The Prevalence of Virulent and Multidrug-Resistant Enterococci in River Water and in Treated and Untreated Municipal and Hospital Wastewater

The aim of this study is to describe the drug resistance and virulence of enterococci in river water sampled downstream (DRW) and upstream (URW) from the wastewater discharge point, to determine the pool of virulent and drug-resistant enterococci in untreated wastewater (UWW) and the extent to which these bacteria are eliminated from hospital wastewater (HWW) and municipal wastewater treated (TWW) by biological and mechanical methods in a wastewater treatment plant (WWTP). A total of 283 strains were identified with the use of culture-dependent methods and PCR, including seven different species including E. faecalis and E. faecium which were predominant in all analyzed samples. Majority of the strains were classified as multidrug resistant (MDR), mostly on streptomycin and trimethoprim. Strains isolated from wastewater and DRW harbored van genes conditioning phenotypic resistance to vancomycin, the highest percentage of vancomycin-resistant strains (57.0%), mostly strains harboring vanC1 genes (27.6%), was noted in TWW. More than 65.0% of the isolated strains had different virulence genes, the highest number of isolates were positive for cell wall adhesin efaA and sex pheromones cob, cpd, and ccf which participate in the induction of virulence. Many of the strains isolated from TWW were resistant to a higher number of drugs and were more virulent than those isolated from UWW and HWW. The enterococci isolated from DRW and wastewater were characterized by similar multidrug resistance and virulence profiles, and significant correlations were observed between these groups of isolates. These findings suggest that pathogenic enterococci are released with TWW and can spread in the river, pose a serious epidemiological threat and a risk to public health

Distribution of <i>Pseudomonas fluorescens</i> and <i>Aeromonas hydrophila</i> Bacteria in a Recirculating Aquaculture System during Farming of European Grayling (<i>Thymallus thymallus</i> L.) Broodstock

Pseudomonas fluorescens and Aeromonas hydrophila bacteria are opportunistic pathogens that occur naturally in the aquatic environment and in the gut flora of healthy fish. Both species can pose a serious threat for fish that are highly sensitive to water pollution. The aim of this study was to determine the extent to which the amount of administered fish feed and fish biomass affect the distribution and abundance of Ps. fluorescens and A. hydrophila bacteria in a recirculating aquaculture system (RAS) during farming of European grayling (Thymallus thymallus L.) broodstock. A total of 68 water samples from the inflow, two rearing tanks and the outflow as well as 17 feed samples were collected and analyzed separately. Bacterial populations were analyzed by the culture-dependent method and a molecular method (fluorescence in situ hybridization, FISH) to detect culturable strains and viable but non-culturable strains, respectively. Fish biomass, feed and 16 water quality parameters (temperature, pH, concentration of dissolved oxygen, oxygen saturation, five-day biochemical oxygen demand (BOD5), total phosphorus, total organic phosphorus and nitrogen, orthophosphates, total nitrogen, nitrite and nitrate nitrogen, ammonia nitrogen, ammonium nitrogen, total suspended solids, and total organic carbon) were the explanatory factors. Statistically significant differences (RM-ANOVA, p &#8804; 0.05) were stated in bacterial abundance in samples from the inflow, rearing tanks and the outflow. Water samples from the RAS were abundantly colonized by non-culturable Ps. fluorescens and A. hydrophila bacteria. Feed was not a source of bacteria, but a redundancy analysis (RDA) revealed that the amount of feed, fish biomass, BOD5, and total suspended solids and total organic carbon were positively correlated in both Ps. fluorescens and A. hydrophila. These parameters also influenced the distribution of both potentially pathogenic bacterial populations and contributed to the bacterial contamination of water in the RAS. Our results are particularly valuable for aquacultures that help to replenish wild stocks and rebuild populations of threatened species in natural aquatic environments

Amaranth Meal and Environmental Carnobacterium maltaromaticum Probiotic Bacteria as Novel Stabilizers of the Microbiological Quality of Compound Fish Feeds for Aquaculture

Fish feed should be characterized by microbiological stability to guarantee the optimal health of farmed fish. The aim of this study was to determine the efficacy of amaranth meal (Amaranthus cruentus) and a highly active environmental strain of probiotic bacteria, Carnobacterium maltaromaticum, as novel supplements that stabilize the quantitative and qualitative composition of microbiota in compound fish feeds for aquaculture, regardless of storage temperature. The total viable counts of mesophilic bacteria at 28 &deg;C (TVC 28 &deg;C), hemolytic mesophilic bacteria (Hem 37 &deg;C), Staphylococcus sp. bacteria, aerobic spore-forming bacteria (ASFB), sulfite-reducing anaerobic spore-forming Clostridium sp. bacteria, yeasts, and molds were analyzed in control feed (CF), in feed supplemented with amaranth meal (AF), and in feed supplemented with amaranth meal and C. maltaromaticum (ACF), stored at a temperature of 4 &deg;C and 20 &deg;C for 98 days. Amaranthus cruentus and C. maltaromaticum significantly reduced bacterial counts in fish feeds, regardless of the temperature and duration of storage. The antibacterial and antifungal effects of the tested additives were statistically significant (p &le; 0.05). The studied novel supplements contribute to the microbiological safety of compound fish feeds. The tested additives could be recognized as the key ingredients of organic, environmentally friendly fish feeds, which guarantee the high quality of fish intended for human consumption