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Crimean-Congo hemorrhagic fever (CCHF) and Rift Valley fever (RVF) are among the list of emerging zoonotic diseases that require special attention and priority. RVF is one of the six priority diseases selected by the Senegalese government. Repeated epidemic episodes and sporadic cases of CCHF and RVF in Senegal motivated this study, involving a national cross-sectional serological survey to assess the distribution of the two diseases in this country throughout the small ruminant population. A total of 2127 sera from small ruminants (goat and sheep) were collected in all regions of Senegal. The overall seroprevalence of CCHF and RVF was 14.1% (IC 95%: 12.5-15.5) and 4.4% (95% CI: 3.5-5.3), respectively. The regions of Saint-Louis (38.4%; 95% CI: 30.4-46.2), Kolda (28.3%; 95% CI: 20.9-35.7), Tambacounda (22.2%; 95% CI: 15.8-28.6) and Kédougou (20.9%; 95% CI: 14.4-27.4) were the most affected areas. The risk factors identified during this study show that the age, species and sex of the animals are key factors in determining exposure to these two viruses. This study confirms the active circulation of CCHF in Senegal and provides important and consistent data that can be used to improve the surveillance strategy of a two-in-one health approach to zoonoses.

Background: Antibiotic resistance (ABR) has emerged as a major threat to health. Properly informed decisions to mitigate this threat require surveillance systems that integrate information on resistant bacteria and antibiotic use in humans, animals, and the environment, in line with the One Health concept. Despite a strong call for the implementation of such integrated surveillance systems, we still lack a comprehensive overview of existing organizational models for integrated surveillance of ABR. To address this gap, we conducted a scoping review to characterize existing integrated surveillance systems for ABR., Methods: The literature review was conducted using the PRISMA guidelines. The selected integrated surveillance systems were assessed according to 39 variables related to their organization and functioning, the socio-economic and political characteristics of their implementation context, and the levels of integration reached, together with their related outcomes. We conducted two distinct, complementary analyses on the data extracted: a descriptive analysis to summarize the characteristics of the integrated surveillance systems, and a multiple-correspondence analysis (MCA) followed by a hierarchical cluster analysis (HCA) to identify potential typology for surveillance systems., Results: The literature search identified a total of 1330 records. After the screening phase, 59 references were kept from which 14 integrated surveillance systems were identified. They all operate in high-income countries and vary in terms of integration, both at informational and structural levels. The different systems combine information from a wide range of populations and commodities -in the human, animal and environmental domains, collection points, drug-bacterium pairs, and rely on various diagnostic and surveillance strategies. A variable level of collaboration was found for the governance and/or operation of the surveillance activities. The outcomes of integration are poorly described and evidenced. The 14 surveillance systems can be grouped into four distinct clusters, characterized by integration level in the two dimensions. The level of resources and regulatory framework in place appeared to play a major role in the establishment and organization of integrated surveillance., Conclusions: This study suggests that operationalization of integrated surveillance for ABR is still not well established at a global scale, especially in low and middle-income countries and that the surveillance scope is not broad enough to obtain a comprehensive understanding of the complex dynamics of ABR to appropriately inform mitigation measures. Further studies are needed to better characterize the various integration models for surveillance with regard to their implementation context and evaluate the outcome of these models., (© 2024. The Author(s).)

The One Health approach calls for collaboration across various sectors and different scales to improve understanding of complex health issues. Regarding epidemiological surveillance, this implies the development of integrated systems that link several surveillance components operating in different domains (human, domestic animals, environment) and involving several actor networks. However, surveillance continues to operate in a very compartmentalized way, with little interaction between sectoral institutions and with the community for the governance and operation of surveillance activities. This is partly explained by the insufficient consideration of the local context and the late involvement of national stakeholders when developing programmes that aimed at strengthening the integration of surveillance. In low- and middle-income countries in particular, there is a strong influence of external partners on the development of intersectoral programmes, including surveillance systems. In this context, we developed and implemented a participatory planning process to support stakeholders of the surveillance system of anthrax in Burkina Faso, in the definition of the One Health surveillance system they wish for and of the pathway to reach it. The workshop produced an action plan that reflects the views and perspectives of representatives of the different categories of stakeholders and beneficiaries of surveillance. In addition, the participation of stakeholders in this participatory co-construction process has also improved their knowledge and mutual understanding, fostering a climate of trust conducive to further collaboration for surveillance activities. However, the quality of the participation raises some questions over the results, and contextual factors may have influenced the process. This underlines the need to include a monitoring and evaluation plan in the process to assess its implementation and ability to produce One Health surveillance modalities that are appropriate, accepted and applied over the long term., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Nana et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Objectives: Collaboration between surveillance programmes is the keystone of One Health surveillance and international organizations call for integrated surveillance systems to manage antibiotic resistance (ABR). In France, the ABR surveillance system covers human, animal, food and the environment sectors, but appears to be fragmented, questioning its level of integration. This study aimed to evaluate collaboration within this system and to formulate recommendations towards more integration., Methods: ECoSur, a semi-quantitative tool, was used to evaluate collaboration between surveillance programmes. A total of 31 attributes were evaluated using information from the literature and 52 interviews with surveillance actors from all four sectors. Evaluation results were visualized via three output figures displaying aspects related to governance and functionality of collaboration. Results were validated by an expert committee., Results: Overall, the French collaborative strategy for ABR surveillance was well formalized and relevant to its objectives. However, a cross-sectoral coordination body was lacking to help with its practical implementation. The environmental sector was largely uncovered, but its integration appeared necessary to meet the strategy objectives. Data sharing and joint data analyses between programmes were insufficient, mainly due to limited resources and data interoperability issues. Collaboration was operational for internal and external communication of the results. Twelve recommendations were suggested to decision makers to foster collaboration within the French surveillance system and feed future strategies against ABR., Conclusions: This first evaluation of collaboration within the French ABR surveillance system produced concrete recommendations to move towards One Health integrated surveillance. Both the approach and the findings could be of interest to other countries., (© The Author(s) 2024. Published by Oxford University Press on behalf of British Society for Antimicrobial Chemotherapy.)

Tackling antimicrobial resistance (AMR) is a goal for many countries. Integrated surveillance of antimicrobial use (AMU) and resistance is a prerequisite for effective risk mitigation. Regular evaluation of any surveillance is needed to ensure its effectiveness and efficiency. The question is how to evaluate specifically integrated surveillance for AMU and AMR. In an international network called CoEvalAMR, we have developed guidelines for selection of the most appropriate tools for such an evaluation. Moreover, we have assessed different evaluation tools as examples using a country case format and a methodology with a focus on the user's experience. This paper describes the updated methodology, which consists of a brief introduction to the case and to the tool separately. Moreover, there are 12 functional aspects and nine content themes which should be scored using a 4-tiered scale. Additionally, four Strengths, Weaknesses, Opportunities, Threats (SWOT) questions should be addressed. Results are illustrated using radar diagrams. An example of application of the updated methodology is given using the ECoSur evaluation tool. No tool can cover all evaluation aspects comprehensively in a user-friendly manner, so the choice of tool must be based upon the specific evaluation purpose. Moreover, adequate resources, time and training are needed to obtain useful outputs from the evaluation. Our updated methodology can be used by tool users to share their experience with available tools, and hereby assist other users in identifying the most suited tool for their evaluation purpose. Additionally, tool developers can get valuable information for further improvements of their tool., Competing Interests: LA works for an organization that gives advice to farmers and meat producing companies. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Alban, Bordier, Häsler, Collineau, Tomassone, Bennani, Aenishaenslin, Norström, Aragrande, Filippitzi, Moura and Sandberg.)

Background: Anthrax is a zoonotic disease that causes frequent outbreaks in livestock and fatal human cases in Burkina Faso. Effective surveillance of this disease calls for the establishment of an integrated surveillance system, in line with the One Health concept. However, despite a strong technical and financial support from international partners, surveillance is still poorly conducted within an integrated approach. Based on stakeholder perspectives, the study has for objective to deepen our understanding of the anthrax surveillance system and to identify the obstacles and levers towards a more integrated approach to anthrax surveillance in Burkina Faso., Methods: The data was collected from a literature review and interviews with surveillance stakeholders. We first conducted a qualitative descriptive analysis of the data to characterize the surveillance system (programmes, actors, collaboration). In a second step, we conducted a thematic analysis of the informants' discourse in order to identify what represents an obstacle or, conversely, a lever for a more integrated approach to anthrax surveillance., Results: The surveillance system of anthrax in Burkina Faso includes three programmes (in the livestock, wildlife and human sectors), which involves 30 actors. These sectoral programmes operate almost independently from one another, although some collaborations are existing for the governance and implementation of surveillance activities. Analysis of the discourse of key stakeholders led to the identification of four categories of factors that may influence the implementation of an integrated surveillance system in the country: knowledge; technical, organizational and social capacities; motivation; intersectoral governance., Conclusions: This study highlights the difficulty of translating One Health governance to the national level and the need to better articulate the visions of all categories of stakeholders. This study also reveals the need to develop specific evaluation systems for integrated policies in order to provide credible evidence of their added value for a better management of zoonotic diseases. Finally, our study underlines the need to act upstream the emergence of zoonoses and allocate more resources to the prevention of zoonoses than to their control., (© 2022. The Author(s).)

Although currently available data indicate that Africa has the lowest usage of antimicrobials in animals in the world (adjusted by animal biomass), data show a high prevalence of antimicrobial resistance in foodborne pathogens isolated from animals and animal products. Apart from the lack of solid data on antimicrobial use in many countries in Africa, different hypotheses could explain this situation. Qualitative interviews of farmers show a lack of knowledge and uninformed use of antimicrobials. Considering the development of animal farming to meet an increasing demand for proteins, this deficiency represents a serious public health issue. We advocate for policies that consider the specific challenges faced by family farmers in Africa, to simultaneously improve access to veterinary drugs while strengthening the regulation of their use. We propose a global approach targeting the agri-food system, offering innovative social and technical interventions on antimicrobial usage, adapted to family farmers.

Regular evaluation of integrated surveillance for antimicrobial use (AMU) and resistance (AMR) in animals, humans, and the environment is needed to ensure system effectiveness, but the question is how. In this study, six different evaluation tools were assessed after being applied to AMU and AMR surveillance in eight countries: (1) ATLASS: the Assessment Tool for Laboratories and AMR Surveillance Systems developed by the Food and Agriculture Organization (FAO) of the United Nations, (2) ECoSur: Evaluation of Collaboration for Surveillance tool, (3) ISSEP: Integrated Surveillance System Evaluation Project, (4) NEOH: developed by the EU COST Action "Network for Evaluation of One Health," (5) PMP-AMR: The Progressive Management Pathway tool on AMR developed by the FAO, and (6) SURVTOOLS: developed in the FP7-EU project "RISKSUR." Each tool was scored using (i) 11 pre-defined functional aspects (e.g., workability concerning the need for data, time, and people); (ii) a strengths, weaknesses, opportunities, and threats (SWOT)-like approach of user experiences (e.g., things that I liked or that the tool covered well); and (iii) eight predefined content themes related to scope (e.g., development purpose and collaboration). PMP-AMR, ATLASS, ECoSur, and NEOH are evaluation tools that provide a scoring system to obtain semi-quantitative results, whereas ISSEP and SURVTOOLS will result in a plan for how to conduct evaluation(s). ISSEP, ECoSur, NEOH, and SURVTOOLS allow for in-depth analyses and therefore require more complex data, information, and specific training of evaluator(s). PMP-AMR, ATLASS, and ISSEP were developed specifically for AMR-related activities-only ISSEP included production of a direct measure for "integration" and "impact on decision making." NEOH and ISSEP were perceived as the best tools for evaluation of One Health (OH) aspects, and ECoSur as best for evaluation of the quality of collaboration. PMP-AMR and ATLASS seemed to be the most user-friendly tools, particularly designed for risk managers. ATLASS was the only tool focusing specifically on laboratory activities. Our experience is that adequate resources are needed to perform evaluation(s). In most cases, evaluation would require involvement of several assessors and/or stakeholders, taking from weeks to months to complete. This study can help direct future evaluators of integrated AMU and AMR surveillance toward the most adequate tool for their specific evaluation purpose., Competing Interests: MB was involved in the development of ECoSur and LN was involved in the development of NEOH. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Sandberg, Hesp, Aenishaenslin, Bordier, Bennani, Bergwerff, Chantziaras, De Meneghi, Ellis-Iversen, Filippizi, Mintiens, Nielsen, Norström, Tomassone, van Schaik and Alban.)

Many One Health surveillance systems have proven difficult to enforce and sustain, mainly because of the difficulty of implementing and upholding collaborative efforts for surveillance activities across stakeholders with different values, cultures and interests. We hypothesize that only the early engagement of stakeholders in the development of a One Health surveillance system can create an environment conducive to the emergence of collaborative solutions that are acceptable, accepted and therefore implemented in sustainable manner. To this end, we have designed a socio-technical framework to help stakeholders develop a common vision of their desired surveillance system and to forge the innovation pathway toward it. We implemented the framework in two case studies: the surveillance of antimicrobial resistance in Vietnam and that of Salmonella in France. The socio-technical framework is a participatory and iterative process that consists of four distinct steps implemented during a workshop series: (i) definition of the problem to be addressed, (ii) co-construction of a common representation of the current system, (iii) co-construction of the desired surveillance system, (iv) identification of changes and actions required to progress from the current situation to the desired situation. In both case studies, the process allowed surveillance stakeholders with different professional cultures and expectations regarding One Health surveillance to gain mutual understanding and to reconcile their different perspectives to design the pathway toward their common vision of a desired surveillance system. While the proposed framework is structured around four essential steps, its application can be tailored to the context. Workshop facilitation and representativeness of participants are key for the success of the process. While our approach lays the foundation for the further implementation of the desired One Health surveillance system, it provides no guarantee that the proposed actions will actually be implemented and bring about the required changes. The engagement of stakeholders in a participatory process must be sustained in order to ensure the implementation of co-constructed solutions and evaluate their effectiveness and impacts., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Bordier, Goutard, Antoine-Moussiaux, Pham-Duc, Lailler and Binot.)

The concept of One Health (OH) promotes the decompartmentalisation of human, animal, and ecosystem health for the more efficient and sustainable governance of complex health issues. This means that traditional boundaries between disciplines and sectors must be transgressed and that all relevant stakeholders must be involved in the definition and management of health problems. International efforts have been made to strengthen collaboration across sectors and disciplines and OH surveillance is strongly encouraged at global, national and local-level to efficiently manage hazards involving humans, animals and ecosystems. This concept is intuitively appealing and would suggest the enhanced performance and cost-effectiveness of surveillance systems, as compared to more conventional approaches. Nevertheless, confusion and uncertainty regarding the practical application, outcomes and impacts prevail. We believe that this is due to the lack of a conceptual and methodological framework which would (i) define the characteristics of OH surveillance, and (ii) identify the appropriate mechanisms for inter-sectoral and multi-disciplinary collaboration, to ensure that the surveillance system performs well, with regard to the objective, the context and the health hazard under surveillance. The objective of the study is to define the organisational and functional characteristics of OH surveillance systems, the context in which they are implemented, as well as the influential factors which may obstruct or support their implementation and performance. To achieve this, a systematic literature review of existing OH surveillance systems was conducted using the Prisma guidelines. The selected systems were assessed according to 38 predetermined variables. These allowed the characterisation of their objectives, organisation, functioning, performance and benefits. Data extraction was conducted using a spreadsheet and a database was built using an electronic multiple-choice questionnaire. The literature search identified a total of 1635 records. After the screening phase, 31 references were kept and 22 additional references retrieved from bibliographies were added. From these 53 selected documents, we retrieved 41 different surveillance systems in line with the definition proposed in this study. The analysis of this database enabled the identification of different dimensions and areas of collaboration. Barriers and levers for the implementation of OH surveillance systems were also identified and discussed. Based on our results, we propose a framework to characterise the organisation of collaboration for the governance and operation of an effective OH surveillance system., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The international community and governmental organizations are actively calling for the implementation of One Health (OH) surveillance systems to target health hazards that involve humans, animals, and their environment. In our view, the main characteristic of a OH surveillance system is the collaboration across institutions and disciplines operating within the different sectors to plan, coordinate, and implement the surveillance process. However, the multisectoral organizational models and possible collaborative modalities implemented throughout the surveillance process are multi-fold and depend on the objective and context of the surveillance. The purpose of this study is to define a matrix to evaluate the quality and appropriateness of multisectoral collaboration through an in-depth analysis of its organization, implementation, and functions. We developed a first list of evaluation attributes based on (i) the characteristics of the organization, implementation, and functionality of multisectoral surveillance systems; and (ii) the existing attributes for the evaluation of health surveillance systems and OH initiatives. These attributes were submitted to two rounds of expert-opinion elicitation for review and validation. The final list of attributes consisted of 23 organizational attributes and 9 functional attributes, to which 3 organizational indexes were added measuring the overall organization of collaboration. We then defined 75 criteria to evaluate the level of satisfaction for the attributes and indexes. The criteria were scored following a four-tiered scoring grid. Graphical representations allowed for an easy overview of the evaluation results for both attributes and indexes. This evaluation matrix is the first to allow an in-depth analysis of collaboration in a multisectoral surveillance system and is the preliminary step toward the creation of a fully standalone tool for the evaluation of collaboration. After its practical application and adaptability to different contexts are field-tested, this tool could be very useful in identifying the strengths and weaknesses of collaboration occurring in a multisectoral surveillance system.

Background: The international community strongly advocates the implementation of multi-sectoral surveillance policies for an effective approach to antibiotic resistance, in line with the One Health concept. To comply with these international recommendations, the Vietnamese government has issued an inter-ministerial surveillance strategy for antibiotic resistance, including an integrated surveillance system. However, one may question the ability and willingness of surveillance stakeholders to implement the collaborations required. To assess the feasibility of operationalising this strategy within the national context, we explored the role of key stakeholders in the strategy, as well as their abilities to comply with it., Methods: We conducted a qualitative approach based on an iterative stakeholder mapping and analysis, in three distinct steps: (1) a description of the structure of the national surveillance strategy (literature review, key informant interviews); (2) an analysis of the key stakeholders' positions regarding the strategy (semi-structured interviews); (3) the identification of factors influencing the operationalisation of the collaborative surveillance strategy (comparison of data collected at the first and second steps)., Results: The mapping of the surveillance system, as well as the characterisation of key stakeholders according to organisational and functional attributes, underlined that inter-sectoral surveillance initiatives do exist, but that the organisation of the national surveillance system remains highly silo-oriented. Based on stakeholder perspectives, we identified seven factors that may influence the implementation of the One Health strategy at national level: governance and operational frameworks, divergence of institutional cultures, level of knowledge, technical capacities, allocation of resources, conflicting commercial interests and influence of international partners., Conclusions: The study suggests that the operationalisation of the collaborative surveillance strategy requires the full adhesion of stakeholders and the provision of appropriate resources. Based on these findings, we have proposed a guidance framework together with recommendations to move towards a more suitable governance and operational model for One Health surveillance of antibiotic resistance in Vietnam. To lever and promote successful inter-sectoral collaboration, a participatory "learning by doing" process could be applied to guide, frame and mentor stakeholders through the identification of appropriate levels of collaboration, depending on the expected positive impacts on the value of surveillance.

Background: OnabotulinumtoxinA has proven its efficacy in reducing the number of headache days in chronic migraine (CM) patients. The usual paradigm includes 31 pericranial injection sites with low dose (5 U) per site. The aim of this study is to present the results obtained using a simpler injection protocol of onabotulinumtoxinA, with injection sites targeted to pericranial myofascial sites of pain., Methods: Observational, open label, real-life, cohort study. We enrolled 63 consecutive patients fulfilling the diagnostic criteria of CM, and refractory to conventional treatments. The patients were injected using a "follow-the-pain" pattern into the corrugator and/or temporalis and/or trapezius muscles. The doses per muscle were fixed. According to the number of muscles injected, the total dose could vary from 70 to 150 U per session. Patients were considered responders if they had a ≥ 50% decrease in number of headache days in at least two consecutive injection cycles., Results: Forty one patients (65.1% in intention to treat analysis) responded to treatment. In 70.7% of responders, the effect size was even higher, with a reduction ≥70% in the number of headache days. The associated cervical pain and muscle tenderness, present in 33 patients, was reduced by ≥50% in 31 patients (94%). Triptan consumption dramatically decreased (81%) in responders. The trapezius was the most frequently injected muscle. We observed no serious adverse event. The mean patient satisfaction rate was 8.5/10., Conclusions: This study provides additional robust evidence supporting the efficacy of onabotulinumtoxinA injections in CM. Furthermore, the paradigm we used, with reduced number of injection sites targeted to pericranial myofascial sites of pain, may provide evidence in favor of the implication of myofascial trigger points in migraine chronicization., Trial Registration: ClinicalTrials.gov Protocol Record I17022 ClinicalTrials.gov Identifier: NCT03175263 . Date of registration: June 7, 2017. Retrospectively registered.

Competing Interests: Competing interests: We have read and understood BMJ policy on declaration of interests and have no relevant interests to declare.

ABHD12 mutations have been linked to neurodegenerative PHARC (polyneuropathy, hearing loss, ataxia, retinitis pigmentosa, and early-onset cataract), a rare, progressive, autosomal, recessive disease. Although ABHD12 is suspected to play a role in the lysophosphatidylserine and/or endocannabinoid pathways, its precise functional role(s) leading to PHARC disease had not previously been characterized. Cell and zebrafish models were designed to demonstrate the causal link between an identified new missense mutation p.T253R, characterized in ABHD12 from a young patient, the previously characterized p.T202I and p.R352* mutations, and the associated PHARC. Measuring ABHD12 monoacylglycerol lipase activity in transfected HEK293 cells demonstrated inhibition with mutated isoforms. Both the expression pattern of zebrafish abhd12 and the phenotype of specific antisense morpholino oligonucleotide gene knockdown morphants were consistent with human PHARC hallmarks. High abhd12 transcript levels were found in the optic tectum and tract, colocalized with myelin basic protein, and in the spinal cord. Morphants have myelination defects and concomitant functional deficits, characterized by progressive ataxia and motor skill impairment. A disruption of retina architecture and retinotectal projections was observed, together with an inhibition of lens clarification and a low number of mechanosensory hair cells in the inner ear and lateral line system. The severe phenotypes in abhd12 knockdown morphants were rescued by introducing wild-type human ABHD12 mRNA, but not by mutation-harboring mRNAs. Zebrafish may provide a suitable vertebrate model for ABHD12 insufficiency and the study of functional impairment and potential therapeutic rescue of this rare, neurodegenerative disease., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Zoonoses are an issue of growing interest in South-East Asia, where environmental factors and socio-economic context favor the endemization of well-known diseases and the emergence of new pathogens at the human-wildlife interface. However, the health status of the region with respect to many zoonotic diseases remains poorly defined, despite the high overall burden of zoonoses on the countries of the area, and the global risk of new biological threats in the region. The first objective of this paper was to provide an update of data on the zoonoses commonly described by the scientific community and reported by governmental institutions and international organizations in continental South-East Asia. The analysis of the available data led to the identification of some trends in the evolution of the diseases, as well as some gaps in knowledge and in the current surveillance and control networks. In light of these findings, we discuss measures for effectively addressing zoonotic disease issues in South-East Asia, such as the allocation of funds for research and for surveillance and control programs, and a multi-sectoral and multi-disciplinary approach at various levels.

Saimiri (Cebidae) groups a complex of species and subspecies, which present a large morphological plasticity. Genetic analysis is complicated by the absence of consensus on classification criteria and the paucity of molecular tools available for the genus. As the squirrel monkey is widely used in biomedical research, breeding centers have been established, but the genetic make up and diversity of many of the existing colonies is unknown precluding a rationale breeding policy. To develop a genetic typing strategy for the Saimiri breeding colony of Pasteur Institute of French Guiana, we have used Cytochrome b, a mitochondrial marker, and nuclear microsatellites. Cytochrome b sequences from wild-caught Saimiri boliviensis, Saimiri sciureus sciureus and S. s. collinsi reference specimens and captive animals identified 11 haplotypes, grouped into three distinct clades. An estimate of genetic variability within each captive morphotype, and of the extent of molecular divergence between the Bolivian, Guyanese and Brazilian breeds was obtained from the analysis of three nuclear microsatellites. Taxon-specific microsatellites enabled typing of F0-F3 animals, but did not differentiate Brazilian from Guyanese animals. Three locus microsatellite analysis of a representative sample from each generation showed no trend for loss of heterozygosity, and identified hybrid animals between Bolivian and the two others sub-species. These data provide novel evidence for taxonomic classification and a rationale strategy to further type the whole colony.

Two recombinant baculoviruses were constructed by inserting via the transfer vector pAcYM1 the genes coding for the structural proteins of dengue (DEN)-2 virus downstream from the polyhedrin promoter of Autographa californica nuclear polyhedrosis virus. The two recombinants differed in truncation of 26 and 71 amino acids, respectively, in the carboxy-terminal sequence of DEN-specific envelope (E) glycoprotein. Recombinant DEN-2 E glycoproteins were processed and transported to the surface of Spodoptera frugiperda Sf9 cells infected with both viruses. We show that about one-third of the E glycoprotein minus its whole C-terminal hydrophobic anchor domain was secreted into an endoglycosidase H-resistant form. The type-specific neutralizing epitopes were conserved in the recombinant proteins as shown with a panel of monoclonal antibodies.