Wastewater and wastewater treatment plants serve as urban reservoirs of pathogenic microorganisms. Wastewaters frequently contain bacteria, antibiotic-resistant bacteria, and developmental stages of parasites with significant zoonotic potential. Five wastewater treatment plants in the central part of Slovakia were investigated to determine the effect of treatment on bacterial community, antibiotic-resistant bacteria, and the occurrence of helminth eggs. Although all monitored chemical factors (chemical oxygen demand, biochemical oxygen demand, N-NH4, total nitrogen, and total phosphorus) in the effluent were in line with the legislative standards for discharge into public waterways, the results of minimal inhibitory concentrations show that reclaimed water harbors E. coli resistant to several commonly used antibiotics (ampicillin, piperacillin, and tazobactam, combine ampicillin and sulbactam, cefotaxime, tetracycline). The presence of endoparasite developmental stages in wastewater and sludge (Ascaris spp., Hymenolepis nana, eggs from the Ancylostomatidae family, Giardia duodenalis) indicates potential health risks for humans and workers at these sites. Treatment such as composting before applying sludge to land is necessary to reduce human pathogens.

Antibiotic resistance of the indicator microorganism Escherichia coli was investigated in isolates from samples collected during the course of one year from two wastewater treatment plants treating municipal and animal wastes in Slovakia, respectively. The genes of antibiotic resistance and virulence factors in selected resistant E. coli isolates were described. A high percentage of the isolates from municipal and animal wastewater were resistant to ampicillin, streptomycin, tetracycline, ceftiofur, ceftriaxone, and enrofloxacin. In the selected E. coli isolates, we detected the following phenotypes: ESBL (20.4% in animal wastewater; 7.7% in municipal wastewater), multidrug-resistant (17% of animal and 32% of municipal isolates), high resistance to quinolones (25% of animal and 48% of municipal samples), and CTX-M (7.9% of animal and 17.3% of municipal isolates). We confirmed an integro-mediated antibiotic resistance in 13 E. coli strains from municipal and animal wastewater samples, of which the Tn3 gene and virulence genes cvaC, iutA, iss, ibeA, kps, and papC were detected in six isolates. One of the strains of pathogenic E. coli from the animal wastewater contained genes ibeA with papC, iss, kpsII, Int1, Tn3, and Cit. In addition, one bla_IMP gene was found in the municipal wastewater sample. This emphasises the importance of using the appropriate treatment methods to reduce the counts of antibiotic-resistant microorganisms in wastewater effluent.

Staphylococcus spp. is the most common cause of mastitis, with a significantly low cure rate. Bacterial characteristics like adhesion and biofilm formation, as well as extracellular factors, can affect the pathogenesis of staphylococcal mastitis. The study’s objectives were to confirm S. aureus, assess their antibiotic resistance, identify methicillin resistance genes, verify biofilm formation, and detect biofilm-associated genes from bovine mastitis samples using multiplex PCR (mPCR). From 215 milk samples, six were confirmed as S. aureus. Most isolates were sensitive to all measured antibiotics. One isolate was identified as an inducible form of MLSB resistance (macrolides, lincosamides, and streptogramin B resistance), while the other two isolates were resistant to penicillins and carboxypenicillins. In S. aureus cultures used for methicillin resistance genotypic analysis by PCR, the mecA and mecC genes were not found. Biofilm formation phenotypes were determined in four strains. An mPCR analysis revealed that all strains of S. aureus carried icaABCD, agrA, srtA, fnbA, clfA, and clfB genes. Only in one isolate was the fnbB gene detected; the bap gene was not detected in any of the isolates. This emphasizes the importance of using appropriate treatment and continuous monitoring of S. aureus to prevent the spread of antibiotic-resistant strains in dairy cow farms.

Nowadays, there is an effort to improve the effectiveness of the composting process, supported by the addition of various supplements to reduce soil nutrition losses and increase soil remediation. The aim of this study was to examine the devitalization effect of natural additives like zeolite-clinoptilolite and its combination with hydrated lime in composted cattle manure on indicator and pathogen bacteria. The composting process was running in three static piles of cattle manure mixed with wheat straw (control, zeolite–lime, and zeolite) for 126 days. Composted manure substrates were determined for physicochemical (temperature, pH, nitrogen and phosphorus content, C/N, organic matter, and moisture) and microbiological analyses (Salmonella spp., indicator bacteria). The effects of additives were reflected in changes in physicochemical factors, e.g., an increase in temperature (<53 °C) or pH (<9.3). According to Pearson correlation, these changes (pH, Nt, Pt) resulted in a significant decrease (p < 0.001) of indicator bacteria (two or three orders) in zeolite pile or zeolite–lime pile. Die-off of Salmonella spp. in the zeolite–lime pile was indicated within 41 days; in other piles, this occurred on day 63. Our results can aid in further optimizing the composting of cattle manure in order to lower environmental pollution and the risk of human infection.

The conditions in livestock housing are suitable for the survival of airborne microorganisms, mainly due to high temperatures, humidity, and the presence of organic material. The total count of airborne bacteria concentrations in cattle farms ranged from 3.01 log₁₀ CFU/mL to 6.90 log₁₀ CFU/mL; for coliform bacteria, they were from 2.18 log₁₀ CFU/mL to 3.34 log₁₀ CFU/mL; and for molds, they ranged from 3.00 log₁₀ CFU/mL to 4.57 log₁₀ CFU/mL. Bacteria resistant to antimicrobial substances and resistance genes can be spread on animal farms. Antimicrobial resistance in ubiquitous Escherichia coli isolated from cattle feces was investigated. Minimum inhibitory concentration (MIC) testing was utilized to identify phenotypic resistance profiles, and the PCR method was employed to detect the presence of resistant genes. A higher percentage of resistance was found to amikacin (65%), tetracycline (61%), streptomycin (56%), ampicillin (55%), and nalidixic acid (45%). Multidrug resistance was determined in up to 64.3% of the isolates studied. The most widespread resistance genes were bla_TEM (85.7%), sul2 (66.7%), tetB (52.38%), and sul1 (47.6%). We found that 4.8% of the E. coli isolates had the bla_CMY gene. We found that, despite phenotypic resistance, E. coli isolates do not necessarily carry genes conferring resistance to that particular antimicrobial agent.

Methicillin-resistant coagulase-negative staphylococci (MRCoNS) are a major cause of infectious diseases, owing to their ability to form biofilms and colonize community and hospital environments. MRCoNS strains were identified using biochemical tests, an MALDI-TOF MS analyzer, and PCR-based 16S rRNA gene confirmation. This study was designed to assess antibiotic resistance and biofilm-forming capacity and to determine the presence of the mecA, mecC, agrA, srtA, icaABCD, bap, fnbAB, and clfAB genes in MRCoNS isolates. From patients undergoing random screening during hospitalization in the Orthopedics Clinic in Slovakia, 28 strains of MRCoNS were identified: S. epidermidis (n = 10), S. hominis (n = 8), S. haemolyticus (n = 4), S. lugdunensis (n = 3), while S. simulans, S. pasteuri, and S. warneri were detected only once. The highest rates of resistance were observed for ampicillin, oxacillin, rifampicin, trimethoprim (100%), and erythromycin (62%). The mecA gene was detected in 12 analyzed isolates. In 12 isolates, MDR, strong efflux pump activity, and strong or moderate biofilm formation were simultaneously detected. Our findings highlight the problems posed by biofilm-forming, resistant CoNS in hospitalized patients and the importance of diagnostics, separation, rapid treatment, and proper hospital hygiene.