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Commercially produced cyanobacteria preparations sold under the name spirulina are widely consumed, due to their traditional use as a nutrient-rich foodstuff and subsequent marketing as a superfood. Despite their popularity, the microbial composition of ponds used to cultivate these bacteria is understudied. A total of 19 pond samples were obtained from small-scale spirulina farms and subjected to metagenome and/or virome sequencing, and the results were analysed. A remarkable level of prokaryotic and viral diversity was found to be present in the ponds, with Limnospira sp. and Arthrospira sp. sometimes being notably scarce. A detailed breakdown of prokaryotic and viral components of 15 samples is presented. Twenty putative Limnospira sp.-infecting bacteriophage contigs were identified, though no correlation between the performance of these cultures and the presence of phages was found. The high diversity of these samples prevented the identification of clear trends in sample performance over time, between ponds or when comparing successful and failed fermentations. [ABSTRACT FROM AUTHOR]

The distinct conjugation machineries encoded by plasmids pNP40 and pUC11B represent the most prevalent plasmid transfer systems among lactococcal strains. In the current study, we identified genetic determinants that underpin pNP40‐ and pUC11B‐mediated, high‐frequency mobilisation of other, non‐conjugative plasmids. The mobilisation frequencies of the smaller, non‐conjugative plasmids and the minimal sequences required for their mobilisation were determined, owing to the determination of the oriT sequences of both pNP40 and pUC11B, which allowed the identification of similar sequences in some of the non‐conjugative plasmids that were shown to promote their mobilisation. Furthermore, the auxiliary gene mobC, two distinct functional homologues of which are present in several plasmids harboured by the pNP40‐ and pUC11B‐carrying host strains, was observed to confer a high‐frequency mobilisation phenotype. These findings provide mechanistic insights into how lactococcal conjugative plasmids achieve conjugation and promote mobilisation of non‐conjugative plasmids. Ultimately, these insights would be harnessed to optimise conjugation and mobilisation strategies for the rapid and predictable development of robust and technologically improved strains. [ABSTRACT FROM AUTHOR]

Bacteriophages (or phages) represent a persistent threat to the success and reliability of food fermentation processes. Recent reports of phages that infect Streptococcus thermophilus have highlighted the diversification of phages of this species. Phages of S. thermophilus typically exhibit a narrow range, a feature that is suggestive of diverse receptor moieties being presented on the cell surface of the host. Cell wall polysaccharides, including rhamnose-glucose polysaccharides and exopolysaccharides have been implicated as being involved in the initial interactions with several phages of this species. Following internalization of the phage genome, the host presents several defences, including CRISPR-Cas and restriction and modification systems to limit phage proliferation. This review provides a current and holistic view of the interactions of phages and their S. thermophilus host cells and how this has influenced the diversity and evolution of both entities. [ABSTRACT FROM AUTHOR]

Pecorino Siciliano PDO is a semi-hard cheese that is produced in wooden vats using raw sheep's milk and its associated autochthonous microbial community. In the present study, we evaluated the microbial ecology of the milk, curd and whey from five Pecorino Siciliano PDO-producing farms in Sicily using a combination of metagenomic and microbiological approaches. We present an overview of the species and strain-level diversity of dairy lactococcal and streptococcal isolates using established genotyping tools and compare the lactic acid bacterial populations present in samples from these farms. Whole genome sequences of representative isolates of Lactococcus spp. and Streptococcus thermophilus were elucidated and the genetic diversity of the strains was established through analysis of predicted phage-resistance systems and prophage-associated regions. The analysis revealed farm-specific dairy lactococcal and streptococcal isolates that possess diverse genotypic features including newly described phage-resistance systems. [ABSTRACT FROM AUTHOR]

Clinical empathy has been studied in a number of health-care disciplines suggesting that higher practitioner empathy leads to improved patient health and wellness and improved patient outcomes. While some aspects of the physical therapist-patient relationship have been described, evidence of quantitative assessment of clinical empathy in physical therapists is scarce. To investigate the level of self-reported clinical empathy in physical therapists and its relationship to practice environment and workplace engagement, the Jefferson Scale of Empathy-Health Provider version (JSE-HP) and the Oldenburg Burnout Inventory (OLBI) were used. Study participants were 123 physical therapists working full time at either an acute care setting, a rehabilitation hospital, or an outpatient clinic. These physical therapists demonstrated a mean JSE-HP score of 118.5 (9.1) and a mean OLBI score of 15.63 (3.5). This mean empathy score was found to be higher than reported empathy level of some health disciplines such as nursing and pharmacy yet lower than others such as mental health workers, psychiatrists, and pediatricians. Practice setting was not found to be a significant factor regarding empathy levels in physical therapists. As reported in previous studies, there was a positive correlation between being female and having higher empathy levels. A positive correlation was found between age and work disengagement. Finally, our hypothesis regarding a negative correlation between empathy and work disengagement was confirmed, suggesting that workplace disengagement may diminish a physical therapist's empathy, which may then negatively affect patient clinical outcomes. [ABSTRACT FROM AUTHOR]

Summary: Available 3D structures of bacteriophage modules combined with predictive bioinformatic algorithms enabled the identification of adhesion modules in 57 siphophages infecting Streptococcus thermophilus (St). We identified several carbohydrate‐binding modules (CBMs) in so‐called evolved distal tail (Dit) and tail‐associated lysozyme (Tal) proteins of St phage baseplates. We examined the open reading frame (ORF) downstream of the Tal‐encoding ORF and uncovered the presence of a putative p2‐like receptor‐binding protein (RBP). A 21 Å resolution electron microscopy structure of the baseplate of cos‐phage STP1 revealed the presence of six elongated electron densities, surrounding the core of the baseplate, that harbour the p2‐like RBPs at their tip. To verify the functionality of these modules, we expressed GFP‐ or mCherry‐coupled Tal and putative RBP CBMs and observed by fluorescence microscopy that both modules bind to their corresponding St host, the putative RBP CBM with higher affinity than the Tal‐associated one. The large number of CBM functional domains in St phages suggests that they play a contributory role in the infection process, a feature that we previously described in lactococcal phages and beyond, possibly representing a universal feature of the siphophage host‐recognition apparatus. [ABSTRACT FROM AUTHOR]

Streptococcus thermophilus strain ST64987 was exposed to a member of a recently discovered group of S. thermophilus phages (the 987 phage group), generating phage‐insensitive mutants, which were then characterized phenotypically and genomically. Decreased phage adsorption was observed in selected bacteriophage‐insensitive mutants, and was partnered with a sedimenting phenotype and increased cell chain length or aggregation. Whole genome sequencing of several bacteriophage‐insensitive mutants identified mutations located in a gene cluster presumed to be responsible for cell wall polysaccharide production in this strain. Analysis of cell surface‐associated glycans by methylation and NMR spectroscopy revealed a complex branched rhamno‐polysaccharide in both ST64987 and phage‐insensitive mutant BIM3. In addition, a second cell wall‐associated polysaccharide of ST64987, composed of hexasaccharide branched repeating units containing galactose and glucose, was absent in the cell wall of mutant BIM3. Genetic complementation of three phage‐resistant mutants was shown to restore the carbohydrate and phage resistance profiles of the wild‐type strain, establishing the role of this gene cluster in cell wall polysaccharide production and phage adsorption and, thus, infection. [ABSTRACT FROM AUTHOR]

The dairy fermentation industry relies on the activity of lactic acid bacteria in robust starter cultures to accomplish milk acidification. Maintenance of the composition of these starter cultures, whether defined or undefined, is essential to ensure consistent and high-quality fermentation end products. To date, limited information exists regarding the microbial composition of undefined starter culture systems. Here, we describe a culture-based analysis combined with a metagenomics approach to evaluate the composition of two undefined mesophilic starter cultures. In addition, we describe a qPCR-based genotype detection assay, which is capable of discerning nine distinct lactococcal genotypes to characterize these undefined starter cultures, and which can be applied to monitor compositional changes in an undefined starter culture during a fermentation. IMPORTANCE This study reports on the development of a combined culture-based analysis and metagenomics approach to evaluate the composition of two undefined mesophilic starter cultures. In addition, a novel qPCR-based genotype detection assay, capable of discerning nine distinct lactococcal genotypes (based on lactococcal cell wall polysaccharide biosynthesis gene clusters), was used to monitor compositional changes in an undefined starter culture following phage attack. These analytical approaches facilitate a multifaceted assessment of starter culture compositional stability during milk fermentation, which has become an important QC aspect due to the increasing demand for consistent and high-quality dairy products. [ABSTRACT FROM AUTHOR]

Background. Standardized tests and outcome measures (STOM) have not been consistently implemented as part of most physical therapists' practice. Incidence of STOM use among physical therapists at Beth Israel Deaconess Medical Center was similar to low levels cited nationally among acute care physical therapists. Targeted knowledge translation (KT) strategies have been suggested to promote the application of research evidence into clinical decision making. Purpose. The purpose of this quality improvement (QI) effort was to implement a series of interventions aimed at increasing both use and interpretation of STOM by physical therapists practicing in acute care. Design. This study used an observational longitudinal design. Methods. A literature review identified current barriers and facilitators to the use of STOM by physical therapists. KT strategies were tailored to the practice setting in order to target barriers and promote facilitators to the use of STOM. Data were collected through retrospective chart review at baseline and then subsequently at 4 periods following the implementation of the QI project. Results. A statistically significant increase in both the use (primary outcome) and interpretation (secondary outcome) of STOM was observed following the implementation of KT strategies. The increase was sustained at all subsequent measurement periods. Limitations. Limitations include the lack of a control group and the small number of setting- and diagnosis-specific STOM available for use by physical therapists practicing in acute care. Conclusions. Implementation of KT strategies was associated with an increase in the frequency of use and interpretation of STOM. Similar QI efforts are feasible in any acute care physical therapy department and potentially other settings. [ABSTRACT FROM AUTHOR]

Predation of starter lactic acid bacteria such as Streptococcus thermophilus by bacteriophages is a persistent and costly problem in the dairy industry. CRISPRmediated bacteriophage insensitive mutants (BIMs), while straightforward to generate and verify, can quickly be overcome by mutant phages. The aim of this study was to develop a tool allowing the generation of derivatives of commercial S. thermophilus strains which are resistant to phage attack through a non-CRISPR-mediated mechanism, with the objective of generating BIMs exhibiting stable resistance against a range of isolated lytic S. thermophilus phages. To achieve this, standard BIM generation was complemented by the use of the wild-type (WT) strain which had been transformed with an antisense mRNA-generating plasmid (targeting a crucial CRISPR-associated [cas] gene) in order to facilitate the generation of non-CRISPR-mediated BIMs. Phage sensitivity assays suggest that non-CRISPR-mediated BIMs exhibit some advantages compared to CRISPRmediated BIMs derived from the same strain. [ABSTRACT FROM AUTHOR]

Despite the persistent and costly problem caused by (bacterio)phage predation of Streptococcus thermophilus in dairy plants, DNA sequence information relating to these phages remains limited. Genome sequencing is necessary to better understand the diversity and proliferative strategies of virulent phages. In this report, whole genome sequences of 40 distinct bacteriophages infecting S. thermophilus were analyzed for general characteristics, genomic structure and novel features. The bacteriophage genomes display a high degree of conservation within defined groupings, particularly across the structural modules. Supporting this observation, four novel members of a recently discovered third group of S. thermophilus phages (termed the 5093 group) were found to be conserved relative to both phage 5093 and to each other. Replication modules of S. thermophilus phages generally fall within two main groups, while such phage genomes typically encode one putative transcriptional regulator. Such features are indicative of widespread functional synteny across genetically distinct phage groups. Phage genomes also display nucleotide divergence between groups, and between individual phages of the same group (within replication modules and at the 3' end of the lysis module)--through various insertions and/or deletions. A previously described multiplex PCR phage detection system was updated to reflect current knowledge on S. thermophilus phages. Furthermore, the structural protein complement as well as the antireceptor (responsible for the initial attachment of the phage to the host cell) of a representative of the 5093 group was defined. Our data more than triples the currently available genomic information on S. thermophilus phages, being of significant value to the dairy industry, where genetic knowledge of lytic phages is crucial for phage detection and monitoring purposes. In particular, the updated PCR detection methodology for S. thermophilus phages is highly useful in monitoring particular phage group(s) present in a given whey sample. Studies of this nature therefore not only provide information on the prevalence and associated threat of known S. thermophilus phages, but may also uncover newly emerging and genomically distinct phages infecting this dairy starter bacterium. [ABSTRACT FROM AUTHOR]

We present the complete genome sequences of four members of a novel group of phages infecting Streptococcus thermophilus, designated here as the 987 group. Members of this phage group appear to have resulted from genetic exchange events, as evidenced by their "hybrid" genomic architecture, exhibiting DNA sequence relatedness to the morphogenesis modules of certain P335 group Lactococcus lactis phages and to the replication modules of S. thermophilus phages. All four identified members of the 987 phage group were shown to elicit adsorption affinity to both their cognate S. thermophilus hosts and a particular L. lactis starter strain. The receptor binding protein of one of these phages (as a representative of this novel group) was defined using an adsorption inhibition assay. The emergence of a novel phage group infecting S. thermophilus highlights the continuous need for phage monitoring and development of new phage control measures. [ABSTRACT FROM AUTHOR]

Brian McDonnell of Deloitte Ireland provides an overview of grant and tax incentives that can be used to help fund investments to meet new carbon targets. [ABSTRACT FROM AUTHOR]

New Zealand is a liberal democracy where state censorship is among the lightest in the world. However, in 1964, the government intervened in the controversy surrounding the publication of a primary schools bulletin called Washday at the Pa. This booklet which showed photographs of the home life of a rural Maori family, engendered strongly differing social views on the representation of Maori, the desirability or otherwise of censoring classroom resources, and the shifting dynamics of race relations in the country. This article scrutinizes the events of 1964 surrounding Washday at the Pa in detail to provide an account of the complex series of actions undertaken by government departments, government ministers, publishers, pressure groups, and commentators on both sides of the debate. It also extends its coverage to subsequent events such as the 2011 re-publication of the bulletin for a twenty-first-century readership. This article argues that the censorship of Washday at the Pa has had long-lasting ramifications and illustrates the evolution of attitudes to the representation of Maori over the decades. [ABSTRACT FROM AUTHOR]

Heavenly Creatures (1994), a dramatized version of a scandalous real-life 1954 Christchurch matricide committed by teenage girls, is one of the most accomplished films ever produced in New Zealand. The article attempts to do justice to this complex film with a detailed analysis. While attention is paid to its place both in the evolution of Peter Jackson and Frances Walsh as film-makers and in the context of New Zealand film history, the primary focus is an analysis of the film text itself, especially in terms of its narrative techniques. Perhaps the film's greatest achievement is its success in taking the audience inside the subjective experience of protagonists Pauline Parker and Juliet Hulme and the article provides insights into just how Jackson and Walsh achieved this considerable feat. It examines the different styles of storytelling that combine in the film's intricate narrational mode and scrutinizes the way Walsh and Jackson adapted Pauline's diaries into the script, demonstrating how the resulting narrative empowers the main characters by recruiting their imaginations to 'co-direct' the film. [ABSTRACT FROM AUTHOR]

This article presents a study that reviewed published literature concerning both the hormone replacement therapy's (HRT) potential risks and benefits, and the safety of prescribing the therapy to endometrial cancer survivors. Depending on the doses and the specific drug use, prescribing HRT to endometrial cancer survivors does not seem to be contraindicated and may even confer modest protection.

Highlights the modeling and control of a pneumatic robot which can be used for position tracking and end-effector force control. Information on previous methods used for controlled motion; Description of the robot utilized in this research; Examination of a pneumatic actuator model; Statistical data compiled in this research.

Bifidobacterium breve is a common and sometimes very abundant inhabitant of the human gut. Genome sequencing of B. breve JCM 7017 revealed the presence of an extrachromosomal element, designated pMP7017 consisting of>190 kb, thus representing the first reported bifidobacterial megaplasmid. In silico characterization of this element revealed several genomic features supporting a stable establishment of the megaplasmid in its host, illustrated by predicted CRISPR-Cas functions that are known to protect the host against intrusion of foreign DNA. Interestingly, pMP7017 is also predicted to encode a conjugative DNA transfer apparatus and, consistent with this notion, we demonstrate here the conjugal transfer of pMP7017 to representative strains of B. breve and B. longum subsp. longum. We also demonstrate the presence of a megaplasmid with homology to pMP7017 in three B. longum subsp. longum strains. [ABSTRACT FROM AUTHOR]

Until the late 2000s, lactococci substantially contributed to the discovery of various plasmid-borne phage defence systems, rendering these bacteria an excellent antiphage discovery resource. Recently, there has been a resurgence of interest in identifying novel antiphage systems in lactic acid bacteria owing to recent reports of so-called 'defence islands' in diverse bacterial genera. Here, 321 plasmid sequences from 53 lactococcal strains were scrutinized for the presence of antiphage systems. Systematic evaluation of 198 candidates facilitated the discovery of seven not previously described antiphage systems, as well as five systems, of which homologues had been described in other bacteria. All described systems confer resistance against the most prevalent lactococcal phages, and act post phage DNA injection, while all except one behave like abortive infection systems. Structure and domain predictions provided insights into their mechanism of action and allow grouping of several genetically distinct systems. Although rare within our plasmid collection, homologues of the seven novel systems appear to be widespread among bacteria. This study highlights plasmids as a rich repository of as yet undiscovered antiphage systems., (© The Author(s) 2024. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Bacteriocins are broad or narrow-spectrum antimicrobial compounds that have received significant scientific attention due to their potential to treat infections caused by antibiotic-resistant pathogenic bacteria. The genome of Bifidobacterium pseudocatenulatum MM0196, an antimicrobial-producing, fecal isolate from a healthy pregnant woman, was shown to contain a gene cluster predicted to encode Pseudocin 196, a novel lantibiotic, in addition to proteins involved in its processing, transport and immunity. Following antimicrobial assessment against various indicator strains, protease-sensitive Pseudocin 196 was purified to homogeneity from cell-free supernatant. MALDI TOF mass spectrometry confirmed that the purified antimicrobial compound corresponds to a molecular mass of 2679 Da, which is consistent with that deduced from its genetic origin. Pseudocin 196 is classified as a lantibiotic based on its similarity to lacticin 481, a lanthionine ring-containing lantibiotic produced by Lactococcus lactis . Pseudocin 196, the first reported bacteriocin produced by a B. pseudocatenulatum species of human origin, was shown to inhibit clinically relevant pathogens, such as Clostridium spp. and Streptococcus spp. thereby highlighting the potential application of this strain as a probiotic to treat and prevent bacterial infections.

Floricoccus penangensis is a Gram-positive coccoid organism that is a member of the lactic acid bacteria. F. penangensis ML061-4 was originally isolated from the surface of an Assam tea leaf, and its genome is herein shown to contain gene clusters predicted to be involved in complex carbohydrate metabolism and biosynthesis of secondary metabolites., Competing Interests: Competing Interests: The authors have declared that no competing interest exists., (© The author(s).)

The field of metagenomics has rapidly expanded to become the go-to method for complex microbial community analyses. However, there is currently no straightforward route from metagenomics to traditional culture-based methods of strain isolation, particularly in (bacterio)phage biology, leading to an investigative bottleneck. Here, we describe a method that exploits specific phage receptor binding protein (RBP)-host cell surface receptor interaction enabling isolation of phage-host combinations from an environmental sample. The method was successfully applied to two complex sample types-a dairy-derived whey sample and an infant fecal sample, enabling retrieval of specific and culturable phage hosts. IMPORTANCE PhRACS aims to bridge the current divide between in silico genetic analyses (i.e., phageomic studies) and traditional culture-based methodology. Through the labeling of specific bacterial hosts with fluorescently tagged recombinant phage receptor binding proteins and the isolation of tagged cells using flow cytometry, PhRACS allows the full potential of phageomic data to be realized in the wet laboratory.

Bifidobacteria are important early colonizers of the human intestinal tract. The relative abundance of bifidobacterial species may be modulated, in part, by bacteriophage activity. Metagenomic studies of these populations is a crucial step in understanding this important interaction. This chapter outlines the technical instructions required to analyze the virome of a bifidobacteria-rich sample, for example, an infant fecal sample.