The formation of precise numbers of neuronal connections, known as synapses, is crucial for brain function. Therefore, synaptogenesis mechanisms have been one of the main focuses of neuroscience. Immunohistochemistry is a common tool for visualizing synapses. Thus, quantifying the numbers of synapses from light microscopy images enables screening the impacts of experimental manipulations on synapse development. Despite its utility, this approach is paired with low-throughput analysis methods that are challenging to learn, and the results are variable between experimenters, especially when analyzing noisy images of brain tissue. We developed an open-source ImageJ-based software, SynBot, to address these technical bottlenecks by automating the analysis. SynBot incorporates the advanced algorithms ilastik and SynQuant for accurate thresholding for synaptic puncta identification, and the code can easily be modified by users. The use of this software will allow for rapid and reproducible screening of synaptic phenotypes in healthy and diseased nervous systems.

Typhoid fever is endemic in many parts of the world and remains a major public health concern in tropical and sub-tropical developing nations, including Fiji. To address high rates of typhoid fever, the Northern Division of Fiji implemented a mass vaccination with typhoid conjugate vaccine (Vi-polysaccharide conjugated to tetanus toxoid) as a public health control measure in 2023. In this study we define the genomic epidemiology of Salmonella Typhi in the Northern Division prior to island-wide vaccination, sequencing 85% (n=419) of the total cases from the Northern and Central Divisions of Fiji that occurred in the period 2017-2019. We found elevated rates of nucleotide polymorphisms in the tviD and tviE genes (responsible for Vi-polysaccharide synthesis) relative to core genome levels within the Fiji endemic S. Typhi genotype 4.2. Expansion of these findings within a globally representative database of 12 382 S. Typhi (86 genotyphi clusters) showed evidence of convergent evolution of the same tviE mutations across the S. Typhi population, indicating that tvi selection has occurred both independently and globally. The functional impact of tvi mutations on the Vi-capsular structure and other phenotypic characteristics are not fully elucidated, yet commonly occurring tviE polymorphisms localize adjacent to predicted active site residues when overlayed against the predicted TviE protein structure. Given the central role of the Vi-polysaccharide in S. Typhi biology and vaccination, further integrated epidemiological, genomic and phenotypic surveillance is required to determine the spread and functional implications of these mutations.

BACKGROUND: Increasing trust and confidence in vaccines is a global priority, as countries have grappled with delivering COVID-19 vaccines, maintaining routine childhood vaccination rates and introducing new vaccines. Community-based vaccine promotion interventions are commonly implemented, but effectiveness evidence is limited. In 2022, supported by the Australian Government and in partnership with Fiji\'s Ministry of Health and UNICEF, we codesigned, delivered and comprehensively evaluated a vaccine education and communication training programme for health workers and community influencers to promote COVID-19 and routine immunisation.
METHODS: The Vaccine Champions programme included three phases: (1) codesign with Fiji stakeholders; (2) vaccine education and communication training for Vaccine Champions and (3) support for Champions to deliver community vaccine discussion sessions over 6 months.The RE-AIM framework evaluation measured programme reach, effectiveness, adoption, implementation and maintenance. Mixed-methods data were collected through interviews, surveys and field notes, integrating qualitative and quantitative data to triangulate findings. Primary outcomes included Champions\' knowledge, communication self-efficacy, trust in COVID-19 vaccines, programme satisfaction and community members\' intention to vaccinate.
RESULTS: We trained 35 Champions (27/35 female), including health workers, faith and community influencers. Half had a health background (17/35). Champions conducted 54 discussion sessions, reaching 1717 community members. Most Champions (22/35) conducted at least 1 session, with 16 running 3 or more. Champions who did not run sessions reported barriers like lack of confidence and competing duties. Training increased Champions\' communication self-efficacy and trust in COVID-19 vaccines. Community member intention to vaccinate increased from 41% (394/960) to 83% (822/991) before and after a session. The programme was well received with interest in continued engagement.
CONCLUSIONS: Training health workers and community Vaccine Champions can promote vaccine confidence. Programmes require government support and engagement for sustainability. Robust evaluation frameworks are needed to build the evidence base.

BACKGROUND: Gram-negative bacteria resistant to carbapenems are also known as critical antimicrobial resistant organisms. Their emergence at Colonial War Memorial Hospital (CWMH), the largest hospital in Fiji, is a major clinical concern. This study was conducted to determine the knowledge, attitudes, and readiness of healthcare workers (HCW) at CWMH regarding management of patients with infections caused by critical antimicrobial resistant organisms.
METHODS: A questionnaire was designed using a Likert scale to assess knowledge, attitudes, and readiness. Two cross-sectional studies were conducted, before and after the implementation of targeted educational activities which were informed by the pre-intervention study findings.
RESULTS: A total of 393 and 420 HCW participated in the pre- and post-intervention studies, respectively. The majority of respondents were female (77.3%) and 18-34 years of age (67%). HCW professional roles included nurses (56.3%), doctors (31.6%), and laboratory personnel (12.2%). In the post-intervention study, significantly more HCW reported having received infection prevention and control (IPC) and antimicrobial resistance education and training (26.8% in pre to 45.5% in post intervention, p < 0.001). The majority of nurses and doctors (> 85% to ≥ 95%) were aware of how AMR organisms spread in healthcare settings and knew the IPC measures to prevent transmission of AMR infections including hand hygiene, standard and transmission-based precautions. Attitudes towards AMR were positive, with 84.2% pre intervention and 84.8% of HCW post intervention expressing their willingness to change their work environment to assist with AMR prevention. Perceived readiness to address the problem showed mixed results. Improvements in laboratory AMR surveillance data availability were noted (29.4-52.4%, p < 0001). Modest improvement in the hospital\'s capacity for outbreak response (44-51.9%, p = 0.01), and treatment of AMR infections (38.9-44.4%, p = 0.01) was reported.
CONCLUSIONS: Our data revealed high levels of staff awareness and knowledge about AMR and IPC. However, readiness for outbreak response and treatment of critical AMR infections requires more attention. Improving AMR prevention and containment in CWMH will likely require sustained and multisectoral interventions with strong administrative commitment.

The enteric nervous system (ENS) consists of an extensive network of neurons and glial cells embedded within the wall of the gastrointestinal (GI) tract. Alterations in neuronal distribution and function are strongly associated with GI dysfunction. Current methods for assessing neuronal distribution suffer from undersampling, partly due to challenges associated with imaging and analyzing large tissue areas, and operator bias due to manual analysis. We present the Gut Analysis Toolbox (GAT), an image analysis tool designed for characterization of enteric neurons and their neurochemical coding using 2D images of GI wholemount preparations. It is developed in Fiji, has a user-friendly interface and offers rapid and accurate segmentation via custom deep learning (DL) based cell segmentation models developed using StarDist, and a ganglion segmentation model in deepImageJ. We use proximal neighbor-based spatial analysis to reveal differences in cellular distribution across gut regions using a public dataset. In summary, GAT provides an easy-to-use toolbox to streamline routine image analysis tasks in ENS research. GAT enhances throughput allowing unbiased analysis of larger tissue areas, multiple neuronal markers and numerous samples rapidly.

In Fiji, 90% of the population has access to basic sanitation; however, there are still persistent health risks from endemic faecal-oral diseases such as typhoid fever. There is a need to assess the contribution of existing sanitation facilities in the faecal pathogen transmission pathway. This study was conducted as part of a larger planetary health study across 29 rural communities within five river catchments. This specific research aimed to characterise latrine front-ends, both infrastructure and usage behaviour, and to assess the faecal contamination levels on various frequently contacted latrine surfaces in rural Fiji. A sanitation survey, along with observation and latrine swab sampling, was conducted in households over three phases: baseline (n = 311) (Aug-Dec 2019), endline (n = 262) (Jun-Sep 2022) and an in-depth front-end study (n = 12) (Oct-Nov 2022). Of 311 households, almost all had pedestal-type latrines, predominately cistern-flush (83%), followed by pour-flush (13%), and then hole-type (pit) latrines (4%). Washable latrine floors had significantly higher E. coli densities (6.7 × 102 CFU/25 cm2) compared to non-washable floors (1.3 × 102 CFU/25 cm2) (p = 0.05), despite washable floors indicating improved latrines. The in-depth front-end analysis found that moist latrine surfaces had significantly elevated E. coli densities (1.2 × 103 CFU/25 cm2) compared to the dry ones (14.3 CFU/25 cm2) (p < 0.001), highlighting the importance of maintaining dry latrine surfaces. Latrine floors and mid-walls were the most frequently contaminated surfaces, emphasising the need to clean and disinfect these surfaces. Only 46% of the households reported always using soap for handwashing after defecation, exacerbating the risk of transmitting faecal pathogens. This study highlights that latrine cleanliness and hygiene are as crucial as latrine infrastructures for the effective disruption of faecal pathogens transmission during latrine use.

UNASSIGNED: Sustainable Development Goal 3.4.1 (SDG3.4.1) targets a one-third reduction in non-communicable disease (NCD) mortality in ages 30-69-years by 2030 (relative to 2015). Directing interventions to achieve this aim requires reliable estimates of underlying cause of death (UCoD). This may be problematic when both cardiovascular diseases (CVD) and diabetes are present due to a lack of consistency in certification of such deaths. We estimate empirically 2013-17 NCD mortality in Fiji, by sex and ethnicity, from CVD, diabetes, cancer, and chronic lower respiratory diseases (CRD), and aggregated as NCD4.
UNASSIGNED: UCoD was determined from Medical Certificates of Cause-of-Death (MCCD) from the Fiji Ministry of Health after pre-processing of mortality data where diabetes and/or hypertension were present in order to generate internationally comparable UCoD. If no potentially fatal complications from diabetes or hypertension accompanied these causes in Part I (direct cause) of the MCCD, these conditions were re-assigned to Part II (contributory cause). The probability of a 30-year-old dying before reaching age 70-years (PoD30-70), by cause, was calculated.
UNASSIGNED: The PoD30-70 from NCD4 over 2013-17 differed by sex and ethnicity: in women, it was 36% (95%CI 35-37%) in i-Taukei and 27% (26-28%) in Fijians of Indian descent (FID); in men, it was 41% (40-42%) in both i-Taukei and FID.PoD30-70 from CVD, diabetes, cancer and CRD in women was: 18%, 10%, 13% and 1·0% in i-Taukei; 13%, 10%, 5·6% and 1·1% in FID; in men was: 28%, 8.4%, 7·6% and 2·2% in i-Taukei; 31%, 8.3%, 3.5% and 3·1% in FID.
UNASSIGNED: To achieve SDG3.4.1 goals in Fiji by 2030, effective population wide and ethnic-specific interventions targeting multiple NCDs are required to reduce PoD30-70 from NCD4: from 36% to 24% in i-Taukei, and 27% to 18% in FID women; and from 41% to 27% in i-Taukei and FID men.
UNASSIGNED: Not applicable.

The southwestern tropical Pacific is a key center for the Interdecadal Pacific Oscillation (IPO), which regulates global climate. This study introduces a groundbreaking 627-year coral Sr/Ca sea surface temperature reconstruction from Fiji, representing the IPO\'s southwestern pole. Merging this record with other Fiji and central tropical Pacific records, we reconstruct the SST gradient between the southwestern and central Pacific (SWCP), providing a reliable proxy for IPO variability from 1370 to 1997. This reconstruction reveals distinct centennial-scale temperature trends and insights into Pacific-wide climate impacts and teleconnections. Notably, the 20th century conditions, marked by simultaneous basin-scale warming and weak tropical Pacific zonal-meridional gradients, deviate from trends observed during the past six centuries. Combined with model simulations, our findings reveal that a weak SWCP gradient most markedly affects IPO-related rainfall patterns in the equatorial Pacific. Persistent synchronous western and central Pacific warming rates could lead to further drying climate across the Coral Sea region, adversely affecting Pacific Island nations.

Primary neuronal cultures are commonly used to study genetic and exogenous factors influencing neuronal development and maturation. During development, neurons undergo robust morphological changes involving expansion of dendritic arbor, formation of dendritic spines, and expression of synaptic proteins. In this chapter, we will cover methodological approaches allowing quantitative assessment of in vitro cultured neurons. Various quantitative characteristics of dendritic arbor can be derived based on immunostaining against anti-microtubule-associated protein 2 followed by dendrite tracing with the SNT plug-in of the FIJI software package. The number and subtypes of dendritic spines can be assessed by double labeling with DiI and Phalloidin iFluor448 followed by laser scanning confocal microscopy analysis. Finally, expression of presynaptic and postsynaptic proteins can be determined by immunohistochemistry and quantification using several available software packages including FIJI and Imaris, which also allows for 3D rendering and statistical displaying of the expression level of synaptic proteins.

Pacific Island Countries (PICs) have exceptionally high rates of obesity and non-communicable diseases. The causes are complex but one prominent factor is the notable shift from traditional plant and seafood diets to diets high in unhealthy processed foods. Literature is sparse on the motives behind food choice decisions of Pacific Islanders. The widely used Food Choice Questionnaire (FCQ) (Steptoe et al., 1995) is a multi-dimensional tool capturing nine potential food choice motives. This study used a hybrid thematic analysis. Fourteen in-depth semi-structured interviews with urban indigenous Fijian mothers were conducted to (1) explore how the nine FCQ motives impact food choice (2) to examine conceptual equivalence of the nine FCQ motives and (3) to identify additional food choice motives, not captured in the FCQ. Mothers identified seven of the nine FCQ motives (i.e., health, mood, convenience, sensory appeal, weight control and familiarity) and three new food choice motives (i.e., food that is filling, religious dietary restrictions and food quality) as important determinants of food choice. Moreover, while choosing foods that promote health and well-being was identified as an important food choice motive, mothers also identified four other motives, that they considered important, which lead to over-eating and unhealthy eating. Food choice is a complex, layered decision, driven by multiple considerations, some more important than others. These findings can inform diet-related interventions and policy within this community. Adapting the FCQ for broader PIC use through further qualitative exploration in PICs, and validating the adapted FCQ with larger samples will enhance its utility in measuring food choice motives in PICs.