Antimicrobial activity of α-mangostin against Staphylococcus species from companion animals in vitro and therapeutic potential of α-mangostin in skin diseases caused by S. pseudintermedius.

Antimicrobial resistance in Staphylococcus species from companion animals is becoming increasingly prevalent worldwide. S. pseudintermedius is a leading cause of skin infections in companion animals. α-mangostin (α-MG) exhibits various pharmacological activities, including antimicrobial activity against G (+) bacteria. This study investigated the antimicrobial activity of α-MG against clinical isolates of Staphylococcus species from companion animals and assessed the therapeutic potential of α-MG in skin diseases induced by S. pseudintermedius in a murine model. Furthermore, the action mechanisms of α-MG against S. pseudintermedius were investigated. α-MG exhibited antimicrobial activity against clinical isolates of five different Staphylococcus species from skin diseases of companion animals in vitro, but not G (-) bacteria. α-MG specifically interacted with the major histocompatibility complex II analogous protein (MAP) domain-containing protein located in the cytoplasmic membrane of S. pseudintermedius via hydroxyl groups at C-3 and C-6. Pretreatment of S. pseudintermedius with anti-MAP domain-containing protein polyclonal serum significantly reduced the antimicrobial activity of α-MG. The sub-minimum inhibitory concentration of α-MG differentially regulated 194 genes, especially metabolic pathway and virulence determinants, in S. pseudintermedius. α-MG in pluronic lecithin organogel significantly reduced the bacterial number, partially restored the epidermal barrier, and suppressed the expression of cytokine genes associated with pro-inflammatory, Th1, Th2, and Th17 in skin lesions induced by S. pseudintermedius in a murine model. Thus, α-MG is a potential therapeutic candidate for treating skin diseases caused by Staphylococcus species in companion animals.

Evaluación in vitro de múltiples agentes antibacterianos para el tratamiento de la blefaritis anterior crónica estafilocócica / In vitro evaluation of multiple antibacterial agents for the treatment of chronic staphylococcal anterior blepharitis

Objetivo Evaluar la eficacia bactericida de varios compuestos utilizados en el tratamiento de la blefaritis anterior estafilocócica crónica mediante un estudio in vitro. Materiales y métodos Se cultivaron cepas comerciales estándar de Staphylococcus aureus (SAu) (ATCC 25923 Culti-Loops) y Staphylococcus coagulasa-negativo (CoNS) (ATCC 12228 Culti-Loops). Se realizaron pruebas de sensibilidad a vancomicina 30μg, netilmicina 30μg, ácido hipocloroso (HOCl) al 0,01% (Ocudox™, Brill®), aceite de hoja de Melaleuca alternifolia (MeAl) (Navyblef® Cuidado diario, NOVAX®) y digluconato de clorhexidina al 1% (DGCH) (Cristalmina™, Salvat®) mediante el método de difusión en disco de agar (Rosco Neo-Sensitabs™). A las 24horas se midieron los halos inducidos con calibradores automáticos. Los resultados se analizaron con las guías EUCAST- y CLSI potency Neo-Sensitabs™. Resultados La vancomicina indujo un halo de 22,37mm y 21,81mm en SAu y CoNS, respectivamente. La netilmicina produjo halos de 24,45mm en SAu y de 32,49mm en CoNS. MeAl indujo halos de 12,65mm en SAu y de 15,83mm en CoNS. Se encontró un halo de 12,11mm en SAu y un halo de 18,38mm en CoNS utilizando HOCl. DGCH produjo halos de 26,55mm y 23,12mm en SAu y CoNS, respectivamente. Conclusión La netilmicina y la vancomicina demostraron actividad antibiótica frente a ambos patógenos, por lo que pueden ser terapias alternativas de rescate para tratar la blefaritis estafilocócica crónica. El DGCH presenta una eficacia frente a ambos comparable a los antibióticos, mientras que el HOCl y la MeAl demuestran menor eficacia (AU)

Objective To evaluate the bactericidal efficacy of several compounds used in the treatment of chronic staphylococcal anterior blepharitis through an in vitro study. Materials and methods Standard commercial strains of Staphylococcus aureus (SAu) (ATCC 25923 Culti-Loops) and coagulase-negative Staphylococcus (CoNS) (ATCC 12228 Culti-Loops) were cultured. Susceptibility tests were performed to vancomycin 30μg, netilmicin 30μg, hypochlorous acid (HOCl) 0.01% (Ocudox™, Brill®), Melaleuca alternifolia leaf oil (MeAl) (Navyblef® Daily Care, NOVAX®) and 1% chlorhexidine digluconate (DGCH) (Cristalmina™, Salvat®) using the agar disk diffusion method (Rosco Neo-Sensitabs®). After 24hours, the induced halos were measured with automatic calipers. The results were analyzed using the EUCAST- and CLSI potency Neo-Sensitabs® guidelines. Results Vancomycin induced a halo of 22.37mm and 21.81mm in SAu and CoNS, respectively. Netilmicin produced halos of 24.45mm in SAu and 32.49mm in CoNS. MeAl induced halos of 12.65mm in SAu and 15.83mm in CoNS. A 12.11mm halo was found in SAu and an 18.38mm halo in CoNS using HOCl. DGCH produced halos of 26.55mm and 23.12mm in SAu and CoNS, respectively. Conclusion Netilmicin and vancomycin demonstrated antibiotic activity against both pathogens, so they can be alternative rescue therapies to treat chronic staphylococcal blepharitis. DGCH has efficacy against both comparable to antibiotics, while HOCl and MeAl show less efficacy (AU)

The bla and mec families of ß-lactam resistance genes in the genera Macrococcus, Mammaliicoccus and Staphylococcus: an in-depth analysis with emphasis on Macrococcus.

ß-Lactamases (Bla) and low-affinity penicillin-binding proteins (PBP2A) are responsible for ß-lactam resistance in the genera Macrococcus, Mammaliicoccus and Staphylococcus. These resistance mechanisms are in most species acquired through mobile genetic elements that carry a blaZ-like ß-lactamase gene for penicillin resistance and/or a mec gene (mecA, mecB, mecC,mecD) encoding a PBP2A for resistance to virtually all classes of ß-lactams. The mecA and mecC genes can be acquired through staphylococcal cassette chromosome mec (SCCmec) elements in Staphylococcus and Mammaliicoccus. The mecB and mecD genes are found in Macrococcus on SCCmec elements, as well as on unrelated mecD-carrying Macrococcus resistance islands (McRImecD) and large mecB-carrying plasmids. This review provides a phylogenetic overview of Macrococcus, Mammaliicoccus and Staphylococcus species and an in-depth analysis of the genetic structures carrying bla and mec genes in these genera. Native bla genes were detected in species belonging to the novobiocin-resistant Staphylococcus saprophyticus group and Mammaliicoccus. The evolutionary relatedness between Macrococcus and Mammaliicoccus is illustrated on the basis of a similar set of intrinsic PBPs, especially, the presence of a second class A PBP. The review further focuses on macrococcal elements carrying mecB and mecD, and compares them with structures present in Staphylococcus and Mammaliicoccus. It also discusses the different recombinases (ccr of SCCmec) and integrases (int of McRI) that contribute to the mobility of methicillin resistance genes, revealing Macrococcus as an important source for mobilization of antibiotic resistance genes within the family of Staphylococcaceae.

Antimicrobial, selective antibiofilm, and antioxidant properties of plasticized PMMA/PVC and zinc oxide nano filler for biomedical applications.

The PMMA/PVC/ZnO-nanocomposites with zinc oxide nanoparticle (particle size < 50nm) was synthesized by solution casting technique. Morphology of the synthesized nano composites have been investigated by FT-IR and XRD techniques. After characterization, synthesized composites were applied for antibacterial, selective antibiofilm and free radical scavenging screening. Antibacterial studies were measured against different bacterial strains. Antibiofilms activities were studied against those bacterial model pathogenic strains which showed highest and minimum sensitivity as a (~94 and ~88 at 160 µg/ml). Antioxidant activity of synthesized nanocomposites were measured by DPPH and showed scavenging capacity with IC50, 110 to > 200 µg/mL. Thus PMMA/PVC/ZnO nanocomposite showed promising antimicrobial activity and antioxidant activity that can be used for biomedical applications.

Effect of Tranexamic Acid against Staphylococcus spp. and Cutibacterium acnes Associated with Peri-Implant Infection: Results from an In Vitro Study.

Tranexamic acid (TXA) is extensively used in orthopedic surgery and traumatology as an antifibrinolytic agent to control intra- and postoperative bleeding and, therefore, indirectly, to reduce postsurgery infection rates. The hypothesis of an additional antibiotic effect against microorganisms associated with periprosthetic joint infection needs to be further evaluated. We aimed to assess whether TXA could reduce bacterial growth using an in vitro model. ATCC and clinical strains of staphylococci and Cutibacterium acnes were tested against TXA in both planktonic and sessile forms. We recorded the percent reduction in the following variables: log CFU/mL by microbiological culture, percentage of live cells by confocal laser scanning microscopy, and, additionally in sessile cells, metabolic activity by the 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide salt (XTT) assay. Variables were compared between groups using the Kruskal-Wallis test, and the results were reported as median (interquartile range [IQR]). Statistical significance was set at a P value of <0.05. Clinical significance was defined as a reduction of ≥25%. TXA at 50 mg/mL led to a slight reduction in CFU counts (4.5%). However, it was at 10 mg/mL that the reduction reached 27.2% and 33.0% for log CFU/mL counts and percentage of live cells, respectively. TXA was not efficacious for reducing preformed 24-h mature staphylococci and 48-h mature C. acnes biofilms, regardless of its concentration. TXA did not exert an antimicrobial effect against bacterial biofilms. However, when bacteria were in the planktonic form, it led to a clinically and statistically significant reduction in bacterial growth at 10 mg/mL. IMPORTANCE The possible use of TXA as an antibiotic agent in addition to its antifibrinolytic effect may play an important role in the prevention of prosthetic joint infection.

Interspecies transfer of plasmid-borne gentamicin resistance between Staphylococcus isolated from domestic dogs to Staphylococcus aureus.

As preconized by the One Health concept, the intimate relationship between pets and owners is a common source for the trade of microorganisms with zoonotic potential, and with them, antimicrobial resistance genes. In this work, we evaluated the presence of antimicrobial resistance genes, that are usually within mobile genetic elements, in a laboratory collection of 79 canine Staphylococcus strains, mostly Staphylococcus pseudintermedius and Staphylococcus coagulans. Resistance to tetracycline was observed in 34% of the strains, followed by resistance to erythromycin (21%) and gentamicin (19%). These phenotypes were partially correlated with the presence of the tetracycline resistance genes tet(M) and tet(K) in 64% and 44% of all strains, respectively; erythromycin resistance genes erm(A) and erm(C) in 53% and 23%; and gentamicin resistance gene aac(6')-aph(2″) in 26% of the strains. At least 45% of the strains harbored high- and/or low-molecular weight plasmids, whose transfer may be facilitated by their widespread biofilm-forming capacity, and absence of restrictive CRISPR systems. We selected eight plasmid-bearing and multidrug resistant strains, which were submitted to plasmid curing by stress with SDS. No strain lost resistance during stressing cultivation but, by conjugation experiments, the S. pseudintermedius strain 27 transferred its plasmid-borne resistance to gentamicin, conferred by the aac(6')-aph(2″) gene, to Staphylococcus aureus. The frequent empirical use of gentamicin to treat skin and ear infections in domestic dogs is likely to select resistant strains. Also, as demonstrated by our study, these strains can serve as gene reservoirs for human pathogens, such as S. aureus.

Trends of Antimicrobial Susceptibility in Clinically Significant Coagulase-Negative Staphylococci Isolated from Cerebrospinal Fluid Cultures in Neurosurgical Adults: a Nine-Year Analysis.

Coagulase-negative staphylococci (CoNS) are the main pathogens in health care-associated ventriculitis and meningitis (HCAVM). This study aimed to assess antimicrobial susceptibility. Moreover, the treatment and clinical outcome were described. All neurosurgical adults admitted to one of the largest neurosurgical centers in China with clinically significant CoNS isolated from cerebrospinal fluid cultures in 2012 to 2020 were recruited. One episode was defined as one patient with one bacterial strain. Interpretive categories were applied according to the MICs. The clinical outcomes were dichotomized into poor (Glasgow Outcome Scale 1 to 3) and acceptable (Glasgow Outcome Scale 4 to 5). In total, 534 episodes involving 519 patients and 16 bacteria were analyzed. Over the 9 years, eight antimicrobial agents were used in antimicrobial susceptibility tests, including six in over 80% of CoNS. The range of resistance rates was 0.8% to 84.6%. The vancomycin resistance rate was the lowest, whereas the penicillin resistance rate was the highest. The linezolid (a vancomycin replacement) resistance rate was 3.1%. The rate of oxacillin resistance, representing methicillin-resistant staphylococci, was 70.2%. There were no significant trends of antimicrobial susceptibility over the 9 years for any agents analyzed. However, there were some apparent changes. Notably, vancomycin-resistant CoNS appeared in recent years, while linezolid-resistant CoNS appeared early and disappeared in recent years. Vancomycin (or norvancomycin), the most common treatment agent, was used in 528 (98.9%) episodes. Finally, 527 (98.7%) episodes had acceptable outcomes. It will be safe to use vancomycin to treat CoNS-related HCAVM in the immediate future, although continuous monitoring will be needed. IMPORTANCE Coagulase-negative staphylococci are the main pathogens in health care-associated ventriculitis and meningitis. There are three conclusions from the results of this study. First, according to antimicrobial susceptibility, the rates of resistance to primary antimicrobial agents are high and those to high-level agents, including vancomycin, are low. Second, the trends of resistance rates are acceptable, especially for high-level agents, although long-term and continuous monitoring is necessary. Finally, the clinical outcomes of neurosurgical adults with coagulase-negative staphylococci-related health care-associated ventriculitis and meningitis are acceptable after treatment with vancomycin. Therefore, according to the antimicrobial susceptibility and clinical practice, vancomycin will be safe to treat coagulase-negative staphylococci-related health care-associated ventriculitis and meningitis.

The Study of Chemical Profile and Antioxidant Properties of Poplar-Type Polish Propolis Considering Local Flora Diversity in Relation to Antibacterial and Anticancer Activities in Human Breast Cancer Cells.

Nine samples of ethanolic extracts of poplar-type propolis (EEP) originated from South-Eastern Poland were analyzed in terms of the diversity of the flora around the apiary. The mineral composition, antioxidant properties, polyphenolic profile (HPTLC), and main polyphenolic constituents (HPLC-DAD) were determined. Only minor differences in chemical composition and antioxidant capacity between tested EEPs were found regardless of their botanical origin. However, the biological activity of the EEPs was more diversified. The tested EEPs showed stronger antibacterial activity against Gram-negative bacteria (Escherichia coli) compared to Gram-positive bacteria (Staphylococcus aureus and Staphylococcus epidermidis). Staphylococci biofilm inhibition occurred as a result of exposure to the action of four out of nine EEPs (P1-P4). Due to the various compositions of individual EEPs, a different MCF-7 cellular response was observed according to inhibition of cells migration and proliferation. Almost every sample inhibited the migration of breast cancer cells at a low concentration (0.04 µg/mL) of propolis. Even at the lowest concentration (0.02 µg/mL), each EEP inhibited the proliferation of MCF-7 cells, however, the level of inhibition varied between samples.

Targeted Antimicrobial Photodynamic Therapy of Biofilm-Embedded and Intracellular Staphylococci with a Phage Endolysin's Cell Binding Domain.

Bacterial pathogens are progressively adapting to current antimicrobial therapies with severe consequences for patients and global health care systems. This is critically underscored by the rise of methicillin resistant Staphylococcus aureus (MRSA) and other biofilm-forming staphylococci. Accordingly, alternative strategies have been explored to fight such highly multidrug resistant microorganisms, including antimicrobial photodynamic therapy (aPDT) and phage therapy. aPDT has the great advantage that it does not elicit resistance, while phage therapy allows targeting of specific pathogens. In the present study, we aimed to merge these benefits by conjugating the cell-binding domain (CBD3) of a Staphylococcus aureus phage endolysin to a photoactivatable silicon phthalocyanine (IRDye 700DX) for the development of a Staphylococcus-targeted aPDT approach. We show that, upon red-light activation, the resulting CBD3-700DX conjugate generates reactive oxygen species that effectively kill high loads of planktonic and biofilm-resident staphylococci, including MRSA. Furthermore, CBD3-700DX is readily internalized by mammalian cells, where it allows the targeted killing of intracellular MRSA upon photoactivation. Intriguingly, aPDT with CBD3-700DX also affects mammalian cells with internalized MRSA, but it has no detectable side effects on uninfected cells. Altogether, we conclude that CBD3 represents an attractive targeting agent for Staphylococcus-specific aPDT, irrespective of planktonic, biofilm-embedded, or intracellular states of the bacterium. IMPORTANCE Antimicrobial resistance is among the biggest threats to mankind today. There are two alternative antimicrobial therapies that may help to control multidrug-resistant bacteria. In phage therapy, natural antagonists of bacteria, lytic phages, are harnessed to fight pathogens. In antimicrobial photodynamic therapy (aPDT), a photosensitizer, molecular oxygen, and light are used to produce reactive oxygen species (ROS) that inflict lethal damage on pathogens. Since aPDT destroys multiple essential components in targeted pathogens, aPDT resistance is unlikely. However, the challenge in aPDT is to maximize target specificity and minimize collateral oxidative damage to host cells. We now present an antimicrobial approach that combines the best features of both alternative therapies, namely, the high target specificity of phages and the efficacy of aPDT. This is achieved by conjugating the specific cell-binding domain from a phage protein to a near-infrared photosensitizer. aPDT with the resulting conjugate shows high target specificity toward MRSA with minimal side effects.

Antibacterial activity and mechanism of slightly acidic electrolyzed water against Shewanella putrefaciens and Staphylococcus saprophytic.

The purpose of this study was to investigate the antimicrobial effect and mechanism of slightly acidic electrolyzed water (SAEW) against Shewanella putrefaciens (S. putrefaciens) and Staphylococcus saprophytic (S. saprophyticus). The results showed that SAEW exhibited strong antimicrobial activity against tested bacteria, which was positively correlated to the available chlorine concentration (ACC) of SAEW. The mortality rate of S. putrefaciens and S. saprophyticus was up to 96% and 85%, respectively, when the ACC of SAEW was 60.0 mg/L. The results of scanning electron microscopy (SEM) showed that the cell morphology and structure were destroyed by SAEW. Besides, the results of confocal laser scanning microscopy (CLSM), leakage of DNA and protein provided evidence that SAEW induced membrane permeabilization in cells. Compared with the control, the intracellular reactive oxygen species (ROS) generated by SAEW was strengthened significantly with the increase of ACC, and the cells were injured to death accordingly.

Novel pyrazoles as potent growth inhibitors of staphylococci, enterococci and Acinetobacter baumannii bacteria.

Background: Methicillin-resistant Staphylococcus aureus, vancomycin-resistant enterococci and Acinetobacter baumannii cause serious antibiotic-resistant infections. Finding new antibiotics to treat these infections is imperative for combating this worldwide menace. Methods & Results: In this study, the authors designed and synthesized potent antimicrobial agents using 4-trifluoromethylphenyl-substituted pyrazole derivatives. In addition to their potency against planktonic bacteria, potent compounds effectively eradicated S. aureus and Enterococcus faecalis biofilms. Human cells tolerated these compounds with good selectivity factors. Furthermore, the authors provide evidence for the mode of action of compounds based on time-kill kinetics, flow cytometry analysis of propidium iodide-treated bacteria and oxygen uptake studies. Conclusion: This study demonstrated 20 novel compounds with potent antibacterial activity that are tolerated by human cell lines.

The clinical significance of staphylococcus pettenkoferi: a retrospective review at a tertiary care medical center.

Staphylococcus pettenkoferi is a recently described coagulase-negative staphylococcal pathogen. We retrospectively reviewed 25 cases in which S. pettenkoferi was identified in routine cultures (12 blood, 13 other). Most were found with commensal flora and considered clinically insignificant, but its significance was uncertain in two cases from non-healing, deep foot wounds.

Frequency, Distribution, and Antimicrobial Resistance of Coagulase-Negative Staphylococci Isolated from Clinical Samples in Dogs and Cats.

This study aimed to investigate the frequency, distribution, and antimicrobial resistance of coagulase-negative staphylococci (CoNS) obtained from clinical samples from dogs and cats and to classify any methicillin-resistant CoNS (MRCoNS). The samples were collected in 2017-2018, and species identification and antimicrobial susceptibility testing were routinely performed using the Vitek2 system. Among 1,056 staphylococci, 185 CoNS (17.5%) were obtained and included 18 species from dogs (n = 116) and 14 species from cats (n = 69). The predominant species were Staphylococcus chromogenes (31.4%), Staphylococcus hominis ssp. hominis (16.2%), Staphylococcus warneri (10.8%), and Staphylococcus epidermidis (8.1%). The primary isolation sites were the skin and urinary tract. High levels of resistance to ß-lactams (65.4%), tetracycline (44.3%), clindamycin (36.8%), and erythromycin (30.8%) were observed. Twenty-five MRCoNS (13.4%), mainly Staphylococcus haemolyticus (n = 8), S. epidermidis (n = 6), and S. hominis ssp. hominis (n = 5), were identified. SCCmec type V (n = 8) was the most common type, followed by SCCmec type IV (n = 6) and SCCmec type III (n = 2), whereas nontypable SCCmec were classified into nine MRCoNS. Some CoNS have been recorded in humans, and these might be transferred to and cause subsequent infections in humans. Moreover, the diversity of SCCmec types and resistant strains suggested that they may serve as a reservoir of resistance genes among staphylococci.

Evaluation of the effects of chlorhexidine digluconate with and without cBD103 or cCath against multidrug-resistant clinical isolates of Staphylococcus pseudintermedius.

BACKGROUND: Because of the increased incidence of multidrug-resistant (MDR) bacteria, the use of disinfectants over antibiotics has been encouraged. However, the interactions between disinfectants and host local immunity are poorly understood. OBJECTIVE: To assess the effects of chlorhexidine digluconate (Chx), with and without selected host defence peptides (HDPs), against MDR Staphylococcus pseudintermedius (MDR-SP). METHODS AND MATERIALS: Ten clinical isolates of MDR-SP were tested, using a modified microbroth dilution method. Four two-fold dilutions of 2% Chx and 1 µg/mL the HDPs synthetic canine ß-defensin 103 (cBD103) or cathelicidin (cCath) were tested alone or in combination. Colony counts after 5, 15, 30 and 60 min, and a minimum inhibitory concentration (MIC) after 24 h were recorded. Friedman followed by Dunn's multiple comparison tests with significance of P < 0.05 were used for statistical analysis. Synergy, additivity/neutrality or antagonism were calculated. RESULTS: Growth was not inhibited by either HDP alone. An MIC of 0.312 µg/mL Chx was achieved for nine of the isolates. One isolate had an MIC of 0.078 µg/mL Chx. A MIC90 (in nine of 10 isolates) of 0.312 µg/mL was seen for Chx in combination with either HDP. Synergy was seen in the combination Chx/cCath used at the highest concentrations of Chx (0.624 µg/mL and 0.312 µg/mL) after 30 and 60 min incubation. Additivity/neutrality was seen for most of the other concentrations and times of incubation. CONCLUSIONS AND CLINICAL IMPORTANCE: These results suggest a synergistic/additive effect between Chx and HDPs in dogs. Further studies evaluating the mechanisms behind this effect are needed.

Biological Activity, Lipophilicity and Cytotoxicity of Novel 3-Acetyl-2,5-disubstituted-1,3,4-oxadiazolines.

Antibiotic resistance is now a global problem, and the lack of effective antimicrobial agents for the treatment of diseases caused by resistant microbes is increasing. The 3-acetyl-2,5-disubstituted-1,3,4-oxadiazolines presented in this article may provide a good starting point for the development of potential new effective antimicrobial agents useful in the treatment of bacterial and fungal infections. Particular attention is drawn to the 1,3,4-oxadiazole derivative marked with the number 29 with 5-nitrofuran-2-yl substituent in its chemical structure. This substance showed a strong bactericidal effect, especially against Staphylococcus spp., and no cytotoxicity to the L929 normal cell line.

Bacterial factors of mastitis in lactating women and its effect on the physical properties and chemical composition of breast milk.

Mastitis is a complication seen in some breastfeeding mothers and is the most common inflammatory lesion of the breast in breastfeeding mothers. In this complication, breast milk undergoes chemical and physical changes. It can lead to a drop in breastfeeding, weight loss, and, consequently, stunted growth of infants. Bacteria are the main cause of breast inflammation. Therefore, in this study, bacterial factors of mastitis were evaluated in lactating women. Also, their effects were considered on the physical properties and chemical composition of mothers' breast milk. For this purpose, 210 breastfeeding mothers referred to health centers were randomly selected, and their milk samples were collected. In addition to collecting mothers' demographic information by a questionnaire, the chemical composition (sugar, protein, and fat) and the physical properties (pH, density, and freezing temperature) of milk were measured. Bacterial evaluations were performed on the milk of these mothers by catalase test, coagulase test, and mannitol salt agar. Data were analyzed by SPSS software, Chi-square, Mann-Whitney U test, and T-test. The results showed that 56 mothers had mastitis, and Staphylococcus aureus and coagulase-negative staphylococci were the main bacteria in the milk of these mastitis mothers. These bacteria caused physical and chemical changes in breast milk so that mothers with Staphylococcus aureus mastitis had less sugar in their milk, and mothers with coagulase-negative staphylococci had less protein in their milk. Therefore, Staphylococcus aureus may reduce milk sugar by consuming milk sugar, and coagulase-negative staphylococci may also target milk protein. But to confirm these results, a larger population of mothers with mastitis is needed. Further studies are also needed to prove this result.

Multi-species host range of staphylococcal phages isolated from wastewater.

The host range of bacteriophages defines their impact on bacterial communities and genome diversity. Here, we characterize 94 novel staphylococcal phages from wastewater and establish their host range on a diversified panel of 117 staphylococci from 29 species. Using this high-resolution phage-bacteria interaction matrix, we unveil a multi-species host range as a dominant trait of the isolated staphylococcal phages. Phage genome sequencing shows this pattern to prevail irrespective of taxonomy. Network analysis between phage-infected bacteria reveals that hosts from multiple species, ecosystems, and drug-resistance phenotypes share numerous phages. Lastly, we show that phages throughout this network can package foreign genetic material enclosing an antibiotic resistance marker at various frequencies. Our findings indicate a weak host specialism of the tested phages, and therefore their potential to promote horizontal gene transfer in this environment.

Sulfonamide-salicylaldehyde imines active against methicillin- and trimethoprim/sulfonamide-resistant Staphylococci.

Background: Increasing resistance has resulted in an urgent need for new antimicrobial drugs. A systematic me-too approach was chosen to modify clinically used sulfonamides to obtain their imines. Methods & results: Twenty-five compounds were synthesized and evaluated for their antibacterial activity. The most active compounds were also investigated against methicillin- and trimethoprim/sulfamethoxazole (SMX)-resistant Gram-positive species. Staphylococci shared the highest susceptibility including resistant strains with minimum inhibitory concentrations from 3.91 µM (≥2.39 µg ml-1). Crucially, the compounds inhibit MRSA and trimethoprim/SMX-resistant Staphylococci without any cross-resistance. Modification of parent sulfonamides turned a bacteriostatic effect into a bactericidal effect. Toxicity for HepG2 and hemolytic properties were also determined. Conclusions: The presence of a dihalogenated salicylidene moiety is required for optimal activity. Based on toxicity, promising derivatives for further investigation were identified.

Alterations in children's sub-dominant gut microbiota by HIV infection and anti-retroviral therapy.

OBJECTIVE: We investigated the impact of human immunodeficiency virus (HIV) infection and anti-retroviral therapy (ART) on the gut microbiota of children. DESIGN: This cross-sectional study investigated the gut microbiota of children with and without HIV. METHODS: We collected fecal samples from 59 children with HIV (29 treated with ART [ART(+)] and 30 without ART [HIV(+)]) and 20 children without HIV [HIV(-)] in Vietnam. We performed quantitative RT-PCR to detect 14 representative intestinal bacteria targeting 16S/23S rRNA molecules. We also collected the blood samples for immunological analyses. RESULTS: In spearman's correlation analyses, no significant correlation between the number of dominant bacteria and age was found among children in the HIV(-) group. However, the number of sub-dominant bacteria, including Streptococcus, Enterococcus, and Enterobacteriaceae, positively correlated with age in the HIV(-) group, but not in the HIV(+) group. In the HIV(+) group, Clostridium coccoides group positively associated with the CD4+ cell count and its subsets. In the ART(+) group, Staphylococcus and C. perfringens positively correlated with CD4+ cells and their subsets and negatively with activated CD8+ cells. C. coccoides group and Bacteroides fragilis group were associated with regulatory T-cell counts. In multiple linear regression analyses, ART duration was independently associated with the number of C. perfringens, and Th17 cell count with the number of Staphylococcus in the ART(+) group. CONCLUSIONS: HIV infection and ART may influence sub-dominant gut bacteria, directly or indirectly, in association with immune status in children with HIV.