Toxin Type Research Articles

Design, performance, processing, and validation of a pooled CRISPR perturbation screen for bacterial toxins.

Unbiased forward genetic screens have been extensively employed in biological research to elucidate functional genomics. In pooled clustered regularly interspaced short palindromic repeats (CRISPR) perturbation screens, various genetically encoded gain-of-function or loss-of-function mutations are introduced into a heterogeneous population of cells. Subsequently, these cells are screened for phenotypes, perturbation-associated genotypes are analyzed and a connection between genotype and phenotype is determined. CRISPR screening techniques enable the investigation of important biological questions, such as how bacterial toxins kill cells and cause disease. However, the broad spectrum of effects caused by diverse toxins presents a challenge when selecting appropriate screening strategies. Here, we provide a step-by-step protocol for a genome-wide pooled CRISPR perturbation screen to study bacterial toxins. We describe technical considerations, pilot experiments, library construction, screen execution, result analysis and validation of the top enriched hits. These screens are applicable for many different types of toxins and are anticipated to reveal a repertoire of host factors crucial in the intoxication pathway, such as receptors, trafficking/translocation factors and substrates. The entire protocol takes 21-27 weeks and does not require specialized knowledge beyond basic biology.

Modeling of Bacillus cereus evolution in pasteurized cow milk locally distributed at point of sale: a case of three companies in Tlemcen region, Algeria

Bacillus cereus is among the most hazardous bacteria that could reduce the shelf life of pasteurized milk and could threaten consumers’ health. These bacteria groups are known for their heat resistance and their ability to produce two types of toxins, diarrheic and emetic. This study aimed to enumerate the B. cereus in raw cow’s milk collected from three companies in the Tlemcen region, then modeled, based on a probabilistic approach, their evolution throughout the process, split into six steps, until the point of sale. At first, the analysis of 242 samples on MYP medium, shows a high prevalence between 92% and 98% of B. cereus with variable concentrations among the three companies. Throughout the storage of milk at refrigeration temperatures (2°C, 4°C and 5°C), no growth of B. cereus was estimated. Otherwise, the pasteurization process reduces the contamination level depending on pasteurization temperature as for company A, the mean concentration decreases from 1.53 log CFU/mL to 1.07 log CFU/mL. In this study, the distribution step at ambient temperature changes significantly the B. cereus concentration to reach a critical concentration during July and August. In conclusion, the quality of raw cow milk must be controlled before reception on the farm and the cold chain must be maintained throughout all process points until consumption.

Antimicrobial and Phylogenomic Characterization of Bacillus cereus Group Strains Isolated from Different Food Sources in Italy.

Background:Bacillus cereus is a widespread environmental Gram-positive bacterium which is especially common in soil and dust. It produces two types of toxins that cause vomiting and diarrhea. At present, foodborne outbreaks due to Bacillus cereus group bacteria (especially Bacillus cereus sensu stricto) are rising, representing a serious problem in the agri-food supply chain. Methods: In this work, we analyzed 118 strains belonging to the Bacillus cereus group, isolated from several food sources, for which in vitro and in silico antibiotic resistance assessments were performed. Results: Many strains showed intermediate susceptibility to clindamycin, erythromycin, and tetracycline, suggesting an evolving acquisition of resistance against these antibiotics. Moreover, one strain showed intermediate resistance to meropenem, an antibiotic currently used to treat infections caused by Bacillus cereus. In addition to the phenotypic antimicrobial resistance profile, all strains were screened for the presence/absence of antimicrobial genes via whole-genome sequencing. There was inconsistency between the in vitro and in silico analyses, such as in the case of vancomycin, for which different isolates harbored resistance genes but, phenotypically, the same strains were sensitive. Conclusions: This would suggest that antibiotic resistance is a complex phenomenon due to a variety of genetic, epigenetic, and biochemical mechanisms.

Emergence of genetic diversity and multi-drug resistant Clostridium perfringens from wild birds

BackgroundClostridium perfringens (C. perfringens) is an important zoonotic microorganism that can cause animal and human infections, however information about the prevalence status in wild birds of this pathogenic bacterium is currently limited.ResultIn this study, 57 strains of C. perfringens were isolated from 328 fecal samples of wild birds. All the isolates were identified as type A and 70.18% of the isolates carried the cpb2 gene. Antimicrobial susceptibility testing showed that and 22.80% of the isolates were classified as multidrug-resistant strains. The MLST analysis of the 57 isolates from wild birds was categorized into 55 different sequence types (STs) and clustered into eight clonal complexes (CCs) with an average of 20.1 alleles and the Simpson Diversity index (Ds) of 0.9812, and revealed a high level of genetic diversity within the C. perfringens populations. Interestingly, the isolates from swan goose were clustered in the same CC while isolates from other bird species were more scattered suggesting that a potential difference in genetic diversity among the C. perfringens populations associated with different bird species.ConclusionC. perfringens exhibits a wide range of host adaptations, varying degrees of antimicrobial resistance, and a high degree of genetic diversity in wild birds. Understanding the prevalence, toxin type, antimicrobial resistance, and genetic diversity of C. perfringens in wildlife populations is essential for developing effective strategies for disease control and management.

Estimating the risks of exposure to harmful algal toxins among Scottish harbour seals

Harmful algal bloom (HAB) toxins consumed by marine predators through fish prey can be lethal but studies on the resulting population consequences are lacking. Over the past approximately 20 years there have been large regional declines in some harbour seal populations around Scotland. Analyses of excreta (faeces and urine from live and dead seals and faecal samples from seal haulout sites) suggest widespread exposure to toxins through the ingestion of contaminated prey. A risk assessment model, incorporating concentrations of the two major HAB toxins found in seal prey around Scotland (domoic acid (DA), and saxitoxins (STX)), the seasonal persistence of the toxins in the fish and the foraging patterns of harbour seals were used to estimate the proportion of adults and juveniles likely to have ingested doses above various estimated toxicity thresholds. The results were highly dependent on toxin type, persistence, and foraging regime as well as age class, all of which affected the proportion of exposed animals exceeding toxicity thresholds. In this preliminary model STX exposure was unlikely to result in mortalities. Modelled DA exposure resulted in doses above an estimated lethal threshold of 1900 µg/kg body mass affecting up to 3.8 % of exposed juveniles and 5.3 % of exposed adults. Given the uncertainty in the model parameters and the limitations of the data these conclusions should be treated with caution, but they indicate that DA remains a potential factor involved in the regional declines of harbour seals. Similar risks may be experienced by other top predators, including small cetaceans and seabirds that feed on similar prey in Scottish waters.

Epidemiology, clinical characteristics, and associated cost of acute poisoning: a retrospective study.

Poisoning is a major public health issue and a leading cause of admission to the emergency department (ED). There is a paucity of data describing the epidemiology and cost of acute poisoning. Therefore, this study investigated the epidemiology, patterns, and associated costs of acute poisoning in emergency department of the largest tertiary care healthcare centre in Qatar. This study was a retrospective review of the health records of patients admitted to the ED due to poisoning between January 2015 and December 2019. Incidence, clinical characteristics, and costs associated with acute poisoning were assessed. Frequency and percentages were calculated for categorical variables and mean and SD for continuous variables. The relationship between sociodemographic characteristics and poisoning profile was assessed using the chi-square test. A micro-costing approach using the cost of each resource was applied for cost calculations. The incidence of acute poisoning was 178 cases per 100,000 patients. Females (56%) and children below 14 years (44.3%) accounted for the largest proportion. Most of the exposures were accidental involving therapeutic agents (64.2%). The mean length of hospital stay was found to be 1.84 ± 0.81 days, and most patients (76.6%) were discharged within the first 8 h. A statistically significant difference was found between age groups and type of toxin (χ2 = 23.3, p < 0.001), cause and route of exposure (χ2 = 42.2, p < 0.001), and length of hospital stay (χ2 = 113.16, p < 0.001). Admission to intensive care units had the highest cost expenditure (USD 326,008), while general wards accounted for the least (USD 57,709). Unintentional poisoning by pharmacological agents is common in infants and children. This study will assist in the development of educational and preventive programmes to minimise exposure to toxic agents. Further studies are required to explore the impact of medical toxicology services, and post discharge monitoring of poisoning.

Multiplex Detection of Seven Staphylococcal Enterotoxins Using Liquid Chromatography–Mass Spectrometry Combined with a Novel Capture Molecule

Food poisoning caused by Staphylococcal enterotoxins (SEs) is prevalent globally, making efficient detection of these toxins very important. Traditionally, liquid chromatography–mass spectrometry required immunosorbent enrichment by magnetic bead-coupled antibodies obtained by animal-specific immunization. However, this method is time-consuming and costly. In this study, two recombinant protein capture molecules were designed based on the principle of toxins binding to Major Histocompatibility Complex (MHCII) and T cell receptor (TCR) molecules. The two capture molecules are called MHCII and MHCII-D10. The design of the MHCII and TCR-D10 was achieved through searching for the binding site protein sequence of Staphylococcal enterotoxins in the relevant literature, and MHCII-D10 was to link MHCII sequence with TCR-D10 sequence using linker (G4S)3 linking peptide. These capture molecules were shown to effectively bind to seven types of toxins and to capture SEs in various matrices. The digestion time, ratio, and temperature were further optimized, reducing the overall digestion time to just 2 h. The specificity, linearity, sensitivity, precision (RSD%), and recovery of the two methods were verified by liquid chromatography–mass spectrometry. When the MHCII and MHCII-D10 captured the toxins, the limit of quantification (LOD) in the 1 × PBS, plasma, and milk matrices ranged from 1.5625 to 100 fmol/µL, with the recovery rate ranging from 18.4% to 96%. The design of these capture molecules eliminates the need for animal-specific immunization, simplifying the pre-detection process and avoiding ethical concerns. This development holds significant promise for clinical diagnosis and reference.

Cyanobacteria and Macroinvertebrate Relationships in Freshwater Benthic Communities beyond Cytotoxicity.

Cyanobacteria are harmful algae that are monitored worldwide to prevent the effects of the toxins that they can produce. Most research efforts have focused on direct or indirect effects on human populations, with a view to gain easy accurate detection and quantification methods, mainly in planktic communities, but with increasing interest shown in benthos. However, cyanobacteria have played a fundamental role from the very beginning in both the development of our planet's biodiversity and the construction of new habitats. These organisms have colonized almost every possible planktic or benthic environment on earth, including the most extreme ones, and display a vast number of adaptations. All this explains why they are the most important or the only phototrophs in some habitats. The negative effects of cyanotoxins on macroinvertebrates have been demonstrated, but usually under conditions that are far from natural, and on forms of exposure, toxin concentration, or composition. The cohabitation of cyanobacteria with most invertebrate groups is long-standing and has probably contributed to the development of detoxification means, which would explain the survival of some species inside cyanobacteria colonies. This review focuses on benthic cyanobacteria, their capacity to produce several types of toxins, and their relationships with benthic macroinvertebrates beyond toxicity.

A high-throughput gut-on-chip platform to study the epithelial responses to enterotoxins

Enterotoxins are a type of toxins that primarily affect the intestines. Understanding their harmful effects is essential for food safety and medical research. Current methods lack high-throughput, robust, and translatable models capable of characterizing toxin-specific epithelial damage. Pressing concerns regarding enterotoxin contamination of foods and emerging interest in clinical applications of enterotoxins emphasize the need for new platforms. Here, we demonstrate how Caco-2 tubules can be used to study the effect of enterotoxins on the human intestinal epithelium, reflecting toxins’ distinct pathogenic mechanisms. After exposure of the model to toxins nigericin, ochratoxin A, patulin and melittin, we observed dose-dependent reductions in barrier permeability as measured by TEER, which were detected with higher sensitivity than previous studies using conventional models. Combination of LDH release assays and DRAQ7 staining allowed comprehensive evaluation of toxin cytotoxicity, which was only observed after exposure to melittin and ochratoxin A. Furthermore, the study of actin cytoskeleton allowed to assess toxin-induced changes in cell morphology, which were only caused by nigericin. Altogether, our study highlights the potential of our Caco-2 tubular model in becoming a multi-parametric and high-throughput tool to bridge the gap between current enterotoxin research and translatable in vivo models of the human intestinal epithelium.

Epidemiology of Enterotoxigenic Escherichia coli among Children and Adults Seeking Care at Hospitals in Two Geographically Distinct Rural Areas in Bangladesh.

Enterotoxigenic Escherichia coli (ETEC) infections undeniably continue to have substantial morbidity and mortality in younger children; however, limited data are available on the disease burden of older children and adults and on ETEC epidemiology by geographical location at the subnational level. Facility-based surveillance over the years was established to identify patients with ETEC diarrhea in two geographically distinct areas in rural Bangladesh, Chhatak in the north and Mathbaria in the southern coastal area. ETEC was highly prevalent in both areas, while the proportions, toxin types and colonization factors varied by location, season and age groups. Children < 5 years old and adults between 20 and 60 years old were at the highest risk of ETEC diarrhea which required urgent care. This study underscores the importance of capturing subnational and seasonal variations in ETEC epidemiology. ETEC vaccine developers and public health stakeholders may need to target adults between 20 and 60 years of age in addition to young children as new vaccines currently under development become licensed and introduction begins.

A microscale 3D organ on a chip for recapitulating reciprocal neuroendocrine crosstalk between the hypothalamus and the pituitary gland

Conventional 2D or even recently developed 3D in vitro culture models for hypothalamus and pituitary gland cannot successfully recapitulate reciprocal neuroendocrine communications between these two pivotal neuroendocrine tissues known to play an essential role in controlling the body’s endocrine system, survival, and reproduction. In addition, most current vitro culture models for neuroendocrine tissues fail to properly reflect their complex multicellular structure. In this context, we developed a novel microscale chip platform, termed the ‘hypothalamic–pituitary (HP) axis-on-a-chip,’ which integrates various cellular components of the hypothalamus and pituitary gland with biomaterials such as collagen and hyaluronic acid. We used non-toxic blood coagulation factors (fibrinogen and thrombin) as natural cross-linking agents to increase the mechanical strength of biomaterials without showing residual toxicity to overcome drawbacks of conventional chemical cross-linking agents. Furthermore, we identified and verified SERPINB2 as a reliable neuroendocrine toxic marker, with its expression significantly increased in both hypothalamus and pituitary gland cells following exposure to various types of toxins. Next, we introduced SERPINB2-fluorescence reporter system into loaded hypothalamic cells and pituitary gland cells within each chamber of the HP axis on a chip, respectively. By incorporating this SERPINB2 detection system into the loaded hypothalamic and pituitary gland cells within our chip platform, Our HP axis-on-chip platform can better mimic reciprocal neuroendocrine crosstalk between the hypothalamus and the pituitary gland in the brain microenvironments with improved efficiency in evaluating neuroendocrine toxicities of certain drug candidates.

Nontyping virulence factors of Clostridium perfringens

The anaerobic, rod-shaped, spore-forming bacterium Clostridium perfringens is known for its production of biochemically active substances. Most of them are proteins that have a pathogenic effect on a wide range of animal tissues and cause a specific syndrome or even a disease in humans or animals. Production of toxins is used to classify isolates of C. perfringens into 7 different toxin types (A–G). Other virulence factors (i.g. beta2-toxin, BEC toxin, sialidases, hyaluronidase etc.) only indirectly or partially participate in the development of the disease, and the function of some substances has not been fully elucidated. The article summarizes basic data on the non-typing virulence factors of C. perfringens.

Metal-organic framework-based adsorbents for blood purification: progress, challenges, and prospects.

Blood purification, such as hemodialysis (HD), plasma exchange (PE), and hemoperfusion (HP), is widely applied in patients with organ failure (such as kidney and liver failure). Among them, HP mainly relies on porous adsorbents to efficiently adsorb accumulated metabolic wastes and toxins, thus improving purification efficiency. Metal-organic frameworks (MOFs), with a high porosity, large surface area, high loading capacity, and tailorable topology, are emerging as some of the most promising materials for HP. Compared with non-metal framework counterparts, the self-built metal centers of MOFs feature the intrinsic advantages of coordination with toxin molecules. However, research on MOFs in blood purification is insufficient, particularly in contrast to materials applied in other biomedical applications. Thus, to broaden this area, this review first discusses the essential characteristics, potential mechanisms, and structure-function relationship between MOFs and toxin adsorption based on porosity, topology, ligand functionalization, metal centers, and toxin types. Moreover, the stability, utilization safety, and hemocompatibility of MOFs are illustrated for adsorbent selection. The current development and progress in MOF composites for HD, HP, and extracorporeal membrane oxygenation (ECMO) are also summarized to highlight their practicability. Finally, we propose future opportunities and challenges from materials design and manufacture to the computational prediction of MOFs in blood purification. It is anticipated that our review will expand the interest of researchers for more impact in this area.

Molecular characterization of common zoonotic protozoan parasites and bacteria causing diarrhea in dairy calves in Ningxia Hui Autonomous Region, China.

Diarrhea caused by zoonotic pathogens is one of the most common diseases in dairy calves, threatening the health of young animals. Humans are also at risk, in particular children. To explore the pathogens causing diarrhea in dairy calves, the present study applied PCR-based sequencing tools to investigate the occurrence and molecular characteristics of three parasites (Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi) and three bacterial pathogens (Escherichia coli, Clostridium perfringens, and Salmonella spp.) in 343 fecal samples of diarrheic dairy calves from five farms in Lingwu County, Ningxia Hui Autonomous Region, China. The total positive rate of these pathogens in diarrheic dairy calves was 91.0% (312/343; 95% CI, 87.9-94.0), with C. perfringens (61.5%, 211/343; 95% CI, 56.3-66.7) being the dominant one. Co-infection with two to five pathogens was found in 67.3% (231/343; 95% CI, 62.4-72.3) of investigated samples. There were significant differences (p<0.05) in the positive rates of Cryptosporidium spp. and diarrheagenic E. coli among farms, age groups, and seasons. Two Cryptosporidium species (C. parvum and C. bovis) and five gp60 subtypes of C. parvum (IIdA15G1, IIdA20G1, IIdA19G1, IIdA14G1, and a novel IIdA13G1) were identified. Two assemblages (assemblage E and zoonotic assemblage A) of G. duodenalis and six ITS genotypes of E. bieneusi (J, Henan-IV, EbpC, I, EbpA, and ESH-01) were observed. Four virulence genes (eaeA, stx1, stx2, and st) of diarrheagenic E. coli and one toxin type (type A) of C. perfringens were detected. Our study enriches our knowledge on the characteristics and zoonotic potential of diarrhea-related pathogens in dairy calves.

Sudden death due to enterotoxemia among Arabian camels (Camelus dromedaries) and associated risk factors.

Sudden death is defined as an unexpected death occurring with no observed antecedent clinical signs. The current study was performed to notice the tangible causes of sudden death among 51 out of 340 she-camels on a private farm in the eastern region of El Khafgi, Saudi Arabia. A retrospective cohort study design was conducted to investigate the sudden death of camels through microscopic examination of fecal matter to identify the gastrointestinal parasites, analysis of whole blood thin films to diagnose blood parasites, blood culturing to recognize bacterial infection as Pasteurella multicida, and macroscopic postmortem examination to identify the gastrointestinal adult worm. The quantity and composition of feed were also analyzed. Afterward, a commercial multiscreen Ag-ELISA kit technique determined the toxins of Clostridium perfringens (C. perfringens). The results revealed that the incidence rate of sudden death was 15%. The sudden death occurred due to C. perfringens enterotoxins detected in the rumen, intestinal content, and intestinal wall. The enterotoxins and Alpha toxins were noticed, but the other toxin types, including Beta and Epsilon, could not be detected. All C. perfringens toxins were discovered to be negative in fecal matter. A significant association was reported between sudden death, she-camels age, and feeding habits as risk factors (p = 0.020 and 0.028, respectively). Risk factor assessment by relative risk (RR) revealed that the odds of RR of sudden death occurring among she-camels aged over two years were higher than those less than two years (2.24 CI 95%, 1.093-4.591). Furthermore, the odds RR of sudden death occurring due to exposure of she-camels to a concentrated ration of 18% were higher twice than those not exposed (2.346 CI 95%, 1.039-5.296). Clostridium perfringens enterotoxaemia should be listed as a cause of sudden death in camels and the alteration in diet with 18% concentration feed changes the intestinal environment, which leads to C. perfringens proliferating and yielding potent toxins. More observations and interferences like regular immunization are recommended to reduce the disease and increase the awareness of the farmers of the importance of risk factors.

Botulinum Toxin Injection for the Treatment of Third, Fourth, and Sixth Nerve Palsy: A Meta-Analysis.

The efficacy of botulinum toxin injection for the treatment of third, fourth, and sixth nerve palsy was evaluated. PubMed, Scopus, EMBASE, Web of Science, and Google Scholar databases were searched. Data about the duration of palsy (acute vs chronic), cause of the palsy, type of toxin used, mean dose, and other background characteristics were collected. Outcome variables were success rate (defined by alleviation of diplopia or reduction in eye deviation) and standardized mean difference of prism diopter and abduction deficit before and after injection. The Joanna Briggs Institute checklist was implemented for the risk of bias assessment. The analysis included 38 articles, comprising 643 patients. The overall treatment success rate in acute and chronic nerve palsy was 79% and 33%, respectively. The success rate was not significantly different between different subgroups of age, type of botulinum toxin, pre-injection prism diopter, etiology of the palsy, duration of follow-up, and mean dose of botulinum toxin injection. However, in both acute and chronic palsy, diabetes etiology was accompanied by the highest success rate. Overall symptomatic response to botulinum injection was 84% (95% CI: 67% to 96%), whereas functional response was observed in 64% (95% CI: 47% to 79%) of the patients. The odds ratio for the success rate of treatment of palsies with botulinum toxin versus expectant management was 2.67 (95% CI: 1.12 to 6.36) for acute palsy and 0.87 (95% CI: 0.17 to 4.42) for chronic palsy. Botulinum toxin can be used for the treatment of acute third, fourth, and sixth nerve palsy, especially in patients with acute palsy and more severe tropia. [J Pediatr Ophthalmol Strabismus. 2024;61(3):160-171.].

Composition, antibiotic resistance, and virulence analysis of microbiota in dormitory drain pipes.

Dormitory washbasins can breed microorganisms that produce odorous gases, polluting the indoor environment. We utilized metagenome sequencing to analyze the microbiota of 40 samples from the drain pipes of dormitory washbasins. Our study aimed to investigate the microbial community structure, antibiotic resistance genes, and virulence factors, and to identify potential influencing factors such as gender, hometown, frequency of hand sanitizer usage, and number of dormitory residents. The analysis revealed 12 phyla and 147 genera, with Proteobacteria and Actinobacteria being the dominant phyla, and Mycobacterium and Nakamurella being the dominant genera. We found that the factors influencing the microbial community structure of the dormitory washbasin drain pipe are complex. The investigated factors have a slight influence on the drain pipe microbial community, with gender exerting a discernible influence. The annotation results revealed the presence of various virulence factors, pathogenic toxins and antibiotic resistance genes, including 246 different toxin types and 30 different types of antibiotic resistance genes. In contrast to the observed differences in microbial composition among samples, the distribution of resistance genes shows relatively small changes among samples. Antibiotics should be a contributing factor in the overall increase of antibiotic resistance genes in drain pipes. Overall, our study provides important insights into the community structure and function of microorganisms in dormitory drainage systems, and can guide efforts to prevent and control microbial pollution.

БОТУЛОТОКСИН. МЕХАНИЗМ ТОКСИЧЕСКОГО ВОЗДЕЙСТВИЯ. ПРИНЦИП ДЕЙСТВИЯ ЛЕКАРСТВЕННЫХ ПРЕПАРАТОВ НА ОСНОВЕ РАЗНЫХ СЕРОТИПОВ БОТУЛОТОКСИНА. ПРИМЕНЕНИЕ В МЕДИЦИНЕ

The paper discusses the most recent understanding of the mechanisms of action of naturally occurring botulinum toxins in the context of chemical analysis, the types of botulism, the conditions of their origin, causes of development, as well as the methods of prevention of the toxic process, according to the type of lesion. The structural and pathogenic mechanisms of these toxins (A, B, C, D, E, F, G) are very similar (due to the property of inhibiting the exocytosis of neurotransmitter and causing muscle paralysis). However, because each type of toxin has a unique receptor, they have different affinities for the three transporter proteins found in SNAR (VAMP, SNAP, and syntaxin). If in the process of pathogenesis BTA (botulotoxin A) cleaves the protein in the synaptose, BTB cleaves synaptobrevin, while the intracellular target of BTC (which is still in the research stage) are two proteins of the transporter complex (synaptotagmin and syntaxin) at the same time, which has a positive effect on some pharmacological features. The existence of the A1 and A2 subtypes of BTA is discussed, and a chemical justification is provided for why even a minor alteration in the amino acid sequence results in a difference in the biological and immunological characteristics of these subtypes. Furthermore, evidence supports the idea that reducing the amount of complexing proteins is a strategy to lessen toxicity, reduce immune-resistance, assure a long-lasting myorelaxant impact, and thereby increase the potential of botulinum toxin therapy. And this can be achieved by reducing the inactive toxins in the mixture, the ways of which are presented. Meanwhile, on the basis of that same mechanism, one of the unique manifestations of A and B botulinum toxins acting as active drugs, the phenomenon of botulinum toxin therapy, is explained, if of course they are completely free of toxic substances and act in minimal amounts. In the meantime, the phenomena of botulinum toxin therapy, one of the distinctive manifestations of A and B botulinum toxins operating as active medications, is explained on the basis of that same mechanism, assuming, of course, that they are completely free of toxic components and act in minimal levels. This is when they are converted from poison to medicine as “wonderful forces of nature that are meant to help people with diseases that were previously thought to be incurable”. The uniqueness of their mechanism of action, the availability of the drug in any zone of injection, the long duration of the pharmacological effect and, most importantly, the absence of systematic and side effects already justify the demand for drugs, based on botulinum toxins, in neurology, surgery, traumatology, nephrology, dentistry, ophthalmology, aesthetic medicine and later also prospective possibilities of their application in other clinical fields.

Recent Advances in Biodecontamination of Aflatoxins in Foodstuffs: Using Biomass (2011-2021)

Background: Aflatoxin is one of the most hazardous toxins produced by Aspergillus spp. and contaminates animal feed and therefore human food. Aflatoxin exposure causes several health problems including liver, kidney, and intestine disorders for human beings. Therefore, an efficient decontamination method for food stuff seems essential. Objective: The aim of this mini-review was to go over the recent advances in the decontamination of aflatoxins in foodstuffs and explain the bio-removal methods based on microorganisms during the last decade. Methods: Literature research was performed using the keywords “aflatoxin”, “bio removal”, “decontamination”, “food”, and “biomass” individually or all together, in Scopus, Web of Science, and Pubmed in the last decade. Results: LABs and yeasts are the most useful and successful microorganisms in this field. These methods are very efficient, specific, environmentally friendly, and cost-effective, also they do not affect the nutritional value of the foodstuffs, unlike the physical or chemical techniques. The bio-removal methods contain aflatoxin adsorption and degradation. Conclusion: Although more studies and researches are required to evaluate the different types of toxin removal using various microorganisms and their practical aspects and the highest efficiency should be determined.

A Chromosome-Level Genome Assembly of the Reef Stonefish (Synanceia verrucosa) Provides Novel Insights into Stonustoxin (sntx) Genes.

Reef stonefish (Synanceia verrucosa) is one of the most venomous fishes, but its biomedical study has been restricted to molecular cloning and purification of its toxins, instead of high-throughput genetic research on related toxin genes. In this study, we constructed a chromosome-level haplotypic genome assembly for the reef stonefish. The genome was assembled into 24 pseudo-chromosomes, and the length totaled 689.74 Mb, reaching a contig N50 of 11.97 Mb and containing 97.8% of complete BUSCOs. A total of 24,050 protein-coding genes were annotated, of which metalloproteinases, C-type lectins, and stonustoxins (sntx) were the most abundant putative toxin genes. Multitissue transcriptomic and venom proteomic data showed that sntx genes, especially those clustered within a 50-kb region on the chromosome 2, had higher transcription levels than other types of toxins as well as those sntx genes scatteringly distributed on other chromosomes. Further comparative genomic analysis predicted an expansion of sntx-like genes in the Percomorpha lineage including nonvenomous fishes, but Scorpaenoidei species experienced extra independent sntx duplication events, marking the clear-cut origin of authentic toxic stonustoxins. In summary, this high-quality genome assembly and related comparative analysis of toxin genes highlight valuable genetic differences for potential involvement in the evolution of venoms among Scorpaeniformes fishes.