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ABSTRACT Antibiotic resistance is a major public health challenge worldwide, whose implications for global health might be devastating if novel antibacterial strategies are not quickly developed. As natural predators of bacteria, (bacterio)phages may play an essential role in escaping such a dreadful future. The rising problem of antibiotic resistance has revived the interest in phage therapy and important developments have been achieved over the last years. But where do we stand today and what can we expect from phage therapy in the future? This is the question we set to answer in this review. Here, we scour the outcomes of human phage therapy clinical trials and case reports, and address the major barriers that stand in the way of using phages in clinical settings. We particularly address the potential of phage resistance to hinder phage therapy and discuss future avenues to explore the full capacity of phage therapy. There is a remarkable potential of phage therapy for the control of antibiotic resistant infections within the One Health approach, thus, the challenges currently faced and the potential solutions in development must be considered.

Background The human-animal bond has been recognized as having positive effects on the health and well-being of both humans and pets. The present study aims to explore the influence of attachment on physical activity (PA), lifestyle, and health outcomes of dog owners (DO), highlighting the mutual benefits resulting from the relationship between DO and dogs. Methods Thirty-eight DO and their dogs participated in this study. Socio-demographic data, the Self-Rated Health (SRH), FANTASTICO Lifestyle Scale, and the Lexington Attachment Pet Scale (LAPS) were assessed. PA was measured in both the DO and the dogs, using an ActiGraph GT3X accelerometer in the context of daily routine. Descriptive statistics and Spearman rank correlation analyses were performed to examine the associations between LAPS, PA levels, socio-demographic variables, lifestyle behaviors, and SRH. Results Significant correlations were found between the dog owners’ light-level PA and the pets’ vigorous level of PA (rho = 0.445, p =  0.01). Furthermore, the importance of the pets’ health (rho = -0.785, p  = 0.02) and the LAPS subscales, namely proximity (rho = 0.358, p  = 0.03), and attachment (rho = 0.392, p  = 0.01), were related to taking the pet for a walk. Regarding lifestyle, DO with a healthier lifestyle had a better self-assessment of their health using the SRH (rho = 0.39, p  = 0.02). Moreover, DO with better lifestyles also exhibited greater concern for their pet’s health (rho = 0.398, p  = 0.01). Conclusions This study emphasizes that individuals who adopt healthier habits tend to perceive themselves as healthier and exhibit greater concern for their pets’ health. The attachment between DO and dogs is important in promoting healthy lifestyle behaviors and engagement in PA. Our results highlight that the presence of a dog is associated with a higher level of PA in DO, depending on the strength of the human-animal bond.

Background A total of 179 Shiga toxin-producing Escherichia coli (STEC) complete genomes were analyzed in terms of serotypes, prophage coding regions, and stx gene variants and their distribution. We further examined the genetic diversity of Stx-converting phage genomes (Stx phages), focusing on the lysis-lysogeny decision and lytic cassettes. Results We show that most STEC isolates belong to non-O157 serotypes (73 %), regardless the sources and geographical regions. While the majority of STEC genomes contain a single stx gene (61 %), strains containing two (35 %), three (3 %) and four (1 %) stx genes were also found, being stx2 the most prevalent gene variant. Their location is exclusively found in intact prophage regions, indicating that they are phage-borne. We further demonstrate that Stx phages can be grouped into four clusters (A, B, C and D), three subclusters (A1, A2 and A3) and one singleton, based on their shared gene content. This cluster distribution is in good agreement with their predicted virion morphologies. Stx phage genomes are highly diverse with a vast number of 1,838 gene phamilies (phams) of related sequences (of which 677 are orphams i.e. unique genes) and, although having high mosaicism, they are generally organized into three major transcripts. While the mechanisms that guide lysis–lysogeny decision are complex, there is a strong selective pressure to maintain the stx genes location close to the lytic cassette composed of predicted SAR-endolysin and pin-holin lytic proteins. The evolution of STEC Stx phages seems to be strongly related to acquiring genetic material, probably from horizontal gene transfer events. Conclusions This work provides novel insights on the genetic structure of Stx phages, showing a high genetic diversity throughout the genomes, where the various lysis-lysogeny regulatory systems are in contrast with an uncommon, but conserved, lytic system always adjacent to stx genes.

Staphylococcus epidermidis is a major causative agent of nosocomial infections, mainly associated with the use of indwelling devices, on which this bacterium forms structures known as biofilms. Due to biofilms’ high tolerance to antibiotics, virulent bacteriophages were previously tested as novel therapeutic agents. However, several staphylococcal bacteriophages were shown to be inefficient against biofilms. In this study, the previously characterized S. epidermidis-specific Sepunavirus phiIBB-SEP1 (SEP1), which has a broad spectrum and high activity against planktonic cells, was evaluated concerning its efficacy against S. epidermidis biofilms. The in vitro biofilm killing assays demonstrated a reduced activity of the phage. To understand the underlying factors impairing SEP1 inefficacy against biofilms, this phage was tested against distinct planktonic and biofilm-derived bacterial populations. Interestingly, SEP1 was able to lyse planktonic cells in different physiological states, suggesting that the inefficacy for biofilm control resulted from the biofilm 3D structure and the protective effect of the matrix. To assess the impact of the biofilm architecture on phage predation, SEP1 was tested in disrupted biofilms resulting in a 2 orders-of-magnitude reduction in the number of viable cells after 6 h of infection. The interaction between SEP1 and the biofilm matrix was further assessed by the addition of matrix to phage particles. Results showed that the matrix did not inactivate phages nor affected phage adsorption. Moreover, confocal laser scanning microscopy data demonstrated that phage infected cells were less predominant in the biofilm regions where the matrix was more abundant. Our results provide compelling evidence indicating that the biofilm matrix can work as a barrier, allowing the bacteria to be hindered from phage infection.

Maintaining a healthy aging process is vital to combating and delaying the adverse health outcomes faced by the growing older adult population, where a key aspect to achieving healthy aging is the preservation of functional fitness. This study aims to present trends on the functional fitness of Portuguese older adults between 2008 and 2018 and build new normative reference standards for the Senior Fitness Battery Tests. Cross-sectional study. Functional fitness was measured with the Senior Fitness Battery Tests in two national representative cohorts of the Portuguese older adult population (≥65 years) using the 2008 (n = 4712) and 2018 (n = 2717) surveys. Changes were assessed according to sex and age group. Sex and age-group normative fitness scores were developed. Overall findings suggested that older adults' point prevalence for arm-curl, 8-ft up-and-go, and 30-s chair sit-to-stand remained stable between 2008 and 2018, with declining trends being observed for the 6-min walk and back-scratch tests. However, results for trends varied when stratified by sex and age categories. Our results suggest a trend toward a stabilization of most of the functional fitness tests over the past decade, but with a critical decline in the 6-min walk test. Thus, this test should be targeted by future health-care policies. Moreover, this investigation provides new and updated normative reference standards for the Senior Fitness Battery Tests that should be used as a tool to promote a healthy aging process among Portuguese older adults.

Most phage-host interaction studies are performed with exponentially growing cells. However, this cell state is not representative of what happens in natural environments. Additionally, most phages fail to replicate in stationary cells. The Staphylococcus epidermidis phage SEP1 is one of the few phages reported to date to be able to infect stationary cells. Here, we unveiled the interaction of SEP1 with its host in both exponential and stationary states of growth at the transcriptomic level. The findings of this study provide valuable insights for a better implementation of phage therapy since phages able to infect stationary cells could be more efficient in the treatment of recalcitrant infections.

The human periodontium health is commonly compromised by chronic inflammatory conditions and has become a major public health concern. Dental plaque, the precursor of periodontal disease, is a complex biofilm consisting mainly of bacteria, but also archaea, protozoa, fungi and viruses. Viruses that specifically infect bacteria - bacteriophages - are most common in the oral cavity. Despite this, their role in the progression of periodontal disease remains poorly explored. This review aims to summarize how bacteriophages interact with the oral microbiota, their ability to increase bacterial virulence and mediate the transfer of resistance genes and suggests how bacteriophages can be used as an alternative to the current periodontal disease therapies.

Citrobacter spp., although frequently ignored, is emerging as an important nosocomial bacterium able to cause various superficial and systemic life-threatening infections. Considered to be hard-to-treat bacterium due to its pattern of high antibiotic resistance, it is important to develop effective measures for early and efficient therapy. In this study, the first myovirus (vB_CfrM_CfP1) lytic for Citrobacter freundii was microbiologically and genomically characterized. Its morphology, activity spectrum, burst size, and biophysical stability spectrum were determined. CfP1 specifically infects C. freundii , has broad host range (>85 %; 21 strains tested), a burst size of 45 PFU/cell, and is very stable under different temperatures (−20 to 50 °C) and pH (3 to 11) values. CfP1 demonstrated to be highly virulent against multidrug-resistant clinical isolates up to 12 antibiotics, including penicillins, cephalosporins, carbapenems, and fluroquinoles. Genomically, CfP1 has a dsDNA molecule with 180,219 bp with average GC content of 43.1 % and codes for 273 CDSs. The genome architecture is organized into function-specific gene clusters typical for tailed phages, sharing 46 to 94 % nucleotide identity to other Citrobacter phages. The lysin gene encoding a predicted D-Ala-D-Ala carboxypeptidase was also cloned and expressed in Escherichia coli and its activity evaluated in terms of pH, ionic strength, and temperature. The lysine optimum activity was reached at 20 mM HEPES, pH 7 at 37 °C, and was able to significantly reduce all C. freundii (>2 logs) as well as Citrobacter koseri (>4 logs) strains tested. Interestingly, the antimicrobial activity of this enzyme was performed without the need of pretreatment with outer membrane-destabilizing agents. These results indicate that CfP1 lysin is a good candidate to control problematic Citrobacter infections, for which current antibiotics are no longer effective.

Physical inactivity has been linked to noncommunicable diseases and increasing global mortality rates, highlighting the importance of assessing physical activity levels across different populations (Gilchrist et al., 2020). The aim of this study was to analyse the levels of physical activity in the university community according to gender (female vs. male), educational background (12th year vs. undergraduate vs. master's vs. doctorate) and occupational status (students vs. teachers vs. non-teaching staff), as a pilot study for the EASIER project, which is developing an APP to reduce sedentary behaviour through a combination of exercise and a digital game. A total of 205 participants (112 females and 93 males) were recruited electronically and completed the International Physical Activity Questionnaire Short Form (IPAQ-SF). Physical activity levels were classified into four categories: sedentary, irregularly active, active, and very active. Among the university community, 76% of the participants were classified as sedentary (n=50) or irregularly active (n=106), while only 24% were classified as active (n=33) or very active (n=16). Regarding educational qualifications, 28% of the participants completed high school, 32% had a bachelor's degree, 12% had a master's degree, and 28% had a doctorate. Additionally, 54% of the participants were students, 24% were teachers, and 23% were non-teaching staff. Descriptive statistics and chi-square test were used to compare physical activity levels between sex, educational background, and employment status, with a significance level set at p≤0.05. The results reported mostly sedentary (17% vs 8%) and irregularly active (25% vs 26%) levels of physical activity for female’s vs males, respectively. Regardless of education or employment status, most participants were classified as irregularly active, both representing more than 50% of the total. Furthermore, no significant differences were found in physical activity levels according to gender (p=0.062), education (p=0.167) or employment status (p=0.059). Effective strategies are needed to promote regular physical activity in the university community, regardless of individual characteristics. Awareness and education about the benefits of physical activity, the availability of appropriate infrastructure, programs, and incentive projects for the integration of physical activity in academic routines are some of the advised strategies. In this sense, the EASIER project, which combines physical exercises and a digital game, presents a promising alternative that can improve levels of physical activity and promote a healthy lifestyle in the university community. Physical inactivity has been linked to noncommunicable diseases and increasing global mortality rates, highlighting the importance of assessing physical activity levels across different populations (Gilchrist et al., 2020). The aim of this study was to analyse the levels of physical activity in the university community according to gender (female vs male), educational background (12th year vs undergraduate vs master's vs doctorate) and occupational status (students vs. teachers vs. non-teaching staff), as a pilot study for the EASIER project, which is developing an APP to reduce sedentary behaviour through a combination of exercise and a digital game. A total of 205 participants (112 females and 93 males) were recruited electronically and completed the International Physical Activity Questionnaire Short Form (IPAQ-SF). Physical activity levels were classified into four categories: sedentary, irregularly active, active, and very active. Among the university community, 76% of the participants were classified as sedentary (n=50) or irregularly active (n=106), while only 24% were classified as active (n=33) or very active (n=16). Regarding educational qualifications, 28% of the participants completed high school, 32% had a bachelor's degree, 12% had a master's degree, and 28% had a doctorate. Additionally, 54% of the participants were students, 24% were teachers, and 23% were non-teaching staff. Descriptive statistics and chi-square tests were used to compare physical activity levels between sex, educational background, and employment status, with a significance level set at p≤0.05. The results reported mostly sedentary (17% vs 8%) and irregularly active (25% vs 26%) levels of physical activity for females vs males, respectively. Regardless of education or employment status, most participants were classified as irregularly active, both representing more than 50% of the total. Furthermore, no significant differences were found in physical activity levels according to gender (p=0.062), education (p=0.167) or employment status (p=0.059). Effective strategies are needed to promote regular physical activity in the university community, regardless of individual characteristics. Awareness and education about the benefits of physical activity and the availability of appropriate infrastructure, programs, and incentive projects for integrating physical activity into academic routines are some of the advised strategies. In this sense, the EASIER project, which combines physical exercises and a digital game, presents a promising alternative that can improve physical activity levels and promote a healthy lifestyle in the university community.

The characteristics of handball played on sand differ from court handball not only in team size, scoring system, duration, rules, and field dimensions but also in surface and playing style. The present study aimed to characterise the intensity of effort through HR and PSE during a training match of the Brazilian Women's Beach Handball team. Nine female athletes from the national team (age 27.3 ± 4.0 years) voluntarily participated in the present study. Athletes body composition was evaluated by Inbody 720 bioimpedance scale (height 167.1 ± 6.5cm, weight 64.4 ± 10.1kg, lean mass 27.5 ± 4.6kg, fat percentage 23.2 ± 5.0%). The Yo-Yo Intermittent Endurance Test - Level 2 (YYIETL2) was applied, and for monitoring and recording HRmax, the Polar H7 Smart Bluetooth connected to Polar Team Software was used. The exertional intensity during the training match (HR and PSE) was analysed by descriptive statistics and T-test for repeated measures (p <0.05). During the 2 sets of the match, HR was continuously monitored and categorised into 4 pre-defined intensity zones (zone 1 (<75% HRmax), zone 2 (75-84,9% HRmax), zone 3 (85-89,9% HRmax) and zone 4 (≥90% HRmax)) and PSE value recorded immediately at the end of each set. The results do not reveal statistically significant differences in the behaviour of HR and PSE between players or sets. However, the athlete's perceived exertion reveals a significant underestimation when compared to the results evaluated by HR monitoring, and there is evidence of PSE differences between outfield players vs goalkeepers (12.57 ± 1.39 vs 11.00 ± 0.00; p <0.025). When analysing the HR zones (fig. 1), there was a higher permanence in zone 3 (85-89.9%HR) in both sets with an average time of 174.6±120.2 seconds in 1st and 248.2±132.1 in 2nd one. When comparing the intensity of effort in the 4 defined zones, it was observed that goalkeepers have a higher percentage in zones 1, 2 and 3, regardless of the set played (being non-existent or residual in zone 4), while field players have higher percentages in zones 2, 3 and 4. Beach handball, played at a national team level, although in training matches as we analysed it, has shown to be a sport with high-intensity demand. The creative and acrobatic actions of this game, such as inflight (aerial plays, alley-oop passes, jump shots, spin shots, goalkeeper saves) and spin shots, may explain some of these results.