Burkholderia pseudomallei, is the causative agent of melioidosis, a disease endemic in South East Asia and northern Australia. Exposure to B. pseudomallei by either inhalation or inoculation can lead to severe disease. B. pseudomallei rapidly shifts from an environmental organism to an aggressive intracellular pathogen capable of rapidly spreading around the body. The expression of multiple virulence factors at every stage of intracellular infection allows for rapid progression of infection. Following invasion or phagocytosis, B. pseudomallei resists host-cell killing mechanisms in the phagosome, followed by escape using the type III secretion system. Several secreted virulence factors manipulate the host cell, while bacterial cells undergo a shift in energy metabolism allowing for overwhelming intracellular replication. Polymerisation of host cell actin into “actin tails” propels B. pseudomallei to the membranes of host cells where the type VI secretion system fuses host cells into multinucleated giant cells (MNGCs) to facilitate cell-to-cell dissemination. This review describes the various mechanisms used by B. pseudomallei to survive within cells.The soil saprophyte,   Burkholderia pseudomallei, is the causative agent of melioidosis, a multifaceted disease endemic to the tropical belt, with hotspots in South East Asia and northern Australia ,103B. psandensis . Furtherandensis ,108. Durandensis . Deletio setting . Howeverhe CPS I . Furtherachieved .O-antigen polysaccharide chain (O-PS) [B. pseudomallei O-antigen was solved and found to be an unbranched heteropolymer of repeating structures of [-3)-β-D-glucopyranose-(1-3)-6-deoxy-α-L-talopyranose-(1-] and is referred to as type II O-PS [BPSL2672-BPSL2688) [B. pseudomallei isolates. Three serotypes of LPS have been observed in B. pseudomallei, smooth typical type A LPS, smooth atypical type B LPS, and rough LPS. The dominant serotype is smooth type A, which accounts for 97% of all screened isolates and consists of the characterised O-antigen moiety, type II O-PS [5 transposon mutant of B. pseudomallei strain 1026b  deficient in type II O-PS was sensitive to killing in 30% non-human serum. Strain SRM117 also displayed a 10-fold reduction in virulence in both hamster and guinea-pig melioidosis models and 100-fold less virulence in an infant diabetic rat model [50 in an intraperitoneal injection model [O-PS resulted in increased uptake into RAW264.7 murine macrophages. However, intracellular counts decreased during the early stages of infection, attributed to the activation of inducible nitric oxide synthase (iNOS), whereas macrophages infected by B. pseudomallei wild-type failed to activate iNOS [O-PS moiety, plays an important role in suppressing the host response during early stages of infection in vitro and deletion results in attenuation in animal models of disease. TLR4 is an important component of this response due to its ability to detect the LPS of Gram-negative pathogens [B. pseudomallei can be recognised by both TLR4 and TLR2, and this has been demonstrated in vitro, but in vivo TLR2 is responsible for the detection of LPS and host response to infection [O-PS, does an increase in TLR4-dependent NF-κB activation occur [in vivo host reliance on TLR2 activation. Differential signalling between murine and human cell line models has been shown with LPS-induced immune activation occurring solely through TLR4 within murine models both in vitro and in vivo, while additional TLR2 activation also occurs in human models [The lipopolysaccharide (LPS) is a glycolipid molecule decorating the outside of the cell wall of Gram-negative bacteria. The LPS consists of three components, the lipid A, the core region, and an n (O-PS) . The str II O-PS ,104. ThePSL2688) . A compr II O-PS . A varia II O-PS . A Tn5 ton model . Furtherate iNOS . These rathogens . B. pseunfection . Furtheron occur , perhapsB. pseudomallei into the host cell, the bacterium comes into contact with many host mechanisms that are utilised to kill and clear foreign material. Phagocytic cells employ and produce both reactive nitrogen intermediates (RNI) and reactive oxygen species (ROS) to control microbial infection [B. pseudomallei is a weak activator of beta-interferon (IFN-β), resulting in reduced iNOS production [B. pseudomallei at a MOI of 10 showed no significant expression of iNOS compared to the high production observed when infected with either Escherichia coli or Salmonella typhi at a MOI of 0.1 [B. pseudomallei is susceptible to chemically generated NO in a macrophage-free system [B. pseudomallei in macrophages.Following the attachment and entry of nfection . B. pseuoduction . iNOS caoduction . InfectiI of 0.1 . Furthere system , highligB. pseudomallei expresses sigma factors, RpoS, and RpoE, and the transcriptional regulator OxyR, which can directly counteract oxidative stress through their downstream effectors. Sigma factors have been shown to mediate the response of B. pseudomallei to oxidative stress [B. pseudomallei, inactivation of rpoS led to multiple effects including increased susceptibility to carbon starvation and oxidative stress with a rpoS mutant unable to induce MNGC formation [B. pseudomallei against menadione and H2O2 and showed reduced intracellular survival in J774A.1 murine macrophages [rpoE mutant also showed differentially present proteins that are involved in oxidative and osmotic stress proteins in addition to chaperones, indicating the role it plays in the survival of B. pseudomallei under different adverse conditions [B. pseudomallei, it exerts a bi-functional role by repressing expression of the KatG catalase under normal growth and activating it under oxidative conditions [oxyR in B. pseudomallei increases the susceptibility to both H2O2 and paraquat, highlighting the essentiality of OxyR in protection against oxidative stress [B. pseudomallei.To protect against oxidative damage, e stress . In B. pormation ,124. Rpoormation . Additiorophages . Proteomnditions . The tranditions ,129. Thenditions . Inactive stress . Taken tkatG (BPSL2865) which encodes for catalase-peroxidase I in B. pseudomallei. Deletion of katG increases susceptibility to killing by different oxidative agents including H2O2, menadione, N-ethylmaleimide (NEM), and hypochlorite [katG gene led to increased resistance to organic hydroperoxide [ahpC gene. Overproduction of AhpC was shown to be protective against both reactive oxygen species and reactive nitrogen species [tert-butyl hydroperoxide (t-BOOH) and demonstrating the role of AhpC in the intracellular survival of B. pseudomallei.There are other genes involved in protection against oxidative stress including chlorite . Interesperoxide . This wa species  explainiB. pseudomallei is also armed with different proteins that can directly protect bacterial DNA. One of these proteins is the putative ferritin DPS-family DNA binding protein (dpsA/BPSL2863) encoded downstream of the katG gene in B. pseudomallei. The gene dpsA is regulated by both OxyR and RpoS, and under oxidative conditions, dpsA is co-transcribed with katG highlighting the multiple responses coordinated by the bacterium to resist oxidative stress [dpsA is important in the resistance of the bacterium against organic hydroperoxide with hypersensitivity to t-BOOH observed with the dpsA deletion mutant [B. pseudomallei also encodes for a spermidine acetyltransferase homologue that can act as a free radical scavenger protecting the DNA [speG, is also indirectly regulated by RpoE under oxidative stress conditions [B. pseudomallei to survive within the lysosomes by mounting a response, detoxifying the environment, and directly protecting the bacterial DNA.In addition to antioxidant effectors that can directly inactivate oxidants, e stress ,130. It n mutant . Further the DNA . This spnditions . This musodC) of B. pseudomallei has been shown to provide some protection against extracellular superoxides [sodC, demonstrating the importance of SodC in the detoxification of extracellular superoxides that may support intracellular growth of B. pseudomallei [B. pseudomallei to adapt to and resist the phagosomal environment by modulating gene and protein expression. However, further studies are required to fully understand the roles of sigma factors and transcriptional regulators in the virulence of B. pseudomallei using in vivo infection models.Superoxide dismutases are enzymes that are able to convert superoxides, one of the most potent reactive oxygen intermediates, into the less damaging hydrogen peroxides for better clearance by other enzymes, including KatG ,134. Alteroxides . Additiodomallei . These sB. pseudomallei has been shown to escape endocytic vesicles of HeLa, RAW264.7, and U937 cells within 15 minutes post-internalisation [B. pseudomallei strain K96243 [B. pseudomallei is homologous to the Hrp2 secretion system identified in Ralstonia solanacearum [Xanthomonas spp [B. pseudomallei has been characterised most extensively, and its components are named the Burkholderia secretion apparatus (Bsa). This secretion system has been observed to share high homology to the inv/spa/prg encoded T3SS of Salmonella Typhimurium and the ipa/mxi/spa T3SS of Shigella flexneri [lisation ,137 , which encodes a structural component of the export apparatus homologous to Salmonella SpaS, showed a severe delay in phagosome escape as demonstrated by bacterial co-localization with the lysosomal-associated membrane protein 1 (LAMP-1) marker within J774.2 murine macrophages [bsaZ mutant to membrane-bound compartments [bsaQ a structural component of the secretion apparatus resulted in delayed escape of the bacterium at 6 hours post-infection with the bsaQ mutant exhibiting greater co-localisation with LAMP-1-positive phagosomes compared to the wild-type [bsaQ mutant to secrete effector proteins. BsaU (BPSS1539) is a part of the T3SS homologous to the InvJ of Salmonella spp., which is responsible for the control of the needle length [bsaU transposon mutant showed significant impairment in vacuolar escape as visualised with LAMP-1 co-localisation staining [B. pseudomallei bsaU mutant [Deletion of genes encoding several structural proteins of the T3SS-3 has been shown to impair bacterial escape. A disruption in the rophages  and tranartments . Insertiild-type . In addie length . A bsaU staining . IntraceU mutant .bsaZ mutant, severe impairment in vacuolar escape was seen with the bipD mutant, including reduced invasion of HeLa human epithelial cells [bipD mutant was also shown to be almost exclusively co-localised with the LAMP-1-associated phagosomes in J774.2 murine macrophages, indicating the inability of the mutant to escape endocytic vesicles [bipD mutant was also unable to induce actin rearrangements and form cellular protrusions. Delay in vacuolar escape was also observed with the inactivation of BipB and BipC, which are homologous to SipB and SipC of Salmonella, respectively [bipB and bipC mutant also showed reduced intracellular survival with the bipB mutant and also display marked reduction in MNGC formation and in vivo virulence [B. pseudomallei Bsa translocators for escape from the phagosome and intracellular spread.The Bsa translocators, BipB, BipC, and BipD, sit at the tip of the Bsa needle complex forming a pore for the delivery of the bacterial effector proteins into host cells . As seenal cells . The bipvesicles . In addiectively ,148. Botirulence . This ilB. pseudomallei in the phagosomes. BopC is an effector protein and was shown to bind to its cognate chaperone BPSS1517 [bopC mutant demonstrated reduction in intracellular survival in addition to delayed bacterial escape from the vacuoles of J774A.1 murine macrophages. Similarly, inactivation of bopA, another Bsa effector also resulted in reduced bacterial survival in phagosomes [bopA mutant with the autophagosomal microtubule-associated protein light chain 3 (LC3) in RAW264.7 murine macrophages highlighted the impaired ability of this mutant to escape the phagosome [B. pseudomallei, BPSS1385 (CHBP) has been shown to be a cycle inhibiting factor (Cif). Cifs are known to cause cytopathic effects in eukaryotic host cells, such as cell cycle arrest and cell death [BPSS1385 in B. pseudomallei have not yet been undertaken, so its exact role in vivo is yet to be elucidated. Taken together, this illustrates the different functions of the T3SS and its effector proteins in facilitating bacterial escape from the endocytic vesicles as well as manipulation of host cell. The importance of the T3SS in B. pseudomallei also makes it an attractive target for therapeutic intervention. This was demonstrated using small molecule ATPase inhibitors, targeting the T3SS ATPase bsaS, which effectively reduced bacterial escape from the phagolysosome, resulting in decreased intracellular bacteria [in vivo remains to be validated, however this highlights the potential for targeting the T3SS in B. pseudomallei.In addition to the translocators, deletion of several genes that encode for effector proteins that subvert host functions has led to the delayed or impaired escape of BPSS1517 . Charactll death . Cifs anll death ,154. Trebacteria . How effB. pseudomallei, three phospholipase C (Plc) are encoded, plc1 (BPSL2403), plc2 (BPSL0338), and plc3 (BPSS0067) [plc2 reduced the intracellular survival, plaque formation, and cytotoxicity in HeLa human epithelial cells, indicating that plc2 is more associated with virulence and intracellular survival compared to plc1 and plc3 [plc2 mutant showed that the reduction of intracellular survival was not due to delayed escape [B. pseudomallei encodes the serine protease inhibitor ecotin (eco/BPSL1054) that is active against elastase, trypsin, and chymotrypsin [Bacterial phospholipases are membrane-bound or secreted hydrolases . In B. pPSS0067) . Deletioand plc3 ,158. Furd escape . B. pseuotrypsin . This alotrypsin . Additiootrypsin . Taken tpurN/BPSL0908) and phosphoribosylformyl-glycinamidine cyclo-ligase (purM/BPSL2818) [purN and purM from B. pseudomallei strain E8 resulted in decreased intracellular survival within HeLa cells, with the latter showing no intracellular replication. Furthermore, gene deletions that affected the histidine (hisF/BPSL3133) and para-aminobenzoate biosynthetic pathways (pabB/BPSL2825) also exhibited reduced intracellular survival in HeLa cells [asd (BPSS1704), which encodes the enzyme aspartate-β-semialdehyde dehydrogenase, important for amino acid biosynthesis, resulted in the generation of a diaminopimelate (DAP) auxotrophic strain that was unable to grow in rich media without supplementation and displayed reduced intracellular replication in both RAW264.7 murine macrophages and HeLa cells [Several metabolic enzymes have also been implicated to be important in the survival of the bacterium inside the host cells . TranspoPSL2818) . The inaLa cells . The reaLa cells . Like otLa cells .Figure B. pseudomallei survival and virulence. In particular, enzymes involved in the biosynthesis of the branched chain amino acids (BCAAs) leucine, isoleucine, and valine have been found to be essential for virulence. Disruption of ilvI (BPSL1196), encoding an acetolactate synthase catalytic subunit involved in 2-oxocarboxylic acid metabolism, was first discovered incidentally and investigated further due to the loss of virulence in a murine melioidosis model. The ilvI transposon mutant was auxotrophic for leucine, isoleucine, and valine. This gene assists the conversion of both pyruvate and 2-oxobutanoate into intermediate metabolites that form the skeleton for these BCAAs. The transposon mutant was unable to persist within mouse models or cause lethal infections to the same extent as wild-type B. pseudomallei strain 576. Interestingly, this gene is non-essential in rich media, but downstream components of the BCAA biosynthesis are necessary for survival (ilvC/BPSL1198) [Other genes encoding for enzymes that participate in amino acid biosynthesis are equally important for PSL1198) ,164.B. pseudomallei following infection of Syrian hamsters showed an upregulation of genes involved in the intermediate steps of the glycolytic pathway , inositol catabolism , glucosamine catabolism (glmS3), and tryptophan catabolism/synthesis . Interestingly, genes involved in oxidative phosphorylation such as cyoA, cyoB, and fdsB, were highly induced, while genes involved in the TCA cycle , serine catabolism/synthesis , and energy metabolism  were highly down regulated [aceA/BPSL2188), in Sprague-Dawley rats (a chronic model of melioidosis), results in the switching of infection from a persistent state to a lethal acute infection if untreated by antibiotics [B. pseudomallei being metabolically flexible and able to utilise alternate energy sources during infection in vivo.Transcriptomic studies of egulated . The swiibiotics . This poB. pseudomallei infection. The gene, BPSL1549, encodes for a toxin or Burkholderia Lethal Factor 1 (BLF1), which shows homology to E. coli toxin cytotoxic necrotizing factor 1 (CNF1) deamidase. This toxin is upregulated during virulence cues such as in the presence of human serum, taurine, and insulin. Purified BPSL1549 caused cell toxicity to J774.2 murine macrophages after 72 hours and a mutant strain, B. pseudomallei ΔBPSL1549 was severely attenuated in BALB/c mice and displayed a 100-fold increase in median lethal dose. BPSL1549 was shown to bind to the eukaryotic transcription initiation factor, eIF4A. BPSL1549 causes deamidation of Gln339 of eIF4A, which results in potent inhibition of host translation and protein synthesis [BPSL1527 or tex, codes for a transcription accessory protein, which has been shown to be involved in toxin regulation in other pathogens. A deletion mutant, Δtex displayed reduced CFU numbers in spleen and lungs 48 hours post-infection and decreased survival in A549 epithelial cells and J774A.1 murine macrophages [tex is directly involved in toxin regulation in B. pseudomallei as studies to determine this have not yet been published.Toxins have also been shown to play a role in ynthesis . A putatrophages . It is nBPSS1823 (mip) and BPSL0918, have also been identified as important for intracellular survival of B. pseudomallei in J774A.1 murine macrophages [cis-trans isomerases and are involved in protein folding and chaperone activity [mip resulted in reduced motility as well as protease production and attenuation in a BALB/c mouse model [B. pseudomallei to establish intracellular infection.Two members of the immunophilin superfamily, rophages ,170. Theactivity . In addiB. pseudomallei is able to propel itself within a host cell by polymerising host cell actin [Burkholderia intracellular motility A (BimA), which was inferred through homology to other auto-secreted proteins in pathogens that polymerise host cell actin, in particular the YadA protein of Yersinia enterocolitica [bimA results in loss of actin-based motility as shown in infected J774.2 murine macrophages, where bacterial cells cluster in the cytoplasm but exhibit a distinct lack of polar actin tails and cellular protrusions [B. pseudomallei towards the host cell membrane and cause cellular protrusion from which cell-to-cell spread is then mediated [B. pseudomallei is capable of polymerising actin in vitro independent of the Arp (actin-related protein) 2/3 complex, the protein complex that regulates and nucleates eukaryotic actin [bimA in the genome, is also required for actin tail formation in HeLa cells, but this deficiency was only partially restored by complementation [ΔbimC-mutant strain, actin tail formation was absent. This indicates that BimC participates to some extent in BimA-mediated actin polymerisation [ll actin ,173 , cathespin K (CTSK), and tartrate-resistant acid phosphatase (TRAP), markers which are hallmarks of functional osteoclasts, natural MNGCs formed from the fusion of mononuclear cells [B. pseudomallei-induced MNGCs are only osteoclast-like [lfpA  was responsible for the increase in expression of osteoblast markers, CTR, CTSK, and TRAP seen in RAW264.7 murine macrophages. Additionally, lfpA is required for optimal virulence in animal models, as deletion of IfpA results in a 4.5-fold-decrease in LD50 and a delayed time-to-death in Syrian hamsters and BALB/c mice [B. pseudomallei, in particular the Cluster 1 T6SS [Following the movement of e formed . This phar cells ,178, altast-like . StudiesB/c mice . The forr 1 T6SS .Vibrio cholerae and Francisella tularensis [B. pseudomallei encodes six T6SS clusters termed clusters 1 through to 6 (tss-5), T6SS-2 (tss-4), T6SS-3 (tss-6), T6SS-4 (tss-3), T6SS-5 (tss-2), and T6SS-6 (tss-1)   [190,191nt broth , with fuer model , indicatrm MNGCs . Hcp prodomallei ,193. Toedomallei  and thatdomallei . T6SS-1 s et al. .Table 2.hcp1, tssAB, and vgrG. The T3SS regulator protein, BprC, mediates expression of tssAB but not of hcp1 and virAG [B. pseudomallei to cysteine and reduced glutathione, present in the cytoplasm of host cells, activated the two-component system sensor kinase VirA, driving expression of T6SS-1 [Activation of the T6SS-1 is mediated by the VirAG two-component system, which senses host intracellular signals, thus activating transcription of genes encoding components of the T6SS-1 such as nd virAG . Work byf T6SS-1 . Expressf T6SS-1 .B. thailandensis have shown that T6SS-6 is important for contact-dependent bacterial competition, with deletion of clpV (encoding an AAA+ ATPase required for recycling the contracted sheath), resulting in a strain unable to persist in mixed population biofilms with Pseudomonas putida [ΔtctR was indistinguishable from wild-type in a Syrian hamster infection model [B. thailandensis and has been observed to be regulated by OxyR. Overexpression of T6SS-2 was observed in the B. thailandensis ΔoxyR mutant [B. pseudomallei may be important for survival in different environmental niches, investigation into this has been limited to date, with focus mainly on the intracellular environment.The precise roles of the remaining five T6SS clusters requires further investigation, with only Cluster 1 shown to be essential for virulence in the mouse model . Studiess putida . A MarR on model , indicatR mutant . This stR mutant ,196. TheB. pseudomallei intracellular spread and MNGC formation is the cyclophilin ppiB. The ppiB gene is essential for establishment of disease in BALB/c mice [ppiB resulted in major disruption of the bacterial proteome ultimately leading to a 67% reduction in MNGC formation [in vitro and in vivo, although further work needs to be done to corroborate this.Another protein important for B/c mice . It was ormation , likely B. pseudomallei has maintained numerous strategies to successfully establish a niche in humans and animals. B. pseudomallei employs many mechanisms for cell entry, vacuole escape, and subsequent cell-to-cell spread. It subverts the immune response to allow for bacterial proliferation, spreads to all systems in the host causing a multitude of symptoms, and can rapidly develop into septic pneumonia which can be fatal.In conclusion, Burkholderia pseudomallei remains a resilient environmental pathogen capable of causing major morbidity in tropical regions of the world. Although studies have now unravelled mechanisms by which B. pseudomallei infects and replicates in the host, there is still much left to understand, such as its ability to persist, hide away effectively, and then remerge to cause infection years later. Unique mechanisms for virulence in B. pseudomallei, such as Arp2/3-independent actin polymerisation by BimA [boaA and boaB [B. pseudomallei KHW strain, the absence of the flagella results in attenuation in BALB/c mice [B. pseudomallei, more investment into genetic tools and characterisation of genes important for pathogenesis and persistence is required. With no currently licensed vaccine and treatment compliance moderate at best, particularly in endemic areas, more work is needed to find novel intervention points, which could be used in conjunction with current antibiotic therapies. This review has highlighted many virulence factors and lifestyle adaptations, which are required by B. pseudomallei to cause infection and survive within the host. Selection of some of these virulence determinants for therapeutic intervention studies, with the focus on synthesis of novel inhibitors, should support the development of new strategies to help melioidosis patients and those living in endemic areas. by BimA  and multand boaB , mean caB/c mice ; in contB/c mice . These i


Centaurium erythraea Rafn  also produces cytotoxic secondary metabolites, and its presence should be carefully monitored. In this study, the complete chloroplast of Centaurium erythraea subsp. majus (Hoffmanns. & Link) M.Laínz  isolate BPTPS121 is described, being the first available plastome belonging to the Centaurium genus. The chloroplast genome (GenBank accession number: ON641347) is 153,107 bp in length with 37.9% GC content, displaying a quadripartite structure that contains a pair of inverted repeat regions , separated by a large single-copy  and small single-copy  regions. A total of 129 genes were predicted, including 37 tRNA genes, eight rRNA genes, and 84 protein-coding genes. The phylogenetic analysis showed that isolate BPTPS121 is placed under the Gentianaceae family, belonging to the Gentianales order. The maximum-likelihood tree supports the already described lineage divergence in the Gentianaceae family, with C. erythraea subsp. majus belonging to the Chironieae tribe positioned below the Exaceae tribe and above the Potalieae and the entire Gentianeae tribes. This study will contribute to conservation, phylogenetic, and evolutionary studies, as well as DNA barcoding applications for food, feed, and supplements safety purposes.Despite having many historically reported ethnomedicinal uses,  Centaurium Hill , and ca. five to Schenkia Griseb.   M.Laínz  under the voucher LISE: 96442 (identified by: Jorge Capelo).The material of nz Laínz  analyzedImmediately after collection, young leaves from the selected specimen were frozen in liquid nitrogen and stored at −80 °C until further processing. Total genomic DNA was extracted from the preserved material using an adaptation of the Doyle and Doyle  methodolERR10047930) were used to assemble the complete chloroplast genome (sequence coverage: 972×) using the GetOrganelle pipeline (v1.7.3.1)  while setting the flags ‘--max-reads’ and ‘--reduce-reads-for-coverage’ to 25 million and one thousand, respectively . The plastome annotation was performed using the GeSeq tool  to compare them with the results obtained by performing a BLAT search on GeSeq .The about 40 million high-quality paired-end reads obtained  G.Don  belonging to the Gentianales order but not from the Gentianaceae family, Nonea vesicaria (L.) Rchb.  belonging to the same lamiids clade but from a different order, and Tilia platyphyllos Scop.  from the malvids clade. The sequences were aligned using MAFFT v7.450  was performed using the concatenated nucleotide sequences coding for the shared proteome (65 coding sequences) extracted from a selected dataset. The dataset was composed of 17 verified and complete chloroplast genomes belonging to the Gentianaceae family available in GenBank (accession date: 26 June 2022), with only one species representing each genus. The selected dataset also included the complete chloroplast genome of C. erythraea subsp. majus isolate BPTPS121 is placed under the Gentianaceae family, belonging to the Gentianales order. The Gentianaceae family has six tribes in the current classification: the Chironieae, Exaceae, Gentianeae, Helieae, Saccifolieae, and Potalieae (Struwe C. erythraea subsp. majus isolate BPTPS121 (ON641347) and Eustoma grandiflorum (MK991810)) is the second most basally positioned tribe, below Exaceae , and above Potalieae  and the entire Gentianeae tribe, with 100/100 percent support (SH-aLRT/UFBoot2). This tree, therefore, supports the already described lineage divergence in the Gentianaceae family .The maximum-likelihood tree showed that e Struwe . These ty Struwe , with coC. erythraea subsp. majus isolate BPTPS121, the first described plastome belonging to the Centaurium genus. This complete genome will contribute to conservation, phylogenetic, and evolutionary studies in the Gentianaceae family. It will also support DNA barcoding applications for food, feed, and supplements safety and quality purposes that target detecting species that produce secondary metabolites with cytotoxic potential.This study describes the chloroplast genome of


After cardiovascular diseases, cancer is the second main cause of death globally. Mushrooms have been demonstrated to contain amalgamation with properties capable of inhibiting carcinogenesis and microbial growth, principally secondary metabolites such as quinolones, steroids, terpenes, anthraquinones, and benzoic acid derivatives among others. This study aimed to substantiate their potency concerning colon cancer carcinogenesis and antimicrobial growth. A systematic search of important literature was performed considering all the articles published until April 2022. Screening was performed by searching the BMC Springer, Elsevier, Embase, Web of Science, Ovid, and MEDLINE databases. In addition, Google Scholar was used to supplement information. Titles and abstracts that matched the established criteria were selected for full-text article scrutiny and subsequently used in the updated present review. Bioactive compounds present in medicinal mushrooms such as ascorbic acid, organic acids, flavonoids, polysaccharides, glycosides, phenols, linoleic acid, grifolin, and tocopherols among other compounds play a key role in suppressing the proliferation of cancerous cells and selectively act as antibacterial and antifungal agents. These metabolites actively scavenge oxygen free radicals, hydroxyl radicals, and nitrite radicals that would otherwise increase the risks of the growth and development of cancerous cells. Mushrooms' bioactive compounds and metabolites actively inhibit nuclear factor-kappa activation, protein kinase B processes, and ultimately the expression of Cyclooxygenases 2 in cancerous cells. Medicinal mushrooms should be considered as alternative natural chemo-preventive agents in the global fight against colon cancer and the evolution of drug-resistant pathogenic microorganisms, as they exhibit robust potency. They have not been reported to exhibit adverse harmful effects compared to synthetic chemotherapies, yet they have been reported to demonstrate significant beneficial effects. Lignosus rhinocerotis has contributed to a better understanding of their molecular biology and expanded avenues for more investigative studies extending to the genome level [Mushrooms have been demonstrated to contain amalgamation with properties capable of inhibiting microbial growth, principally secondary metabolites such as quinolones, steroids, terpenes, anthraquinones, and benzoic acid derivatives . They alme level . Genome me level .After cardiovascular diseases, cancer is the second main cause of death . LocalizMushrooms are taxonomically classified as Basidiomycetes and some as Ascomycetes. They provide nutrients including easily digested proteins, fibre, minerals, carbohydrates, vitamins, and antioxidants. They are therefore largely consumed because of their nutritional benefits as cited by . FurtherThere still exists a dire need to develop new, affordable, and effective anticancer drugs . More thRAS oncogenes [KRAS mutation-negative than KRAS mutation-positive [Although certain cancers have a genetic predisposition, the major external causes of this disease have been linked to lifestyle choices, exposure to pollutants in the environment, and dietary . Due to ncogenes , 19. Howpositive , 19. Howpositive . In Westpositive .The modulation of the composition and metabolic activity of the human G.I tract microbiota, which has an influence on health, is becoming widely understood to be a function of dietary  and other environmental factors . In compIt is with increased dependence on synthetic colon cancer treatment options that this paper undertook to review mushrooms of medical importance containing natural bioactive agents with potency against colon cancer growth and proliferation. Plants' natural constituents may provide a new source of anticancer and antimicrobial treatment with a sufficient novel mode of action. Compared to synthetic agents, phytoconstituents of plant origin are rarely seen to correlate with numerous side effects and have been demonstrated to present overwhelming therapeutic activities to heal numerous infectious diseases . MedicinScreening for important literature was duly performed by searching the BMC Springer, Elsevier, Embase, Web of Science, Ovid, and MEDLINE databases. In addition, Google Scholar was used in supplementing information. Articles in google scholar were first screened for authenticity by interlinking the various articles identified with their corresponding existing publisher. The search yielded a total of 3805 papers that were published between the years 2000 and April 2022. As a result of not meeting the inclusion criteria, a total of 3685 articles were excluded from the study. A total of 120 articles met the inclusion criteria and have adequately been discussed in the present review.For the study, the authors first independently screened only English-language publication titles and abstracts from primary studies, considering all the articles published until April 2022 , 28. Res“Colorectal cancer”, “adenomatous polyps”, “colon cancer”, “colon tumor”, “colorectal tumor”, terms were then combined with either of the following MeSH terms: “Bioactivity”, “biological activity”, “anti-cancer”, “phytochemicals”, “pharmacological activities”, “anti-tumour”. These terms were further combined with either of the following terms “mushroom”, “medicinal mushroom”, “bacteria”, “anti-bacterial”, “antifungal, antiviral”, or “anti-microbial”.The burden of colon tumor pathogenesis is quite multiplex and not well understood. However, its initiation process has been associated with the interactive effects of peril factors like lifestyle, heredity, and environmental factors , 30. EmpRussula virescens exhibited substantive scavenging effects on OH radicals in a dose-dependent mechanism. Derived extracts of Hohenbuehelia serotina, Dictyophora indusiata, and Hypsizygus marmoreus have also been established to demonstrate very significant scavenging potential towards OH radicals, with potency levels reaching 100% [Albatrellus confluence and has been cited to be an important bioactive anticancer agent. It has been demonstrated to inhibit the growth and aggression of cancerous cells by suppression of the ERK1/2 pathway and by induction of apoptosis [Bioactive compounds present in medicinal mushrooms such as ascorbic acid, organic acids, flavonoids, polysaccharides, glycosides, phenols, tocopherols among other compounds , 37 playing 100% . Phytocopoptosis . Bioactipoptosis .Ganoderma lucidum (G. lucidum) water extract (BSGLWE) on colorectal cancer. According to their findings, BSGLWE considerably reduced the viability of colorectal cancer HCT116 cells in a dose- and time-dependent fashion. According to flow cytometry research, BSGLWE interfered with the advancement of the cell cycle at the G2/M phase by downregulating cyclin A2 and cyclin B1 while as well it upregulated the P21 at the level of mRNA. Additionally, BSGLWE caused apoptosis by lowering the protein levels of PARP, Bcl-2, pro-caspase-3, and pro-caspase-9 as well as the mRNA levels of survivin and Bcl-2 [Na et al.  examinednd Bcl-2 .Additionally, through an in vivo experiment, BSGLWE inhibited tumor growth by controlling the expression of genes and proteins linked to cell cycle and death. This effect was further supported by a decrease in Bcl-2, PCNA, and Ki67expression as shown by immunohistochemical labelling . The proG. lucidum. Overall, they reported that BSGLEE efficiently prevents colorectal cancer carcinogenesis by promoting cell cycle arrest, inducing apoptosis, and inhibiting migration. Based on the suppression of apoptosis by reversing microtubule polymerization, Li et al. [G. lucidum polysaccharide could mitigate intestinal barrier impairment caused by paclitaxel. G. lucidum polysaccharide, one of the most studied and representative polysaccharides, is regarded as a prebiotic candidate due to its anti-tumor action [G. lucidum polysaccharide for host cancer prevention, particularly CRC, have been shown in numerous trials.Similarly, Li et al.  investigi et al.  hypothesr action . The benG. lucidum polysaccharide interact has been developed. Using CRC mice as a model, Luo et al. [G. lucidum polysaccharides (GLPs) could significantly alter CRC symptoms by promoting the relative abundances of Enterobacteriaceae, Bacteroides, and Peptostreptococcaceae and lowering those of Desulfovibrionaceae, Oscillospira, Clostridiales and Ruminococcus [G. lucidum polysaccharide intake, was also found to be less helpful at alleviating CRC symptoms [Additionally, a thorough understanding of how the gut microbiota and o et al.  discovernococcus . Guar gusymptoms .G. lucidum polysaccharide could significantly reduce CRC-associated symptoms, including the tumor cell proliferation, inflamed gut barrier, and production of oncogenic signalling molecules, which are connected to the rise in relative abundances of Bacteroidetes and other SCFAs-producing bacteria [Another study established that the combination of jiaogulan saponins and bacteria .G. lucidum that demonstrated anticolorectal cancer properties and purified the ribonuclease protein, (17.4-kDa) using liquid chromatography techniques. They reported that the ribonuclease protein could arrest the cell cycle at G1 phase by controlling the expression of cyclin D1 and P53 in HCT116 and HT29 colorectal cancer cell lines. The G. lucidum ribonuclease protein demonstrated strong anti-proliferative and anti-colony formation actions. The ribonuclease was shown to activate pathways controlled by caspase9 and the unfolded protein response to cause cell death in HCT116 cells. Additionally, the ribonuclease treatment drastically reduced the capacity of HCT116 cells to engage in autophagy, a stress adaptation mechanism to deal with metabolic crises [In another different study, Dan et al.  describec crises .Lentinus edodes  in vitro and their results showed to have anti-proliferative activities against HT-29 and HCT-116 cells. They further reported that the in vitro proliferation assays for adherent HT-29 and HCT-116 carcinoma cells and suspended S-180 sarcoma cells indicated that the three water-soluble polysaccharides had a higher antitumor activity against suspended cells than adherent cells [Jeff et al.  reportednt cells .T. robustus showed immunomodulatory qualities by significantly stimulating thymocytes, splenocytes and macrophages.Polysaccharides from certain Termitomyces mushrooms have anticancer effects. Both water-soluble and insoluble β-glucan obtained from hot water extracts of Cyclooxygenases are proteinous and are correlated with the production of lipid prostaglandins. They are significant for biochemical processes in the body . CyclooxElaphomyces granulates inhibited the COX-2 mechanism of action by about 68%, while the present bioactive compounds: syringic acid and syringaldehyde inhibited the COX-2 mechanism in a dose-dependent manner, with an IC50 of 0.4 μg/mL and 3.5 μg/mL, respectively [C. hookeri and Inonotus obliquus caused downregulation of iNOS and COX-2, and inhibition of mRNA expression of iNOS and COX-2 respectively also. In addition, Agaricoglycerides derived from Grifola frondose induced upregulation of NF-κB and the production of COX-2, iNOS, TNF-α, ICAM-1 and IL-1β [Grifola frondosa. The most significant among these are polysaccharide fractions which stimulate the immune-competent cells and enhance antitumor activities [Crude ethanolic extracts (50 μg/mL) of ectively . Terpenond IL-1β . Numeroutivities .Preventive synthetic COX-2 has been cited to have harmful side effects prompting the exploration of natural inhibitors as alternative forms of chemoprevention interventional therapies in colorectal cancer management. Natural antioxidants abundant in mushrooms such as polyphenols and carotenoids Table , suppresSignalling pathways like mitogen-activated protein kinase (MAPK), phosphatidylinositol 3-kinase (PI3K), nuclear factor-kappa B  modulates the expression of COX-2. The influence of the NFκB process is mediated by P13K via AKT . MushrooLentinus edodes and additionally demonstrates immunomodulating properties through the release of cytokines from immunocytes, and therefore are suitably utilized in prevention of different cancers. Lentinan acts as an intravenous anti-tumor polysaccharide [Lentinan Fig.  is a polccharide .Fig. 1LeBy acting directly on macrophages or indirectly through lentinan-stimulated T cells, lentinan causes an increase in the production of a number of bioactive serum factors linked to immunity and inflammation. These include IL-1, IL-3, CSF, vascular dilation inducer, and acute-phase protein induction, which results in the induction of numerous immunobiological changes in the host .Trametes versicolor and is made up of proteins and β-glucans with 25–38% protein residues. Along with neutral amino acids like leucine and valine and trace levels of basic amino acids like arginine and lysine, it primarily comprises acidic amino acids like glutamic and aspartic acids. Krestin  [The protein-bound polysaccharide (proteoglucan), composed of β -glucan, or maitake D-fraction Figure , is a birondosa) . In patirondosa) , 82. Therondosa) , 81, 82.Schizophyllum commune produces schizophyllan polysaccharide  [S. typhi in the macrophage cells and antibacterial activity in albino mice is correlated with increased autophagic-macrophage activity, causing macrophage-mediated antibacterial mechanism and a systemic antibacterial activity via type 1 interferon [Urgency is needed to curtail and control increasing antimicrobial drug resistance by use of improved antibiotic therapy and minimize hospital cross-infection . In thisbisporus  have all. typhi) . The mecterferon .Coriolus versicolor (C. versicolor) were observed in a different study against Staphylococcus aureus (S. aureus) and Salmonella enterica. The Minimum Inhibitory Concentration (MIC) levels of the tested bacteria ranged from 0.625 to 20 mg mL-1. C. versicolor demonstrated noteworthy activity against both Gram-negative and Gram-positive bacteria. The growth curves of Salmonella enterica and Staphylococcus aureus measured at 630 nm, and precisely confirmed with a macrodilution method demonstrated that the recovered C. versicolor methanolic extract actively inhibited the growth of the two bacterial strains tested in the study [Bacillus subtilis, S. aureus, and Bacillus cereus are largely inhibited by methanolic extracts from Boletusedulis, Agaricus bisporus and Cantharelluscibarius mushrooms [. Further research has similarly established that the antibacterial activity of extracts obtained from Agaricus blazei mushroom exhibited significant inhibition for most Gram-positive bacteria compared to the Gram-negative bacteria tested. The extracts inhibited bacterial strains of Streptococcus pneumoniae, Salmonella typhi, Streptococcus mutans, P. aeruginosa, Streptococcus sobrinus, S. aureus, Listeria monocytogenes, B. cereus, and coliforms [Trametes spp. and Microporus spp. and demonstrated activity against Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Pseudomonas aeruginosa, and S. aureus [The antibacterial characteristics of extract recovered from the fruiting body of he study . Other eushrooms . Furtheroliforms . The mecoliforms . Linoleioliforms . A study. aureus . reportedGanoderma pfeifferi and G. australe extracts have been demonstrated to have potent antibiotic activity against E. coli [G. lucidum demonstrated exceptional antibacterial activity against B. subtilis, E. coli, and Salmonella species. Disk diffusion studies have demonstrated that the chloroform extract of G. lucidum have growth-inhibiting effects on B. subtilis and S. aureus [ E. coli , 96. Met. aureus .G. australate extract against P. aeruginosa and E. coli was 2 mg/ml, whereas it was 0.25 mg/ml for Bacillus species and 1 mg/ml for S. aureus [Ganoderma extract against B. subtilis (3.5mg/ml) and Bacillus species (3.5mg/ml) was also reported. Staphylococcus, Streptococcus, Escherichia, and Vibrio species are susceptible to Polyperin, which was identified from Polystichus sanguineus [Ganoderma applanatum [Lentinula edodes against S. aureus and other bacteria has been attributed to the presence of oxalic acid [The MIC value for . aureus . A high nguineus . Numerouplanatum . The antlic acid .To exist and survive in their natural ecosystems, mushrooms must generate antifungal and antibacterial biomolecules . In thisTrametes spp. and Microporus spp. extracts have been reported to actively inhibit Candida albicans, C. tropicalis, and C. parapsilosis fungi of medical importance [A. bisporus, A. bitorquis, A. essettei, and Oudemansiella canarii methanolic extracts exhibited substantial activity against Candida spp. Grifolin, isolated from Albatrellus dispansus mushroom appeared to be the most significant bioactive molecular compound against the tested phytopathogenic fungi [Ganoderma lucidum (G. lucidum) methanolic extracts showed an activity MIC=0.005mg/mL against Trichoderma viride [. Other authors have further demonstrated that water extracts and ethyl acetate with 5% DMSO of Climadocon pulcherrimus, Agrocybe perfecta, Pycnoporus sanguineus, and Oudemansiella canarii exhibited substantial antifungal potency against Candida krusei [Unlike ethanolic and chloroform extracts, hot water extracts have exhibited the most powerful antifungal activity. The hot water extraction method yields numerous antimicrobial compounds including terpenoids, flavonoids, and tannins . This acportance . It has ic fungi . Ganodera viride . Other aa krusei .Xylaria intracolarata extract and displayed inhibition potential against Aspergillus niger and C. albicans [. Phenolic acids and associated molecular compounds such as cinnamic acids and phydroxybenzoic established in G. lucidum demonstrated potential ability against A. versicolor, Trichoderma viride, Penicillium funiculosum, A. ochraceus, Aspergillus fumigatus, A. niger, P. ochrochloron, and P. verrucosum [.The low molecular weight terpene compound grifolin exhibits overwhelming antifungal potency while other low molecular weight compounds such as rufuslactone, enokipodim, cloratin A, 2-aminoquinoline, and sesquiterpene rufuslactone exhibited minimal but substantial activity compared to grifolin. Cloratin was obtained from albicans , 100. Phrrucosum .A. bisporus have on the other hand been reported in the recent past. The concerns have revolved around their long-term consumption uncooked. Freshly obtained A. bisporus induced tumors in mice upon being fed uncooked for a while. However, upon being air-dried, the mushrooms were fed to mice for 500 days and no carcinogenic tumor effect was observed [Besides the significant benefits and potential of mushrooms of medical importance, fundamental concerns regarding the safety of certain mushrooms such as observed , 106. ThBRAF and KRAS mutations, and molecular characterisation of malignancies have revealed evidence of distinct CRC subtypes that arise through activation of several neoplastic pathways. Epidermal growth factor receptor (EGFR) signalling pathway is constitutively activated as a result of KRAS and BRAF oncogene mutations [KRAS oncogene mutation, which has a significant impact on colonic polyp growth and preclinical tumors [KRAS mutation's predictive significance in metastatic CRC treated with anti-EGFR targeted therapy [Understanding the interactions between tumor molecular alterations and exogenous and endogenous variables can help with the identification of tumor molecular markers in CRC . Microsautations , 108. Moutations . A definutations , 109. Anl tumors . Substan therapy , 112. ThPolyozellus multiplex, Sarcodon imbricatus, and Cortinarius purpurascens, respectively, and have been demonstrated to inhibit EGFR activation in molecular docking, molecular dynamics, and in Absorption, Distribution, Metabolism and Excretion (ADME) studies [Edible mushrooms have been identified as potential EGRF inhibitors . After s studies –116.EGFR and downstream pathways frequently encounter resistance, making it more important than ever to find compounds that may be used in combination with these treatments to give cancer patients a long-lasting response [G. lucidum extract has significant promising efficacy in this regard [EGFR and HER2, a pharmaceutical strategy utilizing tiny tyrosine kinase inhibitors (TKIs) (erlotinib and lapatinib) has been created. While lapatinib suppresses the function of EGFR and HER2-TK activity, erlotinib targets ATP-binding sites to prevent EGFR-TK activity [Tyrosine kinase inhibitors (TKIs) that target response , 118. Itactivity –120. BotMedicinal mushrooms are potential anti-colorectal cancer and antimicrobial agents if explored as chemo-preventive pharmaceutical products because they exhibit robust active potency. From our extensive literature search across different authentic and recognized databases, they have not been reported to exhibit adverse effects compared to synthetic chemotherapies currently in use. Tapping into the wealth of knowledge underlying the use of both edible and medicinal mushrooms in treating various infections across different communities in the world, points out their significant value in human health and wellbeing and should therefore not be ignored. Their demonstrated powerful bioactive metabolites in this research elucidate their inherent potential as suitable sources of the anticarcinoma and antimicrobial explorative pharmaceutical venture.EGFR overexpressing tumors may be best accomplished by combining treatment modalities. A focus of interest should be the development of novel and prospective EGFR inhibitors from edible and medicinal mushrooms and their progenitor mushrooms considering this updated review. We however recommend further research to understand the specific mechanisms of action poised by the metabolites, inhibiting, or promoting metabolic and signalling pathways that would otherwise result in CRC carcinogenesis. We further recommend research to evaluate the potential of these active phytochemicals to reach and cross the biological membrane as this will shed more light on their possible bioavailability or their unfortunate elimination from the human body system. Finally, Medicinal mushrooms should be considered as alternative natural chemo-preventive agents in the global fight against CRC and the evolution of drug-resistant pathogenic microorganisms.In addition, it is fundamental to integrate the information now available on the molecular characteristics of tumors and to define new molecular subtypes of CRC by detecting acquired somatic mutations utilizing targeted sequencing of important cancer genes. To further our understanding of the underlying carcinogenic mechanisms that underlie CRC-associations with known risk factors, it is pivotal to look at inherited genetic variation as well as lifestyle, dietary, and environmental risk factors in relation to various CRC tumor subtypes. We underscore that improving prognosis in CRC patients with


The population of wild felids is large and stable in Romania with many carnivore habitats being protected. Felids can be infected with a wide variety of cardio-pulmonary nematodes and can act as reservoirs of infection for domestic cats. The aim of this study was to evaluate the distribution and species diversity of cardio-pulmonary nematodes in wild felids from Romania.Lynx lynx and 47 Felis silvestris) were legally collected from different locations in Romania and analysed by complete parasitological necropsy. The entire respiratory tract was longitudinally opened and checked for the presence of nematodes. Detected nematodes were collected and morphologically identified to species level.A total of 54 wild felids  followed by Angiostrongylus chabaudi (23.4%) and T. brevior (14.9%). Dirofilaria immitis and Aelurostrongylus abstrusus were each detected in two wildcats (4.3%).Two Eurasian lynxes and 29 European wildcats were positive for cardio-pulmonary nematodes. Eurasian lynxes were infected with two species of cardio-pulmonary nematodes, A. abstrusus in wildcats and a patent infection with T. brevior in Eurasian lynx.The present study expanded the epidemiological knowledge on felid cardiopulmonary nematodes in Romania. We confirmed the presence of The online version contains supplementary material available at 10.1186/s13071-022-05281-z. Felis silvestris catus) [Aelurostrongylus abstrusus  is known as the most important respiratory helminth, with a worldwide distribution, and may pose a health risk to infected animals [Cardio-pulmonary nematodes of wild and domestic felids have become increasingly popular among researchers in recent years, with a significant increase in published studies , 2. Howes catus) –7 even ts catus) , 9. Amon animals . Aeluros animals , 11.Troglostrongylus brevior  have gained attention, in some countries being the most frequent lungworm of domestic cats [Troglostrongylus includes five species that reside in the respiratory system of felids: T. troglostrongylus Vevers, 1922, T. brevior Gerichter 1949, T. subcrenatus Railliet and Henry, 1913, T. assadovi Sadykhov, 1952, and T. wilsoni Stough, 1953 [T. brevior in domestic cats [Troglostrongylus infection was reported in various species such as wildcats , leopard (Panthera pardus), tiger (Panthera tigris), bobcat (Lynx rufus) Canada lynx (Lynx canadensis), and lynx (Lynx lynx) [Troglostrongylus brevior was first described in F. s. lybica and F. chaus in Palestine [T. brevior was identified for the first time in a wildcat [T. brevior and represent important reservoirs with prevalences of up to 71.4% in Italy [T. brevior infection seems to spread among the wild and domestic cats, there is only one report of infection in a Eurasian lynx from Bosnia and Herzegovina [In the last years, infections of domestic and wild felines with tic cats . The gengh, 1953 . Until rtic cats , 12–16. nx lynx) , 18. Troalestine , 21. In  wildcat  and, morin Italy . Even thAngiostrongylus chabaudi Biocca, 1957 , described for the first time in Italy, resides in the pulmonary arteries and the right heart of felids. Since then, it has been reported again in Italy and several other European countries [ountries –24, inclountries . Its lifountries , 27. Eurountries , 25, whiountries , 29.Eucoleus aerophilus and Dirofilaria immitis can infect the trachea and bronchi and the pulmonary arteries, respectively, of wildcats [Crenosoma vismani and Crenosoma sp. were recently reported from the bronchi and bronchioles of Eurasian lynx [In addition, wildcats , 31, whiian lynx , 33. Mixian lynx –32, 34 aian lynx .In Romania, the population of wild felids is large and stable, and many carnivore habitats are protected , 37. OveConsidering the clinical relevance and apparent emergence of pulmonary nematodes, a constant epidemiological monitoring of wild felids is important, as they could represent reservoirs for domestic felids. With this view, the aim of the present study was to evaluate the distribution and species diversity of cardio-pulmonary nematodes infecting wild felids in Romania by parasitological necropsy. The rest of the detected parasites will be included in further studies.n = 1 for lynx and n = 4 for wildcats), the Baermann method [Between 2014 and 2021, a total of 54 wild felids were legally collected from different locations in Romania and submitted to the Department of Parasitology of the University of Agricultural Sciences and Veterinary Medicine of Cluj-Napoca. Most animals included in this study were found as road kills, apart from two wildcats which died in two zoos, one from Reșița and one from Bucharest. The carcasses were kept at − 18 °C until examination. The animal species were identified based on their morphological characteristics, and wildcats were differentiated from domestic cats and hybrids based on the score obtained from the specific morphological characteristics . The appn method  was carrcytochrome c oxidase subunit I (cox1) gene, using LCO1490/HCO2198 primers [® database by means of Basic Local Alignment Search Tool (BLAST) analysis.Genomic DNA was extracted from one nematode specimen collected from the trachea of a Eurasian lynx, using a commercial kit  according to the manufacturer’s instructions. The molecular characterization of the selected nematode was performed by amplification and external sequencing (Macrogen Europe B.V.) of fragments of various sizes of the  primers  and protP values ≤ 0.05.Pulmonary lesions were not evaluated because of the decomposition state of some of the carcasses and long-term freezing. The distribution map was elaborated using ArcMap 10.6.1 software. The statistical analysis was performed using EpiInfo™ 7.2.2.6 software . The frequency, prevalence, and 95% confidence interval (CI) of infection with each species of parasite were calculated globally and according to multiple variables . The differences among groups were assessed by chi-square test and statistically significant for Lynx lynx (n = 7), and wildcats, Felis silvestris (n = 47). Among wildcats, only one carcass was identified as a hybrid, which was also included in the statistical analysis as a wildcat. Overall, 28.6% (n = 2) Eurasian lynxes and 61.7% (n = 29) wildcats were infected with cardio-pulmonary adult nematodes. Co-infections were detected in 14.3% (n = 1) of the Eurasian lynxes and 19.1% (n = 9) of the wildcats. Troglostrongylus sp. was detected in the bronchi of seven European wildcats (14.9%), including a previously published case report [T. brevior isolate from a wildcat in Germany (GenBank accession number: KP641613). The sequence is available in GenBank under the accession no. OM283595. The first-stage larvae recovered from the lung tissue of the infected Eurasian lynx were morphologically consistent with those of Troglostrongylus sp. , A. chabaudi , and D. immitis . Eucoleus aerophilus adults collected from the trachea were identified based on their specific morphology [n = 2) prevalence in Eurasian lynx and 34.0% (n = 16) in wildcats. Infections with A. abstrusus, A. chabaudi and D. immitis were not detected in Eurasian lynxes from Romania  was found for wildcats infected with D. immitis from different regions of Romania, with a higher prevalence in south and south-east Romania. Locations of the positive and negative animals and the distribution map of all pulmonary nematode species are shown in Fig. Besides sus Fig.   coinfected wildcats, representing 31.0% of all infected. A recent study on endoparasites of wildcats in Greece reported polyparasitism with up to five different species of cardio-pulmonary nematodes [A. chabaudi (56.5%), followed by A. abstrusus (43.5%) and T. brevior (34.8%). In contrast to the previously mentioned study [E. aerophilus followed by A. chabaudi and T. brevior. Interestingly, the prevalence of A. abstrusus in wildcats is lower than in domestic cats in Romania [Helminthological surveys on wild felids are rare; however, in the last few years, this topic has gained more interest among researchers , 31. Thehabaudi 5.5%, foll Romania . Diagnos Romania .D. immitis in Romania is known and has been well studied in both domestic and wild animals, including felids [D. immitis in Eurasian lynx (Lynx lynx), which could be related to the lower number of studies or examined animals, its unsuitability as a host, or the lower infection risk associated with the higher altitudes [The presence of g felids , 44–46, g felids . In accoltitudes  where lyTroglostrongylus brevior was previously detected only in one Eurasian lynx from Bosnia and Herzegovina [T. assadovi was described from the same animal host in Azerbaijan [Troglostrongylus spp. were reported more often in bobcats (Lynx rufus) and Canada lynxes (Lynx canadensis) from North America [A. abstrusus [L. lynx [Crenosoma and Angiostrongylus larvae [T. brevior was reported in wildcats in Germany [Troglostrongylus brevior often produces fatal bronchopneumonia in kittens and juvenile cats; however, clinical signs in adult felines are rare [zegovina . Howeverzegovina , 48–51. erbaijan , but no  America –55. Morebstrusus , a paras[L. lynx . More res larvae , even th Germany . Severe  Germany , 58. Troare rare . In the A. abstrusus, T. brevior, and E. aerophilus. All seem to be expanding in distribution [Overall, the significance of our results could have implications for both wildlife and companion animal medicine. As recently reviewed by Morelli et al. , three sribution . Our resA. abstrusus was confirmed in wildcats for the first time in Romania. We confirm the Eurasian lynx as a host of patent infection with T. brevior.In this large-scale survey on the cardio-pulmonary nematodes of wild felids in Romania, we detected a significant variety of cardio-pulmonary nematodes. Infection with Additional file 1: Table S1. Prevalences of cardio-respiratory nematodes in wildcats and lynxes. Table S2. Prevalences of cardio-pulmonary nematodes in wildcats by their sex. Table S3. Prevalences of cardio-pulmonary nematodes in wildcats by their age. Table S4. Prevalences of cardio-pulmonary nematodes in wildcats by their geographical location.Additional file 2: Table S5. The exact location of each examined cat, their age sex, and positivity to lungworms.


A 13-year-old girl in good health presented with a cluster of pigmented lesions on the body that had been present since childhood. Upon examination, multiple brown, evenly pigmented macules were found to be present in the right scapular region, with sharp delineation at the midline ; additioQuestion 1: What is the likely diagnosis in this patient?A.Lentigo solarisB.Partial unilateral lentiginosis (PUL)C.Nevus of ItoD.Speckled lentiginous nevusE.Zosteriform lentiginous nevusAnswers:A.Lentigo solaris – Incorrect. Lentigo solaris, also known as solar lentigo, is characterized by lentigines present on sun-exposed skin and is more commonly seen in older individuals. Histologically, there may be an increased number of melanocytes producing excess melanin, elongation of rete ridges, and solar elastosis within the dermis. This is in contrast to partial unilateral lentiginosis (PUL), which would look similar to lentigo simplex histologically; however, differences may arise in patient age, the clinical distribution of lesions, and histologic findings of solar elastosis.B.Partial unilateral lentiginosis (PUL) – Correct. PUL is a rare disorder of aberrant pigmentation characterized by multiple lentigines with a unilateral segmental pattern and may follow ≥1 dermatomal distributions. These lesions typically present at birth or during childhood, becoming darker and more widespread with age.C.Nevus of Ito – Incorrect. Nevus of Ito is a pigmented patch that is blue to gray-brown in color and is unilaterally located on the neck and shoulder. These nevi are typically present at birth and may mimic Mongolian spots.D.Speckled lentiginous nevus – Incorrect. Speckled lentiginous nevus, also known as nevus spilus, appears as small, dark macules superimposed on a lighter pigmented patch. PUL may commonly be misdiagnosed as nevus spilus; however, PUL appears as brown macules on normal skin, without an associated background of lighter pigmentation.E.Zosteriform lentiginous nevus – Incorrect. Zosteriform lentiginous nevus is from the family of nevus spilus; however, these lesions have more of a dermatomal arrangement.Question 2: All of the following syndromes are strongly associated with multiple lentigines except which of the following?A.NF1B.Sturge-Weber syndromeC.LEOPARD syndromeD.Carney complexE.Peutz-Jeghers syndromeAnswers:A.NF1 – Incorrect. NF1 is associated with axillary freckling. Many authors have suggested that PUL is a form of mosaic NF because of its association with axillary freckling, ipsilateral Lisch nodules, café-au-lait macules, and skeletal abnormalities.B.Sturge-Weber syndrome – Correct. Sturge-Weber syndrome is a vascular disorder that is not associated with lentigines. Although this syndrome may have ocular or central nervous system findings, its characteristic cutaneous finding is a unilateral vascular malformation known as a port-wine stain.C.LEOPARD syndrome – Incorrect. LEOPARD syndrome, also known as multiple lentigines syndrome, describes the association of lentigines, electrocardiographic abnormalities, ocular hypertelorism, pulmonary stenosis, abnormalities of the genitalia, mental retardation, and sensorineural deafness.D.Carney complex – Incorrect. Carney complex, also known as nevi, atrial myxoma, myxoid neurofibroma, ephelides syndrome and lentigines, atrial myxoma, mucocutaneous myxoma, blue nevi syndrome, is characterized by the presence of mucocutaneous lesions, cardiac myxomas, and endocrine and nonendocrine tumors.E.Peutz-Jeghers syndrome – Incorrect. Peutz-Jeghers syndrome is an autosomal dominant syndrome with a hallmark finding of multiple, pigmented macules on the lips that present during childhood.Question 3: What additional workup should be completed for this patient?A.Ophthalmologic examinationB.Infectious disease consultationC.Autoimmune workupD.Punch biopsyE.Oncology consultationAnswers:A.Ophthalmologic examination – Correct. PUL has been shown to be associated with a variety of ophthalmologic findings, including ipsilateral Lisch nodules, which are found in patients with NF1.B.Infectious disease consultation – Incorrect. There have been no reports of infection associated with PUL.C.Autoimmune workup – Incorrect. There have been no reports of autoimmune disease associated with PUL.D.Punch biopsy – Incorrect. A punch biopsy would be nondiagnostic of PUL because this may resemble lentigo simplex histologically.E.Oncology consultation – There have been no reports of malignant transformation of a lentiginous lesion in patients with PUL.None disclosed.


The Uganda National Drug Authority requires phytochemical screening, freedom from microbial contamination, and evidence of safety and efficacy of the constituent plants to register herbal products. Since Uganda has no pharmacopeia, safety, efficacy, and plant processing information are not readily available. We documented the plant materials used to manufacture products in Uganda and established evidence of their safety and efficacy and availability of monographs.  The NDA register of herbal products was reviewed, and a product list was extracted. The herbal products were purchased from local pharmacies, and their labels were studied to identify plant ingredients and drug use. Literature was reviewed to document evidence of the safety and efficacy of the plant materials concerning manufacturer's claims. Also, the WHO and available African Pharmacopeia were searched to establish the availability of the plant monographs. Eucalyptus globulus Labill, being the commonest. Several in vitro and in vivo studies demonstrate efficacy, thus supporting the use of the selected plant species for empirical treatment as stated on the product label. While most plants were safe, some species such as Albizia coriaria Oliv. had dose-dependent toxicities that cannot be predicted in combinations. The WHO, African Pharmacopoeia, and West African Herbal Pharmacopoeia had only 16 plant monographs of the 33 plants of interest. Nevertheless, Aloe vera (L.) Burm.f., Azadirachta indica A.Juss., Zingiber officinale Roscoe, and Allium sativum L. monographs were published by all three pharmacopoeias.  Of the 84 NDA-registered local products, only 18 were obtained from the market; 82% were indicated for respiratory tract disorders. Thirty-three plant materials were listed with  Preclinical evidence of safety and efficacy exists in the literature for most of the plants used to manufacture registered herbal products in Uganda. More specific bioassays and clinical trials are required for the products to provide conclusive evidence of safety and toxicity. Monographs are urgently needed for the Ugandan plants. Traditional herbal medicines have been defined as naturally occurring, plant-derived substances with minimal or no industrial processing. Such substances have been used to treat illness within local or regional healing practices . More thTraditionally, medicinal plant products have been prescribed by traditional healers within communities or self-prescribed. In this regard, assessment of efficacy and safety was based on the history of use and informal knowledge passed on from generation to generation. However, with commercialization, there is a need for scientific proof that the products are effective and safe. Various studies have demonstrated deficiencies in the quality of herbal medicines; from adulteration with conventional medicines to substitution with ineffective or harmful plant materials and contamination with soil, microorganisms, pesticides, aflatoxins, etc. .Consequently, the developed world's regulatory authorities have established policies to regulate herbal and other traditional medicines. The US FDA and the EMA, for instance, have published strict requirements for market authorization of traditional methods; as such, the manufacturer has to demonstrate the efficacy and safety of the product through clinical trials, a quality assurance plan for cultivation and harvesting of the plant materials as well as protocols for good manufacturing practices , 5.In contrast, traditional medicines are hardly regulated in developing countries. In a study to evaluate the regulatory framework of traditional medicines, over 60% of developing countries did not have any framework in place , guideliMany African countries still have weak regulatory frameworks for traditional medicines despite these guidelines. In Uganda, for instance, there are no herbal pharmacopoeial methods for assessing the quality of herbal medicinal products. Consequently, all the locally manufactured herbal products authorized for marketing are not fully registered but rather “notified” . This isUnlike in Western medicines in which pure bioactive compounds are used in pharmaceutical products, traditional medicine utilizes crude forms of whole plants or plant parts. The various plants and herbal products utilized in traditional/herbal medicine differ due to differences in climate and culture , 13. ConThis study aimed to document the plant raw materials used in Uganda for the manufacture of herbal products and establish a database of safety and efficacy information. This information can be added to future herbal pharmacopeia/monographs for routine use in quality assurance and quality assessment by manufacturers and regulators, respectively.This was a cross-sectional study of local herbal products registered and sold in Uganda in 2019 and a review of published literature on the safety and efficacy of the herbal raw materials between 1960 and 2021. The products were bought from the busy cities of Mbarara, Jinja, Wakiso, and Kampala. In addition, most of the manufacturers are located in this area. The pharmacies were accessed conveniently, and the only important factor was the presence of the products of interest.Data were collected mainly by desk review. Drug registers, product labels, and journal articles were considered. The data were collected as mentioned in the following sections.https://www.nda.or.ug/drug-register-downloads/). Then, a list of the notified  local herbal medicinal products was compiled. A sample of each of the products on the notified list was purchased from local pharmacies.The most current drug register (August 2019) of herbal medicinal products was obtained from the NDA website  and the active ingredients . A database of all the herbal plant materials was then developed, and the materials were ranked according to their popularity (frequency of appearance) in the herbal medicinal products.We conducted a literature review to obtain evidence of biological activity concerning the current traditional use (according to the manufacturers' labels) and safety of all the plant species used. We searched databases and search engines such as PubMed, Google Scholar, ResearchGate, Web of Science, and Scopus. The search terms consisted of the scientific plant names, pharmacological/therapeutic activity of interest, and toxicity. Only full-length articles published in English were reviewed.Since there are no Ugandan monographs or pharmacopeia, we were interested in finding out if monographs of the plants used in Uganda have been published in other African countries. The uses and composition of the plants were considered to be similar due to their similarity in culture and climate. We reviewed the African Pharmacopoeia 2014  and the st March 2019, the NDA drug register had 4,148 and 214 conventional products and herbal medicinal products, respectively. All the herbal products are not fully registered but rather notified, and the majority (130) was imported, mainly from India and China. 84 locally manufactured herbal products were notified. Of these, 82% (n = 69) were syrups or solutions indicated for systemic treatment of upper respiratory tract disorders, such as cough, asthma, and flu. For skin infections and wound healing, the others were ointments, creams, balms, or soaps. Three products were indicated for the treatment of peptic ulcers  of the products, followed by Aloe barbadensis, Albizia coriaria, Mangifera indica, and Warburgia ugandensis tied in the third place  of the surveyed locally produced herbal products were for the management of diseases of the respiratory system, with label claims such as dry, wet, and whooping cough, sore throat, bronchial asthma, nasal congestion, flu, common cold, sinusitis, and allergies. Digestive system conditions such as gastric ulcers, flatulence, and constipation are also frequently targeted. Others include immune boosters, mouth wash, antioxidants, wounds, allergies, and a lip balm. Nine of the most frequently used plant materials were reviewed in detail, for evidence of safety and efficacy in managing the conditions on the label claim. These were A. coriaria, M. indica, A. indica, S. aromaticum, A. barbadensis, W. ugandensis, Z. officinale, A. sativum, and E. globulus. The information for the rest of the plant materials is summarized in Half  and testimony by physicians, traditional practitioners, and patients after a long duration of use. Appropriate clinical trials should be conducted for cases where no history of use or biological information has been documented. The WHO has published guidelines for the clinical study of traditional medicines in the WHO African region .To ensure reproducibility of efficacy, the preparation of medicines should be standardized so that each batch contains a defined amount of the active ingredients.Information on safety and efficacy can be obtained from scientific literature, national or international pharmacopeia, and WHO monographs for selected plant materials.Albizia coriaria is used in many local herbal preparations to manage upper respiratory tract infection (URTI) and other infections in Uganda [A. coriaria include Gabogola syrup®, Kwesiima cough mixture®, Muwereza herbal cough remedy®, and Phycof cough syrup®. Upper respiratory tract infections are typically caused by the rhinovirus, adenovirus, influenza, enterovirus, and respiratory syncytial virus [Streptococcus pyogenes. URTI causes a variety of diseases with overlapping presentations, including acute bronchitis, influenza, rhinitis, pharyngitis, tonsillitis, laryngitis, and other respiratory distress syndromes. Symptoms of URTI commonly include cough, sore throat, runny nose, nasal congestion, headache, low-grade fever, facial pressure, sneezing, malaise, and myalgias. Most viral infections are self-limiting [n Uganda , 80. Proal virus . Bacterilimiting . The goalimiting . TherefoA. coriaria had activity against Pseudomonas aeruginosa and Escherichia coli [A. coriaria against Gram-positive cocci, which are the causative agents of URTI. Lupeol has anti-inflammatory activity [Bioactive compounds such as lupeol, lupenone, betulinic acid, and catechin extracted using ethyl acetate from the stem bark of hia coli . These bactivity . Treatmeactivity . The antactivity , but nonactivity .A. coriaria in very small quantities in polyherbal preparations and boil it for several hours because of its known toxicity. Not surprisingly, the DMSO and ethanol extracts of A. coriaria were highly cytotoxic to the human glioblastoma U87.CD4.CXCR4 cells (CC50 = 6.4 and <4 μg/ml respectively) [Albizia spp. has a toxic compound, 4-methoxypyridoxine, which antagonizes vitamin B6 [Herbalists use ctively) . Albiziatamin B6 .Albizia species have shown different toxicities, including several central nervous system disturbances and other effects on the cardiovascular and respiratory systems. For instance, seed methanolic extracts of Albizia greveana and Albizia bernieri are toxic to mice with LD50 of 1.13–2.3 and 52 mg/kg body weight, respectively, the former being more than one toxic [A. coriaria have been shown to have very low cytotoxicity (CC50 >500 μg/ml) against human embryonic lung fibroblast (HELF) cells [A. coriaria were not toxic to the human keratinocyte cell line (HaCaT) IC50 > 512 [Other ne toxic . HoweverF) cells . In cont50 > 512 .A. coriaria in the symptomatic management of respiratory tract conditions. However, specific studies need to be done to assess URTI activity against the causative organisms. Specific toxicity studies on the bark of A. coriaria need to be done to conclude on the safety of the plant.There is scientific evidence for the usefulness of Mangifera indica is also used in many preparations for URTI management from the survey, including Gabogola syrup®, Kwesiima cough mixture®, and Yeco cough doctor®. M. indica is widely used to manage various infections and as an immune booster in Uganda [M. indica [M. indica are important. Extracts from the fruit pulp of M. indica possess antiviral activity against influenza virus H9N2 and chicken embryo fibroblast (CEFs) with low cytotoxicity [M. indica, inhibited herpes simplex virus 2 (HSV-2) replication but did not directly inactivate the virus [M. indica have some bacteriostatic activity against S. aureus albeit lower than the activity of the positive control used, ampicillin [M. indica varieties  from Tamil Nadu (India) against Staphylococcus aureus and Pseudomonas aeruginosa. Acetone, methanol, and water extracts of the leaves on M. indica exhibited lower antibacterial activity against S. aureus, S. pyogenes, S. pneumoniae, Bacillus cereus, E. coli, P. aeruginosa, Proteus mirabilis, Salmonella typhi, and Shigella flexneri using the agar well (cup plate) diffusion method than gentamicin and erythromycin [n Uganda . Various. indica . Howevertoxicity . Mangifehe virus . Extractpicillin . Alok etpicillin  demonstrhromycin .M. indica leaf extract containing 60% mangiferin revealed a no-observed-adverse-effect level in male and female rats at 2000 mg/kg body weight/day. Additionally, no evidence of genotoxicity was found in a bacterial reverse mutation test [In vivo acute toxicological studies of the stem bark aqueous extract of M. indica in rats and mice showed no lethality at the limit dose of 2,000 mg/kg body weight and no adverse effects when given by oral or dermal administration. Deaths occurred when the extract was administered intraperitoneally at 200 mg/kg in mice [M. indica extracts were nonirritating on rabbits' skin and ocular and rectal mucosa [A toxicological evaluation of ion test . In vivo in mice . M. indil mucosa .M. indica. Overall, the antiviral and antibacterial activities of various parts of the plant support its use in URTI management.There were no specific studies found evaluating the expectorant and decongestant activities of Azadirachta indica (neem tree) is used in the following products: Muwereza herbal cough remedy®; Princess aloe lip balm®, a lip balm®; and Rezalin for ulcer®, a syrup for management of gastric ulcers, flatulence, and constipation. To support its use in a cough remedy, A. indica should possess expectorant, decongestant, antiviral, and/or antibacterial activities [tivities .A. indica significantly blocked HSV-1 entry into cells at concentrations ranging from 50 to 100 μg/ml [A. indica is virucidal against Coxsackie virus B-4 and interfered with an early event in the viral replication [The bark extract of lication . Thus, tA. indica showed a concentration-dependent antibacterial activity against bacteria isolated from an adult mouth [Streptococcus mutans [The bark, leaf, seed, and fruit extracts of lt mouth . The eths mutans .Helicobacter pylori, the causative agent of gastric ulcers [H. pylori strains ranged between 25 and 51 μg/ml and 43–68 μg/ml, respectively. The bactericidal activity was time- and concentration-dependent [The methanol fraction of neem oil possesses bactericidal activity against 9 strains of c ulcers . The extependent . This evA. indica promoted wound healing in mice by increasing the rate of wound contraction and levels of hydroxyproline, DNA and protein content, and nitric oxide compared with the vehicle control group. These effects promote wound healing through increased inflammatory response and neovascularization [The wound-healing effect of neem could be beneficial in the lip balm. The water extract of the stem bark of rization .In vivo evaluation of acute toxicity of water extracts of A. indica leaves and seeds in rats revealed an LD50 of 6.2 and 9.4 ml/kg, respectively [Neem oil poisoning is rare in adults. However, one such case was reported in an adult male who presented with vomiting, seizures, metabolic acidosis, and toxic encephalopathy . The patectively .A. indica in URTI management, gastric ulcers associated with H. pylori infection, and wound healing. The use of the oil has the potential to cause adverse events.There is evidence to support the use of S. aromaticum (clove) was found in Kwesiima cough mixture® (a preparation for management of URTI) and Nana herbal mouth wash® . The antibacterial, anti-inflammatory, analgesic, anesthetic, and antiviral activities of S. aromaticum are important for these applications.S. pyogenes and S. mutans are the most common causes of bacterial URTI and tooth decay, respectively [ectively . The essectively . This suectively .S. aromaticum has potent inhibitory activity against both dental caries pathogens with MIC of 0.2 mg/ml. In addition, S. aromaticum bud essential oil was active against S. mutans at 0.05 mg/ml [S. aureus, Pseudomonas aeruginosa, E. coli, S. pyogenes, Corynebacterium species, Salmonella species, and Bacteroides fragilis [The methanolic extract of 05 mg/ml . Furtherfragilis .in vitro study demonstrated cytotoxic properties of clove oil and eugenol toward human fibroblasts and endothelial cells. Clove oil was highly cytotoxic at concentrations as low as 0.03% (v/v) with up to 73% of this effect attributable to eugenol [An  eugenol . Moreove eugenol . However eugenol  evaluateS. aromaticum provide evidence for its value in managing URTI and dental conditions.The antibacterial, anti-inflammatory, analgesic, and anesthetic activities of Aloe barbadensis is a constituent of Nana herbal mouth wash®, Damaleo vera tablets® , Muwereza herbal cough remedy®, and Princess aloe lip balm®.A. barbadensis gel prevent suppressing the induction of T cell-mediated responses such as contact and delayed-type hypersensitivity caused by cutaneous exposure to ultraviolet light [A wound dressing gel containing acemannan, extracted from the internal leaf aloe gel, reduced radiation-induced skin reactions in mice when applied daily for at least 2 weeks, beginning immediately after irradiation . Furtheret light . This efA barbadensis extract stimulated macrophage production of monokines, including interleukin-1 and tumor necrosis factor, resulting in the initiation of immune attack, necrosis, and regression of implanted sarcomas in mice [A. barbadensis stimulated cellular and humoral immune responses in rabbits following immunization against myxomatosis [A. barbadensis extract to boost the immune system.Acemannan from  in mice . In anotomatosis . Furtheromatosis . These sEmodin, an anthraquinone prepared from aloin, was shown to inactivate herpes simplex virus type 1 and type 2, varicella-zoster virus, pseudorabies virus, and influenza virus, not adenovirus and rhinovirus. Electron microscopic examination of the anthraquinone-treated herpes simplex virus demonstrated that the envelopes were partially disrupted . This anA. barbadensis in Damaleo vera tablets for cleansing the GIT.Aloe emodin is also a known stimulant laxative , and thiA. barbadensis on mild damage caused by type-II diabetes on kidney tissue and support its use in Damaleo vera tablets.The generative changes in the kidney tissue were significantly reduced in diabetic rats treated with glibenclamide and aloe leaf gel and pulp extracts compared with the untreated group . The treA. barbadensis in humans. Some include massive intraoperative bleeding after consumption of A. barbadensis tablets [A. barbadensis. In another study, rats fed aloe whole-leaf powder for 90 days showed the adverse effect level to be 2 g/kg BW [50 = 1 g/l), even at concentrations below the recommended dose for human consumption [Several single case reports have been published on the toxicity of  tablets , acute r tablets , severe  tablets , acute h tablets , and Hen tablets . Ernst [ tablets  reported g/kg BW . Aloe insumption . Processsumption .Literature supports the use of aloe extracts for cosmetic purposes, boosting the immune system, and management of URTI and constipation. However, studies also show that aloe extracts can be toxic, although processing can reduce such toxicities.W. ugandensis is a constituent of herbal preparations including Lucas syrup®, Sacco syrup®, Focus herbal cough syrup® used in the management of URTI, and Jenacid negus® for the management of gastric ulcers.W. ugandensis elicited antimicrobial activity against E. coli and S. aureus in the agar well assay, with higher activity against S. aureus than E. coli. This activity was only detectable at high concentrations of the extract, i.e., up to 50 mg/ml of crude extract [W. ugandensis leaves exhibited antimicrobial activity against S. boydii and S. aureus [W. ugandensis was the main antibacterial agent (IC50 = 12.5 μg/ml) against both S. aureus and Bacillus subtilis [The water extracts of  extract . In anot. aureus . The bio. aureus , 135. Thsubtilis .W. ugandensis in the management of viral URTI. Its usefulness in the management of peptic ulcer disease could also not be established from the literature review. Other reported pharmacological activities of W. ugandensis backed by some scientific evidence include antifungal activity, antimycobacterial activity, anti-inflammatory and antioxidant properties, molluscicidal, antifeedant, antiplasmodial, cytotoxic, anthelmintic, and antileishmanial activities [W. ugandensis has anti-inflammatory effects and inhibited both COX-2 and COX-1 in vitro with IC50 values of 13.33 ± 4.36 and 11.05 ± 1.43 μg/ml, respectively [Specific antiviral activity studies could not be found to assess the importance of tivities . The dieectively .W. ugandensis is widely used by herbalists in Uganda for treating various infections, it is also known to be toxic if used in high doses [W. ugandensis in mice gave an LD50 >5000 mg/kg body weight with no mortalities recorded for all dose levels used. The extract was classified as noncytotoxic with 50% cytotoxic concentration (CC50) determined to be >250 μg/ml [W. ugandensis was cytotoxic to intestinal epithelial cells IEC-6, with IC50 values <50 μg/ml [50 = 1.5 μg/ml) and the ethanol (CC50 = 7.6 μg/ml) root were highly cytotoxic to U87CD4CXCR4 cells [While gh doses . The acu50 μg/ml . W. ugan50 μg/ml . Both thR4 cells .W. ugandensis in the management of diseases of the respiratory tract or gastric ulcers. It can be toxic when used in high doses.The evidence in the literature is not enough to draw conclusions on the usefulness of Zingiber officinale (ginger) is locally used as a constituent of syrups for the management of URTI including Yeco cough doctor® and Muwereza herbal cough remedy®.Z. officinale contains various phytochemicals including phenolic compounds, terpenes, polysaccharides, lipids, organic acids, and raw fibers. The pharmacological activities of ginger have however been attributed to phenolic compounds such as gingerols and shogaols [α, IL-6, and IL-1β) and increased the anti-inflammatory cytokines (IL-10 and IL-22) in mouse colitis models [Z. officinale attenuated post-exercise-induced elevation in several proinflammatory cytokines including interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in well-trained male endurance runners [shogaols . Variouss models . Z. offi runners .Z. officinale in herbal cough remedies for their cough-suppressant activity.Intravenous administration of 6-gingerol (at 1.75–3.5 mg/kg) and 6-shogaol (at 1.75–3.5 mg/kg) and oral administration of both (at 70–140 mg/kg) produced an inhibition of spontaneous motor activity, as well as antipyretic and analgesic effects, the latter producing more intense effects than the former. 6-Shogaol also showed an intense antitussive effect in comparison with dihydrocodeine phosphate . This suZ. officinale rhizome was active against common respiratory tract pathogens: S. aureus, S. pyogenes, S. pneumoniae, and Haemophilus influenzae [Z. officinale inhibited biofilm formation and affected the membrane integrity of multidrug-resistant P. aeruginosa, both of which play a role in the mechanism of infection and antimicrobial resistance [S. mutans [The ethanolic extract of fluenzae . Furthersistance . The cru. mutans .Z. officinale dose-dependently inhibited human respiratory syncytial virus (HRSV) and induced plaque formation in human respiratory tract cell lines [β (IFN-β) that possibly contributed to counteracting viral infection. Other studies have demonstrated antiviral activity of Z. officinale in hepatitis C virus (HCV) [The antiviral activity of ginger has been demonstrated in some viruses not specifically implicated in the causation of common URTI. Fresh ll lines . Fresh gus (HCV)  and Chikus (HCV) .Z. officinale rhizome extracts showed no treatment-related signs of toxicity or mortality in any animals at tested doses [Studies on the safety of different ed doses –154. Oneed doses .Z. officinale extracts in preparations for the management of upper respiratory tract diseases. The rhizome has been established to be safe.There is scientific evidence in support of the use of Allium sativum (garlic) is used in Jenacid negus® for the management of peptic ulcer disease and in Yeco cough doctor® for the management of URTI.From the survey, Helicobacter pylori. The MIC90 of the extracts was 5 mg/ml, and the MBC was equal to, or twofold higher than, the MIC. Boiling the garlic extract lowered the efficacy twofold to fourfold the values of MIC and the MBC obtained with fresh extract [μg/ml of thiosulfinate against H. pylori [An aqueous garlic extract was bactericidal against 16 clinical isolates and three reference strains of  extract . Another. pylori .A. sativum juice at 250 and 500 mg/kg in rats produced a gastric ulcer healing effect in acetic acid-induced chronic gastric ulcer. The extracts also produced a gastric antisecretory effect in pylorus-ligated rats; gastric cytoprotective effect in ethanol-induced and indomethacin-induced gastric ulcer; and a significant reduction in stress-induced gastric ulcers and cysteamine-induced duodenal ulcers with higher effectiveness at 250 mg/kg [Orally administered 50 mg/kg . Further50 mg/kg .S. pyogenes, which is commonly implicated in bacterial URTI, with an MBC of 1 g/ml [An ethanolic garlic extract was bactericidal against f 1 g/ml . The antf 1 g/ml ; influenf 1 g/ml , 162; huf 1 g/ml ; and meaf 1 g/ml . Garlic f 1 g/ml . These aThe most commonly reported adverse effects of garlic in randomized clinical trials are garlic breath and gastrointestinal symptoms . AllergiA. sativum in the management of upper respiratory tract conditions and in peptic ulcer disease. The plant can be used with caution as cases of allergic reactions have been reported.Scientific evidence supports the use of Eucalyptus globulus is a constituent of Nana herbal mouth wash®, Princess pain balm®, Phycof cough syrup®, Focus herbal cough syrup®, Lucas syrup®, Kwesiima cough mixture®, Gabogola syrup®, and Sacco syrup®.E. globulus with other oils was effective against fourteen Gram-positive and Gram-negative bacteria strains tested, including some antibiotic-resistant strains [S. pyogenes, S. pneumoniae, Listeria monocytogenes, and S. aureus. MICs ranged from 0.01% to 3%. The study also found the blend effective against H1N1 influenza virus, making it suitable for management of influenza and post-influenza bacterial pneumonia infections [E. globulus has also been demonstrated in the organisms causing dental caries especially S. mutans [Lactobacilli [An essential oil blend of  strains . The orgfections . The ant. mutans  and Lactobacilli , 174.E. globulus exhibited analgesic and anti-inflammatory effects in mice and rat models [E. globulus possesses central and peripheral analgesic effects as well as neutrophil-dependent and neutrophil-independent anti-inflammatory activities [The essential oil of t models . The esstivities .50 of 3811.5 mg/kg. The oil slowed growth of the male rats and damaged the liver and the kidneys at 792 and 1188 mg/kg BW after oral administration, respectively [50 of 1650 mg/kg in mice was determined. In addition, there was no abnormal skin reaction observed when a 5% ointment formulation was administered on the rats' skin, indicating the safety of E. globulus essential oil at a relatively lower concentration [A eucalyptus oil water emulsion had an LDectively . In anotntration .E. globulus is safe for use, at low concentrations, for the management of URTI and dental conditions.Aloe vera (L.) Burm.f. (Asphodelaceae), Azadirachta indica A.Juss. (Meliaceae), Zingiber officinalis Roscoe (Zingiberaceae), and Allium sativum L. (Amaryllidaceae) had monographs published by all three authorities  had monographs published by the WHO, ten (10) by the African Pharmacopoeia, and six (6) by the West African Pharmacopoeia. Nonetheless, horities . The folin vitro and in vivo studies have been carried out to establish the safety and efficacy of the plants used for the management of different conditions. For all plant materials, there is some scientific evidence to support the use of the plants in the management of the conditions on the label claim. However, some of these studies lack specificity for the conditions on the label claim and therefore may not be confidently relied upon to support the use of the plant material. More specific bioassays are required in these cases to provide conclusive evidence. Also, studies in humans are few, and therefore, in most cases, results from animal models and human cell lines are extrapolated to humans. Some plant materials have specific dose-dependent toxicities and interactions that should concern the manufacturers and the regulatory authorities before approval of the products. Other plants' toxicity depends on the preparation method, yet there are no GMP requirements for herbal manufacture in Uganda. Pharmacovigilance efforts should be stepped up to identify toxicity cases and establish efficacy of the finished products. Plant monographs are urgently required to standardize uses, processing, and quality assurance of plant materials.Several