Introduction: Pseudomonas aeruginosa is one of the most common nosocomial pathogens, known with a wide resistance to antimicrobials. Carbapenemases producing Pseudomonas aeruginosa is a growing global public health concern as this pathogen is easily transmissible among patients. Metallo-Beta-lactamases is the most important class of these carbapenemases with their broad-spectrum resistance profile. This study was conducted to investigate the prevalence of MBL-producing P. aeruginosa collected in an Algerian hospital. Methodology: All Metallo-β-lactamase (MBL)-producing P. aeruginosa isolates recovered from patients during a 2 years period (2015-2016) were studied using a combination of phenotypic and molecular typing methods (susceptibility testing, molecular characterization of carbapenemase-encoding genes, multi-locus sequence typing and pulsed-field gel electrophoresis). Results: A total of twenty-six MBL producing P. aeruginosa of 188 isolates were investigated. The burns unit ranked in the first position of the majority of identified cases with 73.07%. About 73.07% of total MBL isolates were mainly isolated from pus samples. The studied isolates were subjected to the molecular typing, in which 4 different Dra1-PFGE patterns and 3 sequences type were assigned (ST244, ST381, and ST1076), and all isolates were revealed positive for VIM-4. Conclusions: We report the third description of blaVIM-4 in Algeria indicating the emergence and spread of carbapenemase-encoding genes among P. aeruginosa in the hospital environment.

Background/Objectives: Pseudomonas aeruginosa causes chronic infections in humans and animals, especially cats and dogs. This bacterium’s ability to adapt and acquire antibiotic resistance traits may complicate and exacerbate antibacterial therapy. This study aimed to evaluate the antibiotic resistance patterns, virulence factors and ability to form biofilms of P. aeruginosa strains isolated from Algerian dogs and cats. Methods: Nineteen samples were collected from healthy and diseased dogs and cats. Isolates were studied for their antibiotic-resistance patterns (disc diffusion method) and biofilm formation (Microtiter assay) and were whole-genome sequenced (MinION). Results: Nineteen P. aeruginosa strains (15 from dogs and 4 from cats) were isolated. Antibiotic-resistance phenotypes were observed against amoxicillin–clavulanic acid (100%); meanwhile, resistance towards ticarcillin was 40% (dogs) and 25% (cats), ticarcillin–clavulanic acid was 13.33% and 25% for dogs and cats, respectively, and imipenem was 75% (cats) and 20% (dogs). Moreover, 95% of strains were biofilm-producers. Different antimicrobial resistance genes (ARGs) were found: beta-lactamase genes, mainly PAO, OXA-494, OXA-50 and OXA-396 and an aminoglycoside gene (aph(3′)-IIb). The main high-risk sequence types (STs) were ST244, 2788, 388 and 1247. A large panel of virulence genes was detected: exoS, exoT, exoY, lasA, toxA, prpL, algD, rhIA and others. Conclusions: The genetic variety in antibiotic-resistance genes of resistant and virulent P. aeruginosa strains in dogs makes public health protection difficult. Continuous monitoring and research in compliance with the One Health policy are needed to solve this problem.

Proteins are crucial molecules of biological processes with particular functions. They are marginally stable biomolecules and are readily prone towards denaturation due to the occurrence of different stressed conditions in solution, dried, or frozen forms. Some factors, such as pH, storage containers, temperature, buffer systems, and repetitive freezing and thawing, play a critical role in maintaining the stability of proteins. This review summarizes the pragmatic methods for the storage of purified proteins, preserving an ample amount of biological activity. It elucidates the simple and empirical means of preventing proteolytic denaturation, microbial contamination, low-temperature storage, and the inclusion of several kinds of stabilizing agents. Various types of protease inhibitors are known to reduce proteolytic degradation with improved protein storage stability. The addition of antimicrobials (e.g., thiomersal and sodium azide) and filtration are used to prevent microbial contamination. Low temperature and freeze-drying (lyophilization) are also known as significant methods for protein storage. Additionally, the inclusion of stabilizing agents, such as osmolytes, reducing agents, ionic stabilizers, specific substrates, ligands, and exogenous proteins, and concentrating diluted proteins are effective strategies for reducing the degradation and enhancing the shelf-life of proteins. Several stabilizing agents, such as -mercaptoethanol, EDTA, EGTA, glycerol, and ethylene glycol, enhance protein stability with less to no damage. Thus, these strategies provide substantial directions for maintaining the integrity of proteins, facilitating both investigational reproducibility and commercial viability of protein-based products.

Background/Objectives : Pseudomonas aeruginosa causes chronic infections in humans and animals, especially cats and dogs. This bacterium's ability to adapt and acquire antibiotic resistance traits may complicate and exacerbate antibacterial therapy. This study aimed to evaluate the antibiotic resistance patterns, virulence factors and ability to form biofilm of P. aeruginosa strains isolated from Algerian dogs and cats. Methods: Nineteen samples were collected from healthy and diseased dogs and cats. Isolates were studied for their antibiotic resistance patterns (disc diffusion method), biofilm formation (Microtiter assay) and were Whole genome sequenced (MinION). Results: Nineteen P. aeruginosa strains (15 from dogs and 4 from cats) were isolated. Antibiotic resistance phenotypes were observed against amoxicillin-clavulanic acid (100%), meanwhile resistance towards ticarcillin was 40% (dogs) and 25% (cats), ticarcillin-clavulanic acid was 13.33% and 25% for dogs and cats respectively and imipenem was 75% (cats) and 20% (dogs). 95% of strains were biofilm producers. Different antimicrobial resistance genes (ARGs) were found: beta-lactamase genes mainly PAO, OXA-494, OXA-50 and OXA-396, aminoglycosides gene (aph(3’)-IIb), fosA for fosfomycin and catB7 for phenicol. The main high risk STs were ST244, 2788, 388 and 1247. A large panel of virulence genes was detected: exoS, exoT, exoY, lasA, toxA, prpL, algD, rhIA and others. Conclusions: The genetic variety in antibiotic resistance genes of resistant and virulent P. aeruginosa strains in dogs makes public health protection difficult. Continuous monitoring and research in compliance with the One Health policy are needed to solve this problem.

Background/Objectives: Staphylococcus aureus and Staphylococcus pseudintermedius are major opportunistic pathogens in both humans and dogs. In pets, the dissemination of methicillin-resistant isolates (MRSA or MRSP) is problematic for the treatment of animals and is a public health issue due to their zoonotic potential. MRSA and MRSP may also harbor virulent genes that increase their dangerousness. This study aimed to assess the prevalence of (MR)SA and (MR)SP in healthy dogs and their owners in Algeria. Methods: Swabs were collected from various body sites of healthy dogs (n = 88) and from the nose of their owners (n = 38). Antimicrobial susceptibility testing was performed by antibiograms according to the disc diffusion method, and clonality was assessed using Pulsed-Field Gel Electrophoresis (PFGE). All methicillin-resistant isolates were short-read whole-genome sequenced using the Illumina technology. Results: 26 S. aureus and 17 S. pseudintermedius isolates were respectively collected from 13 dogs (13/88, 14.8%). No MRSP isolate was detected, while MRSA was found in six dogs (6.8%). Isolates belonged to ST1 (n = 3), ST 80 (n = 1), and ST 22 (n = 2, including the single-locus variant ST7118). All MRSA displayed the immune evasion cluster (IEC) type E. The ST80 isolate presented the Panton–Valentine toxin, and the ST22/ST7118 isolates carried the tst gene coding for the toxic shock syndrome toxin. Conclusions: The epidemiology of MRSA in healthy Algerian dogs mirrors the one in Algerian people. This poses a zoonotic and public health concern due to the virulence and resistance genes displayed by these isolates. Our results indicate the need for developing One Health strategies to avoid a large-scale dissemination of MRSA in Algerian dogs.