Enterococcus Hirae Research Articles

Microbiological Findings and Associated Histopathological Lesions in Neonatal Diarrhoea Cases between 2020 and 2022 in a French Veterinary Pig Practice

This retrospective study described the aetiologies of neonatal diarrhoea cases and their associations with histological findings. A total of 106 diarrhoeic neonatal piglets were selected. Cultures, MALDI typings, PCRs and evaluation of intestinal lesions were performed. A total of 51 cases (48.1%) were positive for only one pathogen and 54 (50.9%) were positive for more than one pathogen. Clostridium perfringens type A was the most frequently detected pathogen (61.3%), followed by Enterococcus hirae (43.4%), rotavirus type A (38.7%), rotavirus type C (11.3%) and enterotoxigenic Escherichia coli (3.8%). Only lesions in the small intestine were correlated with detected pathogens. The detection of rotavirus was associated with an increased probability of observing villous atrophy (p < 0.001), crypt hyperplasia (p = 0.01) and leucocyte necrosis in the lamina propria (p = 0.05). The detection of Clostridium perfringens type A was associated with an increased probability of observing bacilli in close proximity to the mucosa (p < 0.001) and a decreased probability of observing epithelial necrosis (p = 0.04). Detection of Enterococcus hirae was associated with an increased probability of observing enteroadherent cocci (p < 0.001). Multivariate regression logistic models revealed that epithelial necrosis was more likely to occur in Enterococcus hirae-positive piglets (p < 0.02) and neutrophilic infiltrate was more likely to occur in Clostridium perfringens type A- and Enterococcus hirae-positive piglets (p = 0.04 and p = 0.02, respectively).

Comparative evaluation of mutagenic, genotoxic, cytotoxic, and antimicrobial effects of flavour and fragrance aldehydes, ketones, oximes, and oxime ethers

Despite the large number of odoriferous compounds available, new ones with interesting olfactory characteristics are desired due to their potentially high commercial value. Here, we report for the first time mutagenic, genotoxic, and cytotoxic effects, and antimicrobial properties of low-molecular fragrant oxime ethers, and we compare their properties with corresponding oximes and carbonyl compounds. 24 aldehydes, ketones, oximes, and oxime ethers were evaluated for mutagenic and cytotoxic effects in Ames (using Salmonella typhimurium strains TA 98 with genotype hisD3052, rfa, uvrB, pKM101, and TA100 with genotype hisG46, rfa, uvrB, pKM101, concentration range: 0.0781–40 mg/mL) and MTS (using HEK293T cell line concentration of tested substances: 0.025 mM) assays. Antimicrobial evaluation was carried out against Bacillus cereus (ATCC 10876), Staphylococcus aureus (ATCC 6538), Enterococcus hirae (ATCC 10541), Pseudomonas aeruginosa (ATCC 15442), Escherichia coli (ATCC 10536), Legionella pneumophila (ATCC 33152); Candida albicans (ATCC 10231) and Aspergillus brasiliensis (ATCC 16404) with concentration range of tested substances 9.375 – 2.400 mg/mL. Furthermore, 5 representatives of carbonyl compounds, oximes, and an oxime ether (stemone, buccoxime, citral, citral oxime, and propiophenone oxime O-ethyl ether) were evaluated for genotoxic properties in SOS-Chromotest (concentration range: 7.8·10-5 – 5·10-3 mg/mL). All of the tested compounds did not exhibit mutagenic, genotoxic, or cytotoxic effects. Oximes and oxime ethers showed relevant antimicrobial activity against pathogenic species (P. aeruginosa, S. aureus, E.coli, L. pneumophila, A. brasiliensis, C. albicans) in the MIC range 0.075 – 2.400 mg/mL compared to the common preservative methylparaben with the MIC range 0.400–3.600 mg/mL. Our study shows that oxime ethers have the potential to be used as fragrant agents in functional products.

Molecular Characterization of Some Bacillus Species from Vegetables and Evaluation of Their Antimicrobial and Antibiotic Potency.

Numerous natural habitats, such as soil, air, fermented foods, and human stomachs, are home to different Bacillus strains. Some Bacillus strains have a distinctive predominance and are widely recognized among other microbial communities, as a result of their varied habitation and physiologically active metabolites. The present study collected vegetable products (potato, carrot, and tomato) from local markets in Almaty, Kazakhstan. The bacterial isolates were identified using biochemical and phylogenetic analyses after culturing. Our phylogenetic analysis revealed three Gram-positive bacterial isolates BSS11, BSS17, and BSS19 showing 99% nucleotide sequence similarities with Bacillus subtilis O-3, Bacillus subtilis Md1-42, and Bacillus subtilis Khozestan2. The crude extract was prepared from bacterial isolates to assess the antibiotic resistance potency and the antimicrobial potential against various targeted multidrug-resistant strains, including Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus group B, Streptococcus mutans, Candida albicans, Candida krusei, Pseudomonas aeruginosa, Shigella sonnei, Klebsiella pneumoniae, Salmonella enteritidis, Klebsiella aerogenes, Enterococcus hirae, Escherichia coli, Serratia marcescens, and Proteus vulgaris. This study found that the species that were identified have the ability to produce antibiotic chemicals. Additionally, the GC-MS analysis of three bacterial extracts revealed the presence of many antibiotic substances including phenol, benzoic acid, 1,2-benzenedicarboxylic acid and bis(2-methylpropyl), methoxyphenyl-oxime, and benzaldehyde. This work sheds light on the potential of Bacillus to be employed as an antimicrobial agent to target different multidrug-resistant bacterial strains. The results indicate that market vegetables may be a useful source of strains displaying a range of advantageous characteristics that can be used in the creation of biological antibiotics.

Elaboration and general evaluation of chitosan-based films containing terpene alcohols-rich essential oils

Recently, the scientific community is interested in the synthesis of biodegradable and bioactive packaging to replace oil-based ones. Therefore, the present study aims to elaborate an active and biodegradable material using chitosan (CS-film) combined with pelargonium, tea tree, marjoram, and thyme essential oils (EOs), and then evaluate their different properties and biological activities. The obtained data showed an augmentation in CS-film thickness and opacity following the addition of EOs ranging from 17 ± 3 to 42 ± 2μm and from 1.53 ± 0.04 to 2.67 ± 0.09, respectively. Furthermore, a significant decrease in the water vapor transmission rate and moisture content parameters was recorded as regards the treated CS-films. On the other hand, the treatment with EOs engenders random modifications in the physicochemical and mechanical characteristics of the material. Concerning the biological activities, the treated CS-films scavenged around 60% of DPPH radical while the control CS-film exhibited a negligible antioxidant activity. Finally, the CS-films containing pelargonium and thyme EOs exhibited the strongest antibiofilm-forming activity against Escherichia coli, Enterococcus hirae, Staphylococcus aureus, and Pseudomonas aeruginosa with values of inhibition greater than 70%. These encouraging results verify the effectiveness of CS-films containing EOs such as pelargonium and thyme EOs as biodegradable and bioactive packaging.

Probiotic potential of lactic acid bacteria isolated from colostrum of 3 different mammals

ABSTRACT This study aimed to investigate the in vitro properties of lactic acid bacteria isolated from colostrum samples of goat, sheep and cow. Four isolates of lactic acid bacteria identified through 16S rDNA sequencing were screened for probiotic potential including their enzymatic activities (hemolytic, DNase activity, API ZYM profiling and tolerance to the gastrointestinal conditions), their antimicrobial activities, their adhesion properties through biofilm formation capacity, auto-aggregation and co-aggregation with pathogens bacteria and hydrophobicity. They were also tested for their antibiotic susceptibility and antioxidant power. All the strains (ADG1b Lactobacillus gasseri, ZPana Lactiplantibacillus plantarum, S2ana Enterococcus hirae and ZGana Lactobacillus gasseri) demonstrated probiotic properties and they could resist different gastrointestinal conditions. They displayed good antimicrobial activities. Furthermore, they had high antioxidant power, although strain dependent. The results of this study showed that L. gasseri (ADG1b) is the most promising probiotic.

ANTIMICROBIAL ACTIVITY OF ETHANOL EXTRACTS OF $RIBES~NIGRUM$ AND $RIBES~RUBRUM$ LEAVES

Antimicrobial activity of Ribes nigrum L. and Ribes rubrum L. leaf extracts was determined by disk-diffusion method as well as specific growth rate and generation succeeding factor determination. Different Gram-positive (Bacillus subtilis WT-A, Staphylococcus aureus MDC 5233, Enterococcus hirae ATCC 9790) and Gram-negative (Escherichia coli ATCC 25922, Salmonella typhimurium MDC1754, ampicillin resistant E. coli dhpα-pUC18 and kanamycin resistant E. coli pARG25) bacteria and yeasts (Saccharomyces cerevisiae ATCC 9804 and S. cerevisiae ATCC 13007) were used as test-microorganisms. 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay was applied to measure the radical scavenging capacity of extracts obtained from Ribes spp. R. rubrum and R. nigrum extracts antiradical activity expressed with IC50 value of 91.2±1.69 µg‧mL–1 and 66.01±1.65 µg‧mL–1, respectively. The total flavonoid content in plant extracts was determined employing AlCl3 colorimetric assay, and the values were 34.71±0.63 and 49.99±0.86 μg QE‧mg–1 for R. rubrum and R. nigrum extracts, respectively. Total phenolic content of studied extracts was investigated by Folin–Ciocalteu assay. The contents for R. rubrum and R. nigrum extracts were 133.12±6.65 and 167.15±7.29 μg GAE‧mg–1, respectively. Thus, these plants can be considered as potential sources of biologically active substances.

Two cases of bloodstream infections associated with opportunistic bacterial species (Enterococcus hirae and Enterobacter xiangfangensis) in companion animals

BackgroundBloodstream infections are a matter of concern in small animal veterinary practice. Few reports are avaiable, especially regarding the role of opportunistic bacteria in becoming infectious. This report aims to add to the current veterinary literature on two opportunistic bacterial species (Enterococcus hirae and Enterobacter xiangfangensis) associated with bloodstream infections in small animals admitted to the Bologna University Veterinary Hospital.Case presentationIn the first case, a 15-year-old, immunocompromised, cardiopathic dog was admitted to the hospital for anorexia and diarrhea. The patient had a history of previous surgery and hospitalization. After three days, hyperthermia, leukopenia and hyperlactatemia were recorded, and blood culture revealed positivity for Enterococcus hirae, identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The patient’s general conditions progressively worsened, and the patient was euthanized. In the second case, a 2-year-old cat with chronic ocular herpesvirus infection and hypertrophic cardiomyopathy was admitted to the hospital for anorexia and hyperthermia. The cat was hospitalized one week before and received antimicrobial treatment for urinary tract infection by Staphylococcus felis. Hypokalemia and lymphopenia were also diagnosed. The patient progressively improved and was discharged after three days. On the same day, blood culture taken at admission revealed positivity for Enterobacter xiangfangensis, identified using MALDI-TOF MS. After five days, the patient returned with neurological symptoms, hypothermia and bradycardia, and was euthanized.ConclusionsIn small animal veterinary practice, the impact of opportunistic bacterial agents (such as E.hirae and E.xiangfangensis) on bloodstream infections remains unclear. As in human medicine, they can be contracted in every healthcare setting and considered hospital-acquired infections. In this report, we highlighted the threat they pose especially in patients with multiple risk factors. Rapid and accurate diagnostic tools (such as MALDI-TOF MS) could be particularly important for reducing the severity of the infections.

Enterokokların MALDI-TOF MS’de Slanetz ve Bartley ve Kanlı Agar Besiyeri ile Karşılaştırılarak Tanımlanması

Introduction: Matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been used frequently in the last decade as a fast and reliable method for the identification of microorganisms. The fact that only one gram-negative selective medium is recommended in this method causes problems in the diagnosis. There is no information in the scientific literature that Slanetz and Bartley (SB) agar used in the study can be used in MALDI-TOF MS to identify bacteria. Materials and Methods: In this study, it was aimed to compare the identification performance of enterococci isolated from drinking-use and lake waters in Van and its surroundings with blood agar by using MALDI-TOF MS method and to develop a simple and fast sample preparation method that increases this performance. For this purpose, 60 mains and 30 lake water samples were examined for six months. Results: The samples were studied by the membrane filtration method. The isolated enterococci were identified by MALDI-TOF MS method with the isolates taken directly from both blood agar and SB medium, which is a selective medium. The same results were obtained in the species determination in both media. According to these results, 35% of the mains water isolates were Enterococcus casseliflavus, 30% Enterococcus faecalis, 26.7% Enterococcus faecium, 6.6% Enterococcus hirae, 1.7% Enterococcus columbae, and 20% of lake water isolates E. casseliflavus, 40% E. faecalis, 36.7% E. faecium, 3.3% E. hirae, E. casseliflavus in mains water and E. faecalis in lake water are the most frequently isolated enterococci species. Conclusion: Determining that it can also be obtained from selective media such as SB agar, for which there is no data on its suitability for mass spectrometric identification, will contribute to the more effective and rapid use of MALDI-TOF MS in microbiology laboratories. In addition, it has been concluded that all the waters examined are exposed to a significant level of microbial contamination and pose a potential danger to public health. It has been revealed that the consumption of these waters may pose a potential health risk for immunocompromised patients and the elderly, as well as children.

Inhibition of multi-species biofilm formation using chitosan-based film supplemented with essential oils

The biofilm development following microbial attachment on food-grade surfaces represents a main problem in the food sector, leading to significant economic losses. To overcome that, the present investigation aimed to develop biodegradable films that protect against heterogeneous biofilms using chitosan and essential oils (EOs). First, the cross-streak method revealed the absence of antagonism between Staphylococcus aureus, Enterococcus hirae, Escherichia coli, and Pseudomonas aeruginosa strains. The contact angle findings revealed a drop in the electron acceptor (γ+) and qualitative hydrophobicity (θw) associated with an increase in the Lifshitz-van Der Waals component (γLW) and electron donor (γ−) regarding the treated chitosan-based films (CH-films). The environmental scanning electron microscopy (ESEM) images showed that among all the treatments, the CH-film treated with the Pelargonium asperum EO exhibited the weakest antiadhesion activity (73.89 ± 1.42% of the covered surface) while the CH-film treated with the Pelargonium asperum, Thymus satureioides, Cinnamomum zeylanicum, and Eugenia caryophyllus EOs mixture had the strongest activity (9.23 ± 1.66% of the covered surface). From these results, the CH-film containing the EOs mixture presents a high potential for utilization in the food industry as an active coating and packaging that prevent the microorganisms' attachment and the eventual biofilm development.

Redundant potassium transporter systems guarantee the survival of Enterococcus faecalis under stress conditions.

Enterococcus is able to grow in media at pH from 5.0 to 9.0 and a high concentration of NaCl (8%). The ability to respond to these extreme conditions requires the rapid movement of three critical ions: proton (H+), sodium (Na+), and potassium (K+). The activity of the proton F0F1 ATPase and the sodium Na+ V0V1 type ATPase under acidic or alkaline conditions, respectively, is well established in these microorganisms. The potassium uptake transporters KtrI and KtrII were described in Enterococcus hirae, which were associated with growth in acidic and alkaline conditions, respectively. In Enterococcus faecalis, the presence of the Kdp (potassium ATPase) system was early established. However, the homeostasis of potassium in this microorganism is not completely explored. In this study, we demonstrate that Kup and KimA are high-affinity potassium transporters, and the inactivation of these genes in E. faecalis JH2-2 (a Kdp laboratory natural deficient strain) had no effect on the growth parameters. However, in KtrA defective strains (ΔktrA, ΔkupΔktrA) an impaired growth was observed under stress conditions, which was restored to wild type levels by external addition of K+ ions. Among the multiplicity of potassium transporters identify in the genus Enterococcus, Ktr channels (KtrAB and KtrAD), and Kup family symporters (Kup and KimA) are present and may contribute to the particular resistance of these microorganisms to different stress conditions. In addition, we found that the presence of the Kdp system in E. faecalis is strain-dependent, and this transporter is enriched in strains of clinical origin as compared to environmental, commensal, or food isolates.

The effect of essential oils mixture on chitosan-based film surface energy and antiadhesion activity against foodborne bacteria.

In the food sector, the formation of biofilms as a result of microbial adherence on food-grade surfaces causes a major problem resulting in significant economic losses. Thereby, this work aimed to elaborate a biodegradable film using chitosan (CS-film) and reinforce its antiadhesion activity by incorporating pelargonium, clove, thyme, and cinnamon essential oils (EOs). Firstly, the antibacterial activity of these EOs alone and combined against four foodborne bacteria were analyzed by the microdilution method. Synergism was observed in the case of EOs combination. Secondly, the physicochemical characteristics and antiadhesion behavior of the CS-films were assessed by the contact angle method and ESEM, respectively. Results revealed that the EOs mixture treatment impacted considerably the physicochemical characteristics of the CS-film and reduced its qualitative and quantitative hydrophobicity. Moreover, the treated CS-film showed a strong antiadhesion behavior against Enterococcus hirae, Pseudomonas aeruginosa, Escherichia coli, and Staphylococcus aureus with percentages of non-covered surface equal to 97.65 ± 1.43%, 98.76 ± 0.32%, 99.68 ± 0.28%, and 95.63 ± 1.32% respectively. From all these results, the CS-film treated with the mixture of EOs presents a great potential for application as surface coating and food packaging preventing microbial adhesion and thus, avoiding food contamination and spoilage.

Indigenous Lactic Acid Bacteria Isolated from Raw Graviera Cheese and Evaluation of Their Most Important Technological Properties.

The aim of the present study was to characterize LAB isolates from raw-milk cheeses, to evaluate some of their technological properties and to select a few 'wild' LAB strains that could potentially be used as starter cultures. LAB strains were isolated and identified from raw milk, curd, and cheese at 30, 60, and 90 days of ripening. A total of 100 strains were isolated, 20 from each phase of ripening. All isolates were tested for acidification ability, curd formation, and aroma production at 32 °C and 42 °C after 24 and 48 h. Following the acidification test, 42 strains were selected for identification and characterization of their technological properties. A high proportion of lactic acid bacteria and Gram + cocci were found throughout the cheese-making process. Enterococci reached their maximum proportion on the 7th day of ripening while Lactobacilli increased significantly during the first month of ripening. Forty-two strains were identified by phenotypic, biochemical, and molecular techniques. Lactococci were predominant in raw milk and curd while Lactobacilli in the ripening of the cheese. Four LAB strains including one Leuconostoc pseudomenteroides, two Lacticaseibacillus paracasei subsp. paracasei and one Enterococcus hirae, were proposed for their potential use as starters or secondary cultures.

Anti-tumor effect of infant-derived Enterococcus via the inhibition of proliferation and inflammation as well as the promotion of apoptosis.

Probiotic Enterococcus hirae WEHI01 and Enterococcus faecium WEFA23 from infants were previously found to effectively inhibit the development of melanoma. In this study, their immunomodulatory and antitumor mechanisms were systemically studied. In vitro assay showed that E. hirae WEHI01 and E. faecium WEFA23 achieved biphasic immune regulation, which was revealed by the activation of resting spleen lymphocytes and RAW264.7 macrophages, as well as the anti-inflammation effect when immune cells were treated with LPS. The antitumor effects of E. hirae WEHI01 and E. faecium WEFA23 in vitro and vivo were then investigated. CCK8 and the cell scratch assay showed that the conditioned media, which were co-incubated with Enterococcus and spleen lymphocytes, significantly inhibited the proliferation and migration of B16F10, HepG-2 and HT-29 cells. The results of the tumor-bearing mice model experiment showed that E. faecium WEFA23 inhibition of the growth of tumors in mice, and the anti-tumor mechanism involved three aspects, namely tumor proliferation (decreasing expressions of LDHA, VEGF, MMP2, MMP9 and HIF-1α), inhibition of the pro-inflammation state (decreasing expressions of IL-6, TGF-β and IL-17) and the promotion of apoptosis (increasing expression of Bax/Bcl-2, caspase-3 and p53). The results suggest that the two strains of Enterococcus could be promising candidates for treating melanoma with a highly inhibitory effect.

Modulating the Gut Microbiota with Alginate Oligosaccharides In Vitro

Alginate oligosaccharides (AOS) are non-digestible carbohydrates from brown kelp. As such, they are dietary fibers and may have prebiotic potential. Therefore, we investigated the capacity of gut bacteria to utilize AOS with single-strain cultures and as a complex bacterial community. Bifidobacterium adolescentis, Lacticaseibacillus casei and Lacticaseibacillus paracasei showed weak growth (relative to unsupplemented medium; p &lt; 0.05) in the presence of AOS and alginate, while strong growth (p &lt; 0.01) was observed for Bacteroides ovatus when grown with alginate as carbohydrate source. Enterococcus faecium and Enterococcus hirae were for the first time reported to be able to grow on AOS. Further, AOS as substrate was investigated in a complex bacterial community with colonic fermentations in an in vitro gut model. The in vitro gut model indicated that AOS increased short-chain fatty acid (SCFA) levels in donors with a low endogenous SCFA production, but not to the same level as inulin. Bacteroides was found to dominate the bacteria community after in vitro gut simulation with alginate as substrate. Further, stimulation of Bacteroides was observed with AOS in the gut model for two out of three donors with the third donor being more resistant to change. Our results allowed the identification of AOS utilizers among common gut species. The results also demonstrated the capacity of AOS to elevate SCFA levels and positively modulate the gut microbiota during in vitro simulated colon fermentations, although some subjects appear to be resilient to perturbation via substrate changes.

Altered gut microbiome composition by appendectomy contributes to colorectal cancer

Appendectomy impacts the homeostasis of gut microbiome in patients. We aimed to study the role of appendectomy in colorectal cancer (CRC) risk through causing gut microbial dysbiosis. Population-based longitudinal study (cohort 1, n = 129,155) showed a 73.0% increase in CRC risk among appendectomy cases throughout 20 years follow-up (Adjusted sub-distribution hazard ratio (SHR) 1.73, 95% CI 1.49–2.01, P < 0.001). Shotgun metagenomic sequencing was performed on fecal samples from cohort 2 (n = 314). Gut microbial dysbiosis in appendectomy subjects was observed with significant enrichment of 7 CRC-promoting bacteria (Bacteroides vulgatus, Bacteroides fragilis, Veillonella dispar, Prevotella ruminicola, Prevotella fucsa, Prevotella dentalis, Prevotella denticola) and depletion of 5 beneficial commensals (Blautia sp YL58, Enterococcus hirae, Lachnospiraceae bacterium Choco86, Collinsella aerofaciens, Blautia sp SC05B48). Microbial network analysis showed increased correlation strengths among enriched bacteria and their enriched oncogenic pathways in appendectomy subjects compared to controls. Of which, B. fragilis was the centrality in the network of the enriched bacteria. We further confirmed that appendectomy promoted colorectal tumorigenesis in mice by causing gut microbial dysbiosis and impaired intestinal barrier function. Collectively, this study revealed appendectomy-induced microbial dysbiosis characterized by enriched CRC-promoting bacteria and depleted beneficial commensals, signifying that the gut microbiome may play a crucial role in CRC development induced by appendectomy.

Prevalence of macrolide-lincosamide-streptogramin resistant lactic acid bacteria isolated from food samples.

Lactic acid bacteria (LAB) being a reservoir of antibiotic resistance genes, tend to disseminate antibiotic resistance that possibly pose a threat to human and animal health. Therefore, the study focuses on the prevalence of macrolide-lincosamide-streptogramin- (MLS) resistance among LAB isolated from various food samples. Diverse phenotypic and genotypic MLS resistance were determined among the LAB species (n = 146) isolated from fermented food products (n = 6) and intestine of food-producing animals (n = 4). Double disc, triple disc diffusion and standard minimum inhibitory concentration (MIC) tests were evaluated for phenotypic MLS resistance. Specific primers for MLS resistance genes were used for the evaluation of genotypic MLS resistance and gene expressions using total RNA of each isolate at different antibiotic concentrations. The isolates identified are Levilactobacillus brevis (n = 1), Enterococcus hirae (n = 1), Limosilactobacillus fermentum (n = 2), Pediococcus acidilactici (n = 3), Enterococcus faecalis (n = 1). The MIC tests along with induction studies displayed cMLSb, L phenotype, M phenotype, KH phenotype, I phenotype resistance among MLS antibiotics. Genotypic evaluation tests revealed the presence of ermB, mefA/E, msrA/B and msrC genes. Also, gene expression studies displayed increased level of gene expression to the twofold increased antibiotic concentrations. In the view of global health concern, this study identified that food samples and food-producing animals represent source of antibiotic resistant LAB that can disseminate resistance through food chain. This suggests the implementation of awareness in the use of antibiotics as growth promoters and judicious use of antibiotics in veterinary sectors in order to prevent the spread of antibiotic resistance.

Molecular detection and occurrence of vancomycin resistance genes (van A, B, C1, C2/C3) among Enterococcus species isolated from farm ostriches.

Evaluating the prevalence of vancomycin resistance genes (van genes) in enterococcal isolates from food-producing animals is an important public health issue because of the possibility of resistance genes spread to human. The present study aimed to determine the occurrence of vancomycin resistance genes among Enterococcus species obtained from ostrich faecal samples. One hundred and twenty-five faecal samples of apparently healthy ostriches from five different farms were investigated. Genes encoding vancomycin resistance were studied by multiplex-PCR, and susceptibility to six antibiotics was evaluated by disk-diffusion method. In total, 107 Enterococcus spp. isolates were obtained and confirmed by biochemical and molecular tests. Enterococcus faecium was the prevailing species (56 isolates of 107; 52.3%), followed by E. hirae (24 isolates; 22.4%) and E. gallinarum (12 isolates; 11.2%). Of the 107 recovered isolates, 44% harboured at least a type of van genes. vanA, vanC2/3 and vanC1 were identified in 34 (31.7%), 13 isolates (12.1%) and 4 (3.7%) isolates respectively. Additionally, four isolates (E. gallinarum, E. rafinosus) co-harboured the the vanA and vanC1 or vanA and vanC2/3. Enterococcus faecium and Enterococcus hirae strains with the vanA genotype were the most frequent van-carrying enterococci from ostrich faecal samples. Among van-carrying enterococcal isolates, 23.4% were phenotypically resistant to vancomycin. This study revealed a relatively high prevalence (44%) of van-carrying enterococci in ostrich faecal samples. Results of the present study suggest that ostrich faeces could be considered as a reservoir of vancomycin resistance genes, especially vanA containing enterococci that could be potentially transferred to human through the food chain.

Strong anti-viral nano biocide based on Ag/ZnO modified by amodiaquine as an antibacterial and antiviral composite

In this paper, we synthesized Ag/ZnO composite colloidal nanoparticles and the surface of nanoparticles was improved by amodiaquine ligand. The synthesized nanoparticles were characterized using the XRD diffraction pattern, FT-IR Spectroscopy, TEM image, and UV–Vis spectroscopy. The antibacterial, antifungal, and antiviral effects of the synthesized colloid were examined on E.coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Enterococcus hirae bacteria, and Candida Albicans and form spore aspergillus fungi, also influenza, herpes simplex, and covid 19 viruses. The results indicate more than 7 log removal of the bacteria, fungi, and viruses by synthesized colloid with a concentration of 15 μg/L (Ag)/50 µg/ml (ZnO). This removal for covid 19 virus is from 3.2 × 108 numbers to 21 viruses within 30 s. Also, irritation and toxicity tests of the synthesized colloid show harmless effects on human cells and tissues. These colloidal nanoparticles were used as mouthwash solution and their clinical tests were done on 500 people infected by the coronavirus. The results indicate that by washing their mouth and nose three times on day all patients got healthy at different times depending on the depth of the disease. Almost all people with no signs of infection and using this solution as a mouthwash didn’t infect by the virus during the study.

Randomized placebo-controlled trial of feline-origin Enterococcus hirae probiotic effects on preventative health and fecal microbiota composition of fostered shelter kittens.

Diarrhea is the second most common cause of mortality in shelter kittens. Studies examining prevention strategies in this population are lacking. Probiotics are of particular interest but studies in cats are largely limited to healthy adults or those with induced disease. Only one study in domestic cats describes the use of host-derived bacteria as a probiotic. We previously identified Enterococcus hirae as a dominant species colonizing the small intestinal mucosa in healthy shelter kittens. Oral administration of a probiotic formulation of kitten-origin E. hirae (strain 1002-2) mitigated the increase in intestinal permeability and fecal water loss resulting from experimental enteropathogenic E. coli infection in purpose-bred kittens. Based on these findings, we hypothesized that administration of kitten-origin E. hirae to weaned fostered shelter kittens could provide a measurable preventative health benefit. We conducted a randomized, placebo-controlled, blinded clinical trial to determine the impact of a freeze-dried E. hirae probiotic on body weight gain, incidence of diarrhea, carriage of potential diarrheal pathogens, and composition of the intestinal microbiota in weaned fostered shelter kittens. One-hundred thirty kittens completed the study. Fifty-eight kittens received the probiotic and 72 received the placebo. There were no significant differences in age, weight upon initiation of the study, number of days in the study, average daily gain in body weight, or weight at completion of the study. Kittens treated with E. hirae were 3.4 times less likely to develop diarrhea compared to kittens treated with placebo (odds ratio = 0.294, 95% CI 0.109-0.792, p = 0.022). A significant impact of E. hirae was not observed on the presence or abundance of 30 different bacterial, viral, protozoal, fungal, algal, and parasitic agents in feces examined by qPCR. With exception to a decrease in Megamonas, administration of the E. hirae probiotic did not alter the predominant bacterial phyla present in feces based on 16S rRNA gene amplicon sequencing. Decreased incidence of diarrhea associated with preventative administration of E. hirae to foster kittens supports a rationale for use of E. hirae for disease prevention in this young population at high risk for intestinal disease though additional studies are warranted.

Efficacy of Single and Multi-Strain Probiotics on In Vitro Strain Compatibility, Pathogen Inhibition, Biofilm Formation Capability, and Stress Tolerance.

Compatibility of each strain in a multi-strain probiotic (MSP), along with its properties, becomes a strong base for its formulation. In this study, single-strain probiotics (SSPs) and multi-strain probiotics (MSPs) were evaluated in vitro for strain compatibility, microbial antagonism, biofilm formation capacity, and stress tolerance. Bacillus amyloliquefaciens L11, Enterococcus hirae LAB3, and Lysinibacillus fusiformis SPS11 were chosen as MSP1 candidates because they showed much stronger antagonism to Aeromonas hydrophila and Streptococcus agalactiae than a single probiotic. MSP 2 candidates were Lysinibacillus fusiformis strains SPS11, A1, and Lysinibacillus sphaericus strain NAS32 because the inhibition zone produced by MSP 2 against Vibrio harveyi and Vibrio parahaemolyticus was much higher than that produced by its constituent SSPs. MSP1 in the co-culture assay reduced (p &lt; 0.05) A. hydrophila count from 9.89 ± 0.1 CFU mL-1 to 2.14 ± 0.2 CFU mL-1. The biofilm formation of both MSPs were significantly higher (p &lt; 0.05) than its constituent SSPs and the pathogens. The SSPs in both MSPs generally showed resistance to high temperatures (80, 90, and 100 °C) and a wide range of pH (2 to 9). This in vitro assessment study demonstrates that MSP1 and 2 have the potential to be further explored as multi-strain probiotics on selected aquatic species.