Zoonotic bacteria in the vicinity of animal farms as a factor disturbing the human microbiome: a review.

This review is aimed at summarizing the current state of knowledge about the relationship between environmental exposure to the bioaerosol emitted by intensive livestock farming and changes in the microbiome of people living in livestock farm vicinity. The PubMed, Scopus and Web of Science databases were searched by crossing keywords from the following 3 groups: a) "livestock," "animal farms," "animal breeding"; b) "microbiome," "resistome"; c) "livestock vicinity," "farm vicinity," "neighborhoods and health" in 2010-2022. Literature screening did not reveal any paper related to the full microbiome composition in the population studied. In the study, the authors included 7 papers (5 from the Netherlands, 1 from the USA, and 1 from China). The studies confirmed the carriage of Staphylococcus aureus, including methicillin-resistant S. aureus (MRSA), livestockassociated MRSA (LA-MRSA MC398) and multidrug-resistant S. aureus (MDRSA) in the nasal microbiome of adults and children living within 500-2000 m from a livestock farm. Clostridium difficile, including LA-ribotype RT078 carriage, was detected in the intestinal microbiome of adults living within 500-1000 m. Extended-spectrum ß-lactamase (ESBL) producing Enterobacteriaceae were confirmed in the intestinal microbiome of adults living within 500-6200 m. Knowledge on the composition of the microflora of people living in livestock farm vicinity is insufficient to conclude about changes in the microbiome caused by the environmental emission of bioaerosol. The carriage prevalence of the LA-bacteria, including both strains with antimicrobial resistance and antimicrobial resistance genes, confirms the presence of zoonotic bacteria in the human microflora in populations without occupational contact with animals. It cannot be ruled out that zoonotic bacteria, as a component of the microbiome, have a negative impact on people's health. Int J Occup Med Environ Health. 2024;37(2):138-52.

Occurrence and spread of antibiotic-resistant bacteria on animal farms and in their vicinity in Poland and Ukraine-review.

Intensive animal farming emits to the environment very high concentrations of bioaerosol, mainly composed of microorganisms, including antibiotics resistant strains, and their derivatives. Poland is a significant producer of poultry and swine in Europe; Ukraine is located in the immediate vicinity of Poland and the EU. Thus, the review focuses on the presence of potentially pathogenic and antimicrobial-resistant zoonotic bacteria and antimicrobial genes in the environment of farms and food of animal origin in Poland and Ukraine. Existing data confirms presence of these bacteria in the food animal origin chain environment in both countries. However, it is difficult to compare the scale of multidrug-resistant bacteria (e.g. MRSA, ESBL) dissemination in Poland and Ukraine with other EU countries due to lack of more extensive studies and large-scale monitoring in these two countries. A series of studies concerning resistance of pathogenic bacteria isolated from livestock environment have been published in Poland but usually on single farms with a very limited number of samples, and without a genotypic drug resistance marking. From Ukraine are available only few reports, but also disturbing. The risk of antibiotic-resistant bacteria transmission does not only concern animal farming, but also other facilities of animal origin food supply chains, especially slaughterhouses.

[Harmful biological agents intentionally used in enterprises in Poland based on the National Register of Biological Agents]. / Szkodliwe czynniki biologiczne stosowane celowo w zakladach pracy w Polsce na podstawie danych z Krajowego Rejestru Czynników Biologicznych.

BACKGROUND: This work presents intentional use of harmful biological agents based on the National Register of Biological Agents (in Polish abbreviated as KRCB) as of July 2020. MATERIAL AND METHODS: Generally, KRCB is a central database gathering notifications of intentional use of biological agents classified as risk groups 2-4 at work. RESULTS: Notifications were sent by 672 enterprises, including for diagnostic - 63%, research - 30% and industrial purposes - 7%. The largest number of notifications were sent by hospital-based diagnostic laboratories - 34%, laboratories other than hospital-based - 34%, and higher education and research units - 15%. In total, 7077 workers (88% women, 12% men) were exposed to biological agents intentionally used at work. The following bacteria were most frequently used: Escherichia coli (with the exception of non-pathogenic strains) - 4394 (62%) exposed workers, Staphylococcus aureus -4122 (58%), and Pseudomonas aeruginosa - 3726 (53%). Biological agents recognized by the International Agency for Research on Cancer as carcinogenic were used in 107 enterprises (16%), including the following viruses: hepatitis B virus (HBV) - 25 enterprises (4%), hepatitis C virus (HCV) - 24 (4%), human immunodeficiency virus type 1 (HIV-1) - 18 (3%), Epstein-Barr virus (EBV) - 10 (1%), human papilloma virus (HPV) - 5 (1%), human herpes virus type 8 (HHV-8) - 3 (<1%), human T-lymphotropic virus type 1 (HTLV-1) - 3 (<1%), as well as bacteria Helicobacter pylori - 18 (3%) and parasites Schistosoma haematobium - 1 (<1%). CONCLUSIONS: While KRCB is a unique source of information on biological agents intentionally used in workplaces, it also provides an important link in the chain of information on occupational exposure to harmful agents in Poland. Med. Pr. 2021;72(2):131-43.

Occupational exposure to Staphylococcus aureus in the wastewater treatment plants environment.

BACKGROUND: The aim of the study was to assess the occupational exposure to Staphylococcus aureus bacteria, including methicillin- resistant S. aureus (MRSA) and other antibiotic-resistant strains in the municipal wastewater treatment plants (WWTPs) environment. MATERIAL AND METHODS: In 16 WWTPs in Poland, 33 wastewater and 253 air samples were collected in the spring-summer season. The microbiological analysis was carried out using a chromogenic medium. Species identification was carried out using the matrix assisted laser desorption ionization time-of-flight method, while the antibiotic-resistance analysis was performed with an automatic method. RESULTS: Among 2805 bacterial isolates from the air and wastewater, 574 were identified as S. aureus species (20.5%). The presence of S. aureus species was found in 11 WWTPs (69%), among them in 11 WWTPs in raw wastewater and in 1 WWTP additionally in treated wastewater. The concentrations of S. aureus in wastewater ranged 2-1215 colony-forming units per milliliter (CFU/ml). In the air, 2 S. aureus isolates were identified in concentrations of 5 and 10 CFU/m3; both samples were collected at the stage of mechanical wastewater treatment. The results revealed the following trend: the higher the outdoor temperature, the bigger the number of WWTPs with confirmed S. aureus presence. Among 149 S. aureus isolates (2 from the air and 147 from wastewater, including 2 MRSA), 100 isolates were resistant only to penicillin, while 34 isolates showed multi-antibiotic resistance (to penicillin and other drugs). It was found that isolated bacteria were resistant almost strictly to critical and highly important antibiotics in veterinary medicine. CONCLUSIONS: In general, WWTPs workers are occupationally exposed to S. aureus, including MRSA, and other antibiotic- and multi-antibiotic-resistant strains. The highest risk of infection concerns the activities carried out in direct contact with wastewater or devices through which wastewater flows, particularly at the stage of mechanical treatment. A significant source of S. aureus seems to be intensive livestock farming located in the area of the WWTPs under analysis. The study confirms the necessity to disinfect the wastewater discharging into WWTPs. Med Pr. 2020;71(3):265-78.

Combined effect of silver nanoparticles and aluminium chloride, butylparaben or diethylphthalate on the malignancy of MDA-MB-231 breast cancer cells and tumor-specific immune responses of human macrophages and monocyte-derived dendritic cells.

The aim of this study was to assess whether silver nanoparticles (AgNP) or selected cosmetic ingredients may modify functions of various immunocompetent cell populations. To this end, the effect of two AgNP (size of 15 nm or 45 nm), alone and in combination with aluminium chloride, butyl paraben, di-n-butyl phthalate or diethyl phthalate was assessed on: (1) migration and invasion of MDA-MB-231 human breast cancer cells; (2) M1/M2 polarization of phorbol 12-myristate 13-acetate (PMA)-differentiated THP-1 macrophages (M0) and (3) activation/maturation of monocyte-derived dendritic cells (DCs). The results of this study showed that neither any of the test chemicals alone nor the mixtures significantly changed the migration or invasion ability of MDA-MB-231 cells following, both 72-h and 21-day exposure. Analysis of the expression of marker genes for both M1 (IL-1B, CXCL9, TNF) and M2 (DCSIGN, MRC1) polarization revealed that the chemicals/mixtures did not activate M1/M2 differentiation of the M0 macrophages. In addition, no significant changes were observed in the expression of CD86, HLA-DR and CD54 surface markers and phagocytic activity of DCs following 48-h exposure to AgNP alone or in combination with test compounds. Our study suggests that AgNP alone or in combination with tested cosmetic ingredients do not alter function of immunocompetent cells studied.

Airborne Staphylococcus aureus in different environments-a review.

The aim of the literature review was to describe the environments where the presence of airborne Staphylococcus aureus was confirmed and to catalogue the most often used methods and conditions of bioaerosol sampling to identify the bacteria. The basis for searching of studies on S. aureus in the bioaerosol in different environments was PubMed database resources from the years 1990-2019 (May). The review included studies which were carried on in selected environments: hospitals and other health care facilities, large-scale animal breeding, wastewater treatment plants, residential areas, educational institutions, and other public places. The highest concentrations and genetic diversity of identified S. aureus strains, including MRSA (methicillin-resistant S. aureus), have been shown in large-scale animal breeding. The role of the airborne transmission in dissemination of infection caused by these pathogens is empirically confirmed in environmental studies. Commonly available, well-described, and relatively inexpensive methods of sampling, identification, and subtyping guarantee a high reliability of results and allow to obtain fast and verifiable outcomes in environmental studies on air transmission routes of S. aureus strains.

Does the air condition system in busses spread allergic fungi into driver space?

The aim of this study was to establish whether the air-conditioning system in buses constitutes an additional source of indoor air contamination with fungi, and whether or not the fungi concentration depends on the period from the last disinfection of the system, combined with replacement of the cabin dust particle filter. The air samples to fungi analysis using impact method were taken in 30 buses (20 with an air-conditioning system, ACS; 10 with a ventilation system, VS) in two series: 1 and 22 weeks after cabin filter replacement and disinfection of the air-conditioning system. During one test in each bus were taken two samples: before the air-conditioning or ventilation system switched on and 6 min after operating of these systems. The atmospheric air was the external background (EB). After 1 week of use of the system, the fungi concentrations before starting of the ACS and VS system were 527.8 and 1053.0 cfu/m3, respectively, and after 22 weeks the concentrations were 351.9 and 1069.6 cfu/m3, respectively. While in the sample after 6 min of ACS and VS system operating, the fungi concentration after 1 week of use was 127.6 and 233.7 cfu/m3, respectively, and after 22 weeks it was 113.3 and 324.9 cfu/m3, respectively. Results do not provide strong evidence that air-conditioning system is an additional source of indoor air contamination with fungi. A longer operation of the system promoted increase of fungi concentration in air-conditioned buses only.

Inhalable dust, endotoxins and (1-3)-ß-d-glucans as indicators of exposure in waste sorting plant environment.

The aim of the study was to assess the levels of inhalable dust, endotoxins and (1-3)-ß-d-glucans as agents harmful to the respiratory tract of workers of municipal waste sorting plants and interaction between these agents based on the measurements taken in two plants with different processing capacities. The study was conducted in summer season in two waste sorting plants (WSPs) differing in processing capacity. Samples of bioaerosol for inhalable dust (gravimetric method), endotoxins (LAL test in kinetic, chromogenic version) and (1-3)-ß-d-glucans (Glucatell test in kinetic version) were collected from 42 sorting workers using individual aspirators with glass fiber filters during the work shift. Average geometric concentrations (geometric standard deviation; min-max) of inhalable dust, endotoxins and (1-3)-ß-d-glucans were: WSP1: 1.7 mg m-3 (2.2; 0.6-6.9 mg m-3); 15.9 ng m-3 (2.1; 5.4-78.9 ng m-3), 55.1 ng m-3 (1.8; 20.7-188.6 ng m-3) and WSP2: 0.8 mg m-3 (2.2; 0.2-3.8 mg m-3), 9.8 ng m-3 (2.4; 1.6-29.7 ng m-3), 45.0 ng m-3 (3.2, 5.7-212.9 ng m-3), respectively. A significantly higher concentration of inhalable dust was recorded in WSP1 with bigger processing capacity compared to WSP2 (less processing capacity). Significant (p < 0.05) and very high correlations (Spearman rank R > 0.7) were found between the concentrations of all analyzed harmful agents. Processing capacity of waste sorting plants differentially affects the concentrations of inhalable dust, whereas concentrations of endotoxins and glucans are less clearly affected. This suggests that relative concentrations of endotoxin and glucan are depending on the waste sorting capacity.

The capability of fungi isolated from moldy dwellings to produce toxins.

OBJECTIVES: The main objective was analysis and assessment of toxinogenic capabilities of fungi isolated from moldy surfaces in residential rooms in an urban agglomeration situated far from flooded areas in moderate climate zone. MATERIAL AND METHODS: The assessment of environmental exposure to mycotoxins was carried out in samples collected from moldy surfaces in form of scrapings and airborne dust from 22 moldy dwellings in winter season. In each sample 2 mycotoxins were analyzed: sterigmatocystin and roquefortine C produced by Aspergillus versicolor and Penicillium chrysogenum, respectively. Mycotoxins were analyzed by high-performance liquid chromatography (HPLC) in: scrapings from moldy surfaces, mixture of all species of fungi cultured from scrapings on microbiological medium (malt extract agar), pure cultures of Aspergillus versicolor and Penicillium chrysogenum cultured from scrapings on microbiological medium; mycotoxins in the indoor air dust were also analyzed. RESULTS: The production of sterigmatocystin by individual strains of Aspergillus versicolor cultured on medium was confirmed for 8 of 13 isolated strains ranging 2.1-235.9 µg/g and production of roquefortine C by Penicillium chrysogenum for 4 of 10 strains ranging 12.9-27.6 µg/g. In 11 of 13 samples of the mixture of fungi cultured from scrapings, in which Aspergillus versicolor was found, sterigmatocystin production was at the level of 3.1-1683.2 µg/g, whereas in 3 of 10 samples in which Penicillium chrysogenum occurred, the production of roquefortine C was 0.9-618.9 µg/g. The analysis did not show in any of the tested air dust and scrapings samples the presence of analyzed mycotoxins in the amount exceeding the determination limit. CONCLUSIONS: The capability of synthesis of sterigmatocystin by Aspergillus versicolor and roquefortine C by Penicillium chrysogenum growing in mixtures of fungi from scrapings and pure cultures in laboratory conditions was confirmed. The absence of mycotoxins in scrapings and air dust samples indicates an insignificant inhalatory exposure to mycotoxins among inhabitants in moldy flats of urban agglomeration situated far from flooded territories. Int J Occup Med Environ Health 2016;29(5):823-836.

Visible fungi growth and dampness assessed using a questionnaire versus airborne fungi, (1→3)-ß-D-glucan and fungal spore concentrations in flats.

INTRODUCTION: The study aimed at determination of the usefulness of the subjective assessment of selected signs of fungi growth in flats and microclimate parameters to indicate the actual air contamination with culturable fungi, (1→3)-ß-D-glucans and fungal spores. MATERIAL AND METHODS: This analysis covered 22 flats, the inhabitants of which declared in a questionnaire interview the presence of the developed mycelium on solid surfaces in the flat. Air samples for determination of the culturable fungi, (1→3)-ß-D-glucans and (viable and non-viable) fungal spores concentrations indoor and outdoor the flats during the heating period were collected. During bioaerosol sampling microclimate parameters were measured. Predictive models for concentrations of the tested biological agents with regard to various ways to assess fungal contamination of air in a flat (on the basis of a questionnaire or a questionnaire and microclimate measurements) were built. RESULTS: The arithmetic means of temperature, relative humidity, CO2 concentration and air flow velocity in the flats were respectively: 20.5°C, 53%, 1431.6 ppm and 0 m/s. The geometric mean concentrations of airborne fungi, (1→3)-ß-D-glucans and fungal spores in these premises amounted to 2.9×102 cfu/m3, 1.6 ng/m3 and 5.7×103 spores/m3, respectively. The subjective assessment of fungi growth signs and microclimate characteristics were moderately useful for evaluation of the actual airborne fungi and (1→3)-ß-D-glucan concentrations (maximum percent of explained variance (VE) = 61% and 67%, respectively), and less useful in evaluation of the actual fungal spore concentrations (VE < 29%). In the case of fungi, higher usefulness was indicated of the questionnaire evaluation supported by microclimate measurements (VE = 61.2%), as compared to the evaluation only by means of a questionnaire (VE = 46.9%). CONCLUSIONS: Subjective evaluation of fungi growth signs in flats, separately or combined with microclimate measurements, appeared to be moderately useful for quantitative evaluation of the actual air contamination with fungi and their derivatives, but more extensive studies are needed to strengthen those findings.

Assessment of exposure to fungi in the heavily contaminated work environment (a solid waste sorting plant) based on the ergosterol analysis.

OBJECTIVES: This paper reports on the results of the study aimed at application of ergosterol as an quantitative indicator of fungal bioaerosol present in the indoor air in occupational environment heavily contaminated with organic dust as well as its comparison with the culturable method. MATERIAL AND METHODS: The study was conducted in the indoor solid waste sorting plant. Using Andersen impactor adapted to 1 plate at the flow rate of 30 l/min, indoor air was sampled in the workers' breathing zone. Ergosterol was sampled using gelatinous filter (1000 l of air) and then analyzed by means of the spectrophotometric method. Fungi were sampled on malt extract agar (MEA) medium (3 replications: 2 l, 7.5 l, 15 l of air) and analyzed by means of the culturable method. Based on ergosterol analyzes, concentration of fungi was calculated. Results were given as the range assuming min. as 5.1 pg ergosterol/spore and max as 1.7 pg ergosterol/spore. RESULTS: The average concentrations of ergosterol in a working room (arithmetic mean (AM), standard deviation (SD); minimum-maximum (min.-max)) were, respectively: 2.16, 0.72; 0.85-2.92 µg/m3; fungi calculated based on ergosterol - 424.1×103-1272.4×103, 140.1×103- 420.4×103, 167×103-1716.5×103 CFU/m3, and culturable fungi - 13×103, 9.7×103, 1.9×103-34×103 CFU/m3). It was revealed that concentrations of calculated fungi were even 2 orders of magnitude higher than culturable fungi. CONCLUSIONS: The quantitative assessment of moldiness by means of ergosterol measurement seems to be a reliable indicator for environments heavily contaminated with organic dust, where viable and non-viable fungi are present in high proportions. Based on that result, more restrictive (as compared to a similar assessment carried out by means of the culturable method) hygienic recommendations, especially those related to the use of preventive measures protecting the employees' respiratory tract, should have been undertaken.