Anthelminthic and antimicrobial effects of hedge woundwort (Stachys sylvatica L.) growing in Southern Kazakhstan.

The Stachys L. genus has been widely used in traditional medicine in many countries throughout the world. The study aimed to investigate the chemical composition and bioactivity of the hydroethanolic extract (50% v/v) obtained by ultrasonication from the aerial flowering parts of Stachys sylvatica L. (SSE) collected in Almaty region (Southern Kazakhstan). According to RP-HPLC/PDA analysis the leading metabolites of the SSE belonged to polyphenols: chlorogenic acid and its isomers (2.34 mg/g dry extract) and luteolin derivatives (1.49 mg/g dry extract), while HPLC-ESI-QTOF-MS/MS-based qualitative fingerprinting revealed the presence of 17 metabolites, mainly chlorogenic acid and its isomers, flavonoid glycosides, and verbascoside with its derivatives. GC-MS analysis of the volatile metabolites showed mainly the presence of diterpenoids and fatty acid esters. A reduction in the viability of nematodes Rhabditis sp. was obtained for the SSE concentration of 3.3 mg/mL, while 11.1 mg/mL showed activity comparable to albendazole. The SSE exhibited higher activity against Gram-positive (MIC = 0.5-2 mg/mL) than Gram-negative bacteria and yeast (MIC = 8 mg/mL), exerting bactericidal and fungicidal effects but with no sporicidal activity. The SSE showed some antiviral activity against HCoV-229E replicating in MRC-5 and good protection against the cytopathic effect induced by HHV-1 in VERO. The SSE was moderately cytotoxic towards human cervical adenocarcinoma (H1HeLa) cells (CC50 of 0.127 mg/mL after 72 h). This study provides novel information on the SSE extract composition and its biological activity, especially in the context of the SSE as a promising candidate for further antiparasitic studies.

Modified polymeric biomaterials with antimicrobial and immunomodulating properties.

The modification of the surgical polypropylene mesh and the polytetrafluoroethylene vascular prosthesis with cecropin A (small peptide) and puromycin (aminonucleoside) yielded very stable preparations of modified biomaterials. The main emphasis was placed on analyses of their antimicrobial activity and potential immunomodulatory and non-cytotoxic properties towards the CCD841 CoTr model cell line. Cecropin A did not significantly affect the viability or proliferation of the CCD 841 CoTr cells, regardless of its soluble or immobilized form. In contrast, puromycin did not induce a significant decrease in the cell viability or proliferation in the immobilized form but significantly decreased cell viability and proliferation when administered in the soluble form. The covalent immobilization of these two molecules on the surface of biomaterials resulted in stable preparations that were able to inhibit the multiplication of Staphylococcus aureus and S. epidermidis strains. It was also found that the preparations induced the production of cytokines involved in antibacterial protection mechanisms and stimulated the immune response. The key regulator of this activity may be related to TLR4, a receptor recognizing bacterial LPS. In the present study, these factors were produced not only in the conditions of LPS stimulation but also in the absence of LPS, which indicates that cecropin A- and puromycin-modified biomaterials may upregulate pathways leading to humoral antibacterial immune response.

Multi-Host Pathogen Staphylococcus aureus-Epidemiology, Drug Resistance and Occurrence in Humans and Animals in Poland.

Staphylococcus aureus is a drug resistant pathogen with zoonotic potential commonly isolated from humans and animals. The aim of this study was to compare the occurrence of drug resistance, resistance genes, sequence types (STs), and genotypes of S. aureus isolated from humans, livestock, and wildlife in eastern Poland. A high percentage of isolates resistant to penicillin (63%), erythromycin (39%), clindamycin (37%), tetracycline (31%), and methicillin (MRSA-19%), an intermediate resistant to vancomycin (VISA-13%), and a multidrug resistant (MDR-39%) was obtained. Multilocus sequence typing analysis showed the presence of 35 different STs (with dominance ST 15, ST 45, ST 7, and ST 582 in human, and ST 398 and ST 8139 in porcine and cattle isolates, respectively), including 9 new ones that had never been reported before (ST 8133-8141). Identical genotypic patterns were detected among porcine and cattle isolates as well as from humans and cattle. A high percentage of MDR, MRSA, and VISA in humans and livestock combined with the presence of the same genotypes among S. aureus isolated from human and cattle indicates the circulation of strains common in the region and their zoonotic potential. There is a need to develop new strategies to counteract this phenomenon according to the One Health policy.

The Epidemiology of Animal-Associated Methicillin-Resistant Staphylococcus aureus.

Methicillin-resistant Staphylococcus aureus (MRSA) remains an important etiological factor of human and animal infectious diseases, causing significant economic losses not only in human healthcare but also in the large-scale farming sector. The constantly changing epidemiology of MRSA observed globally affects animal welfare and raises concerns for public health. High MRSA colonization rates in livestock raise questions about the meaning of reservoirs and possible transmission pathways, while the prevalence of MRSA colonization and infection rates among companion animals vary and might affect human health in multiple ways. We present the main findings concerning the circulation of animal-associated MRSA (AA-MRSA) in the environment and factors influencing the direction, mechanisms, and routes of its transmission. Studies have shown it that S. aureus is a multi-host bacterial pathogen; however, its adaptation mechanisms enabling it to colonize and infect both animal and human hosts are still rarely discussed. Finally, we elaborate on the most successful strategies and programs applied limiting the circulation of AA-MRSA among animals and humans. Although MRSA strains colonizing animals rarely infect humans, they undergo host-adaptive evolution enabling them to spread and persist in human populations.

Assessment of health safety of pigs taking natural sorbents with feed.

BACKGROUND: The study assessed the effect of smectites (bentonite and zeolite) used as natural sorbents in the diet of pigs on feed digestibility, health parameters, the severity of anatomo-histological changes in organs, and the accumulation of volatile pollutants in organs. MATERIAL AND METHODS: The study was conducted using fattening pigs (crossbreds from multiple breeds) assigned to three groups - a control (C) and two experimental groups (A and B), with 240 pigs in each group (3 replicates × 80). The animals in group C received a standard complete diet, while groups A and B received diets with 1.5% composed smectite sorbents. The feed and faeces were analysed for content of dry matter, crude ash, crude protein, ether extract, and crude fibre. The content of P was determined using a Helios Alpha UV-VIS spectrophotometer. Whole blood was analysed for haematological parameters and serum for biochemical parameters. Tissue samples were collected for analysis of volatile substances and histological analysis. After slaughter, samples of the lungs, liver, kidneys and jejunum were collected for morphological evaluation, and samples of the perirenal fat, liver, kidneys, lungs and brain for headspace gas chromatography (GC) to determine the levels of volatile toxic substances. RESULTS: A statistical increase in the digestibility of crude fibre and an increase in that of P were observed in both experimental groups (A and B) in comparison to the control. The whole blood and serum of the pigs from the control group had statistically significantly higher levels of creatinine, urea, and Mg and a higher WBC count compared to both experimental groups (A and B). CONCLUSIONS: The feed additives were not shown to have a negative effect on the health parameters analysed or on accumulation of pollutants in selected tissues. No significant effect on the digestibility of most nutrients was observed; only an increase in the digestibility of crude fibre and a decrease in P digestibility were noted in the experimental groups.

Phytochemical Profile and Biological Activity of the Ethanolic Extract from the Aerial Part of Crocus alatavicus Regel & Semen Growing Wildly in Southern Kazakhstan.

The composition of the ethanolic extract from the aerial parts of Crocus alatavicus Regel & Semen from southern Kazakhstan spontaneous flora was analyzed together with the determination of its antibacterial, antifungal, antiviral and anticancer activity. The phytochemical profile analysis by high-performance liquid chromatography-electrospray ionization-quadrupole-time of flight-mass spectrometry (HPLC/ESI-QTOF-MS) revealed the presence of multiple kaempferol derivatives. High-performance reverse-phase liquid chromatography combined with a photodiode-array detection (RP-HPLC/PDA) found that kaempferol 3-O-dihexoside and kaempferol 3-O-acyltetrahexoside accounted for 70.5% of the kaempferol derivatives. The minimum inhibitory concentration (MIC) values of the extract for all the tested reference microorganisms were high, reaching 10 mg/mL for yeasts and 20 mg/mL for bacteria. In contrast, antiviral activity was observed at 2 mg/mL, resulting in the inhibition of the HSV-1-induced cytopathic effect and the reduction in virus infectious titer by 1.96 log, as well as the viral load by 0.85 log. Among the tested prostate cancer cell lines, significant cytotoxic activity of the extract was noted only on the LNCaP cell line, with an IC50 value of 1.95 mg/mL. The LNCaP cell line treated with 2 mg/mL of the extract showed a noticeably reduced number of spindle-shaped cells with longer cellular projections, a significant increase in the peak corresponding to the population of apoptotic cells in the sub-G1 phase and a decreased intracellular glutathione (GSH) level, suggesting the prooxidative properties of the extract. The obtained data provide novel information about the flavonoids present in the aerial part of C. alatavicus and suggest its potential application as a source of the compounds active against HSV-1 and metastatic, androgen-sensitive prostate cancer.

The Landscape of Gene Expression during Hyperfilamentous Biofilm Development in Oral Candida albicans Isolated from a Lung Cancer Patient.

The filamentation ability of Candida albicans represents one of the main virulence factors allowing for host tissue penetration and biofilm formation. The aim of this paper was to study the genetic background of the hyperfilamentous biofilm development in vitro in C. albicans isolated from the oral cavity of a lung cancer patient. Analyzed C. albicans isolates (CA1, CA2, CA3) were chosen based on their different structures of mature biofilm. The CA3 isolate possessing hyperfilamentation properties and forming high biofilm was compared with CA1 and CA2 isolates exhibiting low or average biofilm-forming ability, respectively. The detailed biofilm organization was studied with the use of confocal scanning laser microscopy. The whole transcriptome analysis was conducted during three stages of biofilm development (24 h, 48 h, 72 h). In contrast to CA1 and/or CA2 isolate, the CA3 isolate was characterized by a significant upregulation of genes encoding for cell wall proteins (HWP1, PGA13, PGA44, ALS3) and candidalysin (ECE1), as well as being involved in iron metabolism (FRE1, ALS3), sulfur metabolism (HAL21), the degradation of aromatic compounds (HQD2), and membrane transport (DIP5, PHO89, TNA1). In contrast, some genes (SCW11, FGR41, RBE1) in the CA3 were downregulated. We also observed the overexpression of a few genes over time-mainly FRE1, ATX1, CSA2 involved in iron metabolism. This is the first insight into the potential function of multiple genes in the hyperfilamentous biofilm formation in C. albicans, primarily isolated from host tissue, which may have an important clinical impact on cancer patients. Moreover, the presented data can lay the foundation for further research on novel pathogen-specific targets for antifungal drugs.

The Prevalence of Virulence Determinants and Antibiotic Resistance Patterns in Methicillin-Resistant Staphylococcus aureus in a Nursing Home in Poland.

Nursing homes (NH) contribute to the regional spread of methicillin-resistant Staphylococcus aureus (MRSA). Moreover, residents are vulnerable to the colonization and subsequent infection of MRSA etiology. We aimed at investigating the molecular and phenotypic characteristics of 21 MRSA collected from the residents and personnel in an NH (Lublin, Poland) during 2018. All MRSA were screened for 20 genes encoding virulence determinants (sea-see, eta, etb, tst, lukS-F-PV, eno, cna, ebpS, fib, bbp, fnbA, fnbB, icaADBC) and for resistance to 18 antimicrobials. To establish the relatedness and clonal complexes of MRSA in NH we applied multiple-locus variable-number tandem-repeat fingerprinting (MLVF), pulse field gel electrophoresis (PFGE), multilocus sequence typing (MLST) and staphylococcal cassette chromosome mec (SCCmec) typing. We identified four sequence types (ST) among two clonal complexes (CC): ST (CC22) known as EMRSA-15 as well as three novel STs-ST6295 (CC8), ST6293 (CC8) and ST6294. All tested MRSA were negative for sec, eta, etb, lukS-F-PV, bbp and ebpS genes. The most prevalent gene encoding toxin was sed (52.4%; n = 11/21), and adhesins were eno and fnbA (100%). Only 9.5% (n = 2/21) of MRSA were classified as multidrug-resistant. The emergence of novel MRSA with a unique virulence and the presence of epidemic clone EMRSA-15 creates challenges for controlling the spread of MRSA in NH.

Transmission and Long-Term Colonization Patterns of Staphylococcus aureus in a Nursing Home.

Nursing homes might create an environment favorable for the transmission of Staphylococcus aureus because of the presence of hospitalized elderly, overcrowding and close contacts among people. We aimed at identifying risk factors for S. aureus colonization and determining the genetic relatedness of isolates demonstrating transmission among people. We investigated 736 swab samples from 92 residents and personnel for the presence of S. aureus. Swabs from anterior nares and throat were collected quarterly (2018) in a nursing home located in Poland. Genotyping was conducted using the multi-locus variable number of tandem repeats fingerprinting (MLVF) method. We observed high seasonal variation in the proportion of participants colonized with methicillin-resistant Staphylococcus aureus (MRSA) strains (0% to 13.5%). A multivariate analysis revealed that residents aged more than 85 years old are at risk for becoming intermittent S. aureus carriers (p = 0.013). The MLVF analysis revealed a high genetic diversity among methicillin-sensitive S. aureus (MSSA) strains and close genetic relatedness between MRSA strains. We proved the advanced aged were predisposed to intermittent S. aureus carriage. Genotyping revealed the transmission of S. aureus among the participants living in a closed environment. A high genetic relatedness among isolated MRSA suggests its clonal spread in the nursing home.

Molecular Diversity of Staphylococcus aureus Colonizing the Upper Respiratory Tract of Residents and Staff in A Nursing Home.

Elderly people living in nursing homes are a high-risk population for Staphylococcus aureus infection. Multiple comorbidities, a weakened immune system, inadequate hygienic conditions, and crowding might increase the prevalence rates of this opportunistic pathogen. However, the epidemiological aspects, genetic diversity, and transmission of S. aureus in nursing homes are still poorly understood, especially in Poland. This study aimed to determine the genetic relatedness and prevalence of colonization of S. aureus isolated from the anterior nares and the throat of residents and staff in a nursing home located in Lublin, Poland. The study showed a high S. aureus prevalence rate among participants (46.1%), yet there was a low frequency of MRSA strains among residents (1.7%) and staff (0%). The multiple-locus variable-number tandem-repeat fingerprinting (MLVF) analysis demonstrated a high degree of genetic diversity of S. aureus strains colonizing the anterior nares and the throat of the participants. The occurrence of simultaneous colonization with more than one unique S. aureus strain in any one individual as well as the incidence of colonization with the same genetic variant of S. aureus in different individuals was observed. These findings suggest that inter-participant S. aureus transmission might contribute to the development of cross-infections.Elderly people living in nursing homes are a high-risk population for Staphylococcus aureus infection. Multiple comorbidities, a weakened immune system, inadequate hygienic conditions, and crowding might increase the prevalence rates of this opportunistic pathogen. However, the epidemiological aspects, genetic diversity, and transmission of S. aureus in nursing homes are still poorly understood, especially in Poland. This study aimed to determine the genetic relatedness and prevalence of colonization of S. aureus isolated from the anterior nares and the throat of residents and staff in a nursing home located in Lublin, Poland. The study showed a high S. aureus prevalence rate among participants (46.1%), yet there was a low frequency of MRSA strains among residents (1.7%) and staff (0%). The multiple-locus variable-number tandem-repeat fingerprinting (MLVF) analysis demonstrated a high degree of genetic diversity of S. aureus strains colonizing the anterior nares and the throat of the participants. The occurrence of simultaneous colonization with more than one unique S. aureus strain in any one individual as well as the incidence of colonization with the same genetic variant of S. aureus in different individuals was observed. These findings suggest that inter-participant S. aureus transmission might contribute to the development of cross-infections.