Organic acids mitigate Streptococcus agalactiae virulence in Tilapia fish gut primary cells and in a gut infection model.

BACKGROUND: Streptococcus agalactiae, a Gram-positive bacterium, has emerged as an important pathogen for the aquaculture industry worldwide, due to its increased induced mortality rates in cultured fish. Developing interventions to cure or prevent infections based on natural alternatives to antibiotics has become a priority, however, given the absence of scientific evidence regarding their mode of action progress has been slow. METHODS: In this study we aimed to investigate the effect of a mixture of organic acids (natural antimicrobials), AuraAqua (Aq), on the virulence of S. agalactiae using Tilapia gut primary epithelial cells and an in vitro Tilapia gut culture model. Our results show that Aq was able to reduce significantly, in vitro, the S. agalactiae levels of infection in Tilapia gut primary epithelial cells (TGP) when the MIC concentration of 0.125% was tested. RESULTS AND DISCUSSION: At bacterial level, Aq was able to downregulate bacterial capsule polysaccharide (CPS) gene expression, capC, resulting in a significant decrease in bacterial surface capsule production. The decrease in CPS production was also associated with a reduction in the pro-inflammatory IFNγ, IL1ß, TNFα, SOD and CAT gene expression and H2O2 production in the presence of 0.125% Aq (P < 0.0001). The antimicrobial mixture also reduced the levels of S. agalactiae infection in an in vitro gut culture model and significantly reduced the IFNγ, IL1ß, TNFα, SOD, CAT gene expression and H2O2 production in infected tissue. Moreover, genes involved in Tilapia resistance to S. agalactiae induced disease, MCP-8 and Duo-1, were also downregulated by Aq, as a consequence of reduced bacterial levels of infection. CONCLUSION: Conclusively, our study shows that mixtures of organic acids can be considered as potential alternative treatments to antibiotics and prevent S. agalactiae infection and inflammation in the Tilapia fish digestive tract.

Trace Metal Accumulation in Rats Exposed to Mine Waters: A Case Study, Bor Area (Serbia).

Zinc (Zn), copper (Cu), iron (Fe), manganese (Mn), cadmium (Cd), and lead (Pb) levels were measured in the Bor City water supply system (control) and two watercourses exposed to mining wastewaters, i.e., the Lutarica River (one site) and the Kriveljska River (two sites). The same parameters were determined in the brain, heart, lungs, stomach, liver, spleen, kidneys, and testes of male Wistar rats given water from these sources for 2 months. Water Cu, Fe, Cd, and Pb were outside the safe range, excepting the reference site. Significant impacts on intra-organ metal homeostasis were detected, especially in the brain, stomach, kidneys, and testes. The dynamics and magnitude of these changes (versus controls) depended on the target organ, analyzed metal, and water origin. The greatest number of significant intra-organ associations between essential and non-essential metals were found for Cd-Zn, Cd-Cu, and Cd-Mn. A regression analysis suggested the kidneys as the most relevant organ for monitoring water manganese, and the stomach and brain for lead. These results highlight the environmental risks associated with mining wastewaters from the Bor area and could help scientists in mapping the spatial distribution and severity of trace metal contamination in water sources.

Biocides as Biomedicines against Foodborne Pathogenic Bacteria.

Biocides are currently considered the first line of defense against foodborne pathogens in hospitals or food processing facilities due to the versatility and efficiency of their chemical active ingredients. Understanding the biological mechanisms responsible for their increased efficiency, especially when used against foodborne pathogens on contaminated surfaces and materials, represents an essential first step in the implementation of efficient strategies for disinfection as choosing an unsuitable product can lead to antibiocide resistance or antibiotic-biocide cross-resistance. This review describes these biological mechanisms for the most common foodborne pathogens and focuses mainly on the antipathogen effect, highlighting the latest developments based on in vitro and in vivo studies. We focus on biocides with inhibitory effects against foodborne bacteria (e.g., Escherichia spp., Klebsiella spp., Staphylococcus spp., Listeria spp., Campylobacter spp.), aiming to understand their biological mechanisms of action by looking at the most recent scientific evidence in the field.

Antiproliferative and antibacterial potential of tetrahexylammonium bromide-based ionic liquids.

Ionic liquids (ILs) exhibit cytotoxic effects on prokaryotic and eukaryotic cells. In this study, the antibacterial and antiproliferative activities of tetrahexylammonium bromide-based ILs were investigated. In order to evaluate the therapeutic potential of these ionic liquids, firstly microbiological assay using both Gram-positive and Gram-negative bacteria were conducted by employing Disk-Diffusion and 2,3,5-triphenyltetrazolium chlorine (TTC) methods to assess the antimicrobial effects. Likewise, the antitumor effects on 2D and 3D cell culture systems were assessed using the human colon cancer Caco-2 cell line and cytotoxic activity was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Alamar blue assays. Results obtained showed that [CH3(CH2)5]4N(Br) possesses an antibacterial potential, particularly in the case of two bacteria, S. aureus (Gram+) and H. influenzae (Gram-). Preliminary screening of antiproliferative activity showed moderate activity, except for the concentration of 10 mM. Furthermore, regarding the effect of [CH3(CH2)5]4N(Br) on tumor cell aggregation, positive outcomes were noted. [CH3(CH2)5]4N(Br) presents promising and under-explored potential to improve antibacterial or anticancer therapies.

Potential Ecological and Human Health Risks of Heavy Metals in Soils in Selected Copper Mining Areas-A Case Study: The Bor Area.

Surface soil samples were collected near the Open Pit Bor (S1) and Open Pit Cerovo (S2), a grassland along the Borska Reka River (S3) and an unpolluted garden near Slatina village (reference site). Spontaneous plants (dandelion, nettle, coltsfoot, and creeping buttercup) and vegetables (onion, garlic, carrot, parsley, celery, potatoes, dill, and sorrel) were obtained from the former three sites and the reference site, respectively. The samples were analyzed for Zn, Cu, Fe, Mn, and Pb via FAAS. Pollution indices indicated low-to-moderate soil contamination at sites S1, S2, and S3. Cu was the main contaminant of environmental concern, being above the maximum admitted concentration at site S1. Metal levels in spontaneous plants were below phytotoxic levels. Cu content of leafy vegetables and celery roots and Pb content of most vegetables were not safe for human consumption. Metal concentrations tended to be significantly lower in plants than in soils, with only Cu occurring at significantly elevated levels in celery roots and sorrel leaves. Non-carcinogenic risk assessment showed that consumption of carrot roots and especially celery roots grown on unpolluted soils from the Bor area might pose long-term health risks for females and males, with the main contributors being Cu and Fe.

Effects of mine waste water on rat: bioaccumulation and histopathological evaluation.

The highlighting of the bioaccumulation capacity of metals in the internal organs, the mode of distribution at the level of internal organs, the interactions between them, respectively, and the histological changes occurred at the level of the liver and kidneys are the main aspects addressed in the present study. The experiment was performed on 4 groups of Wistar rats: 3 groups which were administered water from rivers located in the vicinity of the Bor mining operation and 1 control group. The determination of the metal content in the administered water samples and in the internal organs was performed using the flame atomic absorption spectrophotometer. Tissue alterations were assessed by histological technique and hematoxylin-eosin staining. The metal retention capacity in the internal organs differs depending on the metal concentration in the administered water sample but also on the organ in which the determination was made. Also, correlations were established between the concentrations of metals at the level of the organs, showing (a) positive and significant correlations-at the level of the heart between Zn and Cu, Fe, and Mn and at the level of the lungs between Mn and Cd-but the most numerous were reported in the testicle; (b) moderate correlations at liver level between Fe and Zn, at spleen level between Cu and Mn and Cd and at the level of the kidneys between Pb and Zn, Cu, and Fe; (c) negative correlations at renal level between Pb and Mn; and (d) insignificant correlations between Pb and Fe. The histological changes identified at the level of the liver and kidney become more obvious, and their aggravation is registered with the increase of the metal content.

MicroRNA profiling in kidney in pigs fed ochratoxin A contaminated diet.

OTA is a toxic metabolite produced by fungus belonging to Aspergillus and Penicillium genera. Kidney is the main target of this toxin; OTA is considered as one of the etiological factors at the origin of the human Balkan endemic nephropathy. microRNA are short non-coding transcrips (18-22 nucleotides in length) regulating key cellular processes. Various miRNAs have been established to play important roles in development of renal carcinoma and urothelial cancer. The objective of this study is to analyse the miRNA profiling in the kidney of piglets experimentally intoxicated with feed contaminated with OTA. Fifteen piglets (five pigs/group) were randomly distributed into 3 groups, fed normal diet (Group 1: control), or diets contaminated with OTA in two concentrations: 50 µg OTA/kg feed (Group 2: 50 µg OTA/kg feed) or 200 µg OTA/kg feed (Group 3: 200 µg OTA/kg feed) for 28 days. At the end of the experiment blood samples were taken for serological analyses. Animals from control group and 200 µg OTA/kg feed were sacrificed and kidney samples were taken for histological and molecular analyses. As resulted from molecular profiling study there are 8 miRNA differentially expressed in OTA kidney vs control kidney, in which five miRNA were overexpressed in the kidney of OTA intoxicated animals: miR-497 (FC = 6.34), miR-133a-3p (FC = 5.75), miR-423-3p (FC = 5.48), miR-34a (FC = 1.68), miR-542-3p (1.65) while three miRNA were downregulated: miR-421-3p (FC = -3.96); miR-490 (FC = -3.87); miR-9840-3p (FC = -2.13). The altered miRNAs as effect of OTA are strongly connected to the engine of cancer, disturbing nodal points in different pathways, as TP53 signalling. This proof-of-concept study proves the actual utility of miRNAs as biomarkers of mycotoxin exposure, including OTA.

Effect of human isolated probiotic bacteria on preventing Campylobacter jejuni colonization of poultry.

This study was performed in order to determine whether human isolated probiotic bacteria can be effective in reducing Campylobacter jejuni infection of chicken intestinal cells, in vitro, and in decreasing its colonization abilities within the chicken gut. Our results show that the probiotic strains Lactobacillus paracasei J. R, L. rhamnosus 15b, L. lactis Y, and L. lactis FOa had a significant effect on C. jejuni invasion of chicken primary cells, with the strongest inhibitory effect detected when a combination of four was administered. In regard to the in vivo effect, using all four strains in one combination prevented mucus colonization in the duodenum and cecum. Moreover, the pathogen load in the lumen of these two compartments was significantly reduced. When probiotics were introduced during the early growth period, the presence of the pathogen in feces was increased (p>0.05), but when they were given during the last week of growth, there was no significant effect. In conclusion, our data indicate that these four new probiotic strains are able to cause modifications in the chicken intestinal mucosa and can reduce the ability of C. jejuni to invade, in vitro, and to colonize, in vivo. These probiotics are now proven to be effective even when introduced in broiler's feed 7 days before slaughter, which makes them cost-effective for the producers.