Rabies virus seroprevalence among dogs in Limpopo National Park and the phylogenetic analyses of rabies viruses in Mozambique

Rabies is considered a neglected disease among many developing Asian and African countries, including Mozambique, where its re-emergence is often attributed to low dog parenteral vaccination coverage. The objectives of this study were two-fold: (1) to assess the level of antibodies against rabies virus in dogs (n = 418) in Limpopo National Park (LNP), and (2) to genetically characterise selected rabies viruses from brain tissue samples collected in 2017 and 2018. To meet the first objective, we used the BioProTM Rabies blocking ELISA antibody kit, and the results were expressed as the percentage of blocking (%PB). Dog sera with PB ≥ 40% were considered positive for antibodies to rabies virus, whereas sera with PB < 40% were negative. Just under ninety percent (89.2%; n = 373) of dogs were seronegative, and the rest (10.8%; n = 45) had detectable levels of rabies virus-specific antibodies. All eight brain tissue samples were positive for rabies virus antigen using a direct fluorescent antibody test and amplified in a quantitative real-time PCR, but only five (n = 4 from dogs and n = 1 from a cat) were amplified in a conventional reverse-transcription PCR targeting partial regions of the nucleoprotein (N) and the glycoprotein (G) genes. All samples were successfully sequenced. Phylogenetically, the rabies viruses were all of dog origin and were very closely related to each other (Africa 1b rabies virus lineage). Furthermore, the sequences had a common progenitor with other rabies viruses from southern Africa, confirming the transboundary nature of rabies and the pivotal role of dogs in maintaining rabies cycles. The study demonstrates the principal application of the BioProTM rabies ELISA antibody for the detection of anti-lyssavirus-specific antibodies in the serum samples of dogs, and most importantly, it highlights the low levels of antibodies against rabies virus in this dog population.The Third Framework Agreement Program “FA 3 III DGD/ITM 2014–2016” Project: “Communities on the move: animal and human health challenges” from the University of Pretoria (UP) and the Directorate-General Development Cooperation and Humanitarian Aid (DGD)/the Institute of Tropical Medicine of Antwerp (ITM) and by Fundo para a Investigação Aplicada e Multissectorial-FIAM.https://www.mdpi.com/journal/pathogensdm2022Centre for Veterinary Wildlife StudiesVeterinary Tropical Disease

Enhanced diagnosis of rabies and molecular evidence for the transboundary spread of the disease in Mozambique

Rabies is a neglected zoonotic disease with veterinary and public health significance, particularly in Africa and Asia. The current knowledge of the epidemiology of rabies in Mozambique is limited because of inadequate sample submission, constrained diagnostic capabilities and a lack of molecular epidemiological research. We wanted to consider the direct, rapid immunohistochemical test (DRIT) as an alternative to the direct fluorescent antibody (DFA) for rabies diagnosis at the diagnostic laboratory of the Central Veterinary Laboratory (CVL), Directorate of Animal Science, Maputo, Mozambique. Towards this aim, as a training exercise at the World Organisation for Animal Health (OIE) Rabies Reference Laboratory in South Africa, we performed the DRIT on 29 rabies samples from across Mozambique. With the use of the DRIT, we found 15 of the 29 samples (52%) to be negative. The DRIT-negative samples were retested by DFA at the OIE Rabies Reference Laboratory, as well as with an established real-time Polymerase chain reaction, confirming the DRIT-negative results. The DRIT-positive results (14/29) were retested with the DFA and subsequently amplified, sequenced and subjected to phylogenetic analyses, confirming the presence of rabies RNA. Molecular epidemiological analyses that included viruses from neighbouring countries suggested that rabies cycles within Mozambique might be implicated in multiple instances of cross-border transmission. In this regard, our study has provided new insights that should be helpful in informing the next steps required to better diagnose, control and hopefully eliminate rabies in Mozambique.The National Research Foundation (NRF) of South Africahttp://www.jsava.co.zaam2017Microbiology and Plant Patholog

Improvement of a liquid phase blocking ELISA for enhanced detection and measurement of antibodies against the SAT3 serotype of FMDV

Foot-and-mouth disease virus (FMDV) is the causative agent of one of the most serious infections of cloven-hoofed animals. An outbreak of FMD not only severely decreases livestock productivity, but also impacts on both the local and export trade of susceptible animals and their products. The Southern African Territories (SAT) types 1-3 are endemic to sub-Saharan Africa and display greater intratypic genomic and antigenic variation than the traditional Euro-Asian types. Southern Africa has an abundant wildlife, especially in National Parks and game reserves. Wildlife, particularly African buffalo (Syncerus caffer), is involved in virus maintenance and epidemiology of the disease. In communities within the proximity of National Parks and game reserves, the wildlife-livestock interface presents a challenge and poses difficulty to livestock disease eradication and control in Africa. In this study, the influence of modifications to the reagents has on the specificity, sensitivity and repeatability of a LPBE, used for the detection of antibodies against FMD, was determined. The sensitivity of the LPBE is dependent on the antigen used in the test and the ability of the sera to cross-react with the antigen. The purified and non-purified virus used as antigen and the capture and detector antibodies were prepared and standardized for this purpose. An attempt was made to reduce the subtype-specificity of the LBPE by including antigens from all the relevant SAT3 strains. A total of 515 sera from FMDV exposed cattle in Mpumalanga during 2011-2012; 1398 sera from unexposed cattle obtained during an FMD survey conducted in the Northern Cape, and 286 sera from FMDV vaccinated cattle next to the Kruger National Park (KNP) were tested with the improved ELISA. A statistical analysis was conducted to compare the results obtained with the newly developed ELISA and the current in house ELISA. The new assay has higher sensitivity for detecting antibodies in vaccinated animals compared to the standard LPBE. The test is specific and suitable for detection of antibodies, and plays a key role toward the control of FMD, specific and suitable for identification and typing of all SAT3 serotype across the range of the genetic variations in the SAT3 serotype of FMDV.Dissertation (MSc)--University of Pretoria, 2016.tm2016Veterinary Tropical DiseasesMScUnrestricte

Enhanced diagnosis of rabies and molecular evidence for the transboundary spread of the disease in Mozambique

Rabies is a neglected zoonotic disease with veterinary and public health significance, particularly in Africa and Asia. The current knowledge of the epidemiology of rabies in Mozambique is limited because of inadequate sample submission, constrained diagnostic capabilities and a lack of molecular epidemiological research. We wanted to consider the direct, rapid immunohistochemical test (DRIT) as an alternative to the direct fluorescent antibody (DFA) for rabies diagnosis at the diagnostic laboratory of the Central Veterinary Laboratory (CVL), Directorate of Animal Science, Maputo, Mozambique. Towards this aim, as a training exercise at the World Organisation for Animal Health (OIE) Rabies Reference Laboratory in South Africa, we performed the DRIT on 29 rabies samples from across Mozambique. With the use of the DRIT, we found 15 of the 29 samples (52%) to be negative. The DRIT-negative samples were retested by DFA at the OIE Rabies Reference Laboratory, as well as with an established real-time Polymerase chain reaction, confirming the DRIT-negative results. The DRIT-positive results (14/29) were retested with the DFA and subsequently amplified, sequenced and subjected to phylogenetic analyses, confirming the presence of rabies RNA. Molecular epidemiological analyses that included viruses from neighbouring countries suggested that rabies cycles within Mozambique might be implicated in multiple instances of cross-border transmission. In this regard, our study has provided new insights that should be helpful in informing the next steps required to better diagnose, control and hopefully eliminate rabies in Mozambique

H7N6 highly pathogenic avian influenza in Mozambique, 2023

ABSTRACTOn 13 October 2023, the National Directorate for Livestock Development in Mozambique was notified of a suspected outbreak of avian influenza in commercial layers. Samples were screened by real-time and conventional RT–PCR and were positive for both H7 and N6. Full genome sequences were obtained for three representative samples. Sequence analysis of the H7 cleavage site confirmed that the viruses were highly pathogenic (i.e. 333- PEPPKGPRFRR/GLF-346). In addition, the H7 and N6 sequences were highly similar (from 99.4-99.5% and 99.6-99.7% for the HA gene and the NA gene, respectively) to the sequences of a H7N6 virus identified in the Republic of South Africa in May 2023 indicating a similar origin of the viruses. The identification of H7N6 HPAIV in Mozambique has important implications for disease management and food security in the region

H7N6 highly pathogenic avian influenza in Mozambique, 2023

On 13 October 2023, the National Directorate for Livestock Development in Mozambique was notified of a suspected outbreak of avian influenza in commercial layers. Samples were screened by real-time and conventional RT–PCR and were positive for both H7 and N6. Full genome sequences were obtained for three representative samples. Sequence analysis of the H7 cleavage site confirmed that the viruses were highly pathogenic (i.e. 333- PEPPKGPRFRR/GLF-346). In addition, the H7 and N6 sequences were highly similar (from 99.4-99.5% and 99.6-99.7% for the HA gene and the NA gene, respectively) to the sequences of a H7N6 virus identified in the Republic of South Africa in May 2023 indicating a similar origin of the viruses. The identification of H7N6 HPAIV in Mozambique has important implications for disease management and food security in the region.</p

A Phylogeographic Analysis of Porcine Parvovirus 1 in Africa

Porcine parvovirus 1 (PPV1) is recognized as a major cause of reproductive failure in pigs, leading to several clinical outcomes globally known as SMEDI. Despite being known since the late 1960s its circulation is still of relevance to swine producers. Additionally, the emergence of variants such as the virulent 27a strain, for which lower protection induced by vaccines has been demonstrated, is of increasing concern. Even though constant monitoring of PPV1 using molecular epidemiological approaches is of pivotal importance, viral sequence data are scarce especially in low-income countries. To fill this gap, a collection of 71 partial VP2 sequences originating from eight African countries (Burkina Faso, Côte d’Ivoire, Kenya, Mozambique, Namibia, Nigeria, Senegal, and Tanzania) during the period 2011–2021 were analyzed within the context of global PPV1 variability. The observed pattern largely reflected what has been observed in high-income regions, i.e., 27a-like strains were more frequently detected than less virulent NADL-8-like strains. A phylogeographic analysis supported this observation, highlighting that the African scenario has been largely shaped by multiple PPV1 importation events from other continents, especially Europe and Asia. The existence of such an international movement coupled with the circulation of potential vaccine-escape variants requires the careful evaluation of the control strategies to prevent new strain introduction and persistence