Seroprevalence of Bas-Congo virus in Mangala, Democratic Republic of the Congo: a population-based cross-sectional study.

BACKGROUND: Bas-Congo virus (BASV), an emerging tibrovirus, was associated with an outbreak of acute haemorrhagic fever in Mangala, Democratic Republic of the Congo, in 2009. In 2012, neutralising antibodies to BASV were detected in the lone survivor and one of his close contacts. However, subsequent serological and molecular surveys were unsuccessful as neither BASV antibodies nor its RNA were detected. In this study, we determined the seroprevalence of BASV infection in Mangala 13 years after the initial outbreak. METHODS: We conducted a population-based serological survey from Jan 17 to Jan 23, 2022. Consenting individuals at least 5 years of age, living in Mangala for at least 4 weeks, and who had no contraindication to venepuncture were enrolled. Participants were interviewed using a pre-tested questionnaire for sociodemographic and clinical characteristics. We supplemented the collected serum samples with 284 archived samples from Matadi and Kinshasa. All samples were tested for antibodies to BASV and other tibroviruses using a pseudovirus-based neutralisation test. FINDINGS: Among the 267 individuals from Mangala, the prevalence of BASV antibodies was 55% (95% CI 49-61; n=147). BASV seropositivity odds significantly increased with age (5·2 [95% CI 2·1-12·9] to 83·9 [20·8-337·7] times higher in participants aged 20 years or older than participants aged 5-19 years). Some occupational categories (eg, farmer or public servant) were associated with seropositivity. Only nine (6%) of 160 samples from Matadi and one (<1%) of 124 samples from Kinshasa had neutralising antibodies to BASV. Moreover, we also detected neutralising antibodies to other tibroviruses-Ekpoma virus 1, Ekpoma virus 2, and Mundri virus-in 84 (31%), 251 (94%), and 219 (82%) of 267 Mangala samples; 14 (9%), 62 (39%), and 120 (75%) of 160 Matadi samples; and six (5%), five (4%), and 33 (27%) of 124 Kinshasa samples, respectively. INTERPRETATION: Human infection with BASV and other tibroviruses seems common in Mangala, although no deadly outbreak has been reported since 2009. Exposure to BASV might be highly restricted to Mangala and the increasing prevalence of neutralising antibodies with age suggests regular contact with the virus in this city. Altogether, our findings suggest that human infection with tibroviruses could be common in the study areas and not associated with deadly haemorrhagic or debilitating syndromes. FUNDING: Japan Agency for Medical Research and Development (AMED) and Japan International Cooperation Agency (JICA) under the Science and Technology Research Partnership for Sustainable Development (SATREPS) and Japan Program for Infectious Diseases Research and Infrastructure from AMED.

Trypanosome spliced leader RNA for diagnosis of acoziborole treatment outcome in gambiense human African trypanosomiasis: A longitudinal follow-up study.

BACKGROUND: Detection of spliced leader (SL)-RNA allows sensitive diagnosis of gambiense human African trypanosomiasis (HAT). We investigated its diagnostic performance for treatment outcome assessment. METHODS: Blood and cerebrospinal fluid (CSF) from a consecutive series of 97 HAT patients, originating from the Democratic Republic of the Congo, were prospectively collected before treatment with acoziborole, and during 18 months of longitudinal follow-up after treatment. For treatment outcome assessment, SL-RNA detection was compared with microscopic trypanosome detection and CSF white blood cell count. The trial was registered under NCT03112655 in clinicaltrials.gov. FINDINGS: Before treatment, respectively 94.9% (92/97; CI 88.5-97.8%) and 67.7% (65/96; CI 57.8-76.2%) HAT patients were SL-RNA positive in blood or CSF. During follow-up, one patient relapsed with trypanosomes observed at 18 months, and was SL-RNA positive in blood and CSF at 12 months, and CSF positive at 18 months. Among cured patients, one individual tested SL-RNA positive in blood at month 12 (Specificity 98.9%; 90/91; CI 94.0-99.8%) and 18 (Specificity 98.9%; 88/89; CI 93.9-99.8%). INTERPRETATION: SL-RNA detection for HAT treatment outcome assessment shows ≥98.9% specificity in blood and 100% in CSF, and may detect relapses without lumbar puncture. FUNDING: The DiTECT-HAT project is part of the EDCTP2 programme, supported by Horizon 2020, the European Union Funding for Research and Innovation (grant number DRIA-2014-306-DiTECT-HAT).

The prevalence of Taenia spp. in pigs slaughtered in Kinshasa.

Taenia hydatigena is a non-zoonotic worm that has dogs and wild canids as definitive hosts. Its presence induces cross reactions in certain diagnostic tests for porcine cysticercosis caused by T. solium, the occurrence of which has a considerable public health and economic impact. In the Democratic Republic of the Congo (DR Congo), T. solium is considered endemic, however, the prevalence of T. hydatigena has not been estimated yet. The objective of the study was therefore to estimate the prevalence of T. hydatigena cysticercosis by serological and molecular diagnostic tools in pigs slaughtered in DR Congo. A total of 480 pigs slaughtered in 6 slaughter slabs in Kinshasa, DR Congo, were examined. The thoracal and abdominal cavity organs were inspected for cysts, which were analyzed using PCR-RFLP. Furthermore, 480 sera were collected, and analyzed for the presence of circulating Taenia spp. cysticercus antigens, using the B158/B60 Ag-ELISA. Upon inspection of the carcass, 41 cysts suspected to be metacestodes of Taenia spp. were collected, from the following viscera: spleen (24/41, 59%), liver (13/41, 32%), intestine (3/41, 7%) and lung (1/41, 2%). Molecular analyses revealed a T. hydatigena prevalence of 0.2% (95% CI: 0.0001-0.0116), based on a single lesion (1/480), taken from the spleen. Out of the 480 sera collected, the presence of circulating Taenia spp. cysticerci antigens was detected in 32 (6.7%; 95% CI: 4.5-11.2). The results of this study revealed that T. hydatigena is present in pigs sold in markets in the city of Kinshasa in DR Congo, albeit at a very low prevalence, thus the impact on the interpretation of the B158/B60 seems low in this setting. Detection of circulating antigens in porcine sera by Ag-ELISA, shows that pigs slaughtered in Kinshasa, DR Congo, were infected with viable cysticerci of Taenia spp. which in turn can infect humans.

Mapping of Antibody Epitopes on the Crimean-Congo Hemorrhagic Fever Virus Nucleoprotein.

Crimean-Congo hemorrhagic fever virus (CCHFV), a nairovirus, is a tick-borne zoonotic virus that causes hemorrhagic fever in humans. The CCHFV nucleoprotein (NP) is the antigen most used for serological screening of CCHFV infection in animals and humans. To gain insights into antibody epitopes on the NP molecule, we produced recombinant chimeric NPs between CCHFV and Nairobi sheep disease virus (NSDV), which is another nairovirus, and tested rabbit and mouse antisera/immune ascites, anti-NP monoclonal antibodies, and CCHFV-infected animal/human sera for their reactivities to the NP antigens. We found that the amino acids at positions 161-320 might include dominant epitopes recognized by anti-CCHFV IgG antibodies, whereas cross-reactivity between anti-CCHFV and anti-NSDV antibodies was limited. Their binding capacities were further tested using a series of synthetic peptides whose sequences were derived from CCHFV NP. IgG antibodies in CCHFV-infected monkeys and patients were reactive to some of the synthetic peptide antigens (e.g., amino acid residues at positions 131-150 and 211-230). Only a few peptides were recognized by IgG antibodies in the anti-NSDV serum. These results provide useful information to improve NP-based antibody detection assays as well as antigen detection tests relying on anti-NP monoclonal antibodies.

Trypanosome SL-RNA detection in blood and cerebrospinal fluid to demonstrate active gambiense human African trypanosomiasis infection.

BACKGROUND: Spliced Leader (SL) trypanosome RNA is detectable only in the presence of live trypanosomes, is abundant and the Trypanozoon subgenus has a unique sequence. As previously shown in blood from Guinean human African trypanosomiasis (HAT) patients, SL-RNA is an accurate target for diagnosis. Detection of SL-RNA in the cerebrospinal fluid (CSF) has never been attempted. In a large group of Congolese gambiense HAT patients, the present study aims i) to confirm the sensitivity of SL-RNA detection in the blood and; ii) to assess the diagnostic performance of SL-RNA compared to trypanosome detection in CSF. METHODOLOGY/PRINCIPAL FINDINGS: Blood and CSF from 97 confirmed gambiense HAT patients from the Democratic Republic of Congo were collected using PAXgene blood RNA Tubes. Before RNA extraction, specimens were supplemented with internal extraction control RNA to monitor the extraction, which was performed with a PAXgene Blood RNA Kit. SL-RNA qPCR was carried out with and without reverse transcriptase to monitor DNA contamination. In blood, 92/97 (94.8%) HAT patients tested SL-RNA positive, which was significantly more than combined trypanosome detection in lymph and blood (78/97 positive, 80.4%, p = 0.001). Of 96 CSF RNA specimens, 65 (67.7%) were SL-RNA positive, but there was no significant difference between sensitivity of SL-RNA and trypanosome detection in CSF. The contribution of DNA to the Cq values was negligible. In CSF with normal cell counts, a fraction of SL-RNA might have been lost during extraction as indicated by higher internal extraction control Cq values. CONCLUSIONS/SIGNIFICANCE: Detection of SL-RNA in blood and CSF allows sensitive demonstration of active gambiense HAT infection, even if trypanosomes remain undetectable in blood or lymph. As this condition often occurs in treatment failures, SL-RNA detection in blood and CSF for early detection of relapses after treatment deserves further investigation. TRIAL REGISTRATION: This study was an integral part of the diagnostic trial "New Diagnostic Tools for Elimination of Sleeping Sickness and Clinical Trials: Early tests of Cure" (DiTECT-HAT-WP4, ClinicalTrials.gov Identifier: NCT03112655).

Purification of Crimean-Congo hemorrhagic fever virus nucleoprotein and its utility for serological diagnosis.

Crimean-Congo hemorrhagic fever virus (CCHFV) causes a zoonotic disease, Crimean-Congo hemorrhagic fever (CCHF) endemic in Africa, Asia, the Middle East, and Southeastern Europe. However, the prevalence of CCHF is not monitored in most of the endemic countries due to limited availability of diagnostic assays and biosafety regulations required for handling infectious CCHFV. In this study, we established a protocol to purify the recombinant CCHFV nucleoprotein (NP), which is antigenically highly conserved among multiple lineages/clades of CCHFVs and investigated its utility in an enzyme-linked immunosorbent assay (ELISA) to detect CCHFV-specific antibodies. The NP gene was cloned into the pCAGGS mammalian expression plasmid and human embryonic kidney 293 T cells were transfected with the plasmid. The expressed NP molecule was purified from the cell lysate using cesium-chloride gradient centrifugation. Purified NP was used as the antigen for the ELISA to detect anti-CCHFV IgG. Using the CCHFV NP-based ELISA, we efficiently detected CCHFV-specific IgG in anti-NP rabbit antiserum and CCHFV-infected monkey serum. When compared to the commercially available Blackbox CCHFV IgG ELISA kit, our assay showed equivalent performance in detecting CCHFV-specific IgG in human sera. These results demonstrate the usefulness of our CCHFV NP-based ELISA for seroepidemiological studies.

A New Variant Among Newcastle Disease Viruses Isolated in the Democratic Republic of the Congo in 2018 and 2019.

Newcastle disease (ND) is a highly transmissible and devastating disease that affects poultry and wild birds worldwide. Comprehensive knowledge regarding the characteristics and epidemiological factors of the ND virus (NDV) is critical for the control and prevention of ND. Effective vaccinations can prevent and control the spread of the NDV in poultry populations. For decades, the Democratic Republic of the Congo (DRC) has reported the impacts of ND on commercial and traditional poultry farming systems. The reports were preliminary clinical observations, and few cases were confirmed in the laboratory. However, data on the phylogenetic, genetic, and virological characteristics of NDVs circulating in the DRC are not available. In this study, the whole-genome sequences of three NDV isolates obtained using the next-generation sequencing method revealed two isolates that were a new variant of NDV, and one isolate that was clustered in the subgenotype VII.2. All DRC isolates were velogenic and were antigenically closely related to the vaccine strains. Our findings reveal that despite the circulation of the new variant, ND can be controlled in the DRC using the current vaccine. However, epidemiological studies should be conducted to elucidate the endemicity of the disease so that better control strategies can be implemented.

Molecular characterization of rabies viruses from two western provinces of the Democratic Republic of the Congo (2008-2017).

Although rabies is enzootic in the Democratic Republic of the Congo, there is very little molecular epidemiological information about the viruses circulating in animals. In this study, a fragment of the rabies virus (RABV) nucleoprotein gene was amplified and sequenced from 21 animal brain samples collected in two western provinces of the country between 2008 and 2017. The samples tested were from cat (n = 1), dog (n = 17), goat (n = 2), and sheep (n = 1). Phylogenetic analysis revealed that the sequences generated were highly similar to each other and belonged to lineage Africa 1b clustering with a single sample identified in a canine in the Republic of Congo in 2014. This is the first molecular epidemiological study of RABV in the DRC and the data generated will assist authorities in the development of effective control strategies for rabies in the country.

Molecular, antigenic, and pathogenic characterization of H5N8 highly pathogenic avian influenza viruses isolated in the Democratic Republic of Congo in 2017.

In May 2017, high mortality of chickens and Muscovy ducks due to the H5N8 highly pathogenic avian influenza virus (HPAIV) was reported in the Democratic Republic of Congo (DR Congo). In this study, we assessed the molecular, antigenic, and pathogenic features in poultry of the H5N8 HPAIV from the 2017 Congolese outbreaks. Phylogenetic analysis of the eight viral gene segments revealed that all 12 DR Congo isolates clustered in clade 2.3.4.4B together with other H5N8 HPAIVs isolated in Africa and Eurasia, suggesting a possible common origin of these viruses. Antigenically, a slight difference was observed between the Congolese isolates and a representative virus from group C in the same clade. After intranasal inoculation with a representative DR Congo virus, high pathogenicity was observed in chickens and Muscovy ducks but not in Pekin ducks. Viral replication was higher in chickens than in Muscovy duck and Pekin duck organs; however, neurotropism was pronounced in Muscovy ducks. Our data confirmed the high pathogenicity of the DR Congo virus in chickens and Muscovy ducks, as observed in the field. National awareness and strengthening surveillance in the region are needed to better control HPAIVs.

Peste des petits ruminants viruses of lineages II and III identified in the Democratic Republic of the Congo.

Understanding the molecular epidemiology and evolution of peste des petits ruminants virus (PPRV), the causative agent of Peste des petits ruminants, can assist in the control of the transboundary spread of this economically important disease. To date, despite having been reported in the majority of northern and central African countries, no molecular epidemiological data on PPRVs are available for the Democratic Republic of the Congo (DRC). This study reports the collection and analysis of 11 samples collected from three provinces of the DRC in 2016 and 2018. Sequence analysis identified two (i.e. II and III) of the four known lineages of PPRV in the country providing important information that will assist in the global eradication of PPR.

Participatory assessment of paid vaccination campaigns for village chickens against Newcastle disease in Kongo Central province.

In the Democratic Republic of Congo (DRC), where state-driven animal vaccination campaigns are organized only in response to epidemics, the organization of a permanent animal vaccination service is urgently needed. A non-governmental organization has set up an experimental paid vaccination service for village chickens against Newcastle Disease (ND) in the Kongo Central province. This mixed-method study presents a participatory assessment of this experiment, identifying socio-economic factors that influence the decision of chicken keepers to adopt vaccination. The study was conducted in four territories of the province. Qualitative and quantitative data were collected through 12 focus group discussions (FGDs) with professionals of animal health and chicken keepers and 160 semi-structured interviews with chicken keepers, sampled by snowball technique. This participatory process has resulted in the design of a grid for assessing animal vaccination service's performance. Here translating the narratives into a preliminary structured assessment, this grid is an output of the study, to be mobilized for future rapid assessments of the vaccination service in a quantitative prospect. The grid consisted of nine criteria, further composed by 16 items, translated into questions to be asked to chicken keepers and vaccinators. In our study area, fieldworkers enumerated four animal vaccination campaigns during a period of 21 years (except those subject to the present assessment). Around 13% of chicken keepers of our sample had participated in ND vaccination programs. Almost 96% of interviewed chicken keepers expressed their willingness to pay for ND vaccination, and 87% of chicken keepers that vaccinated their chickens perceived the vaccine as effective. Vaccinators estimated that 56% of the chicken keepers who were contacted had actually paid for the vaccination of their chickens. The assessment grid highlighted four points in favor of the sustainability of this service, i.e. the general interest of chickens keepers, vaccine efficacy, vaccine availability and ease of use of the vaccine. Two weak points were identified, viz. the poor access of chicken keepers to information and the weak motivation of vaccinators. The vaccine coverage was calculated within the sample at 13.1%. Paid vaccination campaign for village chicken in Kongo Central obtained a performance score of 62.8%, with the highest score in Kwilu-Ngongo (73.1%) and the lowest in Kasangulu (52.4%). Two factors of adoption of vaccination were identified as statistically significant, i.e. chicken housing and territory. Significant differences appeared between territories in access to information for chicken keepers and in vaccinators motivation. The priorities for the improvement of this service appear to be awareness raising among chicken keepers and increasing vaccinators' motivation.

Clinical Evaluation of QuickNaviTM-Ebola in the 2018 Outbreak of Ebola Virus Disease in the Democratic Republic of the Congo.

The recent large outbreaks of Ebola virus disease (EVD) in West Africa and the Democratic Republic of the Congo (DRC) have highlighted the need for rapid diagnostic tests to control this disease. In this study, we clinically evaluated a previously developed immunochromatography-based kit, QuickNaviTM-Ebola. During the 2018 outbreaks in DRC, 928 blood samples from EVD-suspected cases were tested with QuickNaviTM-Ebola and the WHO-approved GeneXpert. The sensitivity and specificity of QuickNaviTM-Ebola, estimated by comparing it to GeneXpert-confirmed cases, were 85% (68/80) and 99.8% (846/848), respectively. These results indicate the practical reliability of QuickNaviTM-Ebola for point-of-care diagnosis of EVD.

Seroprevalence of Rift Valley fever virus in cattle in the Democratic Republic of the Congo.

This study aimed at assessing the serological and virological status of Rift Valley fever virus (RVFV) in cattle from four climatically diverse zones of the Democratic Republic of the Congo (DRC). A total of 1675 sera samples collected between 2014 and 2015 from cattle without clinical manifestation of RVF infection were tested using competitive and capture enzyme ELISA to detect both IgG and IgM. RT-PCR was used for the detection of nucleic acid of RVFV. Out of the 1675 cattle sera tested, 203 were found to be IgG-positive, giving an overall true seroprevalence of 12.37% (95% CI 10.86-14.05). This seroprevalence varied between the four zones with a seroprevalence of 16.16% (95% CI 12.86-20.12), 14.70% (95% CI 11.72-18.29), 10.82% (95% CI 7.19-14.19), and 7.34% (95% CI 5.13-10.41) recorded in cattle sampled in the mountainous, humid savannah, dry savannah, and forest zones, respectively (p < 0.05, χ2 = 17.26). A higher true seroprevalence of 14.58% (95% CI 9.3-22.13) was found in animals aged 1 year compared to 10.43% (95% CI 8.12-13.30) and 13.16% (95% CI 11.19-15.42) in groups aged between 2-3 and > 3 years, respectively, although the difference was not statistically significant (p > 0.05, χ2 = 2.95). Similarly, no statistically significant difference (p > 0.05, χ2 = 0.04) was found between the sexes of the animals. Among the IgG-positive samples screened for anti-RVFV IgM, only 1.47% (3/203) was IgM-positive. One of the IgM-positive samples was positive by RT-PCR. These findings reveal country-wide distribution of RVF in the DRC for the first time.

Molecular identification of Plasmodium species in symptomatic children of Democratic Republic of Congo.

BACKGROUND: Worldwide, the highest malaria mortality is due to Plasmodium falciparum infection. However, other species of Plasmodium (Plasmodium vivax, Plasmodium ovale, Plasmodium malariae, and Plasmodium knowlesi) can also cause malaria. Therefore, accurate identification of malaria species is crucial for patient management and epidemiological surveillance. This study aimed to determine the different Plasmodium species causing malaria in children under 5 years old in two provinces (Kinshasa and North Kivu) of the Democratic Republic of Congo (DRC). METHODS: From October to December 2015, a health-facility based cross-sectional study was conducted in General Reference Hospitals in Kinshasa and North Kivu. Four hundred and seven blood samples were collected from febrile children aged ≤ 5 years. Nested polymerase chain reaction assays were performed for Plasmodium species identification. RESULTS: Out of 407 children, 142 (34.9%) were infected with Plasmodium spp. and P. falciparum was the most prevalent species (99.2%). Among those infected children, 124 had a mono infection with P. falciparum and one with P. malariae. Mixed infections with P. falciparum/P. malariae and P. falciparum/P. vivax were observed in 6 (1.5%) and 8 (2.0%) children, respectively. The prevalence of infection was higher in females (64.8%) than in males (35.2%), p < 0.001. The age-specific distribution of infection showed that children of less than 2 years old were less infected (18.4%) compared to those aged above 2 years (81.6%), p < 0.001. CONCLUSION: Although this study showed clearly that the most prevalent species identified was P. falciparum, the findings demonstrate the existence of non-falciparum malaria, especially P. malariae and P. vivax among children aged ≤ 5 years living both Kinshasa and North Kivu Provinces in DRC.

Do Cryptic Reservoirs Threaten Gambiense-Sleeping Sickness Elimination?

Trypanosoma brucei gambiense causes human African trypanosomiasis (HAT). Between 1990 and 2015, almost 440000 cases were reported. Large-scale screening of populations at risk, drug donations, and efforts by national and international stakeholders have brought the epidemic under control with <2200 cases in 2016. The World Health Organization (WHO) has set the goals of gambiense-HAT elimination as a public health problem for 2020, and of interruption of transmission to humans for 2030. Latent human infections and possible animal reservoirs may challenge these goals. It remains largely unknown whether, and to what extend, they have an impact on gambiense-HAT transmission. We argue that a better understanding of the contribution of human and putative animal reservoirs to gambiense-HAT epidemiology is mandatory to inform elimination strategies.

Determinants of dog owner-charged rabies vaccination in Kinshasa, Democratic Republic of Congo.

Rabies is a preventable fatal disease that causes about 61,000 human deaths annually around the world, mostly in developing countries. In Africa, several studies have shown that vaccination of pets is effective in controlling the disease. An annual vaccination coverage of 70% is recommended by the World Health Organization as a control threshold. The effective control of rabies requires vaccination coverage of owned dogs. Identification of the factors determining dog owners' choice to vaccinate is necessary for evidence-based policy-making. However, for the Democratic Republic of Congo (DRC), the limited data on rabies vaccination coverage makes it difficult for its control and formulation of appropriate policies. A cross-sectional study was conducted in Kinshasa (Lemba commune) with dog-owning households and owned dogs as study populations. The association between dog vaccination and independent factors (household socio-demographics characteristics, dog characteristics, knowledge of rabies and location of veterinary offices/clinics) was performed with Epi-info 7. The Odds Ratio (OR) and p-value < 0.05 were used to determine levels of significance. A total of 166 households owning dogs and 218 owned dogs were investigated. 47% of the dogs had been vaccinated within one year preceding the survey which is higher than the critical coverage (25 to 40%) necessary to interrupt rabies transmission but below the 70% threshold recommended by WHO for control. The determinants of vaccination included socio-economic level of the household (OR = 2.9, p<0.05), formal education level of the dog owner (OR = 4, p<0.05), type of residence (OR = 4.6, p<0.05), knowledge of rabies disease (OR = 8.0, p<0.05), knowledge of location of veterinary offices/clinics (OR = 3.4, p<0.05), dog gender (OR = 1.6, p<0.05) and dog breed (OR = 2.1, p<0.05). This study shows that the vaccination coverage in this area can easily reach the WHO threshold if supplemented by mass vaccination campaigns.

Acute flaccid paralysis surveillance indicators in the Democratic Republic of Congo during 2008-2014.

INTRODUCTION: The last wild poliovirus (WPV) case in Africa was reported in July 2014, thus underscoring the tremendous progress towards polio eradication worldwide. This study aimed to analyze the results of a seven-year surveillance of Acute Flaccid Paralysis (AFP) in the Democratic Republic of Congo (DRC) and to identify potential gaps that need to be addressed. METHODS: Epidemiological and virological data obtained from AFP surveillance among AFP cases less than 15 years from January 2008 to December 2014 in DRC were retrospectively considered and analyzed in this study. RESULTS: Of the 13,749 AFP cases investigated, 58.9% received at least three doses of oral polio vaccine (OPV), 7.3% never received OPV, while the status of 18.3% was unknown. Analysis of surveillance performances showed that all, but two, indicators were below the required WHO-specified targets. Non-polio enterovirus (NPEV) isolation rate was consistently below the minimum requirement at ≥10% and the proportions of stool specimens that reached the laboratory within 72 hours of being sent were always below 15% (WHO target is ≥80%). Virus isolation and differentiation showed that 1.5% of AFP cases were infected by WPVs, 5.5% by Sabin strains, 0.5% by vaccine-derived polioviruses (VDPVs) and 7.2% by NPEVs. CONCLUSION: Our findings indicate that additional efforts are needed to address the timeliness of adequate stool specimens' arrival to the laboratory. It remains essential to maintain high polio vaccine coverage and high AFP surveillance standards to ensure rapid detection and containment of either WPV importation or VDPV re-emergence in DRC.

Development of an Immunochromatography Assay (QuickNavi-Ebola) to Detect Multiple Species of Ebolaviruses.

The latest outbreak of Ebola virus disease (EVD) in West Africa has highlighted the urgent need for the development of rapid and reliable diagnostic assays. We used monoclonal antibodies specific to the ebolavirus nucleoprotein to develop an immunochromatography (IC) assay (QuickNavi-Ebola) for rapid diagnosis of EVD. The IC assay was first evaluated with tissue culture supernatants of infected Vero E6 cells and found to be capable of detecting 103-104 focus-forming units/mL of ebolaviruses. Using serum samples from experimentally infected nonhuman primates, we confirmed that the assay could detect the viral antigen shortly after disease onset. It was also noted that multiple species of ebolaviruses could be detected by the IC assay. Owing to the simplicity of the assay procedure and absence of requirements for special equipment and training, QuickNavi-Ebola is expected to be a useful tool for rapid diagnosis of EVD.

Overview of Animal Rabies in Kinshasa Province in the Democratic Republic of Congo.

INTRODUCTION: Rabies is one of the major public health problems mostly affecting developing countries in Africa and Asia where 99.9% of all rabies related human deaths are recorded each year. In Democratic Republic of Congo, repeated outbreaks have been reported. Despite this, there is little reliable epidemiological data about rabies in the country for the development of effective control strategies. MATERIALS AND METHODS: A retrospective study was carried out in Kinshasa Province during a period of five years (2009-2013) to describe the proportion of rabid animals and the species involved in rabies transmission and maintenance. The survey also aimed at describing the spatial-temporal distribution of rabies. To gather information, the daily registers of institutions involved in rabies diagnosis were reviewed and each rabies case was traced back to area of occurrence for collection of geographic coordinates. RESULTS AND DISCUSSION: A total of 5,053 attacks were registered involving six animal species including dog, cat, monkey, rabbit, rat, and pig. Based on clinical observations, rabies was reported in dogs and cats while data obtained from the laboratory confirmed rabies cases included dogs, cats and a goat. The annual distribution showed a significant decrease of rabies cases from 2009 up to 2011 and a later increase up to 2013. There was no difference in rabies occurrence between seasons (p = 0.721). Rabies cases were three times higher in peri-urban zone than in urban zone OR = 3.4 (95% CI: 2.3-5.1). The positive proportion of rabies was 2.6% (95% CI: 2.1-3) based on clinical evidence and 65.9% (95% CI: 50-79.5) for laboratory confirmed cases. CONCLUSION AND SUGGESTION: This study confirms the endemicity of rabies in Kinshasa where occurrence of rabies cases was related to human population density and lifestyle. In order to control rabies, there is need to set up a surveillance program and implement efficient mass vaccination campaigns of susceptible animals.

Virulence of Trypanosoma congolense strains isolated from cattle and African buffaloes (Syncerus caffer) in KwaZulu-Natal, South Africa.

Trypanosoma congolense and Trypanosoma vivax are major species that infect cattle in north-eastern KwaZulu-Natal (KZN), South Africa. Of the two genetically distinct types of T. congolense, Savannah and Kilifi sub-groups, isolated from cattle and tsetse flies in KZN, the former is more prevalent and thought to be responsible for African animal trypanosomosis outbreaks in cattle. Furthermore, variation in pathogenicity within the Savannah sub-group is ascribed to strain differences and seems to be related to geographical locations. The objective of the present study was to compare the virulence of T. congolense strains isolated from African buffaloes (Syncerus caffer) inside Hluhluwe-iMfolozi Park, and from cattle on farms near wildlife parks (< 5 km), to isolates from cattle kept away (> 10 km) from parks. To obtain T. congolense isolates, blood of known parasitologically positive cattle or cattle symptomatically suspect with trypanosomosis, as well as isolates from buffaloes kept inside Hluhluwe-iMfolozi Park were passaged in inbred BALB/c mice. A total of 26 T. congolense isolates were obtained: 5 from buffaloes, 13 from cattle kept near parks and 8 from cattle distant from parks. Molecular characterisation revealed 80% and 20% of isolates to belong to T. congolense Savannah and Kilifi, respectively. To compare virulence, each isolate was inoculated into a group of six mice. No statistical differences were observed in the mean pre-patent period, maximum parasitaemia or drop in packed cell volume (PCV). Significant differences were found in days after infection for the drop in PCV, the patent period and the survival time. These differences were used to categorise the isolates as being of high, moderate or low virulence. Based on the virulence, 12 of 26 (46%) isolates were classified as highly virulent and 27% each as either of moderate or of low virulence. Whilst 11 of 12 high virulent strains were from buffaloes or cattle near the park, only 1 of 7 low virulent strains was from these animals. All the Kilifi T. congolense types were less virulent than the Savannah types. These results confirmed the higher virulence of T. congolense Savannah type compared to Kilifi type and indicated the prevalence of highly virulent strains to be higher in wildlife parks and in cattle near the parks than on farms further away. The geographical location of these strains in relation to the wildlife parks in the area was discussed.