A Newly Incompatibility F Replicon Allele (FIB81) in Extensively Drug-Resistant Escherichia coli Isolated from Diseased Broilers.

Multiple drug resistance (MDR) has gained pronounced attention among Enterobacterales. The transfer of multiple antimicrobial resistance genes, frequently carried on conjugative incompatibility F (IncF) plasmids and facilitating interspecies resistance transmission, has been linked to Salmonella spp. and E. coli in broilers. In Egypt, the growing resistance is exacerbated by the limited clinical efficacy of many antimicrobials. In this study, IncF groups were screened and characterized in drug-resistant Salmonella spp. and E. coli isolated from broilers. The antimicrobial resistance profile, PCR-based replicon typing of bacterial isolates pre- and post-plasmid curing, and IncF replicon allele sequence typing were investigated. Five isolates of E. coli (5/31; 16.13%) and Salmonella spp. (5/36; 13.89%) were pan-susceptible to the examined antimicrobial agents, and 85.07% of tested isolates were MDR and extensively drug-resistant (XDR). Twelve MDR and XDR E. coli and Salmonella spp. isolates were examined for the existence of IncF replicons (FII, FIA, and FIB). They shared resistance to ampicillin, ampicillin/sulbactam, amoxicillin/clavulanate, doxycycline, cefotaxime, and colistin. All isolates carried from one to two IncF replicons. The FII-FIA-FIB+ and FII-FIA+FIB- were the predominant replicon patterns. FIB was the most frequently detected replicon after plasmid curing. Three XDR E. coli isolates that were resistant to 12-14 antimicrobials carried a newly FIB replicon allele with four nucleotide substitutions: C99→A, G112→T, C113→T, and G114→A. These findings suggest that broilers are a significant reservoir of IncF replicons with highly divergent IncF-FIB plasmid incompatibility groups circulating among XDR Enterobacterales. Supporting these data with additional comprehensive epidemiological studies involving replicons other than the IncF can provide insights for implementing efficient policies to prevent the spreading of new replicons to humans.

The Oxidative Stress Markers' Protective Influence of Sea Buckthorn and Grape Extracts in Atorvastatin-Treated Hyperlipidemic Rats.

Free radicals and reactive oxygen species initiate when the oxidative stress arises. (1) Background: The effect of natural molecules on oxidative stress in hyperlipidemic rats, taking statins, was observed. (2) Methods: One hundred and twelve white Wistar rats, males and females, were divided into seven: Group I received 20 mg of atorvastatin while groups II and III received a combination of 20 mg of atorvastatin and 100 mg of Sea buckthorn and grape extract. Groups IV and V received 100 mg of Sea buckthorn and grape extract, while groups VI and VII received only high-fat diet (HFD) and normal rodents' fodder. After two and six months, rats were euthanized, and blood was gathered to measure the main paraclinical values and total antioxidant capacity (TAC). Also, the liver and kidney were stored for the organs' cytoarchitecture. For statistics, two-way analysis of variance (ANOVA), was performed. (3) Results: HFD produced hyperlipidemia, accompanied by augmented serum and hepatic oxidative stress markers, in addition to a reduction in antioxidant enzyme activities and glutathione levels. Polyphenolic substances proven efficient against HFD caused oxidative stress. (4) Conclusions: Atorvastatin heightened the histological injuries caused by the fatty diet, but these were diminished by taking atorvastatin in combination with 100 mg/kg of plant extracts.

Assessment of the Effect on Periodontitis of Antibiotic Therapy and Bacterial Lysate Treatment.

Periodontitis is an inflammatory process that starts with soft tissue inflammation caused by the intervention of oral bacteria. By modulating local immunity, it is possible to supplement or replace current therapeutic methods. The aim of this study was to compare the effects of an immunostimulatory treatment with the antibiotherapy usually applied to periodontitis patients. On a model of periodontitis induced in 30 rats (divided into three equal groups) with bacterial strains selected from the human oral microbiome (Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum and Streptococcus oralis), we administered antibiotics, bacterial lysates and saline for 10 days. Clinically, no significant lesions were observed between the groups, but hematologically, we detected a decrease in lymphocyte and neutrophil counts in both the antibiotic and lysate-treated groups. Immunologically, IL-6 remained elevated compared to the saline group, denoting the body's effort to compensate for bone loss due to bacterial action. Histopathologically, the results show more pronounced oral tissue regeneration in the antibiotic group and a reduced inflammatory reaction in the lysate group. We can conclude that the proposed bacterial lysate has similar effects to antibiotic therapy and can be considered an option in treating periodontitis, thus eliminating the unnecessary use of antibiotics.

Effect of Sempervivum tectorum Extract on Some Biomarkers of Reproductive Function and Levels of Some Trace Elements in Male Rats Exposed to Aluminum.

Aluminum, a contentious trace element found in the environment, has been demonstrated to have harmful effects on both humans and animals. In contrast, Sempervivum tectorum, an evergreen plant, has been found to offer numerous beneficial effects for both humans and animals. Therefore, this study aims to assess the protective effect of S. tectorum on certain reproductive biomarkers in male rats exposed to aluminum. Thirty-five Wistar rats were randomly divided into five groups: NTC (no-treatment control) received distilled water; NC (negative control) received drinking water containing 1 mg/L aluminum sulfate (AS); E1 received 1 mg/L AS along with an 8% S. tectorum extract; PC (positive control) received only 8% S. tectorum extract; E2 received 1 mg/L AS for three months followed by 8% extract for one month. The study analyzed testosterone, LH, FSH, body weight, and the histological structure of the testis, epididymis, and prostate, as well as the levels of zinc, manganese, copper, and iron in these organs. Significant decreases in body weight, testis, and epididymis size were observed in the aluminum-exposed groups compared to the control, whereas these decreases were not significant in the S. tectorum-treated groups compared to the control. Aluminum exposure led to significant decreases in testosterone and LH levels, with FSH levels showing a nonsignificant decrease in males, which were mitigated significantly by the administration of the plant extract. Histological analysis revealed alterations in the testis, epididymis, and prostate of the AS-exposed groups, including necrosis of seminiferous tubule epithelium and Leydig cells in the testis, and basal epithelial necrosis in the epididymis and prostate. Aluminum levels increased in all organs studied, while levels of zinc, copper, iron, and manganese decreased, showing a negative and significant correlation with aluminum levels. The aqueous extract of S. tectorum demonstrated a protective effect on certain studied biomarkers in male rats affected by aluminum exposure.

Beneficial Effects of Resveratrol and γ-Cyclodextrin on the Hematological and Biochemical Parameters of Healthy Wistar Rats Treated with Cisplatin: A PCA Approach.

It is well known that platinum-based antineoplastic agents, including cisplatin (CP), have side effects that limit their use. Nefrotoxicity, neurotoxicity, and hemolytic anemia are the most common side effects. There are few studies on the reduction in these effects that involves nanoencapsulation; however, almost none involve cyclodextrins (CDs). Changes in the hematological and biochemical parameters of healthy Wistar rats treated with solutions of γ-cyclodextrin/resveratrol/cisplatin (γ-CD/Rv/CP) ternary complexes are investigated for the first time. They are intraperitoneally injected with γ-CD/Rv/CP solutions containing 5 mg CP/kg.b.w. Single shots were administered to six groups of Wistar rats (six individuals for every group) using γ-CD/Rv/CP, γ-CD/CP, γ-CD/Rv complexes, as well as positive- and negative-control groups, respectively. Thirty-two hematological and biochemical parameters were evaluated from blood samples and used as input variables for the principal component analysis (PCA) discrimination of the groups. The best protection was obtained for the γ-CD/Rv/CP ternary complex, which determined closer biochemical values to the control group. These values significantly differ from those of the γ-CD/CP treated group, especially for the IP, UA, and T-Pro kidney-related biochemical parameters. This finding proves the beneficial influence of Rv during CP administration through CD-based carriers.

Pimpinella anisum L. Essential Oil a Valuable Antibacterial and Antifungal Alternative.

Anise (Pimpinella anisum L.) essential oils are intensely investigated worldwide for the beneficial properties, due to the specific bioactive compound's structure. (1) Background: This study characterized the structure of the Pimpinella anisum essential oil and evaluated its antimicrobial properties. (2) Methods: An evaluation of the antibacterial and antifungal activity targeted strains of Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853), Staphylococcus aureus (ATCC 25923), Streptococcus pyogenes (ATCC 19615), and levure Candida albicans (ATCC 10231). Gas chromatography coupled with mass spectrometry (GC/MS) was used for structure identification, and the optical density mass loss was applied for the analysis of different dilutions of aniseed essential oils antimicrobial activity. (3) Results: A total of 13 compounds were identified, of which trans-anethole was in the highest proportion (72.49%), followed by limonene (10.01%), anisole (5%), and α-pinene (3.26%). The results obtained and statistically analyzed, utilizing one-way ANOVA with Bonferroni's multiple comparison test, indicated the antimicrobial activity (p < 0.001) of anise essential oil. (4) Conclusion: Anise essential oil is a promising phyto-remedy with important antimicrobial activity against both Gram-positive and Gram-negative pathogens. Inhibition high percentages were found for the p. aeruginosa and S. aureus strains, but also excellent antifungal activity against C. albicans was ascertained.

The antioxidant effect of Aronia melanocarpa extract in rats oxidative stress induced by cisplatin administration.

BACKGROUND: The reactive oxygen species generated by numerous xenobiotic substances has as consequences the impairment of different organs normal function. Many plants pose antioxidant activity to counteract oxidative stress, among them being the chokeberry (Aronia melanocarpa). The purpose of present study was to determine if the use of A. melanocarpa extract can counteract the oxidative stress induced by cisplatin administration in rats. MATERIAL AND METHODS: The study was made on forty Wistar rats divided in four groups as follows: C (control): receiving i.p. 1 mL of saline solution; E1: receiving cisplatin 20 mg/kg bw, i.p.; E2: receiving cisplatin 20 mg/kg bw, i.p and A. melanocarpa berry 6 % aqueous extract as drinking water, and CB (control blank): i.p 1 mL saline solution and A. melanocarpa 6 % aqueous extract for four weeks. RESULTS: Administration of Cisplatin was followed by the increase of serum superoxide dismutase (+21.18 %, P < 0.05), catalase (+25.44 %, P < 0.001), glutathione peroxidase (+17.88 %, P < 0.05) and thiobarbituric reactive substances (+28.17 %, P < 0.01) but significantly decreased glutathione reductase (-22.35 %, P < 0.001) level comparative to control, pointing out that administration of cisplatin induced oxidative stress in rats. In groups that received A. melanocarpa extract as drinking water, we noted that the levels of the oxidative stress biomarkers tended to be restored almost to normal levels, which could be a possible good antioxidant used in condition to cisplatin use. Also, we noted a significant (P < 0.001) decrease of total antioxidant capacity in liver and kidney of rats exposed to cisplatin, recovered in those that received chokeberry. Studied trace elements important for the stress oxidative enzymes (Cu, Zn, Fe and Mn) were decreased in cisplatin exposed groups compared to control and mainly all were almost to normal level in groups receiving A. melanocarpa. CONCLUSION: A. melanocarpa extract due to its antioxidants content could offer protection against free radicals produced as a consequence of cisplatin use.

The Behavior of Some Bacterial Strains Isolated from Fallow Deer Compared to Antimicrobial Substances in Western Romania.

(1) Background: The resistance levels of Escherichia coli, Salmonella spp., Pseudomonas spp., Staphylococcus spp., etc., isolated from the nasal cavity and the rectum of Dama dama deer from three hunting grounds in Western Romania were assessed. (2) Methods: The analysis was completed using the diffusimetric method, compliant with CLSI reference standards, and with Vitek-2 (BioMérieux, France), on 240 samples. (3) Results: The results were statistically analyzed (by one-way ANOVA) revealing that in four of the ten E. coli strains isolated from animals, 87.5% (p < 0.001) resistance was found. E. coli strains were resistant to cephalexin (100%); seven strains were resistant to cephalothin and ampicillin; six were resistant to cefquinome and cefoperazone; five were resistant to amoxicillin/clavulanic acid; and four were resistant to ceftiofur. However, E. coli was sensitive to amikacin (100%). The most efficient structures were beta-lactams, amikacin, and imipenem, to which all 47 strains studied (100%) were sensitive, followed by nitrofurantoin, to which 45 strains (95.7%) were sensitive, neomycin, to which 44 strains (93.6%) were sensitive, ceftiofur, to which 43 strains (91.5%) were sensitive, and trimethoprim/sulfamethoxazole and marbofloxacin, to which 42 strains (89.4%) were sensitive. (4) Conclusions: In wild animal populations, where a human presence is frequently reported, including a constant presence of domestic animals, despite the perceived low risk of emerging resistance to antimicrobials, resistance is likely to develop frequently.

Antimicrobial Activity of Sempervivum tectorum L. Extract on Pathogenic Bacteria Isolated from Otitis Externa of Dogs.

The present study investigated the antimicrobial activity, total phenolic content, and proanthocyanidin concentration of ethanolic extracts from fresh leaves of Sempervivum tectorum L. The extracts were phytochemically analyzed and evaluated for antimicrobial activity. The broth microdilution method was used to assess antimicrobial activity against pathogenic bacteria isolated from ear swabs taken from dogs with otitis externa. Many compounds were present in the ethanolic aqueous extracts, which exhibited a broad spectrum of antimicrobial activity. They showed strong antibacterial activity against standard clinical Gram-positive strains such as S. aureus and Gram-negative strains such as P. aeruginosa. In our study, the obtained quantity of total phenolic compounds in the ethanol:water extract of leaves was 126.17 mg GAE/g. The proanthocyanidin concentration in the tested Sempervivum tectorum L. extracts was 15.39 mg PAC/g material. The high contents of total phenolics and proanthocyanidin indicated that these compounds contribute to antimicrobial activity. The antimicrobial activity of the tested S. tectorum L. extracts ranged from 1.47 to 63.75 µg/mL, starting with 1.47 µg/mL and 1.75 µg/mL against S. aureus ATCC 25923 and P. aeruginosa ATCC 27853 strains, respectively. Likewise, S. tectorum L. ethanol extract demonstrated a bacteriostatic effect against S. aureus clinical isolate with a median MIC of 23.25 µg/mL and MBC of 37.23 µg/mL; and bactericidal against S. aureus ATCC 25923 with the median MIC of 20.33 µg/mL and MBC of 37.29 µg/mL. In the Gram-negative P. aeruginosa clinical and standard strains, the expressed MIC and MBC values were 24.234 and 20.53 µg/mL for MIC, and 37.30 and 37.02 µg/mL for MBC, respectively.

Effects of Aronia melanocarpa and Hypericum perforatum aqueous extracts on hexavalent chromium induced toxicity in rat's thyrocytes.

BACKGROUND: Hexavalent chromium known as oxidizing agent is able to form reactive oxygen species. Aronia melanocarpa and Hypericum perforatum are two plants known for their antioxidant effects. Our study aimed to establish if CrVI induces apoptosis and structural changes in thyrocytes and if its effect can be counteracted by the administration of both extracts. MATERIALS AND METHODS: Wistar rats divided in five groups: C - distilled water (DW), Cr - 75 mg/L CrVI in DW for 3 months, Cr 2 - 75 mg/L CrVI in DW for 3 months followed by 1 month DW, CrA - 3 months 75 mg/L CrVI in DW and 1 month Aronia 2.5% extract, CrH - 3 months 75 mg/L CrVI in DW and 1 month Hypericum 2.5% extract. Histological assessment and qRT-PCR for evaluation of BAX and Bcl2 protein levels performed on thyroid samples. RESULTS: The Cr and Cr2 groups were those with altered cytoarchitecture: increase in the diameter of many thyroid follicles, a decrease in their number, a decrease in the height of the follicular cells. The histological examination of the CrH group revealed almost recovery of structural architecture. The BAX gene levels were higher in the Cr and Cr2 groups indicating the apoptotic activity of chromium. In extract receiving groups the BAX gene expressions were significantly lower, but the lowest level presented the CrH group. Bcl2 gene expression levels indicate antiapoptotic activity being elevated in the Cr group, followed by CrA, Cr2, and CrH groups. The BAX/Bcl2 ratio which significantly increased in the case of the Cr and Cr2 group compared to the groups that were administered the two plant extracts. CONCLUSION: The results obtained in this study confirm that CrVI has toxic effects on thyroid endocrine cells and H. perforatum has stronger antioxidant properties against the action of hexavalent chromium in thyrocytes than A. melanocarpa.

The Role of Methyl-(Z)-11-tetradecenoate Acid from the Bacterial Membrane Lipid Composition in Escherichia coli Antibiotic Resistance.

Background: The bacterial membrane plays a critical role in the survival of bacteria and the effectiveness of antimicrobial peptides in protecting the host. The lipid constituents of the bacterial membrane are not evenly distributed, and they could be affected by clustering anionic lipids with cationic peptides with multiple positive charges. That could be harmful to bacteria because it prevents lipids from interacting with other molecular components of the cell membrane, disrupts existing natural domains, or creates phase boundary defects between the clustered lipids and the bulk of the membrane. This preliminary quantitative study is aimed at assembling a correlation between antibiotic resistance and bacterial lipid composition in E. coli, based on the function and arrangement of the bilipid coating of the bacterial cell, intimately associated with the path of antimicrobials through membranes. Methods: Fifteen multiresistant E. coli samples are collected from swine with enterocolitis tested for resistance levels using the disc diffusimetric method (Kirby-Bauer disc diffusion). Pathogen identification completed using the API 20E multitest system revealed the E. coli presence in 11 samples. In these samples, bacterial membrane detection of fatty acid methyl esters (FAME) operating a 240 MS Ion Trap (Varian) GC/MS (Agilent Technologies, Santa Clara, CA, USA) was performed, using the MIDI Sherlock recognition software model. Results: Interpreting the descriptive statistical method, the correlation matrix, and regression curves and after ANOVA analysis, we ascertained that the studied E. coli population statistically confirmed different degrees of resistance in most of the samples analyzed in this test. Conclusions: In one case, the methyl-(Z)-11-tetradecenoate acid was observed to have a relationship with the susceptibility evaluation by using the disc diffusimetric method, which has revealed the lowest rate of antimicrobial resistance, so it has importance in further resistance evaluation studies.

Antibacterial Activity of Romanian Propolis against Staphylococcus aureus Isolated from Dogs with Superficial Pyoderma: In Vitro Test.

Staphylococcal infection treatment in dogs is frequently associated with adverse side effects, high costs, prolonged treatment, and resistant strain selection. Staphylococcus aureus is the most frequently isolated staphylococci in cases of canine superficial pyoderma. The number of Staphylococcus strains to exhibit primary resistance to various drugs in vitro is increasing. Propolis has a diverse chemical composition and well-known therapeutic properties against bacterial infections. The current investigation evaluated in vitro the antimicrobial activity of the commercial allopathic antimicrobials, Romanian propolis ethanolic extracts, against clinical Staphylococcus aureus strains isolated from superficial dermatitis clinical samples in dogs and two reference strains: Staphylococcus aureus ATCC 25923 and Staphylococcus aureus ATCC 43300, as the MSSA and MRSA positive controls, respectively, in western Romania. We used the microdilution broth technique to evaluate the susceptibility profile of the bacteria. The minimum inhibitory concentration (MIC) of the Romanian propolis ethanolic extract ranged from 6 to 10 µg/mL for all isolates, determined by the broth microdilution method. The MICs of ethanolic Romanian propolis extracts had a pronounced antibacterial activity. These results indicate that propolis can potentially be used and recommended for in vivo experiments as a promising therapeutic agent against Staphylococcus aureus infections in superficial dermatitis of dogs.

Use of Eisenia fetida as a Biological Risk Marker in a Qualitative Eco Assessment Test of a Romanian Watercourse.

The qualitative trials were conducted by exposing earthworms to diverse contaminants sources using standard earthworms' avoidance tests (considered useful ab initio indicators). For two years, we observed the Jiu River pollution points. We observed soil traits in 15 sampling points from two neighboring Romanian counties where Jiu River flows, by evaluating the risk of pollution on Eisenia fetida earthworms. The ISO: 11268-2:2015 acute static test was used, following the ethological and bodily features of E. fetida earthworms for 14 days, and then the results obtained for clean soils vs. those presumed polluted were statistically analyzed. Results disclosed statistically significant values (p < 0.05) for the two-way ANOVA and Tukey multiple comparisons tests used for the soil samples thought to be polluted. The mortality percentages by location/replica/year/county find out a high statistical correlation documenting observations related to soil samples gathered from 10 m (p < 0.01) and 30 m (p < 0.001). Compared with the control, the statistical analysis of Relative Growth Rate (RGR) (p < 0.05) and Specific Growth Rate (SGR) (p < 0.01) confirmed that, in the case of soil samples collected from 10 and 30 m from the Jiu River's axis, the earthworms did not gain weight, qualitatively attesting the pollution suspicion/presence of chemical factors potentially pernicious for earthworms.

Pathology and Prevalence of Antibiotic-Resistant Bacteria: A Study of 398 Pet Reptiles.

Reptiles are potential reservoirs of bacteria that could be transmitted, thus becoming a zoonotic hazard. (1) Background: This three-year investigation surveyed the pathological status of 398 pet reptiles: chelonians, snakes (venomous/non-venomous), and lizards. The main pathological entities found were related to the skin, the sensory organs, the digestive system, the respiratory system, the cardiovascular system, the urinary system, the genitalia, the osteo−muscular tract, surgical issues, tumors, and intoxications. (2) Methods: In 25 individuals treated with antibiotics, no clinical healing was recorded, for this reason, an antimicrobial resistance profile analysis of the 43 samples gathered was processed. An antibiogram was performed using the VITEK®2 ID-GP (bio-Mérieux, Marcy l'Etoile, France) automated platform, with 22 bacterial strains being isolated. (3) Results: The statistics (ANOVA) revealed that the most common disease category was diseases of the digestive system, followed by diseases of the skin, respiratory system, nervous system, and reproductive system. A significant correlation (p < 0.01) between disease incidence and reptile species was reported, with correlations found between all species and diseases diagnosed. The most common bacteria isolated were Enterococcus faecalis, Pseudomonas aeruginosa, Stenotrophomas (Xanthomonas) maltophilia, Escherichia coli, Klebsiella oxytoca, and Salmonella spp., but Beta-hemolytic Streptococcus, Staphylococcus aureus, Citrobacter spp., and Proteus spp. were also identified. (4) Conclusions: These microorganisms revealed degrees of resistance against penicillins, cephalosporins, macrolides, lincosamides, aminoglycosides, and tetracyclines. The animals can be categorized according to their sensitivity to diseases in the following order (most sensitive to least sensitive): chelonians, venomous snakes, non-venomous snakes, and lizards.

Quercetin Beneficial Role in the Homeostatic Variation of Certain Trace Elements in Dyslipidemic Mice.

Background: Quercetin's role in the homeostasis of certain trace elements in dyslipidemia induced in mice was assessed. Methods: Forty BALB/c mice were allocated into 4 groups as follows: control; HFD, received fat diet; HFD + Q group, received HFD +500 mg/L quercetin; and blank control (Q)-normal food + 500 mg/L quercetin in drinking water. Results: By analyzing the values of total proteins, albumins, cholesterol, triglycerides, HDL cholesterol, LDL cholesterol, and the levels of several trace elements in blood and organs, we perceived a statistically significant increase ( ∗∗ , p < 0.01) of TP, ALB, TC, TGE, and LDL-c. A nonsignificant decrease was ascertained to HDL-c value in the HFD and quercetin groups (p > 0.05). In the HFD group, all analyzed elements in the kidney and spleen increased, also Cu, Li, Mg, Mn, Pb, and of B, Ba, Cr, Cd, Cu, Fe, Li, Mn, Ni, Pb, and Zn in the heart increased, but furthermore, Ag, B, Ba, Cd, Cr, Fe, Ni, and Zn in blood, Ag and Zn in the liver, and Cd in the spleen decreased. In the HFD group who received quercetin, elements (except B) were decreased in kidney and liver, also increased Ag, Ba, Cr, Fe, Li, Ni, Zn in blood, but similarly, Ag, B, Ba, Cd, Cu, Mn, and Pb declined in the spleen and heart. Conclusions: Results proved the quercetin beneficial role.

Aronia melanocarpa Ameliorates Adrenal Cytoarchitecture Against the Hexavalent Chromium-Induced Injury.

Hexavalent chromium is a toxin that penetrates the cell, triggering reactive oxygen species (ROS) production. Aronia melanocarpa, due to its proanthocyanidins, anthocyanins, and phenolic acid contents, is a valuable antioxidant. The aim was to observe the influence of hexavalent chromium Cr(VI) on the adrenal gland, and if this impact can be recovered by the administration of A. melanocarpa. Accordingly, 36 rats were divided into six groups: control; Aronia; Cr receiving Cr(VI) in distilled water for 3 months; CrA receiving a mix of Cr(VI) and A. melanocarpa at 2.5% aqueous extract for 3 months; Cr2 receiving, for 3 months, Cr(VI) in distilled water, and next, for 1 month, only distilled water; and respectively, CrA2 receiving, for 3 months, Cr(VI) in distilled water, followed by 1 month of Aronia at 2.5% extract administration. The adrenal gland samples were examined toward histological and molecular assessment, and results were statistically analyzed (ANOVA). Hexavalent chromium induced changes in the adrenal cortex expressed by focal or diffuse hypertrophies, cytoplasmic vacuolization (due to lipidic accumulation), and cells' shape and size alteration, including necrosis. These structural alterations were carried by Bax and Bcl2 gene expression: the Bax gene expression levels, increased significantly (p < 0.001) in all experimental groups, except the Aronia group, compared with control. In the Cr2, CrA, and CrA2 groups, notable reduction of Bax gene expression (p < 0.001) was reported compared with the Cr group. Regarding the Bcl2 gene expression (p < 0.001), a significant increase was observed in the experimental groups, compared with the control. The Bcl2 expression level had a similar pattern to Bax gene, consequently trying to compensate its overexpression. Aronia administered concomitantly, or after Cr(VI), diminished structural changes and expression of the studied genes, thus reducing the Bax/Bcl2 ratio and suggesting that the active ingredients from Aronia are capable of blocking apoptotic cascade induced by the pathway of Bax and Bcl2 proteins.

Is ECLIA Serum Cortisol Concentration Measurement, an Accurate Indicator of Pain Severity in Dogs with Locomotor Pain?

The purpose of determining serum cortisol level is to reflect the activity of stress axis, ethological alterations, acute and chronic pain, life quality, or psychogenic stress. Although it is stated that stress can produce a measurable influence on the cortisol level, a certified value of this pain biomarker in dogs was not generally accepted yet. This study aimed to investigate if serum cortisol measured follows allopathic treatments only, or it is associated with physiotherapy, point out pain level in dogs with orthopedic disease, which could reveal the healing progress. The diagnostic identified: hip dysplasia, cranial cruciate ligament rupture, or intervertebral disc disease. Ortolani and Barden tests, together with clinical examination, drawer sign, and tibia compression test, were done in dogs exhibiting postures, and motion alteration, and X-Ray confirmed. A total of 30 dogs were grouped in healthy (n = 10) and pain groups (n = 20), the blood sampling is done at the beginning of the investigation, and after ten days of the study. Dogs were handled in two ways: G1-treated with Nonsteroidal Anti-Inflammatory Drugs (NSAIDs) only and respectively, G2-by therapy and physiotherapy. The analysis was performed on a Roche Cobas Analyzer (Roche, USA), serum cortisol being determined by Electrochemiluminescence immunoassay (ECLIA), and statistics using ANOVA, following Tukey's Multiple Comparison Test. The results revealed that, out of ten specimens in the Control group, nine were within the normal limits: 5-65 ng × mL-1 (24.76 ± 19.48678), and one sample under the set limit. In G1, it was observed that the plasmatic P1 values were below the levels of P2, in six situations. In G2, although the status of all subjects improved radically with the removal or evident reduction of pain, confirmed clinically and imagistically, the P2 values in five dogs were higher than the initial P1 values, and in contradiction with the observed clinical reality. Comparing results, the mean difference in G1 was 0.41, and in G2 = 2.54, with an SD for G1 = 13.38, and G2 = 16.66, registering moderate development. Standard deviation illustrated that the values of treated groups were highly spread throughout the interval, and the serum cortisol assay did not generate significant statistical differences between groups in our case. This inferred the doubt whether the used detection method or values registered correctly indicates the pain levels in dog species.

Comparative Evaluation of qnrA, qnrB, and qnrS Genes in Enterobacteriaceae Ciprofloxacin-Resistant Cases, in Swine Units and a Hospital from Western Romania.

Excessive use of antimicrobials and inadequate infection control practices has turned antimicrobial resistance (AMR) into a global, public health peril. We studied the expression of qnrA, qnrB, and qnrS plasmid in ciprofloxacin (CIP)-resistant strains of Escherichia coli in swine and humans from Romania, using the Polymerase Chain Reaction (PCR) technique. Antibiotic Susceptibility Testing (AST) for human subjects (H) on 147 samples and 53 swine (S) was ascertained as well as the isolation of bacterial DNA (E. coli) as follows: bacteriolysis, DNA-binding, rinsing, elution, amplification, and nucleic acids' migration and U.V. visualization stages. From 24 samples of E. coli resistant to CIP collected from H subjects and 15 from S, for PCR analysis, 15 H and 12 S were used, with DNA purity of 1.8. The statistically analyzed results using the Crosstabs function (IBM SPSS Statistics-Ver. 2.1.), revealed the qnrS (417 bp) gene in 13 human subjects (52.0%), as well as in all swine samples studied. The qnrB (526 bp) gene was exposed in 9 of the human patients (36.0%) and in all swine isolates, and the qnrA (516 bp) gene was observed only in 3 of the isolates obtained from human subjects (12.0%) and was not discovered in pigs (p > 0.05). The presence of plasmids qnrA, qnrB, and qnrS in the human samples and of qnrB and qnrS in swine, facilitates the survival of pathogens despite the CIP action. The long-term use of CIP could cause a boost in the prevalence of qnr resistance genes, and resistance in the pigs destined for slaughter, a perturbing fact for public health and the human consumer.

Sea Buckthorn and Grape Antioxidant Effects in Hyperlipidemic Rats: Relationship with the Atorvastatin Therapy.

BACKGROUND: Medications to reduce oxidative stress are preventing cellular damage associated with hyperlipidemia. In this regard, statins (e.g., atorvastatin) act primarily by decrease in low-density lipoprotein-c but, in the last decade, hepatotoxicity, associated with liver injuries in the next months after treatments' initiation, was reported. In this case, associated phytotherapy can be a solution. PURPOSE: To investigate the antioxidant potential and response to free radicals, in the case of hyperlipidemic rats treated with atorvastatin. Sea buckthorn (Hippophae rhamnoides) and a grape extract (antioxivita) efficiency in the oxidative stress were investigated, also being ascertained the rats' organs cytoarchitecture. METHODS: Eighty-four hyperlipidemic Wistar rats were divided into seven groups and orally treated as follows: ATS, atorvastatin (20 mg/kg·bw); ATS + Hr, atorvastatin + H. rhamnoides; ATS + Aox, atorvastatin + grape extract; Hr, H. rhamnoides; and Aox, grape extract (both as 100 mg/kg·bw). HFD and Control received high fat diet and normal fodder only. After two and six months, respectively, rats were euthanized and the heart, liver, and kidneys were gathered. The tissue samples were prepared by homogenization of 0.5 g tissue, in ethanol, kept for 48 hours at 4°C-10°C and then filtered, in order to assess organs' cytoarchitecture and the TAC's values (by using cupric ion reducing antioxidant capacity (CUPRAC) assay). The test tubes were incubated, at room temperature, for 30 minutes, and then analyzed using a spectrophotometer at 450-650 nm. RESULTS: The statistics (ANOVA) revealed that sea buckthorn diminished notably (p < 0.001) the oxidative stress in the heart, liver, and kidney. After six months, the TAC's reduced levels for the heart were significant (p < 0.001) in ATS + Aox. In the case of histology, the liver's cytoarchitecture in ATS revealed abnormal cytoarchitecture. In ATS + Hr, ATS + Aox, Hr, and Aox, cell regeneration improved in different stages, especially for ATS + Hr and ATS + Aox, in comparison with HFD, which exhibited fat degeneration. Kidney's cytoarchitecture revealed cellular healing, especially in ATS + Hr and ATS + Aox.

Gingival proliferative growth - stress and cytoarchitecture related with fixed and mobile orthodontic therapy.

The fixed orthodontic measures taken induce significant stress to the gingival growth process during arch wire maneuvers of aligning and leveling. We observed, for a period of one to four years, fixed orthodontic devices in 80 human subjects. From these, we selected 44 subjects (22 women and 22 men) where the inflammatory process exhibited following the orthodontic fixed treatment, and with vacuum-formed orthodontic retainers (VFR) succeeding to fixed treatment. Samples were collected from each patient and histological and immunohistochemical (IHC) methodology was made to analyze the cytoarchitecture. Statistics were made after one-way analysis of variance (ANOVA), with the Bonferroni's correction. The IHC examination performed in the early stage revealed the presence in the inflammatory infiltrate of CD8-type T-lymphocytes, and of dendritic cells in large numbers. The examination performed in the late stage revealed the presence in the inflammatory infiltrate of CD20-type B-lymphocytes, which are mature cells capable of immunoglobulin synthesis, their activation being an important step in the maturation of the antibody response. The stress generated by arch wires in both genders was significantly higher than in the case of VFR. This observation was pointed out also by the cytohistological investigation outcome but was also based on an original scale conceived by our research team, following gingival hyperplasia evaluation. Also, with statistical significance, the comparative obtained values for men (p=0.01) and for women (p=0.001) illustrate clinical observations, allowing to affirm that, in our case, men were more stressed in bearing arch wire devices (AWD) and VFR, in comparison with women.