Pore-forming toxins of foodborne pathogens

Pore-forming toxins (PFTs) are water-soluble molecules that have been identified as the most crucial virulence factors during bacterial pathogenesis. PFTs disrupt the host cell membrane to internalize or to deliver other bacterial or virulence factors for establishing infections. Disruption of the host cell membrane by PFTs can lead to uncontrollable exchanges between the extracellular and the intracellular matrix, thereby disturbing the cellular homeostasis. Recent studies have provided insights into the molecular mechanism of PFTs during pathogenesis. Evidence also suggests the activation of several signal transduction pathways in the host cell on recognition of PFTs. Additionally, numerous distinctive host defense mechanisms as well as membrane repair mechanisms have been reported; however, studies reveal that PFTs aid in host immune evasion of the bacteria through numerous pathways. PFTs have been primarily associated with foodborne pathogens. Infection and death from diseases by consuming contaminated food are a constant threat to public health worldwide, affecting socioeconomic development. Moreover, the emergence of new foodborne pathogens has led to the rise of bacterial antimicrobial resistance affecting the population. Hence, this review focuses on the role of PFTs secreted by foodborne pathogens. The review highlights the molecular mechanism of foodborne bacterial PFTs, assisting bacterial survival from the host immune responses and understanding the downstream mechanism in the activation of various signaling pathways in the host upon PFT recognition. PFT research is a remarkable and an important field for exploring novel and broad applications of antimicrobial compounds as therapeutics.     

Influence of water activity and temperature on growth and mycotoxin production by Alternaria alternata on irradiated soya beans

The aim of this study was to determine the effects of water activity (a_w) (0.99-0.90), temperature (15, 25 and 30 °C) and their interactions on growth and alternariol (AOH) and alternariol monomethyl ether (AME) production by Alternaria alternata on irradiated soya beans. Maximum growth rates were obtained at 0.980 a_w and 25 °C. Minimum a_w level for growth was dependent on temperature. Both strains were able to grow at the lowest a_w assayed (0.90). Maximum amount of AOH was produced at 0.98 a_w but at different temperatures, 15 and 25 °C, for the strains RC 21 and RC 39 respectively. Maximum AME production was obtained at 0.98 a_w and 30 °C for both strains. The concentration range of both toxins varied considerably depending on a_w and temperature interactions. The two metabolites were produced over the temperature range 15 to 30 °C and a_w range 0.99 to 0.96. The limiting a_w for detectable mycotoxin production is slightly greater than that for growth. Two-dimensional profiles of a_w × temperature were developed from these data to identify areas where conditions indicate a significant risk from AOH and AME accumulation on soya bean. Knowledge of AOH and AME production under marginal or sub-optimal temperature and a_w conditions for growth can be important since improper storage conditions accompanied by elevated temperature and moisture content in the grain can favour further mycotoxin production and lead to reduction in grain quality. This could present a hazard if the grain is used for human consumption or animal feedstuff.     

Detection of warfare agents in liquid foods using the brine shrimp lethality assay

The brine shrimp lethality assay (BSLA) was used for rapid and non-specific detection of biological and chemical warfare agents at concentrations considerably below that which will cause harm to humans. Warfare agents detected include T-2 toxin, trimethylsilyl cyanide, and commercially available pesticides such as dichlorvos, diazinon, dursban, malathion, and parathion. The assay was performed by introducing 50 μL of milk or orange juice contaminated with each analyte into vials containing 10 freshly hatched brine shrimp nauplii in seawater. This was incubated at 28 °C for 24 h, after which mortality was determined. Mortality was converted to probits and the LC(50) was determined for each analyte by plotting probits of mortality against analyte concentration (log(10)). Our findings were the following: (1) the lethal effects of toxins dissolved in milk were observed, with T-2 toxin being the most lethal and malathion being the least, (2) except for parathion, the dosage (based on LC(50)) of analyte in a cup of milk (200 mL) consumed by a 6-y-old (20 kg) was less than the respective published rat LD(50) values, and (3) the BSLA was only suitable for detecting toxins dissolved in orange juice if incubation time was reduced to 6 h. Our results support the application of the BSLA for routine, rapid, and non-specific prescreening of liquid foods for possible sabotage by an employee or an intentional bioterrorist act. Practical Application: The findings of this study strongly indicate that the brine shrimp lethality assay can be adapted for nonspecific detection of warfare agents or toxins in food at any point during food production and distribution.     

Optimization of a rapid QuEChERS sample treatment method for HILIC-MS2 analysis of paralytic shellfish poisoning (PSP) toxins in mussels

A rapid and simple QuEChERS sample treatment was proposed for the development of a selective hydrophilic interaction liquid chromatography-ESI-MS²-based method for the determination of saxitoxins (STXs) in mussel samples. Among different sorbents, ABS Elut-NEXUS phase, composed of polystyrene cross-linked with 50% divinyl benzene and poly(methyl methacrylate), provided the best results. The effects of experimental parameters, including sorbent amount, vortexing time and centrifugation time were investigated and optimized by experimental design. In particular, regression models and desirability functions were applied to find the experimental conditions providing the highest global extraction response. The method was validated under the optimized conditions; detection and quantification limits in the 3–159 μg/kg and 7–436 μg/kg ranges respectively were obtained, except for C2 for which highest values were calculated due to its low ESI ionization efficiency. Finally, the analysis of twenty-eight mussel samples permitted to detect and quantify some of the investigated STXs, proving the applicability of the devised method.     

B型肉毒神经毒素的纯化及胰蛋白酶对其作用分析

目的纯化B型肉毒神经毒素,并比较未激活和用胰蛋白酶激活的B型肉毒神经毒素的分子特性,分析在酸性和碱性条件下B型肉毒毒素复合物的完整性。方法取未激活、胰蛋白酶激活和热处理激活的B型肉毒毒素复合物,采用阴离子交换柱层析纯化神经毒素(分别为A、B和C组)。在酸性条件下溶解激活的B型肉毒毒素复合物(D组)。对各组进行纯度(特异毒性)测定和SDS-PAGE分析。结果柱层析后,神经毒素纯度均比复合物的纯度高,激活的神经毒素比未激活的高2～3倍;在非还原和还原条件下,A组的SDS-PAGE显示神经毒素、重链和轻链3条带;B组显示重链和轻链2条带;C组在非还原条件下显示重链和轻链2条带,在还原条件下只显示轻链1条带。在酸性条件下,B型肉毒毒素复合物由神经毒素与非毒性成分构成,在碱性条件下,神经毒素与非毒性成分分离。结论已成功纯化了B型肉毒神经毒素,经胰蛋白酶处理后,单链裂解为双链,比活性提高。     

几种杀菌剂对芦荟黑斑病菌室内活性测定

通过供试的7种药剂对芦荟黑斑病菌体外抑制作用测试结果表明,所有药剂对其孢子萌发都有不同程度的抑制作用,其中异菌脲、咪鲜胺、嘧霉胺的抑制效果要优于氟硅唑、代森锰锌、百菌清、苯醚甲环唑;7种药剂对芦荟黑斑病菌菌丝体的抑制作用测定结果显示,所有药剂对菌丝生长都有显著的抑制作用,其中咪鲜胺锰络合物可湿性粉剂对菌丝体抑制效果最好,抑菌率达84.24%;异菌脲、氟硅唑、苯醚甲环唑次之。     

Design, synthesis, and pharmacological characterization of polyamine toxin derivatives: potent ligands for the pore-forming region of AMPA receptors

Polyamine toxins, such as philanthotoxins, are low-molecular-weight compounds isolated from spiders and wasps, which modulate ligand-gated ion channels in the nervous system. Philanthotoxins bind to the pore-forming region of AMPA receptors, a subtype of glutamate receptors which are important for memory formation and are involved in neurodegenerative diseases. Previous studies have demonstrated that modification of the polyamine moiety of philanthotoxins can lead to very potent and highly selective ligands for the AMPA receptor, as exemplified with philanthotoxin-56. Much less attention has been paid to the importance of the aromatic head group of philanthotoxins, but herein we demonstrate that modification of this moiety leads to a significant improvement in potency relative to philanthotoxin-56 at cloned AMPA receptors. Interestingly, the incorporation of an adamantane moiety is particularly favorable, and the most potent compound has a Ki value of 2 nM, making it the most potent uncompetitive antagonist of AMPA receptors described to date. Such compounds are potentially useful as neuroprotective agents.     

烟草赤星病菌(Alternaria longipes)产角质酶液体发酵条件的优化

以基础培养基（查彼培养基＋碳酸钙＋苹果角质）为营养优化了长柄链格孢(Alternaria longipes)产角质酶的环境和营养条件。结果表明最佳产酶的环境条件是：培养时间8 d,培养温度25 ℃,pH 5.5;通过正交试验确定最佳产酶的营养条件是：蔗糖3％,干酪素1％,吐温-80 0.3%;酶活高达175.2 U/mL,是基础培养基酶活力的11.0 倍。