Antimicrobial Activities of Extracts of Macrosphyra longistyla against Gram-Positive Oral Biofilm-Formers from School Children in Southwestern Nigeria and Toxicity Studies Using Brine Shrimps

The world will benefit from more effective antimicrobial agents against oral conditions arising from the actions of biofilm forming bacteria. Also, information is lacking on the oral biofilm-forming bacterial diversity in Southwestern Nigeria. In this study, we isolate and characterize oral biofilm producing bacteria in the oral cavities of schoolchildren in Southwestern Nigeria. We also investigate the antimicrobial properties of Macrosphyra longistyla extracts against the biofilm-formers and the toxicity of potent extracts. Samples were obtained from 109 schoolchildren aged 4 - 14 years from Lagos, Oyo and Osun States. Agar well diffusion technique was used in the antimicrobial susceptibility testing. Toxicity testing was done using brine shrimps (Artemia salina). Biofilm-formers in this study are Klebsiella sp., Streptococcus sp., Staphylococcus sp., and Micrococcus sp. Ethanol leaf extracts had the highest activity against all biofilm-producing bacteria. Ethanol stem bark extract, which elicited activity against Klebsiella only, was found to be less toxic than the ethanol leaf extract. Staphylococcus showed >10 mm susceptibility to the ethanol and aqueous extracts of Macrosphyra longistyla. Streptococcus and Micrococcus were susceptible to the antimicrobial actions of the ethanolic leaf extracts. Although the ethanol extracts of the leaves had lower minimum inhibitory concentrations than the ethanol extracts of the stem bark, toxicity studies showed ethanol extracts of the stem-bark to be more toxic than the ethanol extracts of the leaves. In conclusion, ethanolic extracts of Macrosphyra longistyla show potential as sources of antimicrobials against gram-positive, oral biofilm-forming bacteria.

Bacterial Exofactors Modulate Biofilm Growth and Resistivity to Antimicrobial Drugs

Some bacteria have the ability to co-exist, proliferate and survive in a multicellular community, biofilm. Each participating bacteria can form its colonies and encases itself by a self-produced insoluble extracellular matrix substance (EPS). Microcolonies within biofilm are held together by interactions and bonding of the substances present in the EPS with their separation from the water channels. Similar to insoluble EPS, bacterial microcolonies release soluble exofactors that have direct impacts on the survivability, growth and antibacterial resistivity of other microcolonies made of single- or multi-species bacteria in the same biofilm. How the exofactors of microcolonies of one-type bacteria impact on microcolonies of other-type bacteria is still unclear. We studied about the role of exofactors released from Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa, which are common biofilm-forming pathogenic bacteria. Exofactors facilitate to transform the microenvironment where bacteria can acquire alternative lifestyle with a long survival period and resistivity to certain antimicrobial drugs.

Staphylococcus aureus and biofilms:transmission, threats, and promising strategies in animal husbandry

Staphylococcus aureus(S. aureus) is a common pathogenic bacterium in animal husbandry that can cause diseases such as mastitis, skin infections, arthritis, and other ailments. The formation of biofilms threatens and exacerbates S. aureus infection by allowing the bacteria to adhere to pathological areas and livestock product surfaces, thus triggering animal health crises and safety issues with livestock products. To solve this problem, in this review, we provide a brief overview of the harm caused by S. aureus and its biofilms on livestock and animal byproducts(meat and dairy products). We also describe the ways in which S. aureus spreads in animals and the threats it poses to the livestock industry. The processes and molecular mechanisms involved in biofilm formation are then explained. Finally, we discuss strategies for the removal and eradication of S. aureus and biofilms in animal husbandry, including the use of antimicrobial peptides, plant extracts, nanoparticles, phages, and antibodies. These strategies to reduce the spread of S. aureus in animal husbandry help maintain livestock health and improve productivity to ensure the ecologically sustainable development of animal husbandry and the safety of livestock products.

Evolution of Biofilm and Its Effect on Microstructure of Mortar Surfaces in Simulated Seawater

To explore the role of biofilm formation on the corrosion of marine concrete structures, we investigated the attachment of biofilm on mortar surfaces in simulated seawater and the influence of biofilm on the microstructure of mortar surfaces. The results show that the evolution of biofilm on mortar surfaces in simulated seawater is closely related to the corrosion suffered by the mortar, and the process of biofilm attachment and shedding is continuous and cyclical. It is found that the specimens in the absence of biofilm attachment are more severely eroded internally by the corrosive medium in simulated seawater than those in the presence of biofilm attachment. For the specimens without biofilm attachment, after 60 days, gypsum forms,and after 120 days, the number of pores in the mortar is reduced. In contrast, for the specimens in the presence of biofilm attachment, gypsum could only be detected after 90 days, and fewer pores are filled. Therefore, the formation of biofilm could delay the invasion of the corrosive medium into the interior of mortar during the evolution of biofilm on mortar surfaces, mitigating the corrosion of mortars in seawater.

Anti-biofilm and anti-virulence potential of cell free supernatant of Akkermansia muciniphila against Salmonella

Akkermansia muciniphila is one of the commensals residing within the mammalian gut and co-evolving with the host.Numerous studies have demonstrated the benefits of A.muciniphila in ameliorating metabolic disorders,while little is known about the antimicrobial potential of A.muciniphila against pathogens.Here,we examined the antimicrobial and anti-virulence properties of cell free supernatant(CFS)of A.muciniphila against Salmonella Typhimurium.CFS retarded bacterial growth and inhibited the motility of S.Typhimurium SL1344 and S.Typhimurium 14028.CFS dose-dependently reduced cell hydrophobicity and auto-aggregation of both strains.Also,CFS from A.muciniphila significantly attenuated biofilm formation.Compared with untreated bacteria,CFS-treated bacteria significantly decreased adhesion and invasion to Caco-2 cells,and reduced intracellular survival in macrophages.CFS maintained antimicrobial properties after treatment with high temperatures and various proteases,while it lost its antimicrobial activity after pH neutralization.Gas chromatography-mass spectrometry(GC-MS)confirmed that A.muciniphila produced a certain amount of acetate and propionate,and ultra-high-performance liquid chromatography-mass spectrometry(UHPLCMS)identified other organic acids and metabolites in CFS.In summary,CFS from A.muciniphila exhibited anti-biofilm and anti-virulence properties against Salmonella and could be potentially utilized in the food industry for controlling Salmonella contamination and reducing infection.

Charge adaptive phytochemical-based nanoparticles for eradication of methicillin-resistant staphylococcus aureus biofilms

The intrinsic resistance of MRSA coupled with biofilm antibiotic tolerance challenges the antibiotic treatment of MRSA biofilm infections.Phytochemical-based nanoplatform is a promising emerging approach for treatment of biofilm infection.However,their therapeutic efficacy was restricted by the low drug loading capacity and lack of selectivity.Herein,we constructed a surface charge adaptive phytochemical-based nanoparticle with high isoliquiritigenin(ISL)loading content for effective treatment of MRSA biofilm.A dimeric ISL prodrug(ISL-G2)bearing a lipase responsive ester bond was synthesized,and then encapsulated into the amphiphilic quaternized oligochitosan.The obtained ISL-G2loaded NPs possessed positively charged surface,which allowed cis-aconityl-D-tyrosine(CA-Tyr)binding via electrostatic interaction to obtain ISL-G2@TMDCOS-Tyr NPs.The NPs maintained their negatively charged surface,thus prolonging the blood circulation time.In response to low pH in the biofilms,the fast removal of CA-Tyr led to a shift in their surface charge from negative to positive,which enhanced the accumulation and penetration of NPs in the biofilms.Sequentially,the pH-triggered release of D-tyrosine dispersed the biofilm and lipase-triggered released of ISL effectively kill biofilm MRSA.An in vivo study was performed on a MRSA biofilm infected wound model.This phytochemical-based system led to~2log CFU(>99%)reduction of biofilm MRSA as compared to untreated wound(P<0.001)with negligible biotoxicity in mice.This phytochemical dimer nanoplatform shows great potential for long-term treatment of resistant bacterial infections.

Structural insights on anti-biofilm mechanism of heated slightly acidic electrolyzed water technology against multi-resistant Staphylococcus aureus biofilm on food contact surface

Slightly acidic electrolyzed water(SAEW)has proven to be an efficient and novel sanitizer in food and agriculture field.This study assessed the efficacy of SAEW(30 mg/L)at 40℃on the inactivation of foodbome pathogens and detachment of multi-resistant Staphylococcus aureus(MRSA)biofilm.Furthermore.the underlying mechanism of MRS A biofilm under heated SAEW at 40℃treatment on metabolic profiles was investigated.The results showed that the heated SAEW at 40℃significantly effectively against foodbome pathogens of 1.96-7.56(lg(CFU/g))reduction in pork,chicken,spinach,and lettuce.The heated SAEW at 40℃treatment significantly reduced MRS A biofilm cells by 2.41(lg(CFU/cm^(2))).The synergistic effect of SAEW treatment showed intense anti-biofilm activity in decreasing cell density and impairing biofilm cell membranes.Global metabolic response of MRSA biofilms,treated by SAEW at 40℃,revealed the alterations of intracellular metabolites,including amino acids,organic acid,fatty acid,and lipid.Moreover,signaling pathways involved in amino acid metabolism,energy metabolism,nucleotide synthesis,carbohydrate metabolites,and lipid biosynthesis were functionally disrupted by the SAEW at 40℃treatment.As per our knowledge,this is the first research to uncover the potential mechanism of heated SAEW treatment against MRSA biofilm on food contact surface.

Contemporary strategies and approaches for characterizing composition and enhancing biofilm penetration targeting bacterial extracellular polymeric substances

Extracellular polymeric substances(EPS)constitutes crucial elements within bacterial biofilms,facili-tating accelerated antimicrobial resistance and conferring defense against the host's immune cells.Developing precise and effective antibiofilm approaches and strategies,tailored to the specific charac-teristics of EPS composition,can offer valuable insights for the creation of novel antimicrobial drugs.This,in turn,holds the potential to mitigate the alarming issue of bacterial drug resistance.Current analysis of EPS compositions relies heavily on colorimetric approaches with a significant bias,which is likely due to the selection of a standard compound and the cross-interference of various EPS compounds.Considering the pivotal role of EPS in biofilm functionality,it is imperative for EPS research to delve deeper into the analysis of intricate compositions,moving beyond the current focus on polymeric materials.This ne-cessitates a shift from heavy reliance on colorimetric analytic methods to more comprehensive and nuanced analytical approaches.In this study,we have provided a comprehensive summary of existing analytical methods utilized in the characterization of EPS compositions.Additionally,novel strategies aimed at targeting EPS to enhance biofilm penetration were explored,with a specific focus on high-lighting the limitations associated with colorimetric methods.Furthermore,we have outlined the challenges faced in identifying additional components of EPS and propose a prospective research plan to address these challenges.This review has the potential to guide future researchers in the search for novel compounds capable of suppressing EPS,thereby inhibiting biofilm formation.This insight opens up a new avenue for exploration within this research domain.

Evaluation of Azadirachta indica cultivated in Egypt against biofilm formation by Pseudomonas aeruginosa: Insights from molecular docking studies targeting LasR

Background:Azadirachta indica(A.indica),commonly known as neem,is a widely distributed medicinal plant in Asia and Africa and is well known to have a wide spectrum of biological activity.A.indica is considered a skin food that was traditionally used in different cultures to treat a wide range of skin disorders.A.indica was reported to possess antibacterial activity against Pseudomonas aeruginosa(P.aeruginosa)which is considered the most common biofilm model organism.This study aims to investigate the ability of A.indica cultivated in Egypt to inhibit/reduce the biofilm formation by P.aeruginosa.Methods:The microtiter plate assay was used to evaluate the anti-biofilm activity of neem,cultivated in Egypt,leaves against P.aeruginosa as well as the ability to reduce the activity of P.aeruginosa.To investigate the phytocompounds responsible for their bioactivity and to explore potential interactions between their bioactive components and one of the quorum-sensing regulatory proteins of P.aeruginosa involved in biofilm formation,liquid chromatography-mass spectrometric and molecular docking studies were done.Results:Results showed that methanol extract of leaves can reduce the formation of P.aeruginosa biofilm at lower concentrations than those reported in other regions with 1.25 mg/mL as the optimum concentration.The two-way analysis of variance revealed the significance of the extract effect and its concentration on the reduction of biofilm formation(P<0.05).Liquid chromatography-mass spectrometric study revealed the presence of fourteen compounds that belong to limonoids and flavonoids.Molecular docking analysis against LasR,the quorum-sensing regulatory protein,of P.aeruginosa supported these findings.Nimbolinin,a limonoid,has achieved the highest Libdock score of 138.769.Conclusion:It was concluded that A.indica,cultivated in Egypt,leaves can target LasR as a new mechanism of action for biofilm control by A.indica and therefore could be a good source of leads for anti-biofilm medicine.

Numerical Modeling of Mass Transfer in the Interaction between River Biofilm and a Turbulent Boundary Layer

In this article dedicated to the modeling of vertical mass transfers between the biofilm and the bulk flow, we have, in the first instance, presented the methodology used, followed by the presentation of various results obtained through analyses conducted on velocity fields, different fluxes, and overall transfer coefficients. Due to numerical constraints (resolution of relevant spatial scales), we have restricted the analysis to low Schmidt numbers (Sc=0.1, Sc=1, and Sc=10) and a single roughness Reynolds number (Re*=150). The analysis of instantaneous concentration fields from various simulations revealed logarithmic concentration profiles above the canopy. In this zone, the concentration is relatively homogeneous for longer times. The analysis of results also showed that the contribution of molecular diffusion to the total flux depends on the Schmidt number. This contribution is negligible for Schmidt numbers Sc≥0.1, but nearly balances the turbulent flux for Sc=0.1. In the canopy, the local Sherwood number, given by the ratio of the total flux (within or above the canopy) to the molecular diffusion flux at the wall, also depends on the Schmidt number and varies significantly between the canopy and the region above. The exchange velocity, a purely hydrodynamic parameter, is independent of the Schmidt number and is on the order of 10% of in the present case. This study also reveals that nutrient absorption by organisms near the wall depends on the Schmidt number. Such absorption is facilitated by lower Schmidt numbers.

Biofilm analyses and exoproduct release by clinical and environmental isolates of Burkholderia pseudomallei from Brazil

Objective:To characterize biofilm production by clinical(n=21)and environmental(n=11)isolates of Burkholderia pseudomallei and evaluate the production of proteases,hemolysins and siderophores.Methods:Initially,the 32 strains were evaluated for biofilm production in Müller-Hinton broth-1%glucose(MH-1%glucose)and BHI broth-1%glucose,using the crystal violet staining technique.Subsequently,growing(48 h)and mature(72 h)biofilms were evaluated by confocal microscopy.Finally,the production of proteases,hemolysins and siderophores by planktonic aggregates,growing biofilms and mature biofilms was evaluated.Results:All isolates produced biofilms,but clinical isolates had significantly higher biomass in both MH-1%glucose(P<0.001)and BHI-glucose 1%(P=0.005).The structural analyses by confocal microscopy showed thick biofilms,composed of multiple layers of cells,homogeneously arranged,with mature biofilms of clinical isolates presenting higher biomass(P=0.019)and thickness of the entire area(P=0.029),and lower roughness coefficient(P=0.007)than those of environmental isolates.Protease production by growing biofilms was significantly greater than that of planktonic(P<0.001)and mature biofilms(P<0.001).Hemolysin release by planktonic aggregates was higher than that of biofilms(P<0.001).Regarding siderophores,mature biofilms presented higher production than growing biofilms(P<0.001)and planktonic aggregates(P<0.001).Conclusions:Clinical isolates have higher production of biofilms than their environmental counterparts;protease and siderophores seem important for growth and maintenance of Burkholderia pseu­domallei biofilms.

Biofilm removal mediated by Salmonella phages from chicken-related source

Salmonella and their biofilm formation are the primary bacterial causes of foodborne outbreaks and crosscontamination. The objective of the study was to investigate the potential of Salmonella phages as an alternative technology for biofilm removal. In this work, 21 Salmonella phages were isolated from a chicken farm and slaughter plant and the phage(CW1)with the broadest spectrum was characterized. Complete genome sequence analysis revealed that the genomes of phage CW1 is composed of 41 763 bp with 58 open reading frames(ORFs)and a holin-endolysin system and it does not encode any virulence or lysogeny. A phage cocktail consisted of CW1(with the broadest spectrum of 70.49%)and CW11, M4 and M10(with a high lytic activity of more than 67.11%)was established. Treatment with the cocktail reduced the cells in the developing biofilm and mature biofilm by 0.79 lg(CFU/cm~2)and 0.4 lg(CFU/cm~2), respectively. More dead cells and scattered extracellular polymeric substances(EPS)were observed by confocal laser scanning microscopy and scanning electron microscopy. Raman analysis found that carbohydrates and proteins were the identification receptors for scattered EPS. This finding suggests that this phage cockta il has potential applications for the sterilization of Salmonella biofilm during meat processing.

Anti-Biofilm, Anti-Quorum Sensing, and Anti-Proliferative Activities of Methanolic and Aqueous Roots Extracts of Carica papaya L. and Cocos nucifera L.

Objective: This study focused on the antibacterial and anti-proliferative activity of extracts from Carica papaya and Cocos nucifera roots. Methodology: The minimum inhibitory concentration and the minimum bactericidal concentration of the extracts on Escherichia coli, Pseudomonas aeruginosa, Streptococcus mutans, and Staphylococcus aureus were deduced by the microdilution method. The anti-biofilm activity was determined on all four strains and anti-quorum sensing activity by inhibition of violacein production in Chromobacterium violaceum. Anti-proliferative activity on prostate cultured cancer cells was evaluated by MTT assay. Sterols and triterpenes were also assayed in this study. Results: The methanolic extract of Carica papaya showed the best anti-biofilm effect with a percentage inhibition of 66.10 ± 1.79. The methanolic extract of Cocos nucifera had the strongest inhibition on the production of quorum sensing (61.42 ± 0.28). In addition, the methanolic extract of Cocos nucifera roots showed the best cytotoxic effect on prostate cancer LNCaP cell lines (IC50 = 26.98 ± 2.6 μg/mL) and Carica papaya on the PC-3 lines (IC50 = 127.20 ± 5.99 μg/mL). The extracts were also rich in triterpenes and sterols. Conclusion: This study provides support for the ethnomedical use of Carica papaya and Cocos nucifera roots as an antimicrobial and anticancer.

Biofilm formation under high temperature causes the commensal bacteria Bacillus cereus WPySW2 to shift from friend to foe in Neoporphyra haitanensis in vitro model

Although biofilm formation may promote growth,biofilms are not always beneficial to their hosts.The biofilm formation characteristics of Bacillus cereus WPySW2 and its changes at different temperatures were studied.Results show that B.cereus WPySW2 promoted the growth of Neoporphyra haitanensis(an economically cultivated seaweed)at 20℃ but accelerated algal rot at 28℃.Thicker B.cereus WPySW2 biofilms covered the surface of N.haitanensis thalli at 28℃,which hindered material exchange between the algae and surrounding environment,inhibited algal photosynthesis and respiration,and accelerated algal decay.Compared with planktonic bacteria,mature biofilm cells had lower energy consumption and metabolic levels.The biofilm metabolic characteristics of B.cereus WPySW2 changed significantly with temperature.High temperature accelerated biofilm maturation,which made it thicker and more stable,allowing the bacteria to easily adapt to environmental changes and obtain greater benefits from their host.High temperature did not affect the production or increased the abundance of toxic metabolites,indicating that the negative effects of B.cereus WPySW2 on algae were not caused by toxins.This study shows that increased temperature can transform a harmless bacterium into a detrimental one,demonstrating that temperature may change the ecological function of phycospheric bacteria by affecting their morphology and metabolism.

Temporal characteristics of algae-denitrifying bacteria co-occurrence patterns and denitrifier assembly in epiphytic biofilms on submerged macrophytes in Caohai Lake,SW China

Denitrifying bacteria in epiphytic biofilms play a crucial role in nitrogen cycle in aquatic habitats.However,little is known about the connection between algae and denitrifying bacteria and their assembly processes in epiphytic biofilms.Epiphytic biofilms were collected from submerged macrophytes(Patamogeton lucens and Najas marina L.)in the Caohai Lake,Guizhou,SW China,from July to November 2020 to:(1)investigate the impact of abiotic and biotic variables on denitrifying bacterial communities;(2)investigate the temporal variation of the algae-denitrifying bacteria co-occurrence networks;and(3)determine the contribution of deterministic and stochastic processes to the formation of denitrifying bacterial communities.Abiotic and biotic factors influenced the variation in the denitrifying bacterial community,as shown in the Mantel test.The co-occurrence network analysis unveiled intricate interactions among algae to denitrifying bacteria.Denitrifying bacterial community co-occurrence network complexity(larger average degrees representing stronger network complexity)increased continuously from July to September and decreased in October before increasing in November.The co-occurrence network complexity of the algae and nirS-encoding denitrifying bacteria tended to increase from July to November.The co-occurrence network complexity of the algal and denitrifying bacterial communities was modified by ammonia nitrogen(NH_(4)^(+)-N)and total phosphorus(TP),pH,and water temperature(WT),according to the ordinary least-squares(OLS)model.The modified stochasticity ratio(MST)results reveal that deterministic selection dominated the assembly of denitrifying bacterial communities.The influence of environmental variables to denitrifying bacterial communities,as well as characteristics of algal-bacterial co-occurrence networks and the assembly process of denitrifying bacterial communities,were discovered in epiphytic biofilms in this study.The findings could aid in the appropriate understanding and use of epiphytic biofilms denitrification function,as well as the enhancement of water quality.

Virulence Factors and Biofilm Formation in Vancomycin Resistant Enterococcus faecalis and Enterococcus faecium Isolates in Brazil

In this work, we evaluated biofilm formation of Vancomycin Resistant of E. faecalis and E. faecium (VRE) in different culture media and adhesion substrate, as well as cellular hydrophobicity and presence of virulence genes. For this, 35 isolates were collected from a public hospital in Recife, Pernambuco, Brazil and identified by the Matrix-Assisted Laser Desorption Ionization - Time-of-flight - Mass Spectrometry (MALDI-TOF-MS) technique. Biofilm formation was analyzed by the Crystal Violet (CV) method and fluorescence microscopy, cellular hydrophobicity by hydrocarbon interaction and the presence of gelE, esp and asa1 genes by Polymerase Chain Reaction (PCR). 12 isolates were identified as E. faecalis and 23 as E. faecium. Most were obtained in Coronary Units (40.0%) and Intensive Care Unit (31.4%). E. faecium isolates were more resistant to the antibiotics tested than E. faecalis;however, E. faecalis stood out as a biofilm producer. Regarding the presence and gene frequency, it was observed that gelE (54.3%) and esp (54.3%) were the most prevalent, followed by asa1 (22.9%). When comparing the gene frequency, it was observed that gelE and esp were predominant (48.6% for both species), while asa1 was more frequent in E. faecalis (20.0%). The data presented here are worrying, because they reveal the virulence potential of isolates VRE, which contributes to the dissemination and persistence of these pathogens in the hospital environment.

不同填料生物膜对海水养殖尾水的脱氮效能及微生物群落分析

为探究不同填料生物膜对海水养殖尾水氮污染物的处理能力,分别以无填料(C)、牡蛎壳(M)、珊瑚石(S)、弹性填料(T)和悬浮球填料(F)构建5组生物滤池,比较填料生物膜成熟时间、成膜情况及对不同氮污染物的24 h去除能力,同时利用高通量测序技术分析挂膜期间(20、40、60 d)填料生物膜上微生物群落的变化。结果表明:不同填料生物膜成熟时间需要46~50 d,珊瑚石所需时间最短(46 d);扫描电镜显示,弹性填料和悬浮球填料附着生物量最多,以杆状细菌为主;对氨氮、亚硝酸盐氮、硝酸盐氮的24 h去除率最高的填料分别是悬浮球填料(68.66%±6.27%)、珊瑚石(99.99%±0.00%)和悬浮球填料(6.73%±3.41%);高通量测序显示,随着挂膜时间延长,弹性填料生物膜上的细菌丰度显著增加,牡蛎壳和珊瑚石生物膜上的细菌多样性显著下降(P<0.05);在门分类水平上,变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)是不同填料生物膜的主要优势菌群,硝化螺旋菌门(Nitrospirae)、硝化刺菌门(Nitrospinae)的相对丰度随挂膜时间延长不断升高;在属分类水平上,亚硝酸菌属(Nitrosomonas)、硝化螺菌属(Nitrospira)、硝化刺菌属(Nitrospina)和未分类_亚硝化单胞菌科(unclassified_Nitrosomonadaceae)是填料生物膜上具有硝化作用的优势菌属,这4种硝化菌属在挂膜60 d时的相对丰度总和从高到低依次为悬浮球填料(42.53%)>牡蛎壳(30.50%)、弹性填料(29.30%)>珊瑚石(11.74%),假单胞菌属(Pseudomonas)、青枯菌属(Ralstonia)、噬几丁质菌属(Chitinophaga)和草螺菌属(Herbaspirillum)等反硝化菌属在珊瑚石填料生物膜上的相对丰度高于其他填料。研究表明,珊瑚石、悬浮球填料能够实现脱氮菌属的高效富集,对海水养殖尾水具有良好的脱氮能力,是较为理想的生物填料。

百里香酚对犊牛源生物被膜阳性金黄色葡萄球菌耐药性的消除作用

为了了解内蒙古地区犊牛病料中生物被膜阳性金黄色葡萄球菌的流行情况、耐药情况及百里香酚对生物被膜阳性金黄色葡萄球菌耐药性的消除作用。本试验采用生化鉴定和PCR方法,对采集到的104份犊牛病料进行金黄色葡萄球菌的分离鉴定;采用K-B法检测了金黄色葡萄球菌分离株对18种抗菌药物的敏感性;采用改良结晶紫染色法测定分离株的生物被膜形成能力;选取2株生物被膜强阳性菌株,采用微量肉汤稀释法测定百里香酚对分离株的最小抑菌浓度,并进行百里香酚耐药性消除试验。结果显示,在104份犊牛病料中共分离鉴定获得43株金黄色葡萄球菌。药敏试验结果显示,金黄色葡萄球菌分离株对氨苄西林、头孢西丁、卡那霉素、复方新诺明和红霉素5种抗菌药物的耐药率达到50.0%以上;74.4%(32/43)的分离株为多重耐药株。生物被膜形成能力强、中、弱和无生物被膜形成能力的分离株分别占比23.3%(10/43)、16.3%(7/43)、32.6%(14/43)和27.9%(12/43);生物被膜阳性株中83.9%(26/31)为多重耐药株。百里香酚对2株生物被膜强阳性株的最小抑菌浓度均为256 mg/mL,经百里香酚处理后均恢复了对头孢噻肟的敏感性。结果表明,百里香酚对生物被膜阳性金黄色葡萄球菌具有一定的耐药消除作用。

沙门菌噬菌体YT90的分离鉴定及生物学特性分析

[目的]本文旨在分离针对肠炎沙门菌的烈性噬菌体并研究其生物学特性。[方法]从山东烟台某鸭场采集58份粪便样品,以鸭源肠炎沙门菌sdc-11为宿主分离噬菌体,并对噬菌体分离株进行形态学观察,对其温度和pH值稳定性、感染复数(MOI)、一步生长曲线进行测定和分析,测试其对71株不同血清型沙门菌的裂解谱,以及对沙门菌sdc-11生物被膜形成的抑制和对成熟生物被膜的清除作用,最后进行基因组测序分析。[结果]分离获得1株长尾噬菌体,命名为YT90。该噬菌体在30~60℃和pH值4~10的条件下,均表现出较好的增殖能力,最佳MOI值为10^(-5),潜伏期10 min,裂解期90 min,裂解量为每细胞278 PFU;可裂解66.7%(14/21)肠炎沙门菌、66.7%(28/42)鼠伤寒沙门菌和75.0%(3/4)鸡白痢沙门菌;在96孔细胞板上,YT90具有抑制沙门菌sdc-11生物被膜形成的潜力,并能有效清除成熟生物被膜;全基因组测序发现,YT90的基因组全长121240 bp,不含已知的毒力因子和耐药相关基因,不编码整合酶;基于末端酶大亚基构建的进化树分析表明,YT90与沙门菌噬菌体GRNsp50关系最近,属于根西病毒亚科(Guernseyvirinae)新泽西病毒属(Jerseyvirus)。[结论]从鸭场粪便样品中分离获得1株烈性噬菌体YT90,可裂解多种血清型沙门菌,耐受性强、潜伏期短、裂解量大,在防治沙门菌感染方面有较好的临床应用潜力。

爱媛类芽孢杆菌抗菌肽对白色念珠菌生物膜的抑制作用机制

研究爱媛类芽孢杆菌抗菌肽对白色念珠菌生物膜的抑制作用,采用微量二倍稀释法和时间-杀菌曲线测定抑制效果,显微镜观察对白色念珠菌芽管和菌丝形成的影响,2,3-二(2-甲氧基-4-硝基-5-磺酸基苯基)-2H-四唑-5-甲酰苯胺法研究对生物膜、预成型生物膜形成和成熟生物膜清除率的影响,荧光探针观察对生物膜结构和膜内菌体状态的影响,平板涂布法和实时聚合酶链式反应测定生物膜内活菌数量和相关基因表达水平。结果表明,抗菌肽对白色念珠菌的最小抑菌浓度为8.28 AU/mL。抗菌肽能降低白色念珠菌的芽管形成率,阻止菌丝的生成,使其以酵母形式存在。抗菌肽对生物膜的形成和清除产生影响,能在较短时间内破坏生物膜的结构完整性,导致菌体受损和死亡,减少膜内菌落数量。在基因水平上,抗菌肽可降低多个生物膜形成相关的基因(如ALS1、ALS3、HWP1、EFG1、ECE1和UME6)的表达水平,从而抑制生物膜的形成。研究证明抗菌肽能够有效抑制白色念珠菌生物膜形成、成熟膜清除、成膜基因表达等,结果可为白色念珠菌新型抑菌物质的开发奠定理论基础,为食品病原微生物污染防治和食品质量安全保障提供依据。