Application and comparison of point-of-care devices for field evaluation of underlying health status of Guatemalan sugarcane workers.

With chronic disease prevalence on the rise globally, surveillance and monitoring are critical to improving health outcomes. Point-of-care (POC) testing can facilitate epidemiological research and enhance surveillance systems in limited resource settings, but previous research has identified bias between POC devices and laboratory testing. We compared the performance of two POC blood analyzers, the iSTAT handheld (Abbott, Princeton, NJ, USA) and the StatSensor Creatinine (Nova Biomedical, Waltham, MA, USA) to concurrent blood samples analyzed at a local laboratory that were collected from 89 agricultural workers in Guatemala. We measured creatinine and other measures of underlying health status with the POC and the lab blood samples. Pearson correlation coefficients, Bland-Altman plots, no intercept linear regression models and two-sample t-tests were used to evaluate the agreement between the POC and lab values collected across three study days and to assess differences by study day in a field setting. On average there was no observed difference between the iSTAT and lab creatinine measurements (p = 0.91), regardless of study day. Using lab creatinine as the gold standard, iSTAT creatinine results were more accurate compared to the Statsensor, which showed some bias, especially at higher values. The iSTAT had good agreement with the lab for sodium and blood urea nitrogen (BUN), but showed differences for potassium, anion gap, bicarbonate (TCO2), glucose, and hematocrit. In this tropical field setting, the research team devised a protocol to prevent the devices from overheating. In limited resource settings, POC devices carry advantages compared to traditional lab analyses, providing timely results to patients, researchers, and healthcare systems to better evaluate chronic health conditions. Technical challenges due to use of POC devices in high heat and humidity environments can be addressed using a standard protocol for transporting and operating the devices.

Maternal mortality and COVID-19: A nationwide ecological analysis from Ecuador.

BACKGROUND: During the COVID-19 pandemic, distinct population subsets, including pregnant women, have been differentially affected. While over 90% of COVID-19-infected pregnant women experience a benign course, a subset demonstrates marked clinical exacerbation. Symptomatic pregnant individuals, in particular, present a heightened risk of severe disease in comparison to their non-pregnant counterparts. OBJECTIVE: The objective of this study is to systematically evaluate the epidemiological characteristics of COVID-19 in pregnant women, assess related maternal mortalities, ascertain the case fatality rate, and delineate associated risk factors. DESIGN: This is a comprehensive population-based ecological study. METHODOLOGY: A population-based study was conducted to investigate the epidemiological patterns of COVID-19-associated morbidity and mortality in pregnant women in Ecuador from 27 February 2020 to 14 May 2021. RESULTS: A total of 3274 positive COVID-19 cases were identified among pregnant women, with 22 official fatalities, yielding a case fatality rate of 0.67%. The majority of cases were of Mestizo ethnicity (92.66%); however, the highest case fatality rate was noted among indigenous pregnant women (case fatality rate = 1.25%), those aged between 40 and 44 years (case fatality rate = 2.68%), and those with a history of comorbidities (2.08%). Pregnant women residing at lower altitudes (<2500 m) exhibited a higher incidence rate (0.20/100,000) compared to those at higher altitudes (>2500 m), which stood at 0.17/100,000. CONCLUSION: The COVID-19 pandemic has profoundly impacted pregnant women in Ecuador during the first 14 months, particularly those with comorbidities, older age, and of indigenous ethnicity. These factors have heightened their vulnerability and susceptibility to severe COVID-19 infection and subsequent mortality. This underscores the urgency for comprehensive protective measures, including prioritization for vaccination. Further studies are needed to inform tailored prevention strategies and therapeutic interventions for these high-risk groups.

A longitudinal assessment of heat exposure and biomarkers of kidney function on heat shock protein 70 and antibodies among agricultural workers.

Background: Exposure to extreme heat impacts millions of people worldwide and outdoor workers are among the populations most affected by high temperatures. Heat stress induces several biological responses in humans, including the production of heat shock proteins (HSP) and antibodies against HSP (anti-HSP) which may play a central role in the body's cellular response to a hot environment. Objective: This longitudinal study investigated the impact of high temperatures and humidity on the presence of HSP70 and anti-HSP70 and examined relationships with markers of kidney function in an at-risk workforce under conditions of extreme heat and exertion in Guatemala. Methods: We collected ambient temperature and relative humidity data as well as biomarkers and clinical data from 40 sugarcane workers at the start and the end of a 6-month harvest. We used generalized mixed-effects models to estimate temperature effects on HSP70 and anti-HSP70 levels. In addition, we examined trends between HSP70 and anti-HSP70 levels and markers of kidney function across the harvest. Results: At the end of the harvest, temperatures were higher, and workers had, on average, higher levels of HSP70 and anti-HSP70 compared to the beginning of the season. We observed significant increasing trends with temperature indices and HSP70 levels. Maximum temperature was associated with HSP70 increments after controlling for age, systolic and diastolic blood pressure (ß: 0.21, 95% Confidence Interval: 0.09, 0.33). Kidney function decline across the harvest was associated with both higher levels of anti-HSP70 levels at the end of the harvest as well as greater increases in anti-HSP70 levels across the harvest. Conclusions: These results suggest that workplace heat exposure may increase the production of HSP70 and anti-HSP70 levels and that there may be a relationship between increasing anti-HSP70 antibodies and the development of renal injury. HSP70 holds promise as a biomarker of heat stress in exposed populations.

Integration of Total Worker Health ® Training for Occupational Health and Safety Professionals With Efforts to Promote Worker Health, Safety, and Well-being in Mexico.

OBJECTIVE: We sought to test whether a 2-week Total Worker Health (TWH) training mapped to TWH education competencies could be administered to a Mexican audience of occupational safety and health professionals and could lead to positive changes to knowledge and behaviors. METHODS: This study used robust program evaluation methods collected before and after each of the nine training days and at the end of the course. RESULTS: Overall course quality received a mean score of 4.6 (SD = 0.6) and 98.8% of participants agreed that their TWH knowledge increased. All participants intended to make at least one change to their professional practice, most frequently helping companies assess their organizational culture to support health, safety, and well-being. CONCLUSIONS: This TWH training was well received and led to positive self-reported increase in knowledge and abilities to influence workers' health, safety, and well-being.

Quantification of Photobacterium swingsii and characterisation of disease progression in the New Zealand Greenshell™ mussel, Perna canaliculus.

Greenshell™ mussels (Perna canaliculus) are endemic to New Zealand and support the largest aquaculture industry in the country. Photobacterium swingsii was isolated and identified from moribund P. canaliculus mussels following a mass mortality event. In this study, a challenge experiment was used to characterise, detect, and quantify P. swingsii in adult P. canaliculus following pathogen exposure via injection into the adductor muscle. A positive control (heat-killed P. swingsii injection) was included to account for the effects of injection and inactive bacterial exposure. Survival of control and infected mussels remained 100% during 72-hour monitoring period. Haemolymph was sampled for bacterial colony counts and haemocyte flow cytometry analyses; histology sections were obtained and processed for histopathological assessments; and adductor muscle, gill, digestive gland were sampled for quantitative polymerase chain reaction (PCR) analyses, all conducted at 12, 24, 48 h post-challenge (hpc). The most profound effects of bacterial injection on mussels were seen at 48 hpc, where mussel mortality, haemocyte counts and haemolymph bacterial colony forming were the highest. The quantification of P. swingsii via qPCR showed highest levels of bacterial DNA at 12 hpc in the adductor muscle, gill, and digestive gland. Histopathological observations suggested a non-specific inflammatory response in all mussels associated with a general stress response. This study highlights the physiological effects of P. swingsii infection in P. canaliculus mussels and provides histopathological insight into the tissue injury caused by the action of injection into the adductor muscle. The multi-technique methods used in this study can be applied for use in early surveillance programs of bacterial infection on mussel farms.

Tírala Plena: findings from the formative research to inform the initiative "Reaching those most left behind through comprehensive sexuality education for out-of-school young people" in Colombia.

This paper presents the results of formative research conducted from January to June 2020 in the Department of Atlántico, Caribbean region of Colombia, whose findings were used as inputs to design the national strategy for comprehensive sexuality education (CSE) outside school - Tírala Plena - including its curriculum. This is within the framework of the multi-country project coordinated by UNFPA and WHO aimed at generating evidence on the role of facilitators in the delivery of CSE in non-school contexts. The research was carried out in four municipalities in northern Colombia, in rural and marginal urban contexts with conditions of vulnerability for the adolescent population, including a strong presence of migrant populations from Venezuela. A total of 150 male and female adolescents ages 10-17 participated in the formative research. Workshops such as patchwork quilt, body mapping and talking maps were used as methods to gather information. The groups were divided by sex and age (10-13 years old and 14-17 years old). Knowledge, attitudes and social norms regarding adolescent pregnancy, sexually transmitted infections, including human immunodeficiency virus, and gender-based violence, were identified in adolescents (schooled and not schooled, but with minimal or no access to CSE). All of the above enabled us to establish a set of recommendations for the strengthening of the CSE strategy Tírala plena.

Evaporative Cooling Does Not Prevent Vertical Dispersion of Effervescent Seawater Aerosol for Brightening Clouds.

Marine cloud brightening (MCB) is a potential intervention to mitigate the effects of climate change by increasing the reflectance of low-level maritime clouds, including those over the Great Barrier Reef. The technique involves dispersing a plume of submicrometer seawater droplets over the ocean, which evaporate, generating nanosized sea-salt aerosols (SSAs) that disperse through the atmosphere with some fraction incorporated into clouds. Droplet evaporation, which occurs in the immediate vicinity (meters to tens of meters) of the source, has been theorized to produce a negatively buoyant plume hindering the mixing of the sea-salt aerosol to cloud height and compromising the effectiveness of MCB. We characterized in situ for the first time the nearfield aerosol dispersion from a point source of atomized seawater produced using the effervescent technique. We observed consistent vertical mixing of the plume up to 150 ± 5 m height at 1 km downwind. The extent of vertical dispersion was influenced by wind velocity and atmospheric stability. We found no evidence that negative buoyancy due to the evaporation of the 0.068 kg/s water fraction significantly suppressed vertical mixing. Our results can be attributed to the small droplet sizes generated by the effervescent spray technology and associated low flow rates required to generate around 1014 droplets s-1. We estimate that, for a hypothetical implementation producing up to 1016 s-1 similarly sized SSAs, evaporative cooling is unlikely to significantly suppress the vertical dispersion of aerosol for MCB.

Interrater Reliability of the Spanish (Colombia) Version of the Post-COVID-19 Functional Status Scale.

Background: COVID-19 has been one of the most critical public health challenges of recent decades. This disease develops severely in one in five patients, and approximately 5% require admission to a critical care unit. Due to the impact of the sequelae, the Post-COVID-19 Functional Status Scale (PCFS) was developed. This study is aimed at determining the interrater reliability of the Spanish (Colombia) version of the PCFS in adult patients with post-COVID-19 infection. Methods: This is an observational study performed with patients diagnosed with COVID-19. Two evaluators repeated the test-retest of the PCFS scale with knowledge and clinical experience in the care of patients with COVID-19 and had previously applied the test. The PCFS assesses functional limitations at discharge and can be used at 4 and 8 weeks to evaluate practical consequences and determine the degree of disability these patients may have. For interrater reliability, Cronbach's alpha was applied with its respective confidence interval and the Bland-Altman method. A 95% confidence interval (CI) was taken as the basis for the interpretation of the Intraclass Correlation Coefficient (ICC). Results: A total of 112 adult patients participated in the study, aged 51.46 ± 15.94 years. It was evidenced that the survival, constant care, and activities of daily living questions have an ICC of one (1.000) with an ICC (1.000-1.000), demonstrating excellent reliability, while those close to one were instrumental activities, role participation, symptoms, and final score with an ICC 0.918 to 0.984 and an ICC (0.881-0.989). Thus, a homogeneous distribution of the interrater data was evident. Conclusions: Excellent interobserver reliability of the Spanish (Colombia) version of the PCFS in patients with different degrees of functional status was reported.

Perkinsus olseni in green-lipped mussels Perna canaliculus: diagnostic evaluation, prevalence, and distribution.

Perkinsus olseni (Perkinsidae) is a molluscan parasite notifiable to the World Organisation for Animal Health that is reported in several shellfish hosts in New Zealand, including the native green-lipped mussel Perna canaliculus. Green-lipped mussels comprise over half of New Zealand's aquaculture export value and have historically been considered free of serious diseases based on extensive histology-based surveillance. The discovery of P. olseni in green-lipped mussels has raised questions about future disease threats to green-lipped mussels, particularly under changing ocean climatic conditions. Using mussels collected from farmed (n = 358) and wild (n = 236) populations, we aimed to determine the distribution and prevalence of P. olseni in green-lipped mussels around New Zealand, and assess the performance of diagnostic tests, including real-time PCR, conventional PCR, and culture using Ray's fluid thioglycolate medium (RFTM). Prevalence and diagnostic test performance was evaluated using Bayesian latent class analysis with informative priors. The prevalence of P. olseni was 0-3%, except for 1 wild population from a harbour where prevalence was 22%. Real-time PCR had the highest diagnostic sensitivity (87%) compared to 62 and 21% for conventional PCR and RFTM, respectively. Diagnostic specificity was similar among all methods (96-98%). No mortality was observed during the study. Our results suggest that real-time PCR is the diagnostic test best suited for surveillance of P. olseni in subclinically infected green-lipped mussels under New Zealand conditions.

Design of a Bioinspired Underwater Glider for Oceanographic Research.

The Blue Economy, which is based on the sustainable use of the ocean, is demanding better understanding of marine ecosystems, which provide assets, goods, and services. Such understanding requires the use of modern exploration technologies, including unmanned underwater vehicles, in order to acquire quality information for decision-making processes. This paper addresses the design process for an underwater glider, to be used in oceanographic research, that was inspired by leatherback sea turtles (Dermochelys coriacea), which are known to have a superior diving ability and enhanced hydrodynamic performance. The design process combines elements from Systems Engineering and bioinspired design approaches. The conceptual and preliminary design stages are first described, and they allowed mapping the user's requirements into engineering characteristics, using quality function deployment to generate the functional architecture, which later facilitated the integration of the components and subsystems. Then, we emphasize the shell's bioinspired hydrodynamic design and provide the design solution for the desired vehicle's specifications. The bioinspired shell yielded a lift coefficient increase due to the effect of ridges and a decrease in the drag coefficient at low angles of attack. This led to a greater lift-to-drag ratio, a desirable condition for underwater gliders, since we obtained a greater lift while producing less drag than the shape without longitudinal ridges.

Urinary concentrations of metals before and after volcanic eruption: a natural experiment surrounding the eruption of Volcán de Fuego, 2018.

Volcanic eruptions increase environmental heavy metal concentrations, yet little research has been performed on their extrapulmonary human health effects. We fortuitously collected biological samples in a cohort of Guatemalan sugarcane cutters in the area surrounding Volcán de Fuego before and after the June 2018 eruption. We sought to determine whether stratovolcanic activity was associated with changes in urinary concentrations of heavy metals in a cohort of sugarcane workers. In this exploratory analysis, we found significant increases in urinary arsenic, (ß = 1.46, P < 0.0001), cadmium (ß = 1.03, P < 0.0001), and lead (ß = 0.87, P = 0.003) in participants with residential proximity to Volcán de Fuego as compared to participants farther away, suggesting that volcanic activity could be associated with acute heavy metal exposures. This natural experiment is, to our knowledge, the first of its kind and suggests a need for more research into heavy metal exposure-related health impacts of volcanic eruptions.

Pathogenicity and virulence of bacterial strains associated with summer mortality in marine mussels (Perna canaliculus).

The occurrence of pathogenic bacteria has emerged as a plausible key component of summer mortalities in mussels. In the current research, four bacterial isolates retrieved from moribund Greenshell࣪ mussels, Perna canaliculus, from a previous summer mortality event, were tentatively identified as Vibrio and Photobacterium species using morpho-biochemical characterization and MALDI-TOF MS and confirmed as V. celticus, P. swingsii, P. rosenbergii, and P. proteolyticum using whole genome sequencing. These isolates were utilized in a laboratory challenge where mussels were injected with cell concentrations ranging from 105 to 109 CFU/mussel. Of the investigated isolates, P. swingsii induced the highest mortality. Additionally, results from quantitative polymerase chain reaction analysis, focusing on known virulence genes were detected in all isolates grown under laboratory conditions. Photobacterium rosenbergii and P. swingsii showed the highest expression levels of these virulence determinants. These results indicate that Photobacterium spp. could be a significant pathogen of P. canaliculus, with possible importance during summer mortality events. By implementing screening methods to detect and monitor Photobacterium concentrations in farmed mussel populations, a better understanding of the host-pathogen relationship can be obtained, aiding the development of a resilient industry in a changing environment.

Chemical Characterization and Biological Activity of the Essential Oil from Araucaria brasiliensis Collected in Ecuador.

The purpose of this study was to determine the chemical composition, physical properties, enantiomeric composition and cholinesterase inhibitory activity of the essential oil (EO) steam-distilled from the leaves of the plant Araucaria brasiliensis Loud. collected in Ecuador. The chemical composition was determined by gas chromatography coupled to mass spectrometry (GC-MS) analysis on two capillary GC columns (DB5-ms and HP-INNOWax). Thirty-three compounds were identified in the EO; the main compounds were beyerene (26.08%), kaurene (24.86%), myrcene (11.02%), α-pinene (9.99%) and 5,15-rosadiene (5.87%). Diterpene hydrocarbons (65.41%), followed by monoterpene hydrocarbons (21.11%), were the most representative components of the EO. Enantioselective analysis of the EO showed four pairs of enantiomeric compounds, α-pinene, camphene, γ-muurolene and δ-cadinene. In an in vitro assay, the EO showed moderate inhibitory activity towards the enzyme butyrylcholinesterase (BuChE) (95.7 µg/mL), while it was inactive towards acetylcholinesterase (AChE) (225.3 µg/mL). Further in vivo studies are needed to confirm the anticholinesterase potential of the EO.

Latin American Agricultural Workers' Job Demands and Resources and the Association With Health Behaviors at Work and Overall Health.

In the present study, we describe the job demands and job resources (JD-R) experienced by agricultural workers in three Latin American countries and their relationship to proactive health behaviors at work and overall health. Following previous research on the JD-R model, we hypothesized that job demands (H1) would be negatively related to agricultural workers' self-reported overall health. On the other hand, we hypothesized that job resources (H2) would be positively related to agricultural workers' overall health. Furthermore, we hypothesized (H3) that workers' engagement in jobsite health promotion practices via their proactive health behaviors at work would partially mediate the relationship between workers' job resources and job demands and overall health. We also had a research question (R1) about whether there were differences by type of job held. The sample of workers who participated in this study (N = 1,861) worked in Mexico, Guatemala, and Nicaragua for one large agribusiness that produces sugar cane. They worked in two distinct areas: company administration and agricultural operations. We administered employee health and safety culture surveys using survey methods tailored to meet the needs of both types of workers. Stratified path analysis models were used to test study hypotheses. In general, we found support for hypotheses 1 and 2. For example, operations workers reported more physically demanding jobs and administrative workers reported more work-related stress. Regardless, the existence of high job demands was associated with poorer overall health amongst both types of workers. We found that workers in more health-supportive work environments perform more proactive health behaviors at work, regardless of their role within the organization. However, hypothesis 3 was not supported as proactive health behaviors at work was not associated with overall health. We discuss future research needs in terms of evaluating these hypotheses amongst workers employed by small- and medium-sized agribusinesses as well as those in the informal economy in Latin America. We also discuss important implications for agribusinesses seeking to develop health promotion programs that meet the needs of all workers.

Environmental metal exposures and kidney function of Guatemalan sugarcane workers.

BACKGROUND: Exposure to environmental metals can cause nephrotoxicity. There is an international epidemic of chronic kidney disease of unknown cause (CKDu). Whether metal exposures contribute to kidney dysfunction in populations at risk for CKDu remains unresolved. OBJECTIVE: Urinary metals (arsenic, cadmium, nickel, and uranium) were examined in 222 sugarcane cutters in Guatemala at three time points over 1 year. METHODS: We explored the relationships between metal concentrations and markers of kidney function using multivariable linear mixed-effect models. RESULTS: Arsenic, cadmium, and nickel were detected in the majority of the 340 urine samples and were generally within limits previously considered to be nonnephrotoxic. Nevertheless, higher urine cadmium was inversely associated with estimated glomerular filtration rate (eGFR) (ß: -4.23, 95% confidence interval [CI]: -6.92, -1.54) and positively associated with neutrophil gelatinase-associated lipocalin (NGAL) (ß: 2.92, 95% CI: 1.20, 4.64). Higher urine arsenic was also inversely associated with eGFR (ß: -4.36, 95% CI: -7.07, -1.64). SIGNIFICANCE: Our findings suggest that exposures to metals, including cadmium and arsenic, might contribute to kidney toxicity seen in workers at risk for CKDu. These findings are consistent with the potential for metal nephrotoxicity at lower than expected levels in the setting of manual work in a very hot environment.

Body Composition, Anemia, and Kidney Function among Guatemalan Sugarcane Workers.

Rates of anemia among agricultural workers, who are also at risk for kidney injury and chronic kidney disease of unknown cause (CKDu), are unknown. We evaluated body composition through the sum of three skinfolds among 203 male sugarcane cutters and assessed the relationship of variables related to nutrition, anemia (hemoglobin < 13 g/dL), and elevated hemoglobin A1c (HbA1c ≥ 5.7%) with estimated glomerular filtration rate (eGFR) using linear regression. Eleven percent of workers were at the level of essential body fat (2-5%). Anemia was present among 13% of workers, 70% of which were normochromic normocytic, a type of anemia suggesting potential underlying chronic disease. Anemia was more common among those with lower BMI and fat free mass. The prevalence of elevated HbA1c was 21%. A moderate negative correlation was found between hemoglobin and HbA1c (Pearson's r = -0.32, p <0.01) which suggests that HbA1c values should be interpreted with caution in populations that have high rates of anemia. Twelve percent of workers had reduced kidney function with an eGFR < 90 mL/min/1.73 m2. On average, the eGFR was 18 mL/min per 1.73 m2 lower [(95% CI:-24, -12), p <0.01)] for those with anemia than those without, and 8 mL/min per 1.73 m2 lower among those with elevated HbA1c [(95% CI: -13, -2), p < 0.01]. Results will inform future studies examining the role of anemia in the evaluation of CKDu and interventions to improve nutrition for workers in low-resource settings.

Sugarcane Workweek Study: Risk Factors for Daily Changes in Creatinine.

INTRODUCTION: Agricultural workers laboring in thermally stressful environments are at increased risk for kidney injury and chronic kidney disease of unknown origin (CKDu), and their environmental and occupational exposures have been considered to be important risk factors. This study examined the effects of repeated kidney stress from the simultaneous strain of work and other factors experienced by workers in Guatemala during a typical workweek. METHODS: We collected data from 107 sugarcane workers across 7 consecutive work shifts. Data included information on daily occupational, meteorological, environmental, and lifestyle factors. We used multivariable linear mixed models to evaluate associations of these factors with percent change in creatinine. RESULTS: We observed that increasing wet bulb globe temperature (ß = 2.5%, 95% confidence interval [CI] = 0.3%, 4.7%) and increasing diastolic blood pressure (ß = 6.2%, 95% CI = 0.9%, 11.6%) were associated with increases in creatinine across the shift, whereas consumption of water from chlorinated dormitory tanks as compared to artesian well water (ß = -17.5%, 95% CI = -29.6%, -5.4%) and increasing number of rest breaks (ß = -5.8%, 95% CI = -9.0%, -2.6%) were found to be protective against increases in creatinine. Workers reporting drinking tank water had lower concentrations of urine creatinine-corrected arsenic, lead, uranium, and glyphosate compared to workers reporting the use of well water or municipal water. CONCLUSION: These results reinforce the need to focus on preventive actions that reduce kidney injury among this worker population, including strategies to reduce heat stress, managing blood pressure, and examining water sources of workers for nephrotoxic contaminants.

International Total Worker Health: Applicability to Agribusiness in Latin America.

Total Worker Health® (TWH) is a framework for integrating worker and workplace safety, health, and well-being, which has achieved success in European and US settings. However, the framework has not been implemented in Latin America or in agricultural sectors, leaving large and vulnerable populations underrepresented in the implementation and evaluation of these strategies to improve safety and promote health and well-being. This study presents a case study of how a TWH approach can be applied to a multinational Latin American agribusiness. We describe the process and adaptation strategy for conducting a TWH assessment at multiple organizational levels and in multiple countries. We follow this with a description of a TWH leadership training that was conducted based on the results of the assessment. Finally, we describe our methods to make corporate recommendations for TWH policies and programs that were informed by the TWH assessment and leadership trainings. With this case study we aim to demonstrate the importance and feasibility of conducting TWH in Latin America.

Intracellular bacteria in New Zealand shellfish are identified as Endozoicomonas species.

Kaimoana (shellfish, seafood) is an important food source and a significant social and cultural component of many New Zealand communities, especially the indigenous Maori. Over the past decade a decline has been detected in shellfish health and an increase in mortality events around New Zealand. Intracellular bacteria termed Rickettsia-like organisms (RLOs) have been observed in New Zealand bivalve molluscs during shellfish mortality events. Affected bivalves include cockles Austrovenus stutchburyi, ringed dosinia Dosinia anus, green-lipped mussels Perna canaliculus, pipi Paphies australis, toheroa Paphies ventricosa, tuatua Paphies subtriangulata, deepwater tuatua Paphies donacina and scallops Pecten novaezelandiae. RLOs are an informal morphology-based classification of intracellular bacteria, with the exact identification often unknown. Using shellfish collected during mortality events from 2014 to 2019 and apparently healthy samples collected in 2018 and 2019, we aimed to identify RLOs in New Zealand shellfish. Bacterial 16S rRNA gene sequences from RLO-infected shellfish showed >95% identity to published Endozoicomonas species. In situ hybridization confirmed the presence of the sequenced gene in the gill epithelium and digestive epithelium of all study species. A genus-specific quantitative PCR, targeting the 16S rRNA gene was developed to detect Endozoicomonas spp. in shellfish tissue. Prevalence of Endozoicomonas spp. in samples from mortality events and healthy shellfish analysed by quantitative PCR was high. Samples collected from mortality events, however, had a significantly higher load of Endozoicomonas spp. than the healthy samples. These results give us a greater understanding of these intracellular bacteria and their presence in populations of New Zealand shellfish.