Perspectives on digital testing services for sexually transmitted and blood-borne infections from Two-Spirit, gay, bisexual, transgender, and other queer Black, Indigenous, people of colour living in Ontario, Canada.

OBJECTIVES: Increased sexually transmitted and blood-borne infections (STBBI) testing can reduce the burden of disease among Two-Spirit, gay, bisexual, transgender, and other queer Black, Indigenous, people of colour (2SGBTQ+ BIPOC). However, this population encounters barriers, such as discrimination, when accessing in-person STBBI testing services. Digital STBBI testing, such as self-testing/collection kits ordered online and digital requisitions, may address some of these barriers. Our aim was to understand acceptability of free digital STBBI testing among 2SGBTQ+ BIPOC living in Ontario, Canada. DESIGN: We approached this analysis using Implementation Science and Critical Race Theory. We conducted interviews and focus groups with 21 2SGBTQ + BIPOC individuals from 2020-2021. Participants were asked about their perceptions of the benefits and drawbacks of digital STBBI testing, populations that would benefit from using these services, and recommendations for how these services may be implemented in Ontario. Interviews and focus groups were transcribed verbatim and analyzed using reflexive thematic analysis. RESULTS: Six themes emerged. Digital STBBI testing services: (1) May reduce oppression experienced by 2SGBTQ + BIPOC when testing in-person; (2) Should address the unique needs that 2SGBTQ + BIPOC experience due to other intersecting identities they possess; (3) Should adapt their services to suit the varying cultural contexts and living circumstances of 2SGBTQ + BIPOC; (4) Should be accessible to 2SGBTQ + BIPOC who hold diverse or no documentation; (5) Should be offered in multiple languages; (6) May be inaccessible to those without Internet access or devices. CONCLUSION: Digital STBBI testing is one strategy that may reduce discrimination experienced by 2SGBTQ + BIPOC when getting tested in-person. However, digital STBBI testing services may not address all the needs of 2SGBTQ + BIPOC. Racism and other forms of oppression embedded into in-person and digital testing services will need to be addressed to meet the needs of this diverse population.

'Sometimes white doctors are not very friendly or inclusive': a Critical Race Theory analysis of racism within and beyond sexual health settings.

Many Two-Spirit, gay, bisexual, transgender, and other queer Black, Indigenous, people of colour in Canada encounter racism when testing for sexually transmitted and blood-borne infections. Our objective in this study was to understand how racism shapes testing experiences for these communities in Ontario, Canada. Four peer researchers conducted recruitment and data collection in consultation with a community advisory board. Focus groups and interviews took place with 21 participants and their narrative accounts were analysed using reflexive thematic analysis. Participants identified three interrelated issues when testing: (1) experiencing judgement and discomfort due to racism; (2) lack of community and cultural indicators in testing spaces; and (3) barriers to accessing testing centres and services. Systemic racism was linked to each of these barriers, including increased distance to testing centres due to racial segregation. Participant accounts signal the need for antiracist testing spaces and practices. Key implications include the need for antiracism training for health service providers and others working with Two-Spirit, gay, bisexual, transgender, and other queer Black, Indigenous, people of colour, and the organisations that serve them, in order to make testing spaces safer. Dismantling systemic racism is imperative to achieve health equity for members of these communities.

Diisopropylfluorophosphate-induced status epilepticus drives complex glial cell phenotypes in adult male mice.

Organophosphate pesticides and nerve agents (OPs), are characterized by cholinesterase inhibition. In addition to severe peripheral symptoms, high doses of OPs can lead to seizures and status epilepticus (SE). Long lasting seizure activity and subsequent neurodegeneration promote neuroinflammation leading to profound pathological alterations of the brain. The aim of this study was to characterize neuroinflammatory responses at key time points after SE induced by the OP, diisopropylfluorophosphate (DFP). Immunohistochemistry (IHC) analysis and RT-qPCR on cerebral tissue are often insufficient to identity and quantify precise neuroinflammatory alterations. To address these needs, we performed RT-qPCR quantification after whole brain magnetic-activated cell-sorting (MACS) of CD11B (microglia/infiltrated macrophages) and GLAST (astrocytes)-positive cells at 1, 4, 24 h and 3 days post-SE. In order to compare these results to those obtained by IHC, we performed, classical Iba1 (microglia/infiltrated macrophages) and GFAP (astrocytes) IHC analysis in parallel, focusing on the hippocampus, a brain region affected by seizure activity and neurodegeneration. Shortly after SE (1-4 h), an increase in pro-inflammatory (M1-like) markers and A2-specific markers, proposed as neurotrophic, were observed in CD11B and GLAST-positive isolated cells, respectively. Microglial cells successively expressed immuno-regulatory (M2b-like) and anti-inflammatory (M2a-like) at 4 h and 24 h post-SE induction. At 24 h and 3 days, A1-specific markers, proposed as neurotoxic, were increased in isolated astrocytes. Although IHC analysis presented no modification in terms of percentage of marked area and cell number at 1 and 4 h after SE, at 24 h and 3 days after SE, microglial and astrocytic activation was visible by IHC as an increase in Iba1 and GFAP-positive area and Iba1-positive cells in DFP animals when compared to the control. Our work identified sequential microglial and astrocytic phenotype activation. Although the role of each phenotype in SE cerebral outcomes requires further study, targeting specific markers at specific time point could be a beneficial strategy for DFP-induced SE treatment.

Decreased microglial Wnt/ß-catenin signalling drives microglial pro-inflammatory activation in the developing brain.

Microglia of the developing brain have unique functional properties but how their activation states are regulated is poorly understood. Inflammatory activation of microglia in the still-developing brain of preterm-born infants is associated with permanent neurological sequelae in 9 million infants every year. Investigating the regulators of microglial activation in the developing brain across models of neuroinflammation-mediated injury (mouse, zebrafish) and primary human and mouse microglia we found using analysis of genes and proteins that a reduction in Wnt/ß-catenin signalling is necessary and sufficient to drive a microglial phenotype causing hypomyelination. We validated in a cohort of preterm-born infants that genomic variation in the Wnt pathway is associated with the levels of connectivity found in their brains. Using a Wnt agonist delivered by a blood-brain barrier penetrant microglia-specific targeting nanocarrier we prevented in our animal model the pro-inflammatory microglial activation, white matter injury and behavioural deficits. Collectively, these data validate that the Wnt pathway regulates microglial activation, is critical in the evolution of an important form of human brain injury and is a viable therapeutic target.

Oxytocin receptor agonist reduces perinatal brain damage by targeting microglia.

Prematurity and fetal growth restriction (FGR) are frequent conditions associated with adverse neurocognitive outcomes. We have previously identified early deregulation of genes controlling neuroinflammation as a putative mechanism linking FGR and abnormal trajectory of the developing brain. While the oxytocin system was also found to be impaired following adverse perinatal events, its role in the modulation of neuroinflammation in the developing brain is still unknown. We used a double-hit rat model of perinatal brain injury induced by gestational low protein diet (LPD) and potentiated by postnatal injections of subliminal doses of interleukin-1ß (IL1ß) and a zebrafish model of neuroinflammation. Effects of the treatment with carbetocin, a selective, long lasting, and brain diffusible oxytocin receptor agonist, have been assessed using a combination of histological, molecular, and functional tools in vivo and in vitro. In the double-hit model, white matter inflammation, deficient myelination, and behavioral deficits have been observed and the oxytocin system was impaired. Early postnatal supplementation with carbetocin alleviated microglial activation at both transcriptional and cellular levels and provided long-term neuroprotection. The central anti-inflammatory effects of carbetocin have been shown in vivo in rat pups and in a zebrafish model of early-life neuroinflammation and reproduced in vitro on stimulated sorted primary microglial cell cultures from rats subjected to LPD. Carbetocin treatment was associated with beneficial effects on myelination, long-term intrinsic brain connectivity and behavior. Targeting oxytocin signaling in the developing brain may be an effective approach to prevent neuroinflammation - induced brain damage of perinatal origin.

γ-Aminobutyric acid receptor alpha 1 subunit loss of function causes genetic generalized epilepsy by impairing inhibitory network neurodevelopment.

OBJECTIVE: In humans, mutations of the γ-aminobutyric acid receptor subunit 1 (GABRA1) cause either mild or severe generalized epilepsy. Although these epilepsy-causing mutations have been shown to disrupt the receptor activity in vitro, their in vivo consequences on brain development and activity are not known. Here, we aim at unraveling the epileptogenesis mechanisms of GABRA1 loss of function. METHODS: We generated a gabra1-/- zebrafish mutant line displaying highly penetrant epileptic seizures. We sought to identify the underlying molecular mechanisms through unbiased whole transcriptomic assay of gabra1-/- larval brains. RESULTS: Interestingly, mutant fish show fully penetrant seizures at juvenile stages that accurately mimic tonic-clonic generalized seizures observed in patients. Moreover, highly penetrant seizures can be induced by light stimulation, thus providing us with the first zebrafish model in which evident epileptic seizures can be induced by nonchemical agents. Our transcriptomic assay identified misregulated genes in several pathways essential for correct brain development. More specifically, we show that the early development of the brain inhibitory network is specifically affected. Although the number of GABAergic neurons is not altered, we observed a drastic reduction in the number of inhibitory synapses and a decreased complexity of the GABAergic network. This is consistent with the disruption in expression of many genes involved in axon guidance and synapse formation. SIGNIFICANCE: Together with the role of GABA in neurodevelopment, our data identify a novel aspect of epileptogenesis, suggesting that the substratum of GABRA1-deficiency epilepsy is a consequence of early brain neurodevelopmental defects, in particular at the level of inhibitory network wiring.

Seroprevalence of brucellosis and associated risk factors in camels and its herders in selected districts of Somali Pastoral Region, Eastern Ethiopia.

Brucellosis poses a major public and animal health problem in many parts of the world, particularly in pastoral settings, however, seroepidemological studies are scarce. A cross-sectional study was conducted from December 2021 to April 2022 to estimate the prevalence of brucellosis and to identify the associated risk factors for camels and occupational individuals from three purposively selected districts of the Somali pastoral region in Eastern Ethiopia. Serum samples were serially diluted using the Rose Bengal Plate Test (RBPT) as a screening test and a competitive Enzyme-Linked Immunosorbent Assay (cELISA) test as a confirmatory test. From a total of 450 camels and 250 human serum samples tested, the overall seroprevalence was confirmed to be 2.9 % (95 % CI, 1.5-4.9) in camels and 2.0 % (95 % CI, 0.2-3.7) in humans. In camels, abortion and retained fetal membrane (RFM) were significant risk factors for Brucella seropositivity (p<0.05). However, in humans, RFM disposal differed significantly (p<0.05). The fact that brucellosis is found in both camels and humans highlights the importance of implementing a coordinated One Health approach to control and eliminate the disease. This would ensure improved public health and increased livestock productivity.

Participatory epidemiology on major camel calf health and management problems in pastoral and semi-pastoral zones of Somali region, Ethiopia.

A cross-sectional study was conducted from May 2017 to March 2019. A participatory epidemiological appraisal was used to gain a rapid overview of the range of camel calf health problems and traditional management, a measure of the importance that people place on each of them, and to identify and prioritize economically important diseases in target zones. The most important constraints to camel production were identified to be the widespread prevalence of diseases such as camelpox, contagious ecthyma, calf scour, ticks, and nonspecific pneumonia; poor management and husbandry practices such as restrictive colostrum feeding, lack of concentrate and salt supplementation and inappropriate housing; shortage of feed; and scarce seasonal variation in water. Additionally, the livestock herders not only showed their knowledge of common camel calf diseases for affected organs and symptoms but also indicated the seasonality of disease occurrences with strong agreement (W = 0.899, P< 0.003) among the informants of all focus group discussions. The overall prevalence of mange, tick infestation, and bacteria-induced diarrhea in the study area was found to be 36.3%, 36%, and 74%, respectively. Sarcoptes scabie var. cameli was the only identified mite species from mange-infested calves, while Hyalomma, Rhipicephalus, and Amblyoma were the most commonly identified tick species. Similarly, the overall prevalence of diarrhea was 74% among this about 34.6% was caused by E. coli while 38.9% was affected by Salmonella and E. coli. Therefore, based on these findings, five diseases have been prioritized as the most significant calf diseases in the area (Camelpox, contagious ecthyma, and causes of pneumonia among camel calves). Improving veterinary health infrastructure and capacity, and increasing community awareness on camel health constraints are also recommended to enhance optimal camel calf rearing.

Prevalence and risk factors of bovine tuberculosis in cattle in selected districts of Fafan pastoral settings, Eastern Ethiopia.

Cross-sectional study was conducted from October 2021 to August 2022 to investigate the prevalence and associated risk factors of bovine tuberculosis in cattle in selected districts of the pastoral settings of Fafan zone, Somali region, eastern Ethiopia. A comparative intradermal tuberculin test was performed using purified protein derivatives. Animal-related characteristics, and the owner's knowledge on the importance of BTB were collected using a structured questionnaire. The prevalence was 11.24 % (95 % CI, 8.61-14.35) and 43.3 % (95 % CI, 33.27-53.75) at the individual and herd levels, respectively. There were statistically significant differences in the proportions of positive reactor animals according to body condition score (P = 0.000), age (P = 0.048), seasonal migration (P = 0.038), parity number (P = 0.005), and reproductive status (P = 0.037). Animals with poor body condition scores had a significantly higher likelihood of testing positive, with their odds being 11.4 times greater (COR = 11.408, CI = 3.43-37.94, P < 0.001). In multivariate logistic regression, poor body condition score remained significantly associated with the odds of a positive reaction to tuberculosis (AOR = 0.137, CI = 0.053-0.356, P < 0.001). Similarly, the analysis showed that seasonal migration (AOR = 2.882, CI = 1.155-7.191, P = 0.023) and parity number (AOR = 11.64, CI = 1.818-74.464, P = 0.010) were significant predictors of bovine tuberculosis infection in cattle. According to the questionnaire, 14.2 % (17 of 120) and 13.3 % (16 of 120) of the respondents were knowledgeable about bovine tuberculosis and its transmission from animals to humans, and vice versa, respectively. The general judgment of herders' understanding of bovine tuberculosis transmission methods to humans was very low. The study findings showed a high prevalence of bovine tuberculosis in the study area, emphasizing the need for an effective control and prevention strategy.

Determination of prevalence of subclinical mastitis, characterization of intra-mammary infection-causing bacteria, and antibiotic susceptibility in dairy camels in Jigjiga City, Somali region, Ethiopia.

Background: Subclinical mastitis in camels, an inflammation of the udder without visible signs, can reduce milk quality and raise bacteria levels. Regular monitoring of camel milk is crucial for consumer safety. Methods: A cross sectional study was conducted in Jigjiga city, Ethiopia to investigate the prevalence and characteristics of subclinical mastitis in she-camels. The study included 244 lactating she-camels from three privately-owned camel dairy farms, and a questionnaire survey was conducted with 60 camel owners. Results: The overall prevalence of subclinical mastitis in she-camels was 10.6% (26/244), with no significant difference among the studied dairy farms. Risk factors that influenced the result of California Mastitis Test (CMT) included age and udder and leg hygiene. The study revealed that S. aureus was the most prevalent bacterium among the isolated bacteria, with a prevalence rate of 34.5%. This was followed by S. agalactiae, S. dysgalactiae, and Pasteurella multocida, with prevalence rates of 29.8, 19.4, and 16.2%, respectively. Among the isolated bacteria, 84.5% were sensitive to Erythromycin, 60% to Streptomycin, 44.7% to Oxytetracycline, and 36.7% to Tetracycline. Interviews with camel owners revealed that 66.7% used mixed herd grazing methods and reported feed shortage. Treatment practices for sick camels included modern veterinary drugs, traditional medicines, or a combination of both. The owners of camel dairy farms did not maintain proper hygiene practices during milking, such as not using soap when washing hands. Conclusion: Addressing camel mastitis necessitates access to alternative drugs, comprehensive herder training, and enhanced management practices.

Brief review on ebola virus disease and one health approach.

Ebola virus disease (EVD) is a severe and highly fatal zoonotic disease caused by viruses in the family Filoviridae and genus Ebolavirus. The disease first appeared in Zaire near the Ebola River in 1976, now in the Democratic Republic of the Congo. Since then, several outbreaks have been reported in different parts of the world, mainly in Africa, leading to the identification of six distinct viral strains that cause disease in humans and other primates. Bats are assumed to be the main reservoir hosts of the virus, and the initial incidence of human epidemics invariably follows exposure to infected forest animals through contact or consumption of bush meat and body fluids of forest animals harboring the disease. Human-to-human transmission occurs when contaminated body fluids, utensils, and equipment come in contact with broken or abraded skin and mucous membranes. EVD is characterized by sudden onset of 'flu-like' symptoms (fever, myalgia, chills), vomiting and diarrhea, then disease rapidly evolves into a severe state with a rapid clinical decline which may lead potential hemorrhagic complications and multiple organ failure. Effective EVD prevention, detection, and response necessitate strong coordination across the animal, human, and environmental health sectors, as well as well-defined roles and responsibilities evidencing the significance of one health approach; the natural history, epidemiology, pathogenesis, and diagnostic procedures of the Ebola virus, as well as prevention and control efforts in light of one health approach, are discussed in this article.

Camel gastrointestinal helminths in selected districts of Fafan zone, eastern Ethiopia.

A cross-sectional study was conducted to estimate the incidence and prevalence of helminths in camels in the Jigjiga and Gursum districts of Fafan zone, Somali regional state of Ethiopia. Fecal samples were collected from individual animals and analyzed using a McMaster fecal flotation method. Fecal samples were mixed with water and centrifuged to remove excess debris prior to mixing with flotation solution and performing the McMaster. For each sample, the number and types of parasite eggs present were recorded. 77.3% of examined camels were found harboring gastrointestinal parasites. Trichostrongylid spp. (68.06%) were the predominant parasite followed by Strongyloides spp. (25.6%), Trichuris spp. (15.5%) and Monezia spp. (8.4%). Risk factors for gastrointestinal parasite prevalence included age, body condition score and fecal quality (P < 0.05). Camels from the Gursum district had a significantly higher mean egg count than camels from the Jigjiga district (868.9 ± 1064.2 vs 351 ± 422.4; F = 20.8, P < 0.001). Moreover, there was a statistically significant difference in mean egg count between the sexes (F = 5.9, P = 0.02), with females (724.6 ± 960.6) having higher egg counts than males (373.4 ± 470.6). This study indicates that gastrointestinal helminths are highly prevalent and may affect the health and productivity of the camels in pastoral areas of Fafan zone.

Gastrointestinal nematodiasis of goats in Somali pastoral areas, Ethiopia.

Livestock, mainly goats, are crucial for animal protein, household income, economic security, and wealth creation in the pastoral areas of eastern Ethiopia. However, gastrointestinal parasitosis poses a substantial challenge in this sector. A cross-sectional study was conducted in the Gursum district of the Somali region, Ethiopia, to investigate the prevalence and intensity of gastrointestinal nematodes in goats and their associated risk factors. A total of 384 goat fecal samples were collected and examined using flotation and McMaster egg counting techniques for GIT nematodes. Coprological cultures have also been conducted for nematode identification. Fecal samples showed an overall parasite prevalence of 54.17%, with identified nematodes including Haemonchus (24%), Strongyloides (10.4%), Trichostrongles (6.5%), Nematodirus (6%), Oesophagostomum (5.5%) and Trichuris (1.87%). Older and poor body condition animals had higher chances of hosting nematodes than younger (OR = 0.245; CI = 0.144-0.417) and good body condition animals (OR = 0.069; CI = 0.030-0.157), according to multivariate logistic regression analysis. Quantitative examination of eggs revealed light 75(36.06%), moderate 99(47.60%), and heavy infection (n = 34, 16.35%). Analysis of the different study variables indicated that the age and body condition of the animals and the season of the year had a statistically significant association with the prevalence of GIT nematode infections (P-value <0.05). The high prevalence and intensity of GIT nematodiasis in goats from the study area warrants immediate attention and the implementation of strategic control and prevention measures.

Participatory and Conventional Investigation of Tick Infestation in Camels and Cattle of Somali Pastoral Areas, Eastern Ethiopia.

Ticks are a common parasite that affect many animals by causing slowed growth, reduced milk output, and financial losses for industries that depend on animal hides and skins. From June to December 2017, participatory and conventional investigations on tick infestation in camels and cattle were conducted in Kebribayah and Afdem districts of Ethiopia's Somali Regional State. The aim of this study was to determine the prevalence and density of ticks in these animals and establish strategic control measures to enhance livestock productivity and livelihoods in pastoral areas. The current study found that the prevalence of tick infestation in Kebribayah and Afdem districts was 83.3% and 86.8%, respectively. Rhipicephalus pulchellus (48.9%) was identified as the most common tick species in camels and cattle, followed by Amblyomma gemma (26.3%), Hyalomma truncatum (11.6%), Amblyomma lepidum (6.7%), and Amblyomma variegatum (6.5%). Among the variables considered, age and body condition score were significant risk factors (p < 0.001). Tick density varied depending on the recorded months and seasons (p < 0.001), with the highest mean tick density occurring in November (32.69 ± 21.750) and during the wet season (28.56 ± 19.750). Livestock owners in Kebribayah and Afdem ranked topical acaricide application as the most effective tick control method, followed by ivermectin injections, with the traditional hand removal method being the least effective. These rankings were consistent across both districts, and there was moderate agreement among livestock keepers from both regions regarding the best method. Afdem livestock keepers had slightly weak agreement on high tick burden in spring (W = 0.475, p = 0.127), and Kebribayah livestock keepers showed slightly strong agreement in tick burden across seasons (W = 0.700, p = 0.038), with spring having a significantly higher burden than winter. Consequently, participatory appraisal indicated that ticks were important and prevalent ectoparasites in the study area. Finally, strategic tick control appropriate for specific management and production environments should be implemented biannually in wet seasons.

Cisnormativity as a structural barrier to STI testing for trans masculine, two-spirit, and non-binary people who are gay, bisexual, or have sex with men.

Trans masculine, two-spirit, and non-binary people who are gay, bisexual or otherwise have sex with men (TGBM) are under-tested for sexually transmitted infections (STI) and may face complex, intersectional barriers that prevent them from accessing STI testing. As part of a study on gay, bisexual and other men who have sex with men's (GBM) experiences of current STI testing systems in Ontario, Canada, this paper reports on the findings from TGBM participants' experiences with in-person STI testing in a range of venues (i.e. Family doctors, walk-in clinics, and community-based organizations) to explore testing barriers specific to TGBM. Using a community-based research approach, between June 2020 and December 2021 peer researchers who identified as GBM conducted focus groups and interviews with 38 cis and trans GBM, 13 of whom identified as TGBM. Data were analyzed following grounded theory. When questioned about past experiences with testing, TGBM participants reported several barriers to STI testing within current testing models in Ontario due to cisnormativity and heteronormativity. Cisnormativity is the assumption that everyone identifies as the gender they were assigned at birth, and those who do not are considered "abnormal", while heteronormativity is when it is assumed that everyone is heterosexual. From our research we identified three overarching themes concerning testing barriers among TGBM participants: (1) non-inclusive clinic environments, (2) lack of provider knowledge and competency, and (3) legal documentation. Inherent cis and heteronormativity in healthcare institutions appear to be factors shaping the historical under-testing for STI in the TGBM population. These findings suggest the relevance of implementing trans-specific clinical practices that reduce the stigma and barriers faced by TGBM in STI testing contexts, including: hosting all-gender testing hours, opening more LGBTQ+ clinics, offering training in transgender health to testing providers, and conducting a review of how gender markers on health documents can be more inclusive of trans, two-spirit, and non-binary communities.

Bixafen, a succinate dehydrogenase inhibitor fungicide, causes microcephaly and motor neuron axon defects during development.

Succinate dehydrogenase inhibitors (SDHIs), the most widely used fungicides in agriculture today, act by blocking succinate dehydrogenase (SDH), an essential and evolutionarily conserved component of mitochondrial respiratory chain. Recent results showed that several SDHIs used as fungicides not only inhibit the SDH activity of target fungi but also block this activity in human cells in in vitro models, revealing a lack of specificity and thus a possible health risk for exposed organisms, including humans. Despite the frequent detection of SDHIs in the environment and on harvested products and their increasing use in modern agriculture, their potential toxic effects in vivo, especially on neurodevelopment, are still under-evaluated. Here we assessed the neurotoxicity of bixafen, one of the latest-generation SDHIs, which had never been tested during neurodevelopment. For this purpose, we used a well-known vertebrate model for toxicity testing, namely zebrafish transparent embryos, and live imaging using transgenic lines labelling the brain and spinal cord. Here we show that bixafen causes microcephaly and defects on motor neuron axon outgrowth and their branching during development. Our findings show that the central nervous system is highly sensitive to bixafen, thus demonstrating in vivo that bixafen is neurotoxic in vertebrates and causes neurodevelopmental defects. This work adds to our knowledge of the toxic effect of SDHIs on neurodevelopment and may help us take appropriate precautions to ensure protection against the neurotoxicity of these substances.

Damage-Induced Calcium Signaling and Reactive Oxygen Species Mediate Macrophage Activation in Zebrafish.

Immediately after a wound, macrophages are activated and change their phenotypes in reaction to danger signals released from the damaged tissues. The cues that contribute to macrophage activation after wounding in vivo are still poorly understood. Calcium signaling and Reactive Oxygen Species (ROS), mainly hydrogen peroxide, are conserved early wound signals that emanate from the wound and guide neutrophils within tissues up to the wound. However, the role of these signals in the recruitment and the activation of macrophages is elusive. Here we used the transparent zebrafish larva as a tractable vertebrate system to decipher the signaling cascade necessary for macrophage recruitment and activation after the injury of the caudal fin fold. By using transgenic reporter lines to track pro-inflammatory activated macrophages combined with high-resolutive microscopy, we tested the role of Ca²âº and ROS signaling in macrophage activation. By inhibiting intracellular Ca²âº released from the ER stores, we showed that macrophage recruitment and activation towards pro-inflammatory phenotypes are impaired. By contrast, ROS are only necessary for macrophage activation independently on calcium. Using genetic depletion of neutrophils, we showed that neutrophils are not essential for macrophage recruitment and activation. Finally, we identified Src family kinases, Lyn and Yrk and NF-κB as key regulators of macrophage activation in vivo, with Lyn and ROS presumably acting in the same signaling pathway. This study describes a molecular mechanism by which early wound signals drive macrophage polarization and suggests unique therapeutic targets to control macrophage activity during diseases.

A Fast, Simple, and Affordable Technique to Measure Oxygen Consumption in Living Zebrafish Embryos.

In all animal species, oxygen consumption is a key process that is partially impaired in a large number of pathological situations and thus provides informative details on the physiopathology of the disease. In this study, we describe a simple and affordable method to precisely measure oxygen consumption in living zebrafish larvae using a spectrofluorometer and the MitoXpress Xtra Oxygen Consumption Assay. In addition, we used zebrafish larvae treated with mitochondrial respiratory chain inhibitors, antimycin A or rotenone, to verify that our method enables precise and reliable measurements of oxygen consumption.

Organophosphorus diisopropylfluorophosphate (DFP) intoxication in zebrafish larvae causes behavioral defects, neuronal hyperexcitation and neuronal death.

With millions of intoxications each year and over 200,000 deaths, organophosphorus (OP) compounds are an important public health issue worldwide. OP poisoning induces cholinergic syndrome, with respiratory distress, hypertension, and neuron damage that may lead to epileptic seizures and permanent cognitive deficits. Existing countermeasures are lifesaving but do not prevent long-lasting neuronal comorbidities, emphasizing the urgent need for animal models to better understand OP neurotoxicity and identify novel antidotes. Here, using diisopropylfluorophosphate (DFP), a prototypic and moderately toxic OP, combined with zebrafish larvae, we first showed that DFP poisoning caused major acetylcholinesterase inhibition, resulting in paralysis and CNS neuron hyperactivation, as indicated by increased neuronal calcium transients and overexpression of the immediate early genes fosab, junBa, npas4b, and atf3. In addition to these epileptiform seizure-like events, DFP-exposed larvae showed increased neuronal apoptosis, which were both partially alleviated by diazepam treatment, suggesting a causal link between neuronal hyperexcitation and cell death. Last, DFP poisoning induced an altered balance of glutamatergic/GABAergic synaptic activity with increased NR2B-NMDA receptor accumulation combined with decreased GAD65/67 and gephyrin protein accumulation. The zebrafish DFP model presented here thus provides important novel insights into the pathophysiology of OP intoxication, making it a promising model to identify novel antidotes.

One Health education in Kakuma refugee camp (Kenya): From a MOOC to projects on real world challenges.

Today, the world counts millions of refugees but only a fraction of them have access to higher education. Despite the multiple public health problems in refugee camps and the need to build local capacities to prevent and combat them, University level courses in public health are largely unavailable for refugees. This paper describes the development, implementation and evaluation of an innovative two-module blended-learning programme on One Health in Kakuma refugee camp (Kenya). This programme combines: (I) Interdisciplinary and multi-expert MOOC on "Global Health at the Human-Animal-Ecosystem interface"; (II) peer-to-peer learning involving students from University of Geneva Master of science in Global Health and research collaborations around specific and locally-relevant problems; (III) online mentoring and lecturing by experts from the Institute of Global Health of the University of Geneva in Kakuma. A total of 67 refugees applied to Module 1; 15 started the Module 1 in October 2017, of these 14 completed it and 6 passed the exams, finally five students started the Module 2 in October 2018 which they all passed in February 2019. Five student-led collaborative projects were developed focusing on the conception of a community-based monitoring system for prevalent diseases in the camp. With such a pedagogic approach, the programme provides an overview on Global Health challenges at the human-animal-ecosystem interface and the importance of the One Health approach, and introduces students to scientific research through interdisciplinary and international collaborations and innovation. The high number of applicants and positive feedback from students in Kakuma show the interest in One Health education in the camp. This learning experience ultimately aims at building local knowledge and capacity fostering "One Health" champions to reinforce local and national health system. This framework for One Health education could be potentially scaled up to other camps in Africa and the world.