<abstract><title style='display:none'>Abstract</title>Research on the susceptibility of the spores of anaerobic bacteria such as <italic>Clostridium sporogenes</italic> or <italic>Clostridioides difficile</italic> is vital for assessing the sporicidal activity of disinfectants. The diverse susceptibility of anaerobic bacteria spores may lead to different disinfection parameters being determined by laboratories that prepare spore suspensions to test sporicidal effectiveness. The tests were performed using the suspension method according to PN-EN 13704:2018-09. In order to assess the susceptibility of the <italic>C. sporogenes</italic> spores, the criterion established for the <italic>C. difficile</italic> ribotype 027 spores was used in accordance with PN‑EN 17126:2019-01. The susceptibility of the <italic>C. sporogenes</italic> spores to glutardialdehyde corresponded to the susceptibility ranges established for the <italic>C. difficile</italic> ribotype 027 spores. The <italic>C. sporogenes</italic> spore suspension was susceptible to low concentrations of peracetic acid (0.01%). A disinfectant containing peracetic acid as the active substance showed high sporicidal activity at a low concentration (1%), a short contact time (15 minutes), and a high organic load (3.0 g/l bovine albumin + 3.0 ml/l sheep erythrocytes), as compared to a disinfectant with glutardialdehyde, which was sporicidal at a higher concentration (2.5%), at a longer contact time
(60 minutes) and lower organic conditions (3.0 g/l bovine albumin). There is a need to define the minimum susceptibility criteria for the <italic>C. sporogenes</italic> spores to the reference substances most often found in disinfectants with sporicidal activity. Excessive susceptibility of the <italic>C. sporogenes</italic> spores to reference substances may result in low-performance parameters of disinfection products with sporicidal activity and lead to ineffective disinfection in practice.<fig id="j_pjm-2022-031_fig_001" position="float" fig-type="figure">
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</fig></abstract>

Susceptibility of <italic>Clostridium sporogenes</italic> Spores to Selected Reference Substances and Disinfectants

<abstract><title style='display:none'>Abstract</title>Hepatitis B infection is still a global concern progressing as acute-chronic hepatitis, severe liver failure, and death. The infection is most widely transmitted from the infected mother to a child, with infected blood and body fluids. Pregnant women, adolescents, and all adults at high risk of chronic infection are recommended to be screened for hepatitis B infection. The initial analysis includes serological tests that allow differentiation of acute and chronic hepatitis. Molecular assays performed provide detection and quantification of viral DNA, genotyping, drug resistance, and precore/core mutation analysis to confirm infection and monitor disease progression in chronic hepatitis B patients. All patients with chronic hepatitis B should be treated with antiviral medications and regularly monitored for efficient treatment. The current treatment is based on nucleos(t)ide analogs and pegylated interferons that save lives by decreasing liver cancer death, liver transplant, slow or reverse the progression of liver disease as well as the virus infectivity.</abstract>

Hepatitis B Virus: From Diagnosis to Treatment

<abstract>
<title style='display:none'>Abstract</title>
This study aims to evaluate associations between the immunochromatographic rapid test technique and <italic>Trichomonas vaginalis</italic> (TV) infection in patients with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) in Taiwan. All patients received post-prostate massage urine (VB3) Trichomonas rapid tests. The demographic characteristics and urogenital symptoms of CP/CPPS were recorded. Routine urinalysis of VB3 was also performed, and laboratory examination results of semen were recorded if available. A total of 29 patients with TV infection and 109 without TV infection were enrolled, which reflected that the prevalence in patients with TV infection was approximately 21%. Patients with TV infection displayed a significantly higher frequency of suprapubic/lower abdominal pain (<italic>p</italic> = 0.034), semen leukocyte &gt; 5/high-power field (HPF) (<italic>p</italic> = 0.020), and an inflammatory type (category IIIA) (<italic>p</italic> = 0.005) than patients without TV infection. A higher prevalence of TV infection was found in patients with category IIIA (47.37%). No significant difference was found in the symptom duration and other clinical symptoms. In conclusion, the high prevalence of TV infection was revealed in CP/CPPS patients using the VB3 rapid Trichomonas test, especially in CP/CPPS patients with category IIIA. Thus, rapid TV testing might be vital for CP/CPPS patients in the hospital.</abstract>

Detection of <italic>Trichomonas vaginalis</italic> Infection in Chronic Prostatitis/Chronic Pelvic Pain Syndrome Patients by Rapid Immunochromatographic Test

<abstract>
<title style='display:none'>Abstract</title>
Alpha-mangostin (α-mangostin) was discovered as a potent natural product against Gram-positive bacteria, whereas the underlying molecular mechanisms are still unclear. This study indicated that α-mangostin (at 4 × MIC) rapidly killed <italic>Staphylococcus aureus</italic> planktonic cells more effectively (at least 2-log10 CFU/ml) than daptomycin, vancomycin and linezolid at 1 and 3 h in the time-killing test. Interestingly, this study also found that a high concentration of α-mangostin (≥4×MIC) significantly reduced established biofilms of <italic>S. aureus</italic>. There were 58 single nucleotide polymorphisms (SNPs) in α-mangostin nonsensitive <italic>S. aureus</italic> isolates by whole-genome sequencing, of which 35 SNPs were located on both sides of the <italic>sarT</italic> gene and 10 SNPs in the <italic>sarT</italic> gene. A total of 147 proteins with a different abundance were determined by proteomics analysis, of which 91 proteins increased, whereas 56 proteins decreased. The abundance of regulatory proteins SarX and SarZ increased. In contrast, the abundance of SarT and IcaB was significantly reduced (they belonged to SarA family and <italic>ica</italic> system, associated with the biofilm formation of <italic>S. aureus</italic>). The abundance of cell membrane proteins VraF and DltC was augmented, but the abundance of cell membrane protein UgtP remarkably decreased. Propidium iodide and DiBaC4(3) staining assay revealed that the fluorescence intensities of DNA and the cell membrane were elevated in the α-mangostin treated <italic>S. aureus</italic> isolates. In conclusion, this study reveals that α-mangostin was effective against <italic>S. aureus</italic> planktonic cells by targeting cell membranes. The anti-biofilm effect of α-mangostin may be through inhibiting the function of SarT and IcaB.
<fig id="j_pjm-2023-021_ufig_001" position="float" fig-type="figure">
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</fig>
</abstract>

Mechanisms of Rapid Bactericidal and Anti-Biofilm Alpha-Mangostin <italic>In Vitro</italic> Activity against <italic>Staphylococcus aureus</italic>

<abstract><title style='display:none'>Abstract</title><italic>Aspergillus fumigatus</italic> is one of the ubiquitous fungi with airborne conidia, which accounts for most aspergillosis cases. In immunocompetent hosts, the inhaled conidia are rapidly eliminated. However, immunocompromised or immunodeficient hosts are particularly vulnerable to most <italic>Aspergillus</italic> infections and invasive aspergillosis (IA), with mortality from 50% to 95%. Despite the improvement of antifungal drugs over the last few decades, the therapeutic effect for IA patients is still limited and does not provide significant survival benefits. The drawbacks of antifungal drugs such as side effects, antifungal drug resistance, and the high cost of antifungal drugs highlight the importance of finding novel therapeutic and preventive approaches to fight against IA. In this article, we systemically addressed the pathogenic mechanisms, defense mechanisms against <italic>A. fumigatus</italic>, the immune response, molecular aspects of host evasion, and vaccines’ current development against aspergillosis, particularly those based on AFMP4 protein, which might be a promising antigen for the development of anti-<italic>A. fumigatus</italic> vaccines.</abstract>

The Pathogenesis of <italic>Aspergillus fumigatus</italic>, Host Defense Mechanisms, and the Development of AFMP4 Antigen as a Vaccine

<abstract><title style='display:none'>Abstract</title>Silent genes are DNA sequences that are generally not expressed or expressed at a very low level. These genes become active as a result of mutation, recombination, or insertion. Silent genes can also be activated in laboratory conditions using pleiotropic, targeted genome-wide, or biosynthetic gene cluster approaches. Like every other gene, silent genes can spread through horizontal gene transfer. Most studies have focused on strains with phenotypic resistance, which is the most common subject. However, to fully understand the mechanism behind the spreading of antibiotic resistance, it is reasonable to study the whole resistome, including silent genes.<fig id="j_pjm-2021-040_fig_001" position="float" fig-type="figure"><graphic xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="graphic/j_pjm-2021-040_fig_001.jpg"/></fig></abstract>

Silent Genes: Antimicrobial Resistance and Antibiotic Production

<abstract>
<title style='display:none'>Abstract</title>
This study aimed to elucidate the influence of various culture medium components, including carbon sources, nitrogen sources, inorganic salts, suspension agents, and temperature, on the mycelial growth characteristics of <italic>Phallus dongsun</italic>. Employing single-factor experiments and response surface methodology within glass Petri dishes, the research identified that carrot powder, soybean powder, and ZnSO4 notably enhanced the proliferation of aerial mycelium, significantly augmenting the growth rate of <italic>P. dongsun</italic> mycelium. The resultant mycelium was observed to be dense, robust, and fluffy in texture. In particular, ZnSO4 markedly accelerated the mycelium growth rate. Furthermore, xanthan gum was found to effectively modulate the medium’s viscosity, ensuring a stable suspension and facilitating nutrient equilibrium. The optimal cultivation temperature was determined to be 25°C, with mycelial growth ceasing below 5°C and mycelium perishing at temperatures exceeding 35°C. The optimal medium composition was established as follows: wheat starch 5 g/l, carrot powder 5 g/l, soybean powder 7.50 g/l, glucose 10 g/l, ZnSO4 0.71 g/l, NH4Cl 0.68 g/l, xanthan gum 0.5 g/l, and agar 20 g/l. Under these optimized conditions, the mycelium of <italic>P. dongsun</italic> exhibited a rapid growth rate (1.04 ± 0.14 mm/day), characterized by a thick, dense, and well-developed structure. This investigation provides a theoretical foundation for the conservation, strain selection, and breeding of <italic>P. dongsun</italic>.
</abstract>

Biological Characteristics of the Mycelium and Optimization of the Culture Medium for <italic>Phallus dongsun</italic>

<abstract>
<title style='display:none'>Abstract</title>
Serological testing can be a powerful complementary approach to achieve timely diagnosis of severe acute respiratory coronavirus 2 (SARS-CoV-2) infection, along with nucleic acid detection. Immunoglobulin (Ig) A antibodies are less frequently utilized to detect SARS-CoV-2 infection than IgM and IgG antibodies, even though IgA antibodies play an important role in protective immunity against SARS-CoV-2. This review discusses the differences in kinetics and assay performance between IgA and IgM antibodies and the factors influencing antibody responses. It highlights the potential usefulness of analyzing IgA antibodies for the early detection of SARS-CoV-2 infection. The early appearance of IgA and the high sensitivity of IgA-based immunoassays can aid in diagnosing coronavirus disease 2019. However, because of cross-reactivity, it is important to recognize the only moderate specificity of the early detection of SARS-CoV-2 IgA antibodies against spike antigens. Either the analysis of antibodies targeting the nucleocapsid antigen or a combination of antibodies against the nucleocapsid and spike antigens may strengthen the accuracy of serological evaluation.
</abstract>

Potential Usefulness of IgA for the Early Detection of SARS-CoV-2 Infection: Comparison With IgM

<abstract>
<title style='display:none'>Abstract</title>
To establish a rapid detection method for norovirus GII.2 genotype, this study employed reverse transcription recombinase polymerase amplification (RT-RPA) combined with CRISPR/Cas12a and lateral flow strip (RT-RPA-Cas12a-LFS). Here, the genome of norovirus GII.2 genotype was compared to identify highly conserved sequences, facilitating the design of RT-RPA primers and crRNA specific to the conserved regions of norovirus GII.2. Subsequently, the reaction parameters of RT-RPA were optimized and evaluated using agar-gel electrophoresis and LFS. The results indicate that the conserved sequences of norovirus GII.2 were successfully amplified through RT-RPA at 37°C for 25 minutes. Additionally, CRISPR/Cas12a-mediated cleavage detection was achieved through LFS at 37°C within 10 minutes using the amplification products as templates. Including the isothermal amplification reaction time, the total time is 35 minutes. The established RT-RPA-Cas12a-LFS method demonstrated specific detection of norovirus GII.2, yielding negative results for other viral genomes, and exhibited an excellent detection limit of 10 copies/μl. The RT-RPA-Cas12a-LFS method was further compared with qRT-PCR by analyzing 60 food-contaminated samples. The positive conformity rate was 100%, the negative conformity rate was 95.45%, and the overall conformity rate reached 98.33%. This detection method for norovirus GII.2 genotype is cost-effective, highly sensitive, specific, and easy to operate, offering a promising technical solution for field-based detection of the norovirus GII.2 genotype.
</abstract>

Establishment of a Nucleic Acid Detection Method for Norovirus GII.2 Genotype Based on RT-RPA and CRISPR/Cas12a-LFS

<abstract>
<title style='display:none'>Abstract</title>
Interferon-alpha (IFN-α) is a first-line drug for treating chronic hepatitis B (CHB). Guanylate-binding protein 1 (GBP1) is one of the interferon-stimulating factors, which participates in the innate immunity of the host and plays an antiviral and antibacterial role. In this study, we explored how GBP1 is involved in IFN-α antiviral activity against HBV. Before being gathered, HepG2-NTCP and HepG2 2.15 cells were transfected with the wild-type hGBP1 plasmid or si-GBP1, respectively, and followed by stimulation with Peg-IFNα-2b. We systematically explored the role of GBP1 in regulating HBV infection in cell models. Additionally, we also examined GBP1 levels in CHB patients. GBP1 activity increased, and its half-life was prolonged after HBV infection. Overexpression of GBP1 inhibited the production of HBsAg and HBeAg, as well as HBs protein and HBV total RNA levels, whereas silencing of GBP1 inhibited its ability to block viral infections. Interestingly, overexpressing GBP1 co-treatment with Peg-IFNα-2b further increased the antiviral effect of IFN-α, while GBP1 silencing co-treatment with Peg-IFNα-2b partly restored its inhibitory effect on HBV. Mechanistically, GBP1 mediates the anti-HBV response of Peg-IFNα-2b by targeting HBs. Analysis of clinical samples revealed that GBP1 was elevated in CHB patients and increased with Peg-IFNα-2b treatment, while GBP1 showed good stability in the interferon response group. Our study demonstrates that GBP1 inhibits HBV replication and promotes HBsAg clearance. It is possible to achieve antiviral effects through the regulation of IFN-α induced immune responses in response to HBV.
</abstract>

Guanylate-Binding Protein 1 (GBP1) Enhances IFN-α Mediated Antiviral Activity against Hepatitis B Virus Infection

<abstract>
<title style='display:none'>Abstract</title>
Chikungunya virus (CHIKV) causes a debilitating fever and joint pain, with no specific antiviral treatment available. Halogenated secondary metabolites from plants are a promising new class of drug candidates against chikungunya, with unique properties that make them effective against the virus. Plants produce these compounds to defend themselves against pests and pathogens, and they are effective against a wide range of viruses, including chikungunya. This study investigated the interactions of halogenated secondary metabolites with nsP2pro, a therapeutic target for CHIKV. A library of sixty-six halogenated plant metabolites screened previously for ADME properties was used. Metabolites without violation of Lipinski’s rule were docked with nsP2pro using AutoDock Vina. To find the stability of the pipoxide chlorohydrin-nsP2pro complex, the GROMACS suite was used for MD simulation. The binding free energy of the ligand-protein complex was computed using MMPBSA. Molecular docking studies revealed that halogenated metabolites interact with nsP2pro, suggesting they are possible inhibitors. Pipoxide chlorohydrin showed the greatest affinity to the target. This was further confirmed by the MD simulations, surface accessible area, and MMPBSA studies. Pipoxide chlorohydrin, a halogenated metabolite, was the most potent against nsP2pro in the survey.<fig id="j_pjm-2024-020_fig_001" position="float" fig-type="figure">
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</fig>
</abstract>

Halogenated Secondary Metabolites from Higher Plants: Potent Drug Candidates for Chikungunya Using <italic>in silico</italic> Approaches

<abstract>
<title style='display:none'>Abstract</title>
Negative Pressure Wound Therapy (NPWT) has been widely adopted in wound healing strategies due to its multimodal mechanism of action. While NPWT’s positive impression on wound healing is well-established, its effect on bacterial load reduction remains equivocal. This study investigates NPWT’s efficacy in reducing bioburden using an <italic>in vitro</italic> porcine skin model, focusing on the impact of <italic>Staphylococcus aureus</italic> and <italic>Staphylococcus epidermidis</italic>. Custom-made negative pressure chambers were employed to apply varying negative pressures. Porcine skin was cut into 5 × 5 cm squares and three standardized wounds of 6 mm each were created using a biopsy punch. Then, wounds were infected with <italic>S. aureus</italic> and <italic>S. epidermidis</italic> bacterial suspensions diluted 1:10,000 to obtain a final concentration of 1.5 × 104 CFU/ml and were placed in negative pressure chambers. After incubation, bacterial counts were expressed as colony-forming units (CFU) per ml. For <italic>S. aureus</italic> at 120 hours, the median CFU, mean area per colony, and total growth area were notably lower at −80 mmHg when compared to −250 mmHg and −50 mmHg, suggesting an optimal negative pressure for the pressure-dependent inhibition of the bacterial proliferation. While analyzing <italic>S. epidermidis</italic> at 120 hours, the response to the negative pressure was similar but less clear, with the minor CFU at −100 mmHg. The influence of intermittent negative pressure on the <italic>S. epidermidis</italic> growth showed notably lower median CFU with the interval therapy every hour compared to the <italic>S. aureus</italic> control group. This study contributes valuable insights into NPWT’s influence on the bacterial load, emphasizing the need for further research to reformulate its role in managing contaminated wounds.
<fig id="j_pjm-2024-018_fig_001" position="float" fig-type="figure">
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</fig>
</abstract>

Negative Pressure Level and Effects on Bacterial Growth Kinetics in an <italic>in vitro</italic> Wound Model

<abstract><title style='display:none'>Abstract</title>Urinary tract infections (UTIs) belong to the most common community-acquired and nosocomial infections. A main etiological factor of UTIs is uropathogenic <italic>Escherichia coli</italic> (UPEC). This review describes the current state of knowledge on the resistance of UPEC to antibiotics recommended for the treatment of UTIs based on the available literature data. Nitrofurantoin and fosfomycin are recommended as first-line therapy in the treatment of uncomplicated cystitis, and the resistance to these antimicrobial agents remains low between UPEC. Recently, in many countries, the increasing resistance is observed to trimethoprim-sulfamethoxazole, which is widely used as the first-line antimicrobial in the treatment of uncomplicated UTIs. In European countries, the resistance of UPEC to this antimicrobial agent ranges from 14.6% to 60%. The widespread use of fluoroquinolones (FQs), especially ciprofloxacin, in the outpatients is the cause of a continuous increase in resistance to these drugs. The resistance of UPEC to FQs is significantly higher in developing countries (55.5–85.5%) than in developed countries (5.1–32.0%). Amoxicillin-clavulanic acid is recommended as first line-therapy for pyelonephritis or complicated UTI. Resistance rates of UPEC to amoxicillin-clavulanic acid are regionally variable. In European countries the level of resistance to this antimicrobial ranges from 5.3% (Germany) to 37.6% (France). Increasing rates of UPEC resistance to antimicrobials indicate that careful monitoring of their use for UTI treatment is necessary.</abstract>

Antibiotic Resistance Among Uropathogenic <italic>Escherichia coli</italic>

<abstract><title style='display:none'>Abstract</title><italic>Brucellae</italic> are Gram-negative, small rods infecting mammals and capable of causing disease called brucellosis. The infection results in abortion and sterility in domestic animals (sheeps, pigs, rams etc). Especially dangerous for humans are: <italic>Brucella melitensis, Brucella suis, Brucella abortus</italic>, and <italic>Brucella canis</italic> that trigger unspecific symptoms (flu-like manifestation). <italic>Brucella</italic> rods are introduced via host cells, by inhalation, skin abrasions, ingestion or mucosal membranes. The most important feature of <italic>Brucella</italic> is the ability to survive and multiply within both phagocytic and non-phagocytic cells. <italic>Brucella</italic> does not produce classical virulence factors: exotoxin, cytolisins, exoenzymes, plasmids, fimbria, and drug resistant forms. Major virulence factors are: lipopolysaccharide (LPS), T4SS secretion system and BvrR/BvrS system, which allow interaction with host cell surface, formation of an early, late BCV (<italic>Brucella</italic> Containing Vacuole) and interaction with endoplasmic reticulum (ER) when the bacteria multiply. The treatment of brucellosis is based on two-drug therapy, the most common combinations of antibiotics are: doxycycline with rifampicin or fluoroquinolones with rifampicin. Currently, also other methods are used to disrupt <italic>Brucella</italic> intracellular replication (tauroursodeoxycholic acid or ginseng saponin fraction A).</abstract>

<italic>Brucella</italic> – Virulence Factors, Pathogenesis and Treatment

<abstract><title style='display:none'>Abstract</title>The growing resistance of microorganisms towards antibiotics has become a serious global problem. Therapeutics with novel chemical scaffolds and/or mechanisms of action are urgently needed to combat infections caused by multidrug resistant pathogens, including bacteria, fungi and viruses. Development of novel antimicrobial agents is still highly dependent on the discovery of new natural products. At present, most antimicrobial drugs used in medicine are of natural origin. Among the natural producers of bioactive substances, Actinobacteria continue to be an important source of novel secondary metabolites for drug application. In this review, the authors report on the bioactive antimicrobial secondary metabolites of Actinobacteria that were described between 2011 and April 2018. Special attention is paid to the chemical scaffolds, biological activities and origin of these novel antibacterial, antifungal and antiviral compounds. Arenimycin C, chromopeptide lactone RSP 01, kocurin, macrolactins A1 and B1, chaxamycin D as well as anthracimycin are regarded as the most effective compounds with antibacterial activity. In turn, the highest potency among selected antifungal compounds is exhibited by enduspeptide B, neomaclafungins A-I and kribelloside D, while ahmpatinin iBu, antimycin A1a, and pentapeptide 4862F are recognized as the strongest antiviral agents.</abstract>

Secondary Metabolites of Actinomycetes and their Antibacterial, Antifungal and Antiviral Properties

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 eISSN: 2544-4646Print ISSN: 1733-1331   The Polish Journal of Microbiology (PJM), founded...

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