualitative screening tests for G6PD deficiency  are adequate for screening before giving primaquine for radical cure, but a quantitative point of care G6PD test to detect <70% normal enzyme activity is needed for tafenoquine.Radical curative efficacy depends on the total 8-aminoquinoline dose given; higher primaquine doses (7 mg base/kg rather than 3.5 mg base/kg) are required in parts of Southeast Asia and Oceania. The currently recommended dose of tafenoquine (300 mg) is sub-optimal. In low transmission settings, elimination of vivax malaria is possible with current tools.Plasmodium vivax is more geographically dispersed than Plasmodium falciparum, with transmission occurring over a wider range of temperatures than for P. falciparum [P. vivax is the predominant cause of human malaria in Asia and the Asia-Pacific regions where, with large populations and a declining incidence of P. falciparum infections, over 80% of the global P. vivax burden occurs [P. vivax is also prevalent in the horn of Africa, Madagascar, and parts of Central and South America, but it has been eradicated from Europe, Russia, North America, and most of the Middle East [lciparum , and at lciparum . P. vivan occurs . P. vivadle East ,4.P. vivax malaria relies on clinical suspicion, a reliable blood diagnostic, and access to efficacious effective schizonticidal (blood stage) and hypnozoiticidal  drug regimens. Currently, P. vivax malaria is treated with chloroquine or artemisinin combination therapy (ACT) for the blood stage infection. While chloroquine has been standard treatment for vivax malaria for some 70 years, the emergence and global spread of chloroquine resistance in P. falciparum has meant that different treatments are now required for P. falciparum and the other human malarias. Use of ACTs (except artesunate-sulfadoxine-pyrimethamine) allows again a unified treatment for all malaria, and it provides a safety net in case of misdiagnosis or an unidentified mixed infection with P. falciparum. Despite declining susceptibility, chloroquine is still effective for P. vivax infection in most of the world, except for Indonesia, Sabah and Oceania where resistance is high grade [Clinical management of gh grade ,6. ChlorP. vivax has been considered benign, although severe infections may sometimes occur [P. vivax infection [P. vivax malaria is similar to that of P. falciparum malaria [P. vivax malaria in pregnancy is associated with miscarriage and low birth weight [Historically, es occur –10. Concnfection ,12. The  malaria . P. vivah weight ,15.P. vivax malaria treatment is how to prevent relapses. These result from dormant liver stage parasites (hypnozoites) that activate weeks or months after the primary infection to cause a recurrent P. vivax infection (relapse). The intervals to and frequency of symptomatic relapses vary geographically and depend also on age and the intensity of transmission [P. vivax infections are a major cause of morbidity in vivax malaria, particularly in young children in whom repeated relapse may cause severe anaemia, malnutrition, and growth delay [The great challenge for smission –18. Freqth delay , 20. ThuP. vivax malaria has been the microscopy observation of asexual parasites on a thick or thin blood film. Accurate microscopy requires trained laboratory staff supported with adequate laboratory supplies and well-maintained microscopes, which are not always available in resource-limited settings. The limit of detection for expert microscopy is about 50 parasites/uL which coincides approximately with the pyrogenic density [P. vivax infections, but overall, it is suitably sensitive and specific for clinical diagnosis. Rapid diagnostic tests (RDTs) have been developed which detect antigens common to all malarias  or those specific to P. vivax . First-generation RDTs had lower sensitivities for P. vivax parasitaemia [P. falciparum infections. The sensitivity and specificity of P. vivax RDTs have both been improved significantly. From 2008 to 2018, the proportion of P. vivax, P. falciparum, and pan-malaria RDTs meeting the current WHO performance threshold for procurement increased from approximately 20% to 90% [P. vivax infections.For over a century, the gold standard for diagnosing  density . The diasitaemia  and coulsitaemia  with hig% to 90% . During % to 90% . Access PCR allows low densities of malaria parasites to be detected reliably in blood samples. For low-volume blood samples such as filter paper blood spots, lower limits of reliable detection are typically in the range of 1 to 10 parasites/uL. In recent years, high-volume ultrasensitive PCR methods (uPCR) have been developed which amplify a larger amount of DNA, and these provide lower limits of detection around 28 parasites per mL (based on genus) with a second species-specific probe based on the 18S RNA gene . These uP. falciparum infections as relapses can be with genotypes similar or identical to the initial infection, or they can be with completely different genotypes resulting from heterologous hypnozoite activation [Molecular genotyping is not needed for clinical management, although it is useful in clinical trials where, together with time-to-event information, it can help distinguish relapse from reinfection . The inttivation , 31. NevP. vivax infections). Despite these limitations, ITNs do provide partial benefit and are an adjunct method to prevent primary P. vivax infections and interrupt transmission [Insecticide-treated nets (ITNs) are the mainstay of malaria control in most of the malaria-endemic world. Between 2017 and 2019, over 25,000,000 ITNs were distributed annually in malaria-endemic regions outside sub-Saharan Africa . The effsmission . Long-lasmission ,38, and smission .P. vivax endemic regions [P. vivax is endemic, though even within a single country, resistance patterns may vary [The use of insecticides for malaria prevention focuses mainly on indoor residual spraying (IRS) and long-lasting ITNs, although there are a variety of other approaches. Organochlorides, organophosphates, carbamates, and pyrethroids are recommended by WHO for IRS. From 2000 to 2009, over 3,200 metric tonnes of insecticide were used against malaria vectors in  regions . Widespr regions ,42. Resi regions  where P.may vary . The fulP. vivax vaccine, and none is on the near horizon. In contrast to P. falciparum, where hundreds of clinical trials and nearly 20 preclinical trials are registered, only 1 clinical trial and 2 preclinical trials are registered for P. vivax [P. vivax are irradiated sporozoites [P. vivax Duffy binding protein (PvDBP), P. vivax merozoite surface proteins , P. vivax apical membrane antigen (PvAMA1), and the liver stage antigen P. vivax circumsporozoite protein (PvCSP) [P. vivax vaccine trial  is listed on the global WHO malaria clinical vaccine projects “Rainbow tables”  [P. vivax faces different challenges than with P. falciparum. P. vivax has greater genetic diversity [There is no generally available P. vivax . Potentirozoites  and the  (PvCSP) . Transmi (PvCSP)  and PvDB (PvCSP)  and asex (PvCSP) ,51. Curr1816113) . Vaccineiversity  and cannP. falciparum, the more dangerous malaria parasite, both to prevent severe disease, and also because nearly all the drugs which work against P. falciparum  are also effective against P. vivax. Intermittent presumptive treatment for infants and pregnant women (IPTi and IPTp) and seasonal malaria chemoprevention (SMC) are interventions which aim to prevent P. falciparum blood stage infections in populations that live in areas of moderate to high P. falciparum transmission. These are typically areas where P. vivax is either uncommon or absent. The only suppressive chemoprophylactic strategy specifically targeting P. vivax for endemic populations is a weekly chloroquine dose of 5 mg base/kg (300 mg base adult dose) given during pregnancy and lactation [P. vivax infection—and then it should be given. The role of IPTp with dihydroartemisinin-piperaquine in parts of Indonesia and Papua New Guinea, where transmission and resistance to chloroquine are both high, is under investigation with promising results in clinical trials [Chemoprophylaxis is recommended to protect travellers from nonendemic areas entering a malaria-endemic area. Recommendations are largely focused on actation  as 8-amil trials .P. falciparum has been the primary approach for malaria prevention , although they do not prevent the establishment of hypnozoites. The 8-aminoquinolines (primaquine and tafenoquine) have good activities against the pre-erythrocytic stages of all malarias, as well as blood stage activities against P. vivax, and so can be used for chemoprophylaxis. These drugs prevent later relapses as they kill hypnozoites (whereas IPTi or SMC with schizonticides only delay multiplication of the blood stage). Atovaquone-proguanil is also considered to provide highly effective causal prophylaxis against P. vivax but it does not prevent relapses. Recent observations in travellers showed a similar rate of P. vivax infections (40% to 50%) up to 1 year after returning to a nonendemic area following either atovaquone-proguanil, mefloquine, or doxycycline prophylaxis. This compares with a rate of only 1% to 2% in travellers who had received primaquine [P. vivax and Plasmodium ovale are prevented presumptive antirelapse therapy (PART) with a radical curative dose of 8-aminoquinolines (primaquine and tafenoquine) may be given following departure from an endemic area to persons receiving suppressive chemoprophylaxis  prescribed for the previous clinical reatment .For over 60 years, the mainstay of antirelapse treatment has been a 14-day primaquine regimen. Divergent policies and practices, concerns over haemolysis, and the unavailability of G6PD testing have contributed to low rates of primaquine uptake for radical cure in malaria-endemic countries .The radical curative efficacy of primaquine depends on the total dose given. There have been relatively few randomised trials in which the radical curative efficacy of primaquine has been assessed with a necessary ≥4-month follow-up (1 year is preferable). In South America, 7-day regimens are used. The radical curative efficacy of the 7-day low-dose  primaquine regimen was approximately 80% –92. In SIn early clinical trials, it was observed that chloroquine potentiated the radical curative efficacy of primaquine . The mecPrimaquine is contraindicated in pregnancy, is not recommended in infants <6 months, and has also been withheld from lactating women, although a recent study shows that the dose ingested by the breastfeeding infant is very small  and therP. vivax radical cure is scaled up. Patients who have a test result indicating G6PD deficiency should not receive the standard primaquine regimen. Instead, in areas where the less severe G6PD variants  are prevalent, they should receive a once weekly 0.75 mg base/kg dose for 8 weeks [P. vivax malaria, particularly in young children who may be ill every second month, has often been underappreciated [P. vivax malaria, so at a population level, the risk of haemolysis in patients is lower than would be predicted from prevalence surveys in healthy individuals [G6PD deficiency is the most common human enzymopathy. Gene frequencies average 8% to 10% across much of the tropical world . The ris 8 weeks ,96. The reciated . Importaividuals . In a reividuals . The resividuals ,111. Theividuals .P. vivax relapse rates are high, and also in South America where both radical cure regimens performed poorly [Tafenoquine is a slowly eliminated 8-aminoquinoline  which allows single-dose treatment. As with primaquine, tafenoquine is generally well tolerated and abdominal discomfort is lessened if given with food. During the long development of tafenoquine, concerns were raised about the potential for vortex keratopathy, electrocardiograph QT prolongation, and psychiatric reactions. The clinical evidence to date has largely resolved these issues –115 althd poorly . Tafenoqd poorly , but morAs described, tafenoquine’s advantage in being slowly eliminated , and thus effective in a single dose, is also its Achilles heel. If given to an individual who is G6PD deficient, the drug exposure and resulting haemolysis will persist until all the susceptible erythrocytes have been destroyed. Some female heterozygotes, who would be undetected by the standard screens, could still theoretically lose up to 70% of their red cells. This is why the higher G6PD activity threshold (>70% G6PD activity) has been chosen for tafenoquine eligibility. The gold standard method for G6PD quantitation is spectrophotometry requiring relatively expensive and sophisticated laboratory techniques, so it is not feasible in nearly all healthcare settings. Thus, for safe use of tafenoquine, new quantitative point-of-care G6PD tests have been developed. Studies to validate G6PD quantitative tests are completed ,121 and P. vivax malaria show that dosing regimens in 4 weight bands were safe and efficacious and reached the target AUC0-∞ [P. vivax malaria. These data prompted a product label change to Krintafel (the US regulatory approved form of tafenoquine for treatment) in early 2020 [P. vivax malaria treatment. If followed this would prevent the use of tafenoquine in areas with chloroquine-resistant P. vivax parasites where national malaria programmes recommend ACTs for vivax malaria. Clearly, further clinical trials are needed to identify the correct dose of tafenoquine for radical cure [The current exclusions to tafenoquine apply to approximately 25% of the population in endemic areas, depending on the G6PD allelic frequency, and fertility and lactation rates . Approvit AUC0-∞ . As a ret AUC0-∞  in preverly 2020 . Now, oncal cure  and resoP. vivax transmission is obviously a risk factor, and having had vivax malaria without radical treatment is clearly the major risk. Pregnant women cannot receive 8-aminoquinolines so are at significant risk if they have vivax malaria during pregnancy. If primaquine could be allowed during lactation , this sP. vivax infections can be assessed by measurement of symptom resolution, fever clearance, and parasite clearance. The lower parasite densities in P. vivax mean that parasite rate clearance estimates are more difficult to assess, although sequestration is not a confounder in measuring parasite clearance rates because all parasite stages are in the circulation [P. vivax gametocyte clearance parallels asexual stage clearance after a blood schizonticide [As in falciparum malaria, the treatment responses in culation . Quantitculation . These ponticide . The majonticide . Lumefanonticide ,125. Thionticide . With hionticide .P. vivax malaria causes substantial preventable morbidity. Better diagnosis, easier G6PD testing, and use of ACTs as an alternative to chloroquine are improving the clinical management of P. vivax malaria, but the challenge now is to make these more widely available. Primaquine radical cure is under used. Tafenoquine provides single-dose radical cure, but the optimum dose and method of using tafenoquine safely still need to be determined. The tools needed for P. vivax elimination are available. Malaria elimination activities conducted in remote settings [P. vivax incidence considerably, but continuing to sustained elimination will require substantial commitment and resources.Frequent recurrent settings –128 have


Salvia spinosa L. on mitochondrial function along with its protection against streptozotocin-induced nephropathy in diabetic mice. After the injection of streptozotocin (STZ) and verification of the establishment of diabetes, mice (n = 30) were randomly divided into the following groups: control group, diabetic-control, S. spinosa-treated diabetic , and metformin-treated diabetic group (500 mg/kg). After four weeks of treatment, the mice were weighed. Blood and kidney tissues were examined for biochemical and histological evaluation. Hematoxylin and eosin staining was used for evaluating renal pathologic damage. Oxidative damage in the kidney was assessed by the evaluation of lipid peroxidation and glutathione oxidation. Furthermore, differential centrifugation was used to obtain the isolated mitochondria, and mitochondrial toxicity endpoints  were determined in them. S. spinosa remarkably reduced the blood urea and creatinine concentrations, and also normalized kidney weight/body weight coefficient in the diabetic mice. S. spinosa ameliorated the incidence of glomerular and tubular pathological changes in histological analyses. Moreover, the oxidative and mitochondrial damages were notably attenuated in renal tissues of S. spinosa-treated mice. These results indicate that the methanolic extract of S. spinosa modulates the nephropathy in the diabetic mice by the amelioration of oxidatively induced mitochondrial damage and provides a reliable scientific base, suggesting S. spinosa as a promising alternative remedy against DN.Mitochondrial oxidative damage is a crucial factor in the pathogenesis of diabetic nephropathy (DN), which is among the most prevalent problems of diabetes, and there hasn't been an effective treatment for DN yet. This study planned to investigate the effects of  Diabetes mellitus (DM) leads to profound pathological complication including diabetic nephropathy , in approximately 25–35% of patients with both main types of DM , 2. The Regarding the complex pathogenesis of diabetes, the persistent hyperglycemia leads to an excessive reactive oxygen species (ROS) generation, oxidative stress, and mitochondrial dysfunction , 5. EvenIn clinical practice, controlling excessive oxidative stress, hyperglycemia, dyslipidemia, and hypertension are the important strategies to improve albuminuria and ultimately treat DN , 10. NowMedicinal herbs have gained significant importance in the last few decades, and available literature shows that huge numbers have demonstrated considerable antioxidant effects on diseases including DN , 5, 10.Salvia spinosa L. is a plant belonging to the Salvia genus, in the Lamiaceae family, which is native to Iran. According to the previous reports, Salvia species are rich in bioactive compounds such as phenolic compounds, terpenoids, isoprenoids, triterpenoids, and sesterterpenoids -2, 5- diphenyltetrazoliumbromide (MTT) assay. Mitochondrial succinate dehydrogenase reduces MTT and produces formazan as a chromophoric product. After the isolation of mitochondria from the kidneys of mice, MTT (0.4%) was added to the isolated mitochondria and incubated at 37°C (30 minutes). The produced formazan was dissolved in dimethyl sulfoxide (DMSO), and the absorbance was measured with Epoch™ Microplate Spectrophotometer  at 570 nm .As previously explained, glutathione levels were evaluated by using DTNB as the indicator by spectrophotometry. Afterwards, 0.1 ml of isolated renal mitochondria was added to phosphate buffers and DTNB (0.04%) (pH 7.4). The yellow color was read at 412 nm on a UV spectrophotometer  .Malondialdehyde (MDA) was measured based on the method used by Zamani et al. for the evaluation of lipid peroxidation . BrieflyProtein concentration was assessed in tissue homogenates by the Bradford technique. The samples were mixed with Coomassie blue; 10 minutes later, absorbance was determined at 595 nm  .post hoc Tukey's test. P < 0.05 was considered statistically significant.All statistical analyses were performed using GraphPad Prism software, version 6. The results were expressed as mean ± standard deviation. The assays were performed in at least triplicate, and the mean was used for statistical analysis. Comparison between groups was made using the one-way ANOVA test, followed by the P < 0.001). Metformin dramatically decreased FBG compared with diabetic mice (P < 0.001). S. spinosa at 200 mg/kg significantly attenuated FBG compared with diabetic mice (P < 0.01); but it was not as effective as metformin-treated diabetic mice (Fasting blood glucose (FBG) is shown in tic mice .P < 0.001 in comparison with control). On the other hand, treated diabetic mice with S. spinosa extract and metformin showed a significant weight loss compared to diabetic mice (P < 0.001). There were no significant differences in metformin and S. spinosa (at 100 and 200 mg/kg)-treated diabetic mice groups  compared to control mice. S. spinosa and metformin decreased BUN concentrations remarkably compared to diabetic mice (P < 0.05). Interestingly, there are no significant changes in BUN concentrations in diabetic mice given S. spinosa (at 200 mg/kg) compared to the metformin group (P < 0.01) (As shown in  < 0.01) .P < 0.01). The S. spinosa treatment considerably reduced sCr in diabetic mice so that the high doses of S. spinosa (100 and 200 mg/kg) attenuated sCr in diabetic mice, almost as well as metformin-treated mice. Then, these data demonstrated that S. spinosa had a protective effect on kidney function in diabetic mice , as a marker of GFR estimation, increased significantly in diabetic mice as compared to control mice . Treatment with S. spinosa at all concentrations increased this content significantly compared with diabetic mice (P < 0.001). Interestingly, there were no noteworthy differences between S. spinosa and metformin-treated mice (Glutathione (GSH) is a main intrinsic antioxidant in tissues, and a decrease in its levels indicates oxidative damage. ted mice .μM in control group mice to 35.90 ± 1.85 μM in untreated diabetic mice  (P < 0.001). Four-week treatment with S. spinosa significantly inhibited lipid peroxidation damage compared to the diabetic untreated group (P < 0.001). Intriguingly, significant changes were not seen between diabetic mice treated with S. spinosa (200 mg/kg) and metformin-treated mice (Elevation of MDA (product of lipid peroxidation) is a main marker for oxidative stress. Similarly, in the current experience, diabetes led to an increase in MDA levels from 10.55 ± 0.88 ted mice .S. spinosa effects on the activity of mitochondrial complex II (succinate dehydrogenase) via the MTT test. After the isolation of renal mitochondria, a significant reduction was observed in the mitochondrial metabolism of MTT to formazan in diabetic mice, compared with normal control mice (P < 0.001). S. spinosa clearly improved mitochondrial function in diabetic mice (P < 0.05). No significant difference was observed in S. spinosa-treated mice (100 and 200 mg/kg) in comparison with metformin-treated mice  (P < 0.05 compared with diabetic mice). According to our results, S. spinosa represented a protective effect on mitochondrial glutathione contents, specially at 100 and 200 mg/kg doses. Furthermore, the glutathione concentrations of metformin-treated mice were significantly higher than those of diabetic mice (P < 0.05). (The GSH concentrations of diabetic mice were approximately 50% of control groups. As shown in < 0.05). .P < 0.05). Interestingly, S. spinosa-treated diabetic mice  significantly decreased mitochondrial lipid peroxidation in diabetic mice (P < 0.05 compared with untreated diabetic mice). The significant differences were not shown between S. spinosa- and metformin-treated groups , thereby forming superoxide (O2−), leading to other reactive oxygen species (ROS). Minimal levels of ROS are detoxified by cellular antioxidants such as glutathione, catalase, and superoxide dismutase [S. spinosa led to a remarkable amelioration of mitochondrial toxicity [S. spinosa reduced the level of oxidative stress and led to mitochondrial protecting effects in the presence of diabetes-induced mitochondrial damage.The renal cells have a high susceptibility to the hyperglycemia caused by diabetes. Although the complex mechanisms underlying mitochondrial dysfunction in diabetic nephropathy are not fully assumed , hyperglismutase . Hypothetoxicity . In addiS. spinosa treatment. Similar to our results, Trujillo et al. showed that antioxidant plants such as turmeric had nephron-protective effects due to the inhibition of mitochondrial dysfunction and prevention of oxidative stress [S. spinosa can probably alleviate diabetic nephrotoxicity via protecting mitochondria, by reducing oxidative stress. S. spinosa combined with antihyperglycemic agents could be highly effective in the treatment of diabetic nephropathy. Nevertheless, the lack of phytochemical analysis is a limitation of our study. As previous data suggest, the quality of plant materials is associated with the geographic and ecological conditions and phytochemical analysis should be carried out in subsequent studies.In addition, the ROS along with oxidized proteins, lipids, nucleic acids, and carbohydrates contribute to cellular dysfunction and compromised integrity, and eventually to apoptosis (programmed cell death) . These le stress , as welle stress , 29. ThuSalvia spinosa effectively decreased oxidative stress and renal injury in STZ-induced diabetic mice. S. spinosa reduced the levels of sCr and BUN and improved the histopathological state in the diabetic mice. Moreover, S. spinosa significantly prevented mitochondrial dysfunction. The results suggest that in the treatment of diabetic nephropathy, S. spinosa is a potential therapeutic agent. Therefore, it is necessary to carry out further exploration for authentic data to justify clinical application of this plant against diabetic complications, besides other blood glucose-lowering protocols.In summary, advanced oxidative stress is a probable mechanism, which contributes to the diabetic complications, including nephropathy. To the best of our knowledge, our study has shown for the first time that


Vibrio parahaemolyticus has not been established and, therefore, understanding about fibrillar adhesins remain limited in V. parahaemolyticus. This in silico analysis can aid in the systematic identification of Ig-like-folded and fibrillar adhesin-like proteins in V. parahaemolyticus, opening new avenues for disease prevention by interfering in microbial interaction between V. parahaemolyticus and the host.Immunoglobulin-like (Ig-like) fold domains are abundant on the surface of bacteria, where they are required for cell-to-cell recognition, adhesion, biofilm formation, and conjugative transfer. Fibrillar adhesins are proteins with Ig-like fold(s) that have filamentous structures at the cell surface, being thinner and more flexible than pili. While the roles of fibrillar adhesins have been proposed in bacteria overall, their characterization in  Vibrio parahaemolyticus is a marine-oriented pathogen, which can be detected in estuarine, coastal environments and seafood. V. parahaemolyticus has resulted in significant economic loss in animal production and increased spread of foodborne diseases [diseases . Its virdiseases . The intdiseases  fold domdiseases .Vibrio. VesB is a serine protease in Vibrio cholerae with an Ig-like fold domain at C-terminal of the protein; the deletion of the Ig-like domain resulted in the degradation of the protein [Vibrio fischeri, the mutant of Vibrio Ig-like protein (Vig) was created, and the Ig-like protein was associated with roles in influencing luminescence produced by V. fischeri and the ability of the symbiont to colonize the squid [Vibrio phage ØVC8 interactions with the human intestine, and are related to the lytic activity of the phage in preventing V. cholerae O1 colonization [Vibrio adhesins which play essential roles in adhesion and invasion of pathogens into host cells [Vibrio.Ig-like domains play critical roles in  protein . The rol protein . In Vibrhe squid . Chitinahe squid . Ig-foldhe squid . Ig-likenization . Ig-likest cells . Comparest cells . AlthougVibrio, their characterization in V. parahaemolyticus has not been established. Therefore, this study aimed to uncover the characteristics and functions of Ig-like proteins and fibrillar adhesins in V. parahaemolyticus systematically, giving insights into promising targets for preventing virulence and infection of V. parahaemolyticus.To sum up, although roles of Ig-like domains have been revealed in V. parahaemolyticus RIMD 2210633, V. parahaemolyticus CHN25, V. cholerae NCTC 9420 and Escherichia coli K-12 MG1655 genomes. The heatmap of Pfam hits was generated to present the distributions of multiple Pfams in different genomes  were the Pfam domains which only existed in the E. coli but not in Vibrio species. E. coli shared only 1.5% similarity with V. parahaemolyticus RIMD 2210633 in Ig-like fold domains, suggesting their different adhesion and signalling mechanisms. V. cholerae possesses the least Ig-like fold domains, and only 7.6% of domains were distinctive in this species. V. parahaemolyticus RIMD 2210633 and V. parahaemolyticus CHN25 shared the most similarity in Ig-like fold domains.There were 64, 63, 33 and 42 Pfam  hits identified in  genomes . Via TabE. coli may have the most different signaling and adhesion mechanism from V. cholerae and V. parahaemolyticus; meanwhile, V. parahaemolyticus RIMD 2210633 and V. parahaemolyticus CHN25 may possess the most similar mechanisms of Ig-like fold domains. The single-copy orthologues of these four genomes were OG0000002, OG0000003, OG0000004, OG0000005, OG0000006 and OG0000008.OrthoFinder assigned 121 Ig-like containing proteins  to 40 orthogroups, and 50% of all proteins were in orthogroups with three or more proteins and were contained in the largest 17 orthogroups. There were seven orthogroups present in all genomes, and six of these consisted entirely the single-copy proteins. The results from Orthogroup analysis was in consist with Ig-like fold domain distribution analysis , that E.Four proteins were identified as fibrillar adhesin-like proteins: WP_005477759.1, WP_005480168.1, WP_005489282.1 and WP_005490731.1. The physico-chemical properties of four fibrillar adhesin-like proteins was analyzed and the results are presented in Pseudomonas aeruginosa, Azotobacter vinelandii, Marinomonas primoryensis, V. cholerae and Leptospira interrogans. Most ligands of these homologues were related to CA  and ZN . The protein interface results indicated promising protein–protein interfaces were present that can be designed as drug targets; residues were coloured according to the Z-score of dL (likelihood difference) score used in BindML tool, from red  to grey  and blue (non-interface).Based on mmseq2 searching algorithm, structural homologues of these four fibrillar adhesin-like proteins have been demonstrated . InteresE. coli [To verify the effects of known virulence inhibitors against E. coli ,14,15,16V. parahaemolyticus, as well as predict their tertiary structures and binding interfaces. Following this, the phylogenetic ortholog analysis of Ig-like fold containing proteins was examined in a comparative genome manner. Fibrillar adhesins were identified by Pfam domains and HMM search. This study has led to the discovery of four fibrillar adhesins in V. parahaemolyticus.This study aimed to identify fibrillar adhesins from Ig-like fold-containing proteins in E. coli K-12 MG1655, none of it was present in V. parahaemolyticus RIMD 2210633, V. parahaemolyticus CHN25, or V. cholerae NCTC 9420; this domain is related with pili assembly and attachment mediation to different receptors. V. parahaemolyticus and V. cholerae were also found to have divergent Ig-like folds, domains of RET_CLD1 (Pfam ID: PF17756), Big_11 (Pfam ID: PF18200), Cadherin (Pfam ID: PF00028) and fn3 (Pfam ID: PF00041) existed only in V. cholerae; interestingly these four domains are also related to adhesion or ligand binding functions. The phylogenetically restricted domains and proteins have sparked interest due to the possibility that they may have evolved with distinct binding specificities. The orthogroups analysis assisted in narrowing down the distinctive Ig-like fold proteins in V. parahaemolyticus.While the domain of PapD_C (Pfam ID: PF02753) generated 10 hits in Streptococcus gordonii colonization at multiple sites within the host [Streptococcus mutans, SpaP binds to salivary agglutinin glycoprotein and the proline-rich protein of the acquired pellicle on the tooth surface [Staphylococcus aureus adhesion to host cells via N-terminal Ig-like domain specifically binding to N-acetylneuraminic acid [V. parahaemolyticus has been found. Therefore, this study filled in the knowledge gap and identified four fibrillar adhesin-like proteins, three of which have not been characterized in V. parahaemolyticus. Notably, Gram-negative bacteria have been suggested with less fibrillar adhesins as they may use more pili or outer membrane proteins for colonization [Due to the increased binding possibility of target sites to host cells, multiple domains and tandem sequence repeats being contained within a single protein may be adhesins ,17. Bactthe host . As a ty surface . SraP menic acid . Howevernization .V. parahaemolyticus. AprA has been reported as a virulence factor in P. aeruginosa which is the homologue of protein WP_005477759.1 [2+) and ZN . Ca2+ and Zn2+, working as determinants in the host environment, play critical roles in regulating host colonization and bacterial virulence [A. vinelandii and another adhesin in M. primoryensis were identified as the homologues while searching against the PDB database. AlgE6 is a calcium-dependent mannuronan C-5 epimerase, and its structure is elongated parallel β-roll with a shallow [M. primoryensis was responsible for positioning and gaining access to oxygen and nutrients; the Ig-like fold domain region in this adhesion protein bound with Ca2+ and helped the adhesin protein project into medium [V. parahaemolyticus, enabling bacteria to enter seafood, such as prawn (Macrobrachium rosenbergii) [Leptospira, which were identified as homologues to WP_005490731.1; they were involved in adhesion to host cells via extracellular matrix binding and immune evasion [V. parahaemolyticus. However, proteins of WP_005477759.1, WP_005489282.1 and WP_005490731.1 have not been characterized yet, and they should be verified further in wet labs.Prediction of 3D structures and protein interfaces among structural homologues helped us understand fibrillar adhesin functions in 477759.1 ,22. The irulence . The pre shallow . The 0.6o medium . WP_0054nbergii) . LigA an evasion ,27, suggV. parahaemolyticus with the host. Other natural compounds that are promising resources and have not been examined could be further investigated. This study helped set the foundation of analysis for the identification of virulence inhibition targets and novel active compounds.The molecular docking analysis suggested that obacunone, rutin, 8-oxocoptisine and limonin were highly suitable active compounds defending against fibrillar adhesins, revealing applications of these natural plants or activate compounds in the virulence inhibition of V. parahaemolyticus and other veterinary pathogens. Genetic regulators or repressors have been proposed to defend against biofilms, however, there exists a three-dimensional matrix outside bacterial communities which has high tolerance against disinfectants; the chemicals in disinfectants would be difficult to diffuse into the gel like matrix and attack the bacteria. The Ig-like domain was shown to have carbohydrate (lectin)-binding activity, acting as a surface glycan; therefore, it was used to develop methods for detecting E. coli biofilms with recombinant antibodies, thereby locating and destroying the biofilm matrix [Salmonella isolates [Vibrio phage and help develop or modify phage cell surface proteins to easily bind V. parahaemolyticus. The identification of fibrillar adhesins in V. parahaemolyticus help identify those in other pathogens, which therefore will prevent pathogen colonization, quorum sensing and virulence expansion.Ig-like fold proteins have been shown to be effective in regulating m matrix . Antimicisolates . Ig-likeV. parahaemolyticus. The results underline characteristics and unveil the orthologous relationship of Ig-like folded proteins in V. parahaemolyticus. There were three novel fibrillar adhesin-like proteins that have been identified in V. parahaemolyticus within this study. The molecular docking analysis set a foundation for discovering adhesion sites of virulence inhibitors and paved the way for finding out prior drugs for targeting cell attachment and virulence expansion for V. parahaemolyticus and other pathogens.This study examined Ig-like-folded proteins and novel fibrillar adhesin-like proteins in V. parahaemolyticus RIMD 2210633, V. parahaemolyticus CHN25, V. cholerae NCTC 9420 and E. coli K-12 MG1655. Ig fold domains in bacteria were identified which belong to CL0159, the Ig-like fold superfamily [−5; (2) searching a domain score of above 200; (3) the protein sequence identity of over 70%; (4) predicted domains with more than 15 amino acids. Sequence hits were summarized from HMMER searching results.Ig proteins were defined based on Pfam database across erfamily . The selAmino acid sequences of Ig-fold containing proteins were examined for orthogroup clustering via OrthoFinder using DIAMOND as the all-versus-all alignment tool. This examination of homologues was based on the MCL algorithm.Proteins that met standards as follows were considered as the fibrillar adhesin-like proteins ,31: (1) http://cello.life.nctu.edu.tw/cello2go/, accessed on 2 December 2021), which is a web platform to describe gene ontology (GO)-type categories and subcellular localizations based on machine-learning algorithm and BLAST homology searching approaches. The virulence of fibrillar adhesin-like proteins was predicted using VirulencePred  that applied a bi-layer cascade support vector machine (SVM) as the algorithm method. The theoretical physico-chemical properties were examined via ProtParam tool available at ExPASy server.The subcellular localization of fibrillar adhesin-like proteins was predicted using CELLO2GO .Amino acid sequences of the promising fibrillar adhesin-like proteins were loaded into the PDB database, which aims to predict 3D protein structures and biological functions and obtains their structural homologues. The protein interface prediction was performed via BindML , which is an integrated traditional Chinese medicine (TCM) database, and the sdf database was generated via PubChem  via PubChem ID. The chemical compounds were decorated by removing the ligands and water motifs, reviewing this and optimizing the mutation sites, and adding hydrogen through the Pymol 2.3 and UCSF Chimera 1.14rc software. 3D structural information of these four identified fibrillar adhesin proteins used the best-fit molecules based on the PDB database. The binding ability sites, interaction between compounds and fibrillar adhesins were performed using AutoDock vina .The chemical compounds of virulence inhibitors against