Challenges to improved animal rabies surveillance: Experiences from pilot implementation of decentralized diagnostic units in Chad.

Better surveillance is desperately needed to guide rabies prevention and control to achieve the goal of zero dog-mediated human rabies by 2030, defined by the World Health Organization (WHO) and partners in 2015. With the help of funding from the Vaccine Alliance (GAVI) learning agenda, we implemented animal rabies surveillance based on One Health communication, improved accessibility of diagnostic testing and facilitated sample transport to increase case detection in three regions of Chad. Through the project, rabies surveillance, previously only available in N'Djaména, was extended to selected provincial rural and urban areas. Nine decentralized diagnostic units (DDU) were established, hosted by veterinary district agencies (VDA) in four different administrative regions. Four additional VDAs in the study area were reinforced with facilitation of sample collection and transport. Staff from all these 13 veterinary facilities were trained in sample collection and diagnostics. DDUs performed Rapid Immunodiagnostic Tests (RIDT) providing a preliminary result before samples were sent to the central laboratory in N'Djamena for confirmation with the standard Florescent Antibody Test (FAT). Within the project period from June 2016 to March 2018, 115 samples were reported by veterinary facilities in the study areas compared to 63 samples received from outside the study area, the vast majority of them originating from the capital city N'Djaména (N=61). Eighty nine percent of all 178 samples reported to IRED during the project period tested positive. Most of the samples originated from dogs (92%). Other confirmed rabies positive animals observed were cats, a donkey and a pig. Although surveillance of animal rabies was the focus, four human saliva samples were also submitted for diagnosis. We observed high differences in reporting rates between the four study regions. This could be attributable to differences in rabies epidemiology but are also influenced by the distance to the central laboratory in N'Djaména, the cultural background and the level of public awareness. The possibility for local testing through RIDT was very welcomed by local veterinary staff and preliminary insights suggest a positive influence on One Health communication and PEP initiation. However, these aspects as well as the relative impact of local testing on sample collection in comparison to reinforcement of sample collection and transport alone, need to be further investigated. Challenges encountered related to poor infrastructure (buildings, appliances, materials) and low logistic capacity (lacking means and material for transport and communication) of veterinary services in Chad. In addition, veterinary personnel lack experience in data management. Together with staff turnover, this leads to a need for repeated training. Major shortcoming of the approach was the high cost per sample and limited sustainability beyond the project timeframe.

Predictors of free-roaming domestic dogs' contact network centrality and their relevance for rabies control.

Free roaming domestic dogs (FRDD) are the main vectors for rabies transmission to humans worldwide. To eradicate rabies from a dog population, current recommendations focus on random vaccination with at least 70% coverage. Studies suggest that targeting high-risk subpopulations could reduce the required vaccination coverage, and increase the likelihood of success of elimination campaigns. The centrality of a dog in a contact network can be used as a measure of its potential contribution to disease transmission. Our objectives were to investigate social networks of FRDD in eleven study sites in Chad, Guatemala, Indonesia and Uganda, and to identify characteristics of dogs, and their owners, associated with their centrality in the networks. In all study sites, networks had small-world properties and right-skewed degree distributions, suggesting that vaccinating highly connected dogs would be more effective than random vaccination. Dogs were more connected in rural than urban settings, and the likelihood of contacts was negatively correlated with the distance between dogs' households. While heterogeneity in dog's connectedness was observed in all networks, factors predicting centrality and likelihood of contacts varied across networks and countries. We therefore hypothesize that the investigated dog and owner characteristics resulted in different contact patterns depending on the social, cultural and economic context. We suggest to invest into understanding of the sociocultural structures impacting dog ownership and thus driving dog ecology, a requirement to assess the potential of targeted vaccination in dog populations.

Comparative Study of Free-Roaming Domestic Dog Management and Roaming Behavior Across Four Countries: Chad, Guatemala, Indonesia, and Uganda.

Dogs play a major role in public health because of potential transmission of zoonotic diseases, such as rabies. Dog roaming behavior has been studied worldwide, including countries in Asia, Latin America, and Oceania, while studies on dog roaming behavior are lacking in Africa. Many of those studies investigated potential drivers for roaming, which could be used to refine disease control measures. However, it appears that results are often contradictory between countries, which could be caused by differences in study design or the influence of context-specific factors. Comparative studies on dog roaming behavior are needed to better understand domestic dog roaming behavior and address these discrepancies. The aim of this study was to investigate dog demography, management, and roaming behavior across four countries: Chad, Guatemala, Indonesia, and Uganda. We equipped 773 dogs with georeferenced contact sensors (106 in Chad, 303 in Guatemala, 217 in Indonesia, and 149 in Uganda) and interviewed the owners to collect information about the dog [e.g., sex, age, body condition score (BCS)] and its management (e.g., role of the dog, origin of the dog, owner-mediated transportation, confinement, vaccination, and feeding practices). Dog home range was computed using the biased random bridge method, and the core and extended home range sizes were considered. Using an AIC-based approach to select variables, country-specific linear models were developed to identify potential predictors for roaming. We highlighted similarities and differences in term of demography, dog management, and roaming behavior between countries. The median of the core home range size was 0.30 ha (95% range: 0.17-0.92 ha) in Chad, 0.33 ha (0.17-1.1 ha) in Guatemala, 0.30 ha (0.20-0.61 ha) in Indonesia, and 0.25 ha (0.15-0.72 ha) in Uganda. The median of the extended home range size was 7.7 ha (95% range: 1.1-103 ha) in Chad, 5.7 ha (1.5-27.5 ha) in Guatemala, 5.6 ha (1.6-26.5 ha) in Indonesia, and 5.7 ha (1.3-19.1 ha) in Uganda. Factors having a significant impact on the home range size in some of the countries included being male dog (positively), being younger than one year (negatively), being older than 6 years (negatively), having a low or a high BCS (negatively), being a hunting dog (positively), being a shepherd dog (positively), and time when the dog was not supervised or restricted (positively). However, the same outcome could have an impact in a country and no impact in another. We suggest that dog roaming behavior is complex and is closely related to the owner's socioeconomic context and transportation habits and the local environment. Free-roaming domestic dogs are not completely under human control but, contrary to wildlife, they strongly depend upon humans. This particular dog-human bound has to be better understood to explain their behavior and deal with free-roaming domestic dogs related issues.

Field Postmortem Rabies Rapid Immunochromatographic Diagnostic Test for Resource-Limited Settings with Further Molecular Applications.

Functional rabies surveillance systems are crucial to provide reliable data and increase the political commitment necessary for disease control. To date, animals suspected as rabies-positive must be submitted to a postmortem confirmation using classical or molecular laboratory methods. However, most endemic areas are in low- and middle-income countries where animal rabies diagnosis is restricted to central veterinary laboratories. Poor availability of surveillance infrastructure leads to serious disease underreporting from remote areas. Several diagnostic protocols requiring low technical expertise have been recently developed, providing opportunity to establish rabies diagnosis in decentralized laboratories. We present here a complete protocol for field postmortem diagnosis of animal rabies using a rapid immunochromatographic diagnostic test (RIDT), from brain biopsy sampling to the final interpretation. We complete the protocol by describing a further use of the device for molecular analysis and viral genotyping. RIDT easily detects rabies virus and other lyssaviruses in brain samples. The principle of such tests is simple: brain material is applied on a test strip where gold conjugated antibodies bind specifically to rabies antigens. The antigen-antibody complexes bind further to fixed antibodies on the test line, resulting in a clearly visible purple line. The virus is inactivated in the test strip, but viral RNA can be subsequently extracted. This allows the test strip, rather than the infectious brain sample, to be safely and easily sent to an equipped laboratory for confirmation and molecular typing. Based on a modification of the manufacturer's protocol, we found increased test sensitivity, reaching 98% compared to the gold standard reference method, the direct immunofluorescence antibody test. The advantages of the test are numerous: rapid, easy-to-use, low cost and no requirement for laboratory infrastructure, such as microscopy or cold-chain compliance. RIDTs represent a useful alternative for areas where reference diagnostic methods are not available.

The importance of dog population contact network structures in rabies transmission.

Canine rabies transmission was interrupted in N'Djaména, Chad, following two mass vaccination campaigns. However, after nine months cases resurged with re-establishment of endemic rabies transmission to pre-intervention levels. Previous analyses investigated district level spatial heterogeneity of vaccination coverage, and dog density; and importation, identifying the latter as the primary factor for rabies resurgence. Here we assess the impact of individual level heterogeneity on outbreak probability, effectiveness of vaccination campaigns and likely time to resurgence after a campaign. Geo-located contact sensors recorded the location and contacts of 237 domestic dogs in N'Djaména over a period of 3.5 days. The contact network data showed that urban dogs are socially related to larger communities and constrained by the urban architecture. We developed a network generation algorithm that extrapolates this empirical contact network to networks of large dog populations and applied it to simulate rabies transmission in N'Djaména. The model predictions aligned well with the rabies incidence data. Using the model we demonstrated, that major outbreaks are prevented when at least 70% of dogs are vaccinated. The probability of a minor outbreak also decreased with increasing vaccination coverage, but reached zero only when coverage was near total. Our results suggest that endemic rabies in N'Djaména may be explained by a series of importations with subsequent minor outbreaks. We show that highly connected dogs hold a critical role in transmission and that targeted vaccination of such dogs would lead to more efficient vaccination campaigns.