[Community-Based Testing and Factors Associated with HIV among Men Who Have Sex with Men (MSM) in Haiti between 2015 and 2018]. / Dépistage communautaire et facteurs associés au VIH d'hommes ayant des relations sexuelles avec des hommes (HSH) en Haïti entre 2015 et 2018.

Men who have sex with men (MSM) are an HIV key population in Haiti. However, little data exists on that population and on factors associated with this infection. Our study carried out the factors associated with HIV-positive screening among MSM in a community-based rapid testing program in Haiti between 2015 and 2018. Among the 1416 MSM screened, a third reported that it was their very first HIV test and 7.0% had an HIV-positive test. With a median age of 25 years old [21-29], over half of them were living in urban areas (60.7%) and were in financial precarious conditions (68.6%). Multivariate analysis showed that two factors were significantly associated with an HIVpositive result: having had an STI in the last 12 months, strengthened by psychoactive drug use; transactional sex practice in the last 12 months, strengthened by the age between 18 and 20 years old. These results should be taken into account when developing and implementing targeted and comprehensive HIV prevention programs and services for young MSM in Haiti.

Les hommes ayant des relations sexuelles avec des hommes (HSH) constituent une population clé du virus de l'immunodéficience humaine (VIH) en Haïti. Cependant, peu de données existent sur cette population et les facteurs associés à cette infection. Notre étude s'intéresse aux facteurs liés à un test rapide positif au VIH chez les HSH dans le cadre d'un dispositif de dépistage communautaire en Haïti entre 2015 et 2018. Parmi les 1 416 HSH dépistés, un tiers déclaraient leur premier test VIH et 7,0 % avaient un résultat positif. Avec un âge médian de 25 ans [21­29], plus de la moitié d'entre eux vivaient en milieu urbain (60,7 %) et étaient en situation de précarité financière (68,6 %). Une analyse multivariée a montré que deux facteurs étaient significativement associés à un résultat positif au VIH : avoir eu une infection sexuellement transmissible dans les 12 derniers mois, facteur accentué lorsqu'il est combiné à une consommation de produits psychoactifs ; la pratique du sexe transactionnel dans les 12 derniers mois, facteur accentué par l'appartenance à la classe d'âge des 18­20 ans. Ces résultats doivent se traduire dans la mise en place de futurs programmes et services de prévention du VIH vers les jeunes HSH en Haïti.

Bonamia exitiosa transmission among, and incidence in, Asian oyster Crassostrea ariakensis under warm euhaline conditions.

Previously reported in Australia, New Zealand, and more recently in Europe, the protistan parasite Bonamia exitiosa was also reported in the mid-Atlantic region of the USA after causing serious mortalities there in the Asian oyster Crassostrea ariakensis. At the time, this oyster was being considered for introduction, and the potential consequences of introducing this species were being assessed using field and laboratory studies. B. exitiosa emerged as the most serious disease threat for this oyster species, especially under warm euhaline conditions and for oysters <50 mm in size. To better evaluate how quickly this parasite may be able to spread among C. ariakensis, we investigated B. exitiosa transmission and incidence in C. ariakensis. During a first trial, potential direct transmission of B. exitiosa was evaluated by cohabitating infected C. ariakensis with uninfected C. ariakensis under in vivo quarantine conditions. In a second experiment, B. exitiosa incidence was estimated in situ by determining its prevalence in C. ariakensis deployed in an enzootic area after 4, 7, 14, 21 and 28 d of exposure. Results suggest that under warm euhaline conditions B. exitiosa can be transmitted among C. ariakensis without requiring any other parasite source and that parasite incidence may be at least as high as 40% after only 4 d exposure to an enzootic area. These results underscored the severity of the bonamiasis disease threat to C. ariakensis and provided further evidence that efforts to build an aquaculture industry based on C. ariakensis in the eastern USA might have been thwarted by parasitic disease.

Shellfish tissues evaluated for Perkinsus spp. using the Ray's fluid thioglycollate medium culture assay can be used for downstream molecular assays.

Ray's fluid thioglycollate medium (RFTM) culture assay is the standard, recommended method for surveillance of Perkinsus spp. infections in marine molluscs. In this assay, shellfish tissues are incubated in RFTM, stained with Lugol's iodine solution to render Perkinsus spp. cells blue-black, and evaluated microscopically to rate infection intensities. A limitation of this assay, however, is the lack of pathogen species specificity. Generally, identification of Perkinsus spp. requires DNA sequence analysis of parallel or additional samples since the exposure to iodine is believed to hamper DNA amplification from samples processed by the RFTM assay. However, we show that P. marinus DNA can be successfully amplified by PCR from Crassostrea virginica tissues cultured in RFTM and stained with Lugol's iodine. The beneficial consequence is that, where necessary, DNA sequence data may be obtained from RFTM-cultured tissues, allowing the identification of the Perkinsus sp. responsible for an observed infection. This would obviate further sampling, representing gain of time and reduction in cost, where a Perkinsus sp. is unexpectedly observed in new host(s) or location(s) but where parallel samples are not available for molecular diagnostics. Laboratories without molecular diagnostic tools for Perkinsus spp. may fix presumptive Perkinsus sp.-positive culture material in 95% ethanol for transport to, and subsequent analysis by, a laboratory that does have this capacity.

Real-time PCR investigation of parasite ecology: in situ determination of oyster parasite Perkinsus marinus transmission dynamics in lower Chesapeake Bay.

Perkinsus marinus is a severe pathogen of the oyster Crassostrea virginica on the East Coast of the United States. Transmission dynamics of this parasite were investigated in situ for 2 consecutive years (May through October) at 2 lower Chesapeake Bay sites. Compared to previous studies where seasonal infection patterns in oysters were measured, this study also provided parasite water column abundance data measured using real-time PCR. As previously observed, salinity and temperature modulated parasite transmission dynamics. Using regression analysis, parasite prevalence, oyster mortalities and parasite water column abundance were significantly positively related to salinity. Perkinsus marinus weighted prevalence in wild oysters and parasite water column abundance both were significantly related to temperature, but the responses lagged 1 month behind temperature. Parasite water column abundance was the highest during August (up to 1,200 cells/l) and was significantly related to P. marinus weighted prevalence in wild oysters, and to wild oyster mortality suggesting that parasites are released in the environment via both moribund and live hosts (i.e. through feces). Incidence was not significantly related to parasite water column abundance, which seems to indicate the absence of a linear relationship or that infection acquisition is controlled by a more complex set of parameters.

Needle in a haystack: involvement of the copepod PARACARTIA grani in the life-cycle of the oyster pathogen Marteilia refringens.

Marteilia refringens is a major pathogen of the European flat oyster, Ostrea edulis Linnaeus. Since its description, the life-cycle of this protozoan parasite has eluded discovery. Attempts to infect oysters experimentally have been unsuccessful and led to the hypothesis of a complex life-cycle involving several hosts. Knowledge of this life-cycle is of central importance in order to manage oyster disease. However, the exploration of M. refringens life-cycle has been previously limited by the detection tools available and the tremendous number of species to be screened in enzootic areas. In this study, these two restrictions were circumvented by the use of both molecular detection tools and a mesocosm with low biodiversity. Screening of the entire fauna of the pond for M. refringens DNA was systematically undertaken using PCR. Here, we show that the copepod Paracartia (Acartia) grani is a host of M. refringens. Not only was DNA of M. refringens consistently detected in P. grani but also the presence of the parasite in the ovarian tissues was demonstrated using in situ hybridization. Finally, successful experimental transmissions provided evidence that P. grani can be infected from infected flat oysters.

Molecular evidence for the existence of two species of Marteilia in Europe.

Marteilia refringens is one of the most significant pathogens of bivalve molluscs. Previous sequencing of the small subunit ribosomal RNA gene of M. refringens isolates derived from the infected mussels (Mytilus edulis and Mytilus galloprovinciallis) and the oyster (Ostrea edulis) in Europe did not reveal genetic polymorphisms despite indications from epizootiological data that distinct types may exist. We investigated the existence of polymorphisms in the internal transcribed spacer region of the ribosomal RNA genes. The sequences of this region proved to be clearly dimorphic among Marteilia from five sampling sites. The distribution of the two genetic types, named "O" and "M", appeared to be linked to the host species, oysters and mussels, respectively. We therefore support the recognition of two species of Marteilia in Europe and propose that the "O" type corresponds to M. refringens and the "M" type to M. maurini.

Claire ponds as an experimental model for Marteilia refringens life-cycle studies: new perspectives.

Since its first description, the paramyxean parasite Marteilia refringens (Grizel et al.) has been recognised as a significant pathogen of the European flat oyster Ostrea edulis L. The existence of a complex life-cycle involving several hosts was postulated early on by many authors, although it remains unsolved. Recent developments in the DNA-based diagnosis of M. refringens provides new prospects for the detection of the parasite in potential hosts. However, this screening remains impeded by the number of species living in the vicinity of oyster beds. We report here on the use of semi-closed oyster ponds (so called 'claire' in Marennes-Oléron Bay) as a study model for the life-cycle of M. refringens. Claires are located in an endemic area for M. refringens and transmission of the disease to healthy oysters has been shown to be effective during the course of this study. The environmental characteristics of the claires strongly limit the number of species compared with intertidal areas and oyster beds. Consequently, extensive sampling of a limited number of species cohabiting with oysters was possible. These were preserved for future screening of M. refringens. The experimental model should bring new insights to the life-cycle of M. refringens, as it enables us to propose new conceptual schemes of M. refringens transmission. The role of species as potential hosts is discussed regarding their biology and geographical distribution.

DNA Probes As Potential Tools for the Detection of Marteilia refringens.

Since its first description, the paramyxean parasite Marteilia refringens has been recognized as a significant pathogen of bivalve mollusks. The existence of a complex life cycle was postulated by many authors. Here we report the development of DNA-based detection assays as powerful tools to elucidate the Marteilia refringens life cycle. After alignment of the Marteilia refringens ribosomal DNA small subunit sequence with those of various eukaryotic organisms, polymerase chain reaction primers were designed. Specific primers were used to amplify DNA extracted from purified Marteilia refringens and infected hosts. The specificity of amplified fragments was confirmed by Southern blotting with an oligoprobe. For in situ hybridization, four probes were tested for specific detection of 18S rRNA isolated from Marteilia refringens and other eukaryotic cells by Northern blotting. The most specific probe, Smart 2, was successfully used to detect Marteilia refringens by in situ hybridization in infected oysters and mussels.