MosquirixTM malaria vaccine: an evaluation of patients' willingness to pay in Cameroon.

OBJECTIVES: The purpose of this study was to determine the average price that a patient living in Cameroon would be willing to pay for the MosquirixTM vaccine and the factors influencing the proposed price. STUDY DESIGN: Cross-sectional study Methods: Data were collected using a semi-open questionnaire in 5 hospitals in Cameroon. This study included all persons over 18 years who came for consultation in one of the 5 selected hospitals during the study period (from 02th to 14th April 2018 and from 02th to 22th July 2018). The factors associated with the price of the vaccine proposed by the patient were determined by linear multiple regression analysis. The average price was determined based on the patient's income and the percentage of that income proposed for the purchase of the vaccine. RESULTS: We collected data from 1,187 participants aged 18 to 80 years. The average price that Cameroonian patients were willing to pay for the MosquirixTM vaccine was 1,514±475 XAF (2.3±0.73 Euro). The minimum and maximum purchase price of the vaccine were 1,178 XAF (1.8 Euro) and 1,850 XAF (2.8 Euro) respectively. We also noted that patients were willing to spend an average of 1.34% of their income on the vaccine. This percentage of income was significantly (lt;0.001) associated with the respondents' income, the fact that they had been consulted at least once for malaria in the 12 months preceding the survey (lt;0.001) and the fact that the respondent had at least one under- five year child (lt;0.001). CONCLUSION: Factors associated with the average price are elements that should be strongly considered by policy makers to introduce this vaccine in Cameroon. This pilot study can serve as a framework for a potential national population-based study.

The Gulf of Ambracia's Common Bottlenose Dolphins, Tursiops truncatus: A Highly Dense and yet Threatened Population.

The common bottlenose dolphin (Tursiops truncatus) is the only cetacean present in the semiclosed waters of the Gulf of Ambracia, Western Greece. This increasingly degraded coastal ecosystem hosts one of the highest observed densities in the Mediterranean Sea for this species. Photo-identification data and tissue samples collected through skin-swabbing and remote biopsy sampling techniques during boat-based surveys conducted between 2006 and 2015 in the Gulf, were used to examine bottlenose dolphin abundance, population trends, site fidelity, genetic differentiation and toxicological status. Bottlenose dolphins showed high levels of year-round site fidelity throughout the 10-year study period. Dolphin population estimates mostly fell between 130 and 170 with CVs averaging about 10%; a trend in population size over the 10 years was a decline of 1.6% per year (but this was not significant). Genetic differentiation between the bottlenose dolphins of the Gulf and their conspecifics from neighbouring populations was detected, and low genetic diversity was found among individuals sampled. In addition, pesticides where identified as factors posing a real toxicological problem for local bottlenose dolphins. Therefore, in the Gulf of Ambracia, high dolphin density does not seem to be indicative of favourable conservation status or pristine habitat.

Paraffin waxes in the North-Western Mediterranean Sea: A comprehensive assessment in the Pelagos Sanctuary, a Specially Protected Area of Mediterranean Importance.

Paraffin waxes are widely recognized as emerging marine pollutants, even their classification by the recent monitoring programs and the knowledge of their occurrence, and sources of contamination in marine ecosystems are poorly defined and reported. Wax presence and distribution have been evaluated in different environmental compartments in the Pelagos Sanctuary (Mediterranean Sea) floating on the sea surface and stranded on beaches, focussing on their characterization, accumulation areas and pollution inputs. More than 2500 yellow paraffin residues were detected and analysed in the study area showing a prevailing dimension smaller than 5 mm. The Genoa Canyon and the waters facing Gorgona Island resulted in the more polluted areas representing two distinct hotspots of wax accumulation potentially related to the high density of tanker vessels sailing to and from the harbour of Genova and Livorno. Higher concentrations of beached particles were found along the Tuscan coast (11 items/100 m) and on Pianosa Island (110 items/m2). This study gives valuable insights into paraffin wax pollution in the Pelagos Sanctuary, emphasizing the need for harmonized monitoring and detection methods to elucidate the potential impacts on marine organisms. Moreover, mitigating actions are crucial to prevent and curb the waxes pollution of marine ecosystems.

Comparison of the proportion of patients potentially treated with an anti-osteoporotic drug using the current criteria of the Belgian national social security and the new suggested FRAX criteria.

To assess the number of anti-osteoporosis treatments that would be reimbursed by the Belgian social security if either FRAX or the current criteria were used to determine access to reimbursement. This is a retrospective study based on data from 1,000 women randomly selected from an outpatient hospital specialized in bone metabolism in Belgium. Proportions of potentially refunded treatments between FRAX and current criteria were compared. Out of the 1,000 women files, 890 have sufficient information to assess FRAX. In Belgium, current criteria include a bone mineral density (BMD) T score below -2.5 at the lumbar spine, the femoral neck or the total hip and/or at least a prevalent vertebral fracture. Using these criteria, 167 women (18.8 %) would have access to reimbursement. Using the criteria based on the validated Belgian FRAX tool, only 116 women (13.0 %) would have access to reimbursement, meaning that access to reimbursement based on FRAX criteria would reduce by 30 % the anti-osteoporosis drug expenses covered by the national social security. Interestingly, only 65 women out of the 116 (56.0 %) selected with the FRAX criteria were also selected with the current criteria of the national social security. A substantial proportion of individuals that would potentially receive a reimbursement for their treatment using the FRAX criteria do not have access to any refund for their treatment with the current criteria. Since patients identified with the FRAX tool are those with the highest risk profile for future fractures, reappraisals of treatment reimbursement guidelines are expected in Belgium.

Impact of the marine litter pollution on the Mediterranean biodiversity: A risk assessment study with focus on the marine protected areas.

In this paper a novel methodology to assess the risk of marine litter (ML) pollution in the Mediterranean Sea is implemented. In this approach, the hazard component is estimated using a state-of-the-art 3D modeling system, which allows the simulation of floating and sinking ML particles; the exposure component is defined from biodiversity estimates; and the vulnerability is related to ML ingestion rates of each species. The results show that the hot-spots for the ML risk concentrate in the coastal regions, and are mainly conditioned by the biodiversity in the region. A dedicated analysis on the marine protected areas shows that the risk therein is controlled by the proximity to ML sources and that their present-day protection levels are not effective in the case of ML pollution. Only a reduction of ML at the sources could reduce the impact of ML pollution in protected areas.

Assessment of PCDD/Fs, dioxin-like PCBs and PBDEs in Mediterranean striped dolphins.

Bio-accumulation of high levels of persistent organic pollutants represent a serious conservation concern for Mediterranean marine odontocetes. In this study, blubber samples from 10 striped dolphins (Stenella coeruleoalba) stranded along the Italian coasts during 2015-2016 were analyzed. All specimens showed dl-PCBs > PBDEs ≫ PCDD/Fs. Median concentrations were 1820 ng/g l.w., 456 ng/g l.w. and 23.9 pg/g l.w., respectively. dl-PCBs accounted for 93.3% of total TEQs. PBDE concentrations suggest that the Mediterranean basin may be considered a hotspot for organobromine compounds. OCDD did not represent the greatest contributor to PCDD/Fs profile, most likely due to a change in dioxin environmental sources in the last two-three decades. Despite international regulations, the present study emphasized that POP exposure levels in Mediterranean striped dolphins have not declined significantly in recent years. Toxicological and risk assessment studies on this sentinel species may provide an early indication of potential adverse health effects on Mediterranean ecosystems.

Relevance of current PCB concentrations in edible fish species from the Mediterranean Sea.

Legal restrictions and bans have led to a steady decrease in PCB environmental concentrations. Yet, in recent years PCBs have been found at very high levels in the Mediterranean Sea, for instance, in some apex predators. This work aimed to investigate current PCB (eighteen congeners: #28,52,77,81,101,105,114,118,123,126,138,153,156,157,167,169,180,189) concentrations in the Mediterranean Sea and their relevance today, focusing on their occurrence in edible fish species typically consumed in the Mediterranean diet. In spring 2017, a total of 48 fish samples from the Northern Thyrrenian Sea were collected: 16 specimens of sardine (Sardina pilchardus), 16 of anchovy (Engraulis encrasicolus) and 16 of bogue (Boops boops). PCBs were quantified in the muscle of the animals by means of GC-QqQ-MS. They were found in all samples at the greatest concentrations (ng/g w.w.) in sardine (4.15-17.9, range), and very similar values between anchovy (1.01-7.08) and bogue (1.46-7.22). WHO-TEQ PCB values followed the same order, i.e. sardine (0.410-1.24, range in pg/g w.w.) > anchovy (0.0778-0.396) ~ bogue (0.0726-0.268). These concentrations lied below the European limits of 75 ng/g (w.w.) for the six indicator PCBs and 6.5 pg/g WHO-TEQ for dioxins and dioxin-like PCBs in muscle meat of fish. Additionally, estimated weekly intakes (EWI, in pg WHO-TEQ/Kg/week) for sardine (1.2), anchovy (0.29) and bogue (0.35) scored below the safe value proposed by EFSA of 2 pg WHO-TEQ/Kg/week. When comparing with data reported for the same species in previous Mediterranean studies, values found here were lower than those surveyed in the late 90s and early 2000s; however, they were often not notably different from concentrations reported in last years. This builds up on the concept of a current slow decrease of PCBs in the Mediterranean Sea, likely linked to new inputs and/or remobilization of burdens, and reinforces the need of continous monitoring of these legacy contaminants still ubiquitous today.

A new digestion approach for the extraction of microplastics from gastrointestinal tracts (GITs) of the common dolphinfish (Coryphaena hippurus) from the western Mediterranean Sea.

Plastic ingestion is one of the main impacts of marine litter on organisms. The occurrence of microplastics (MPs < 5 mm) in the stomachs of Mediterranean species was already reported in several studies. In this context, the present study aims to develop a new approach of digestion for the identification of MPs in the gastrointestinal tracts (GITs) of marine organisms. The new approach combines two digestion protocols, including potassium hydroxide (KOH) and nitric acid (HNO3), to remove most organic and inorganic materials. This digestion allows recording small MPs that are difficult to find via routinely stomach content analysis and also to minimize the overestimation of the phenomenon trough the control of airborne contamination. The new approach was tested on a voracious pelagic opportunistic predator, the common dolphinfish, a fishery resource exploited in several Mediterranean areas. The results showed that a large amount of ingested meso- and microplastics, such as fragments or sheets, was recorded in GITs (F = 65.5 %). The FTIR analysis on litter samples allowed to identify polyethylene, polypropylene and polystyrene as dominant constituent polymers of microplastics. These results confirmed that our novel combined digestion protocol represents a reliable approach to detect MPs in opportunistic pelagic predators.

Lessons learned from an intercalibration exercise on the quantification and characterisation of microplastic particles in sediment and water samples.

An intercalibration exercise on the characterisation of microplastics in marine sediment and water samples was carried out among five laboratories involved in the implementation of the Marine Strategy Framework Directive (MSFD) in their country. The samples were prepared by mixing cleaned natural sediment and sea water with microplastics sets made of particles of various polymers, shapes and colours. Overall, the errors on total counts were under 25% in absolute value. The risk of non-detection and loss of particles is greater than the risk of contamination during sample analysis. Significant differences are observed among particle types. It appears difficult to obtain reliable and comparable data on the colour of microplastics. A comparison of the errors with regards to the protocols used led to recommend NaCl [1.2 g/cm3] density separation for sediment and one filtering step (200 µm). The operators' experience appears as a key factor for the quality of the results.

Evaluation of PCDD/Fs, dioxin-like PCBs and PBDEs in sperm whales from the Mediterranean Sea.

Numerous studies to date have reported concentrations of Persistent Organic Pollutants (POPs) in different marine mammal species worldwide. Yet data on sperm whales are scarce from rich and unique biodiverse areas such as the Mediterranean Sea. This work aimed to assess levels of dioxin-like polychlorinated biphenyls (dl-PCBs), polybrominated diphenyl ethers (PBDEs), and polychlorodibenzo-p-dioxins and furans (PCDD/Fs) in blubber of sperm whales stranded along the Italian coast between 2008 and 2016. POP mean concentrations (dl-PCBs: 6410 ng/g l.w.; PBDEs: 612 ng/g l.w.; PCDD/Fs: 57.8 pg/g l.w.) were mostly in line with what has been previously reported on the same species in the Mediterranean environment and tended to be higher than those reported from other geographical regions. The relative abundance followed the order dl-PCBs > PBDEs ≫ PCDD/Fs. Interestingly, the non-ortho dl-PCB pattern (126 > 169 > 77) was similar to that described in other studies worldwide and different from what is described in its main prey. This could be linked to particular metabolic activities in sperm whales against these highly toxic contaminants. Total TEQs ranged from 275 to 987 pg/g l.w. and showed the pattern Σnon-ortho-dl-PCBs > Σortho-dl-PCBs > PCDDs > PCDFs, with PCBs' contribution about 96%. These findings highlight the high abundance of PCBs still found in the Mediterranean environment despite having been banned for decades. All sperm whales analyzed in this study surpassed the threshold of 210 pg WHO-TEQ/g l.w. proposed as starting point of immunosuppression in harbour seals; a level of contamination that may have contributed to an impairment of their immune system.

Effects of copper on larvae of the marbled crab Pachygrapsus marmoratus (Decapoda, Grapsidae): Toxicity test and biochemical marker responses.

The importance of trace elements in ecotoxicological investigations is a well-known issue when monitoring polluted areas such as commercial harbors. Copper represents one of the most common metal contaminants, often detected in these areas as it is widely employed in various fields and has many sources of inflow in the marine environment. Pachygrapsus marmoratus is a widespread intertidal crab species that has been extensively studied in ecology, ethology and population genetics. Ecotoxicological studies have also been performed, exclusively on the adult stage. In the present study we investigated the mortality and biochemical (oxidative stress and neurotoxicity) responses of P. marmoratus larvae exposure to environmental relevant concentration of copper. Results showed dose-dependent responses in terms of larval mortality, with a calculated LC50 value of 0.5 mg/L of Cu2+. The LC50 concentration was used as the starting point for subsequent biochemical response evaluation. Results also demonstrated dose-dependent activation of antioxidant systems assuming a compensatory antioxidant activity to prevent higher cellular damage when larvae were exposed to the highest concentrations of copper. Moreover, a significant enhancement of neurotransmitter activities was observed, assuming a possible direct interaction of copper with the enzymes or an increase of free copper ion aliquot into the cells.

Tools and constraints in monitoring interactions between marine litter and megafauna: Insights from case studies around the world.

Adverse impacts of marine litter is documented on >1400 species, including marine megafauna (fish, birds, sea turtles and mammals). The primary impacts include ingestion and entanglement, and there is increasing concern about chemical contamination via ingestion. Numerous survey approaches and monitoring programs have been developed and implemented around the world. They may aim to provide data about parameters such as species distribution and interactions with anthropogenic activities. During the Sixth International Marine Debris Conference, a session was dedicated to the tools and constraints in monitoring interactions between litter and megafauna. In the present paper, we summarize 7 case studies which discuss entanglement and ingestion including macro- and micro-debris in several taxa and across multiple geographic regions. We then discusses the importance of tools and standardizing methods for assessment and management purposes, in the context of international environmental policies and marine litter strategies.

Quality-control ranges for antimicrobial susceptibility testing by broth dilution of the Brachyspira hyodysenteriae type strain (ATCC 27164T).

There are no approved standards for antimicrobial susceptibility testing of the fastidious spirochete Brachyspira hyodysenteriae. An interlaboratory study was performed to establish MIC quality control ranges for six antimicrobial agents for the type strain of B. hyodysenteriae using broth dilution. The results showed that B. hyodysenteriae B78T ATCC 27164T is a suitable quality control strain. This is a first step toward standardization of methods regarding this anaerobe.

Development of new-tools to investigate toxicological hazard due to endocrine disruptor organochlorines and emerging contaminants in Mediterranean cetaceans.

The possibility that certain Mediterranean cetaceans are subject to toxicological hazard due to organochlorines and emerging contaminants, such as polybrominated diphenyl ethers (PBDEs) with endocrine disrupting capacity, was investigated using non-lethal methods. The need for new biomarkers for EDCs and for a "cell model" to explore the different susceptibilities to several classes of ECDs, including emerging contaminants, led us to culture fibroblasts of different cetacean species ("dolphins in test tubes"). We then explored interspecies and gender susceptibility to OC-EDCs and PBDEs using qualitative and semi-quantitative evaluation of target proteins, such as CYP1A and CYP2B in cultured cetacean fibroblasts (Stenella coeruleoalba, Tursiops truncatus and Balaenoptera physalus), by western blot and immunofluorescence techniques.

Use of biomarkers to investigate toxicological effects of produced water treated with conventional and innovative methods.

The aim of this study was to develop and apply a multi-biomarker system to assess the toxicological effects of produced water (PW) from a Mediterranean off-shore oil platform. The selected bioindicator organism, mosquitofish (Gambusia affinis), was exposed in the laboratory to high concentrations of different PW: PW before treatment (BT), after conventional treatment (ACT) and after innovative treatment with zeolites in a prototype system (AIT). A set of biomarkers (benzo(alpha)pyrene monooxygenase, ethoxyresorufin-o-deethylase, vitellogenin, porphyrins, PAH bile metabolites, esterases, catalase, micronuclei) and PAH concentrations were measured in the experimental organism. The methodology proved to be appropriate and biomarker responses (CYP 1A1, PAH bile metabolites, micronuclei, esterases, porphyrins) affected by BT were less affected by ACT. PW treated with zeolites (AIT) had the lowest toxicological impact. The results obtained applying this multi-biomarker approach suggest that the system using zeolites is effective for treating produced water.

Endocrine Disruptors in Mediterranean top marine predators.

BACKGROUND, AIMS AND SCOPE: Man-made Endocrine Disruptors (EDs) range across all continents and oceans. Some geographic areas are potentially more threatened than others: one of these is the Mediterranean Sea. Levels of some xenobiotics are much higher here than in other seas and oceans. In this paper we review the final results of a project in which the hypothesis that Mediterranean top predator species (such as large pelagic fish and marine mammals) are potentially at risk due to EDs was investigated. METHODS: In a four-year survey on the Mediterranean population of swordfish (Xiphias gladius), the potential toxicological effects of organochlorine compounds (OCs) on specimens of swordfish and tuna fish (Thunnus thynnus thynnus), caught in the spawning seasons from 1999 to 2002 in the Straits of Messina, Sicily (Italy), were investigated using vitellogenin (Vtg), Zona radiata proteins (Zrp), and cytochrome P4501A (CYP1A) activities (EROD, BPMO). Tissues (skin and blubber) were obtained from Stenella coeruleoalba, Tursiops truncatus, Delphinus delphis and Balaenoptera physalus from the western Ligurian Sea, between Corsica and the French-Italian coast, and Ionic Sea using biopsy darts launched with a crossbow. Benzo(alpha)pyrene monoxigenase (BPMO) activity was mesured in biopsies and cholrinated hydrocarbon levels were detected. RESULTS AND DISCUSSION: We illustrate the need to develop and apply sensitive methodological tools, such as biomarkers (Vitellogenin, Zona Radiata proteins and CYP1A activities) for evaluation of toxicological risk in Xiphias gladius and Thunnus thynnus thynnus), and nondestructive biomarkers (CYP1A activities and fibroblast cell culture in skin biopsy), for the hazard assessment of threatened marine mammals species (Stenella coeruleoalba, Tursiops truncatus, Delphinus delphis and Balaenoptera physalus) exposed to EDs. CONCLUSION: The present research shows that: a) Vtg and Zrp can be used as diagnostic tools for fish stocks hazard assessment in the Mediterranean Sea; b) that CYP1A1 (BPMO) induction in cetaceans skin biopsy may be an early sign of exposure to EDs such as OCs and a potential alert for transgenerational effects. RECOMMENDATION AND OUTLOOK: This research represents a warning signal of the potential reproductive alterations in marine top predators and suggest the need for continuous monitoring to avoid reductions in population and biodiversity in the Mediterranean Sea.

Nondestructive biomarkers in ecotoxicology.

The aim of this article is to attempt a concise review of the state of the art of the nondestructive biomarkers approach in vertebrates, establishing a consensus on the most useful and sensitive nondestructive biomarker techniques, and proposing research priorities for the development and validation of this promising methodology. The following topics are discussed: the advantages of the use of nondestructive strategies in biomonitoring programs and the research fields in which nondestructive biomarkers can be applied; the biological materials suitable for nondestructive biomarkers and residue analysis in vertebrates; which biomarkers lend themselves to noninvasive techniques; and the validation and implementation strategy of the nondestructive biomarker approach. Examples of applications of this methodology in the hazard assessment of endangered species are also presented.

Are the mediterraneantop predators exposed to toxicological risk due to endocrine disrupters?

Man-made endocrine disrupting chemicals (EDCs) range across all continents and oceans; some geographic areas are potentially more threatened than others: one of these is the Mediterranean Sea. This basin has limited exchange of water with the Atlantic Ocean and is surrounded by some of the most heavily populated and industrialized countries in the world. Accordingly, levels of some xenobiotics are much higher here than in other seas and oceans. In this research the unexplored hypothesis that Mediterranean top predator species (such as large pelagic fish and marine mammals) are potentially at risk due to EDCs is investigated. Here we illustrate the development of sensitive biomarkers (Vitellogenin, Zona Radiata proteins) for evaluation of toxicological risk in top marine predators (Xiphias gladius, Thunnus thynnus thynnus), and nonlethal techniques, such as nondestructive biomarkers (BPMO activities in skin biopsy), for the hazard assessment of threatened species exposed to EDCs, such as marine mammals (Stenella coeruleoalba, Tursiops truncatus, Delphinus delphis, and Balaenoptera physalus).

Daily physiological iron requirements in children.

Median daily iron absorption was determined in iron-replete males and females between 2 and 19 years of age from the upper and lower socioeconomic strata of the Venezuelan population. A comparison was made with iron absorption of well-nourished children, on the basis of hematological and anthropometric reference values from the U.S. The median absorption level, which was calculated from the increase in total body iron due to growth and the daily losses through exfoliation and menstruation, was also used to estimate the requirements of 95% of the population. When the requirements were expressed in terms of body weight, no significant difference in iron absorption was observed between the three Venezuelan and one U.S. groups, ranging from about 30 to 38 micrograms/kg/day in both sexes between 4 and 16 years of age. However, when the requirements were expressed without division by the weight factor, the requirements of the better nourished groups were somewhat higher than those of the Venezuelan lower socioeconomic population with, in addition, a threefold variation over the 5 to 16 year age range. These findings suggest that the total iron requirements of children at a certain age may be most adequately expressed in terms of the optimal body weight for that age.

Eradication of endemic Brachyspira pilosicoli infection from a farrowing herd: a case report.

Brachyspira pilosicoli and B. innocens were isolated repeatedly from a herd of 60 sows which mostly produced feeder pigs but also raised some fattening pigs. Postweaning diarrhea had been a severe problem in this herd for years. The B. pilosicoli eradication plan was based on the general guidelines for elimination of B. hyodysenteriae, with some modifications. The eradication measures were run in August 1997. In-feed medication with 200 p.p.m. tiamulin lasted for 18-30 days, depending on the age group. The piggery unit was emptied, cleaned, disinfected and dried, and all worn surfaces were repaired. The animals were removed to temporary sheds situated 0-100 m from the piggery unit. Only the sows and the boar returned to the piggery unit. All other pigs were sold from the sheds within 3 months after the eradication. Immediately after the eradication, the clinical postweaning diarrhea disappeared. The success of the program was monitored four times bacteriologically, and the last control sampling was in December 1999, 7 months after the total withdrawal of antimicrobial feed additives. The primary cultures from the last three samplings were also analysed with B. pilosicoli-specific PCR. All the samples were negative for B. pilosicoli. However, B. innocens could be isolated from each batch of samples. The analysis of B. innocens isolates by pulsed-field gel electrophoresis indicated that at least one genotype persisted in the herd. The clinical and laboratory findings suggest that the eradication of B. pilosicoli had succeeded in this herd.