Molecular Epidemiology Of Rabies Virus Research Articles

Current status and molecular epidemiology of rabies virus from different hosts and regions in Malawi

Although rabies is endemic in Malawi, there have been no studies in which rabies virus was systematically investigated and characterized in multiple animal hosts in that country. In order to provide molecular epidemiological data on rabies virus in Malawi, 683 suspected rabies case reports from 2008 to 2021 were examined, and 46 (dog = 40, cow = 5, and cat = 1) viable rabies-positive brain samples archived at the Central Veterinary Laboratory (CVL), Lilongwe, Malawi, were analyzed genetically. The results showed an increase in the submission of brain samples from 2008 to 2010, with the highest number of submissions observed in 2020. Of the 683 case reports analyzed for the period under review, 38.1% (260/683) (CI: 34.44 – 41.84) were confirmed by direct fluorescent antibody test. Among the confirmed cases, 65.4% (170/260) (CI: 59.23 – 71.09) were canine rabies. Further, phylogenetic analysis revealed that sequences from different animal hosts clustered together within the Africa 1b lineage, suggesting that the strains circulating in livestock are similar to those in domestic dogs. This finding supports the hypothesis that canine rabies is spilling over to livestock and emphasizes the need for further studies to provide data for effective control of rabies in Malawi.

Rabies surveillance in Senegal 2001 to 2015 uncovers first infection of a honey-badger.

Despite the establishment of Rabies surveillance in animals and humans since 2008, there is a lack of data on its circulation, dynamic of transmission and real burden in Senegal. To better understand the molecular epidemiology of rabies virus in Senegal, we investigated the genetic diversity of 18 new characterized Senegalese rabies virus sequences collected over 14 years, including a honey-badger-related isolate. Phylogeographic analyses demonstrated that the Senegalese isolates belong to a monophyletic cluster into the Africa-2 clade and supported two RABV introductions in Senegal from West-African neighbour countries, 36-40 years ago. Our study is noteworthy for reporting on the first characterization of an African honey-badger-related rabies virus that did not have the N-glycosylation site NKT at position 338-G of the glycoprotein. The identified amino acid polymorphisms found in the Senegalese rabies virus sequences are worthy of further investigation. Although a strong multidisciplinary stepwise cooperation is important for the successful elimination of Rabies in dog populations in Senegal by 2030, the establishment of surveillance in wildlife could be necessary to avoid future re-introductions into domestic hosts.

Whole-genome sequencing and phylogenetic analysis of rabies viruses from Jordan.

Human fatalities caused by rabies are rarely reported in Jordan; however, domestic animals are more likely to fall victim to rabies compared to wild animals, at least this is the case in Jordan due to the presence of canine rabies. In this study, twelve brain samples from domestic and wild animals suspected of being infected with rabies virus from different regions of Jordan were collected during 2019. Seven of them tested positive using the fluorescent antibody test and real-time SYBR RT-PCR assay. Five specimens were from stray dogs and two from foxes. The whole genome sequences were obtained from the positive samples. Sequence analysis showed that one dog virus from Al Quwaysimah city located in Amman governorate, was closely related to an Israeli strain belonging to a Cosmopolitan ME1a clade. The genomes of the remaining six viruses (four from dogs and two from foxes) collected from different areas of Jordan were genetically-related to each other and clustered together with sequences from Iran and Turkey; all belong to Cosmopolitan ME2 clade. These sequences were analyzed with six other Jordanian rabies virus nucleoprotein (N) gene sequences available in the public database, five of them belong to ME1a clade and one belongs to ME1b clade. Rabies virus whole genome data is scarce across the Middle East. This study provides a better understanding of the molecular epidemiology of rabies virus in the region.

Molecular study of rabies virus in slaughtered dogs in Billiri and Kaltungo local government areas of Gombe state, Nigeria

Rabies is one of the most dreadful diseases and a major viral zoonosis which has been shown to cause an almost 100% fatality rate in infected victims. It is characterized by acute progressive encephalitis in mammals. This study determined the genotypic characteristics of rabies virus in dogs slaughtered for human consumption based on sequence of a fragment of nucleoprotein gene. Brain tissues were collected from 50 dogs slaughtered in Billiri and Kaltungo Local Government Areas of Gombe State, Nigeria. Direct fluorescent antibody test (DFAT) was used to screen for the presence of rabies virus antigen. Viral RNA isolated from DFAT positive brain tissues were subjected to the reverse transcription polymerase chain reaction (RT-PCR) followed by sequencing of the amplicons. Maximum Likelihood (ML) was used to construct a phylogenetic tree for sequences obtained with 1000 bootstrap replicates. The DFAT detected rabies antigen in 3 (6%) of the 50 dog brain tissues, from which 1 (2%) was positive by RT-PCR. ML phylogeny approach of the nucleotide sequences inferred members as originating lyssavirus genus and dog species. Essentially, MK234794 in this study displayed 99.3% sequence similarity with other related rabies viruses in the Africa 2 cluster (Nigeria, Cameroon, Chad and Niger). Interestingly, MK234794 showed no cluster relation with the Africa 1a, 1b, 3 and Africa 4 clades, respectively. This indicates there is in-country and trans-boundary circulation of the rabies viruses with no co-circulation between the Africa lineages, especially as dogs are continuously being traded due to consumption of dog meat in West Africa. This finding has given additional insight into the molecular epidemiology of rabies virus in Nigeria, therefore providing more baseline information for future design of rabies control programs in the country.

Genomic sequencing, evolution and molecular epidemiology of rabies virus.

Advances in sequencing techniques, improved computational methods of sequence interrogation and more accurate collection of epidemiological data through Global Positioning System (GPS) technology are improving our ability to monitor rabies outbreaks and better understand the processes that affect viral spread, evolution and host restriction. Whole-genome sequencing of rabies viruses (RABVs), using a range of different methodological approaches, is becoming more widespread and permits evolutionary and epidemiological studies at an unprecedented rate. Such studies are yielding insights into the fundamental processes of viral evolution, including molecular mechanisms of host adaptation and viral emergence in novel hosts. In addition, sequence data are revealing the importance of both landscape features and anthropomorphic activities as drivers of rabies spread; knowledge that is crucial for disease control efforts. This review summarises the state of the RABV genomics field and suggests how the above-mentioned approaches can be used to further understand and develop intervention strategies for rabies in the future.

Molecular epidemiology of rabies virus circulating in domestic animals in India.

Rabies is a neglected viral zoonotic disease affecting humans, domestic and wild animals and is endemic in most parts of the India. Dog mediated rabies is more predominant than other forms of rabies and molecular epidemiology is poorly understood in both reservoir and susceptible hosts. In the present study, a total of 140 rabies suspected brain samples from different species of animals from different geographical regions of India were used. The samples were parallelly tested by direct fluorescent antibody test, reverse transcriptase PCR and real-time PCR. Thirty positive samples were subjected for partial nucleoprotein gene sequencing and phylogenetic analysis. On sequence and phylogenetic analysis, it was observed that all Indian rabies viruses belonged to classical rabies virus of genotype 1 of Lyssavirus and formed two distinct groups. The majority of isolates were in group-1 and are closely related to arctic/arctic like lineage, whereas group-II isolated are closely related to cosmopolitan lineage. These results indicated there is simultaneous existence of two distinct lineages of rabies viruses in Indian subcontinent. Further whole genome studies are needed for better understanding of molecular epidemiology of rabies virus circulating in animals for control and prevention of rabies in India.

Molecular Epidemiology of Rabies Virus in the Territory of the Russian Federation

In this study, we report the phylogenetic relationship of RABV circulating in Russia. A total of 81 new isolates obtained from different regions of Russia in 2008 - 2016 were partially sequenced. Evolutionary analysis based on 100 representative sequences of rabies virus N-gene fragments (1110 nt) with sample time span ranging from 1977 to 2016 was made by the Bayesian coalescent analysis. Temporal estimates for major evolutionary events were done. The division of steppe rabies virus group C to discrete subgroups was revealed. According to our data, this most widespread in the Russian Federation group emerged in the beginning of the 1940th. The presence of steppe rabies virus was detected in the Far East (Primorsky Krai). The most likely cause of this event is anthropogenic introduction.. For the first time, representatives of arctic rabies subgroup A2 were isolated in the north of European Russia and FranzJosef Land. Evolutionary analysis suggested relatively recent emergence of all rabies virus groups currently circulating in the territory of the Russian Federation.

Molecular epidemiology of rabies virus in Nepal

Molecular epidemiology of rabies virus in Nepal

Molecular Epidemiology of Rabies Virus Isolates in Ethiopia

Rabies is a subject of public health concern in Ethiopia. The current rabies profile in Ethiopia shows 12 exposure cases/ 100,000 population and 1.6 rabies deaths /100,000 populations primarily because of canine rabies virus transmission exacerbated by uncontrolled stray dog population.The objective of this piece of work was to study the molecular epidemiology of rabies virus based on nucleotide sequence analysis of rabies virus (RABV) isolates originating from three selected Regional States of Ethiopia. Active surveillance of rabies suspected domestic animals that have demonstrated signs of encephalitis and abnormal behavioral change were subjected for Lyssavirus antigen detection using direct fluorescent antibody test (DFAT) positive and negative results. Ten DFAT positive brain samples comprising of three different animal species (Canine, Feline and Bovine) were sampled during 2013-2014 and subjected to reverse transcriptase PCR analysis. Ten brain tissue samples were collected from a Northern Regional State and Central and Western Regions of Ethiopia. The nucleotide sequences of these samples were studied to obtain an understanding of the epidemiological relationships of the RABV isolates in the various Ethiopian geographical locations. All samples submitted were positive by RT-PCR and no samples were negative. Ten amplified cDNA samples were sequenced and the sequence data was phylogenetically analyzed as indicated in the phylogenetic tree with RABV isolates from other countries (Fig.1). These results also indicate the postal transportation of FTA card is workable for the quality preservation of RNA samples under the specific period in time.

Molecular epidemiology of rabies virus isolated of herbivores from Brazilian Amazon

Amostras do vírus da raiva (n = 17) isoladas de bovinos (n = 11), equinos (n = 4) e bubalinos (n = 2) procedentes do Pará (n = 7), Tocantins (n = 6) e Rondônia (n = 4) foram submetidas à técnica de RT-PCR para amplificação parcial dos genes da Nucleoproteína (N) e Glicoproteína (G). As sequências nucleotídicas obtidas foram analisadas pelo método de reconstrução filogenética Neighbor-Joining com o modelo evolutivo Kimura 2-parâmetros. Todas as 17 amostras pertenceram ao cluster A, que se encontrou na linhagem associado com morcego hematófago Desmodus rotundus. A análise filogenética baseada nos genes N e G, sugere a presença de cinco sublinhagens (A1-A5) e sete sublinhagens (A1-A7), respectivamente. Quando se compara ambas as filogenias, as sublinhagens A1 até A3 mostram composição e distribuição geográfica concordante, já a diversidade observada na composição das sublinhagens restantes é atribuída ao uso de sequências de diferentes alinhamentos. A glicoproteína mostrou marcadores moleculares nas sublinhagens A2, A3, A4 e A7, o que fornece elementos para melhor compreensão da epidemiologia molecular da raiva das linhagens circulantes na Amazônia Brasileira.

Molecular epidemiology of rabies virus isolated of herbivores from Brazilian Amazon

Molecular epidemiology of rabies virus isolated of herbivores from Brazilian Amazon

Molecular epidemiology of rabies virus in Nepal

Molecular epidemiology of rabies virus in Nepal

Molecular epidemiology of rabies virus in Poland

The paper describes a phylogenetic study of 58 Polish isolates of rabies virus collected between 1992 and 2010. Sequences of the nucleoprotein (N) and glycoprotein (G) genes approximately 600 bp long were compared with reference sequences (GenBank) of European rabies viruses from neighbouring countries. The study confirmed a very high level of homology (94.4–100 %) of the Polish rabies virus strains irrespective of the date of isolation. Two variants of rabies virus: NEE (Northeastern Europe variant) and CE (Central Europe variant), depending on the geographical place of isolation, were circulating in Poland from 1992 to 2010. The Polish rabies virus isolates showed high similarity to European RABV strains, especially those collected in Ukraine and Romania. They were clearly different from vaccine strains SAD B19 and SAD Bern, which have been used for oral vaccination of foxes against rabies in Poland since 1993.

Molecular Epidemiology of Rabies Virus in Vietnam (2006–2009)

This study was aimed at determining the molecular epidemiology of rabies virus (RABV) circulating in Vietnam. Intra vitam samples (saliva and cerebrospinal fluid) were collected from 31 patients who were believed to have rabies and were admitted to hospitals in northern provinces of Vietnam. Brain samples were collected from 176 sick or furious rabid dogs from all over the country. The human and canine samples were subjected to reverse transcription-polymerase chain reaction analysis. The findings showed that 23 patients tested positive for RABV. Interestingly, 5 rabies patients did not have any history of dog or cat bites, but they had an experience of butchering dogs or cats, or consuming their meat. RABV was also detected in 2 of the 100 sick dogs from slaughterhouses. Molecular epidemiological analysis of 27 RABV strains showed that these viruses could be classified into two groups. The RABVs classified into Group 1 were distributed throughout Vietnam and had sequence similarity with the strains from China, Thailand, Malaysia, and the Philippines. However, the RABVs classified into Group 2 were only found in the northern provinces of Vietnam and showed high sequence similarity with the strain from southern China. This finding suggested the recent influx of Group 2 RABVs between Vietnam and China across the border. Although the incidence of rabies due to circulating RABVs in slaughterhouses is less common than that due to dog bite, the national program for rabies control and prevention in Vietnam should include monitoring of the health of dogs meant for human consumption and vaccination for workers at dog slaughterhouses. Further, monitoring of and research on the circulating RABVs in dog markets may help to determine the cause of rabies and control the spread of rabies in slaughterhouses in Vietnam.

Molecular characterization of Indian rabies virus isolates by partial sequencing of nucleoprotein (N) and phosphoprotein (P) genes

Rabies is endemic and an important zoonosis in India. There are very few reports available on molecular epidemiology of rabies virus of Indian origin. In this study to know the dynamics of rabies virus, a total of 41 rabies positive brain samples from dogs, cats, domestic animals, wildlife, and humans from 11 states were subjected to RT-PCR amplification of N gene between nucleotide N521-N1262 (742 bp) and P gene between nucleotide P239-P750 (512 bp). The N gene could be amplified from 30, while P gene from 41 samples, using specific sets of primers. The N gene-based phylogenetic analysis indicated that all Indian virus isolates are genetically closely related with a single cluster under arctic/arctic-like viruses. However, two distinct clusters were realized in P gene-based phylogeny viz., Rabies virus isolates of Punjab and Rabies virus isolates of remaining parts of India (other than Punjab). All the Indian rabies virus isolates were closely related to geography (>95% homology), but not to host species.

Molecular epidemiology of rabies virus in Romania provides evidence for a high degree of heterogeneity and virus diversity

Rabies has been present in Romania since antiquity and remains a continual threat to veterinary and public health. The current epizootic of rabies in Romania is fox mediated. However, little is known about the genetic diversity among rabies virus isolates from this part of the Balkan region. In this study, we analyzed a cohort of 64 rabies viruses isolated from infected animals in Romania collected between 2005 and 2008. Partial nucleoprotein sequence data (322 bp) were obtained and compared to a panel of available sequence data of Eurasian classical rabies virus (genotype 1) isolates. The Romanian sequences were identified as belonging to the cosmopolitan lineage of rabies viruses that have been reported throughout the world. The sequences showed a high degree of heterogeneity and six different lineages were identified. Single isolates clustered with the formerly established Eastern European group, the North-Eastern European group or with isolates from the Southwest of Russia. However two lineages were unique to Romania. This sequence diversity is unprecedented in European countries and could be a result of trans-border movement or remnants of previous epidemic waves.

Molecular epidemiology of rabies virus isolated from different sources during a bat-transmitted human outbreak occurring in Augusto Correa municipality, Brazilian Amazon

We genetically characterize rabies virus (RABV) strains isolated from human cases, domestic and wild animals during a human outbreak of bat-transmitted rabies in Augusto Correa municipality, Pará state, Brazilian Amazon in 2005. Partial nucleotide sequences of the N gene (491 bp) were obtained for all strains, and phylogenetic analysis grouped these into two major clades (Pará and Central-Southeast) and identified them as bat-related viruses genotype I, Desmodus rotundus antigenic variant 3 (AgV3). A molecular clock was used to estimate the time of emergence for each RABV isolate. The molecular data from this study suggest the association of vampire bats with human and domestic animal cases reported in the outbreak, the circulation of at least two predominant lineages in the Pará state, and also a geographic association to lineages dispersion.

Molecular Epidemiology of Rabies in Vietnam

The present study was done to determine the molecular epidemiology of rabies virus (RV) in Vietnam. The nucleoprotein (N) and glycoprotein (G) genes of RVs were amplified from the brains of ten rabid dogs of Ho Chi Minh City, Vietnam. The nucleotide sequences of these genes were compared with those of other Asian strains to find the possible relationship among them. Phylogenetic analysis revealed that the Asian N gene segregated into three main branches, namely South-East Asia 1 (SEA 1), South-East Asia 2 (SEA 2) and Indian subcontinent (ISC) genotypes. The SEA 1 genotype comprised RVs from Malaysia, Vietnam and Thailand. The SEA 2 genotype contained strains from the Philippines, and the ISC genotype comprised strains from Sri Lanka and India. Phylogenetically G genes of RVs from Vietnam and Thailand were clustered together. Our study suggests that Vietnamese and Thai RVs are closely related and might have originated from a common ancestor.

Molecular epidemiology of rabies virus isolates from South Korea

A molecular epidemiological study was performed on 13 Korean virus isolates, which were collected from wild and domestic animals diagnosed as rabid between 1998 and 2004. Seven samples were from domestic animals such as dogs and cattle infected by rabid raccoon dogs ( Nyctereutes procyonoides koreensis), and the rest of the six samples were from raccoon dogs in the wild. The study was carried out based on the comparison of nucleotide and amino acid sequences of nucleoprotein (N) and glycoprotein (G) coding regions and nucleotide sequence of the G-L intergenic (Ψ) non-coding region of the isolates. The similarities of nucleotide and amino acid sequence were at least 97.8 and 98.5%, respectively, between all Korean isolates. Phylogenetic analyses of the isolate showed that they formed a monophyletic group closely related to the Arctic strains but distant from other Asian strains, including Chinese strains. The fact that the raccoon dog is the main epidemic carrier of rabies in Korea and the results of these studies supported the conclusion of previous studies (Kuzmin et al.) that the raccoon dogs take part in the circulation of rabies virus within their natural territories in the Far East. The Korean isolates can be divided into two subgroups. All the topology of the most likelihood tree of Korean isolates using nucleotide and amino acid sequences of N, G and G-L region reflected not the species but the year of isolation and geographical location of the virus isolates. This study presents the detailed description of the molecular epidemiology of rabies virus in Korea.

Genetic typing of feline rabies virus isolated in greater Bangkok, Thailand.

To study the molecular epidemiology of rabies virus that is prevalent among cats in greater Bangkok, Thailand, a total of 17 rabies virus isolates from cats were characterized and compared with 120 rabies virus isolates from dogs. Analyses were performed on the genetic polymorphism in the rabies virus nucleoprotein (N) gene. Rabies virus N gene of isolates was amplified by reverse transcriptionpolymerase chain reaction. The diversity of N gene was revealed by the restriction fragment length polymorphism (RFLP) method. The rabies virus isolates from cats could be classified into 5 types, designated as Dd I-Hf I, Dd II-Hf II, Dd III-Hf I, Dd IV-Hf I, and Dd IV-Hf III. Type Dd I-Hf I was encountered more frequently than the others. It was apparent that no less than five rabies virus types presented in the areas of Bangkok. Moreover, all five RFLP patterns were typical of those which had been observed in dogs. Our findings suggest that there had been viral transmission between the dogs and the cats.