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.

Surveys on Pet-Reptile-Associated Multi-Drug-Resistant Salmonella spp. in the Timișoara Metropolitan Region-Western Romania.

The number of reptiles owned as pets has risen worldwide. Additionally, urban expansion has resulted in more significant human encroachment and interactions with the habitats of captive reptiles. Between May and October 2022, 48 reptiles from pet shops and 69 from households were sampled in the Timișoara metropolitan area (western Romania). Three different sample types were collected from each reptile: oral cavity, skin, and cloacal swabs. Salmonella identification was based on ISO 6579-1:2017 (Annex D), a molecular testing method (invA gene target), and strains were serotyped in accordance with the Kauffman-White-Le-Minor technique; the antibiotic susceptibility was assessed according to Decision 2013/652. This study showed that 43.28% of the pet reptiles examined from households and pet shops carried Salmonella spp. All of the strains isolated presented resistance to at least one antibiotic, and 79.32% (23/29) were multi-drug-resistant strains, with the most frequently observed resistances being to gentamicin, nitrofurantion, tobramycin, and trimethoprim-sulfamethoxazole. The findings of the study undertaken by our team reveal that reptile multi-drug-resistant Salmonella is present. Considering this aspect, the most effective way of preventing multi-drug-resistant Salmonella infections requires stringent hygiene control in reptile pet shops as well as ensuring proper animal handling once the animals leave the pet shop and are introduced into households.

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.

Occurrence of Chlamydia spp. in Conjunctival Samples of Stray Cats in Timișoara Municipality, Western Romania.

Despite the widespread public health concern about stray cats serving as reservoirs for zoonotic agents, little is known about the effect of urban and peri-urban landscapes on exposure risk. We conducted this study to monitor the presence of Chlamydia spp. in stray cats, with or without conjunctivitis, living in Timișoara Municipality, Western Romania, using staining and PCR methods. A total of 95 cats were enrolled, and conjunctival samples were harvested from 68 clinically healthy cats and another 27 cats presenting with clinical signs of conjunctivitis. Overall, we found that 65.3% (62/95) of the cats tested positive for Chlamydia spp. by PCR. Chlamydia spp. were detected in 45/95 conjunctival samples using a standard Giemsa stain, compared with 62/95 using PCR (Cohen's kappa index = 0.308; p = 0.0640). Of the cats that tested positive by PCR, 72.6% (45/62) were asymptomatic, and another 27.4% (17/62) expressed clinical signs of conjunctivitis. We found no significant difference between (p > 0.05) the distribution of infection and the recorded epidemiological data (sex, breed, age, territorial distribution, or sampling season). However, the Chlamydia spp. detection frequency was significantly higher in asymptomatic than in symptomatic cats (p = 0.0383). The obtained results increase the level of concern and awareness about the possible zoonotic potential of this pathogen and highlight that urban stray cats can be essential sources of feline chlamydiosis.

The Variation of Serotonin Values in Dogs in Different Environmental Conditions.

Serotonin is considered to be the neurotransmitter that controls several types of behavior: aggressiveness, impulsivity, food selection, stimulation, sexual behavior, reaction to pain, and emotional manifestations. The aim of this study was to determine the serotonin values in 43 dogs, divided into three different experimental variants: (1) between two groups of medium (n = 6) and small (n = 4) breed shelter dogs; (2) in dogs with (n = 15) and without (n = 10) owners after administration of pre-spaying/neutering anesthesia; (3) in different behavioral states (n = 8) classified as follows: M1-happy, M2-aggressive, M3-calmed status, post-exposure to a stressful situation, compared to the reference time referred to as M0. There were no significant differences (p ≥ 0.05) regarding the serotonin values between the two groups of medium and small breed shelter dogs. Following anesthesia, the average mean serotonin values were significantly lower (p ≤ 0.003), by 63.85 ng/mL, in stray dogs compared to dogs with owners. No significant differences (p ≥ 0.05) were found when comparing the reference time M0 to M1, M2, and M3. The differences decreased significantly (p ≤ 0.05), by 89.61 ng/mL, between M1 and M2 and increased significantly (p ≤ 0.008), by 112.78 ng/mL, between M2 and M3.

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.

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.

Antimicrobial Drug-Resistant Salmonella in Urban Cats: Is There an Actual Risk to Public Health?

The present study was undertaken to investigate the presence of Salmonella spp. in the faeces of client-owned cats in urban areas and to evaluate the risk that is posed to public health. Fresh faecal samples were collected directly from the rectums from 53 diarrhoeic and 32 non-diarrhoeic cats. The samples were individually screened for the presence of Salmonella spp. using standard methods and, in the case of positive findings, the resulting typical colonies were then biochemically confirmed using the VITEK®2 automated system. Subsequently, all of the Salmonella spp. isolates were molecularly tested for the presence of the invA gene. All of the isolates were serotyped using the slide agglutination technique according to the White-Kauffmann-Le Minor scheme. The phenotypic antimicrobial susceptibility profile of the isolated strains was obtained from the VITEK®2 system using specific cards from the Gram-negative bacteria. A total of 16 of the samples (18.82%) tested positive for Salmonella spp. according to conventional and molecular testing methods. Serotyping of the Salmonella isolates showed the presence of three serotypes, namely S. enteritidis (n = 9; 56.3%), S. typhimurium (n = 4; 25%), and S. kentucky (n = 3; 18.8%). All of the tested strains showed strong resistance towards cefazolin, cefepime, ceftazidime, and ceftriaxone. Additionally, resistance (listed in descending order of strength) was observed to trimethoprim/sulfamethoxazole (11/16; 68.8%), ampicillin (10/16; 62.5%), ampicillin/sulbactam (9/16; 56.3%), gentamicin (9/16; 56.3%), nitrofurantoin (8/16; 50.0%), and amikacin (5/16; 31.3%). No resistance was expressed against ciprofloxacin, ertapenem, imipenem, levofloxacin, piperacillin/tazobactam, and tobramycin. The results of this study highlight a substantial public health issue and medical concern, especially in vulnerable people, such as children, the elderly, and immunocompromised individuals.

Antibiotic Susceptibility Profile of Pseudomonas aeruginosa Canine Isolates from a Multicentric Study in Romania.

Treating infections caused by Pseudomonas aeruginosa is increasingly difficult due to high antibiotic resistance, materialized through the presence of multiple resistance strains, as well as due to rapid development of resistance throughout treatment. The present survey was conducted to investigate the antibiotic susceptibility profile of Pseudomonas aeruginosa pathogens in two University Veterinary hospitals from different geographical regions of Romania (i.e., Southwest Timișoara county and Northeast Iași county) involved in superficial canine infections. A total of 142 swab specimens were collected from dogs with superficial infections (superficial skin infections, otitis externa, and perianal abscess) to assess the presence of Pseudomonas aeruginosa, based on phenotypic and molecular characterization. According to their confirmed morphological and molecular features, 58 samples (40.84%; 58/142) were positive for Pseudomonas aeruginosa (according to their confirmed morphological and molecular features). Antibiotic susceptibility testing for 12 antibiotics was conducted using the Kirby-Bauer disc diffusion method. Drug resistance was observed in the case of all tested antibiotics. The susceptibility rate of P. aeruginosa strains that were tested in this study was in the following order: ceftazidime (53.44%; 31/58), followed by aztreonam (51.72%; 30/58), amikacin (44.82%; 26/58), azithromycin (41.37%; 24/58), gentamicin (37.93%; 22/58), cefepime (36.20%; 21/58), meropenem (25.86%; 13/58), piperacillin-tazobactam (25.86%; 13/58), imipenem (22.41%; 13/158), ciprofloxacin (17.24%; 10/58), tobramycin (8.62; 5/58), and polymyxin B (1.72; 1/58). The results highlight the importance of antibiotic susceptibility testing in Pseudomonas aeruginosa isolates from dogs with superficial infections to use an adequate treatment plan to manage the skin condition and other pathologies (otitis externa and perianal abscesses).

Boar Semen Contamination: Identification of Gram-Negative Bacteria and Antimicrobial Resistance Profile.

Bacterial contamination of boar semen occurs with some frequency in artificial insemination centers and may have a negative effect on the quality of the semen as well as on the sows' reproductive capacity. Normally, the source of bacterial contamination in pig seminal doses is the own boar. However, distilled water or laboratory equipment used to elaborate the seminal doses can be an important source of bacterial contamination. This study focused on the identification of gram-negative bacteria in boar semen, and impact on the quality of ejaculates obtained from boar, as well as on the establishment of antimicrobial resistance patterns of isolated gram-negative bacteria. Semen samples were collected from 96 boars, ranging in age from 12-36 month, from three artificial insemination centers from the North-West of Romania. Bacterial species were identified by two methods: matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry and API 20 E (BioMerieux, France). The main bacteria isolated from the doses diluted semen were gram-negative bacteria (47.91%), with a majority of the contaminant bacteria belonging to the family Enterobacteriaceae: Seratia marcescens 19.56%, Proteus mirabilis 15.21% and Escherichia coli 10.86% and to the family Pseudomonaceae: Ralstonia picketii 17.39%, Burkholderia cepacia 10.86%, Pseudomonas aeruginosa 8.69%, and Pseudomonas fluorescens 4.34%, respectively. More than half of these isolates (56.52%) were resistant to gentamycin and 58.69% were resistant to penicillin. These antibiotics are very frequently added in sperm diluent in the centers for the processing of sperm from boars in Romania. Regular monitoring for bacterial contamination is an important aspect of a control program.

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.