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Detection and molecular characterization of lumpy skin disease and bovine papular stomatitis viruses in lumpy skin disease-suspected outbreaks in Tanzania

Virology Journal volume 21, Article number: 276 (2024) Cite this article

Abstract

Background

Lumpy Skin Disease (LSD) is endemic in sub-Saharan countries and is currently a global threat to the cattle industry. Information on the circulating Capripoxvirus lumpyskinpox, formerly known as Lumpy Skin Disease Virus (LSDV), and other poxviruses infecting cattle is very scant in Tanzania. The current study aimed to confirm and characterize LSDV and other poxviruses infecting cattle, from LSD suspected outbreaks in Tanzania.

Methods

A total of 24 samples were collected from four LSD suspected outbreaks reported in Tanzania between February and May 2023. Samples were screened for LSDV genome by real-time PCR and then subjected to a high-resolution multiplex melting (HRM) assay where 10 samples were positive for Capripoxvirus (CaPV) and one sample was Parapoxvirus (PPV) positive. Four LSDV genes; RPO30, GPCR, EEV glycoprotein and B22R and the partial B2L gene of PPVs were analyzed.

Results

All targeted LSDV genes from the Tanzanian isolates showed 100% similarity and isolates clustered with commonly circulating LSDV field isolates. Furthermore, the single nucleotide polymorphism (SNP) at position 240 (A-> G) of the EEV gene differentiates the Tanzanian LSDVs from the group of ancient Kenyan LSDV isolates while the B22R sequences of the Tanzanian LSDV isolates differed from the LSDV Neethling and LSDV KSGP-0240 derived vaccines. Sequence analysis of the partial B2L gene of the Tanzanian parapoxvirus bovinestomatitis, formerly known as Bovine papular stomatitis virus (BPSV) showed a different BPSV strain circulating compared to publicly available sequences.

Conclusion

These findings confirm the presence of LSDV in Tanzania, which suggesting the need for establishing an effective control program and continuous monitoring. The presence of a typical profile for Tanzania BPSV is an indication that, although never reported before, BPSV is established in the country therefore this virus should be included in the differential diagnosis of LSDV.

Introduction

Lumpy Skin Disease (LSD) is an economically important viral disease of cattle and water buffalo that threatens the cattle industry globally [1]. The disease is caused by the Capripoxvirus lumpyskinpox, formerly known as lumpy skin disease virus (LSDV) belonging to the genus Capripoxvirus (CaPV), Chordopoxvirinae subfamily, together with Capripoxvirus sheeppox and Capripoxvirus goatpox, formerly known as sheeppox virus and goatpox virus, respectively [2]. LSD is characterized by high fever, lymphadenopathy, nasal and ocular discharges, loss of appetite and, drop in milk production in lactating animals in early stages of infection followed by development of integumentary lesions over different parts of the body in advanced stages [3]. The characteristic LSD lesions are multiple, well circumscribed to coalescing, 0.5–5 cm in diameter, firm, flat-topped papules and nodules which can further develop into cone-shaped central core within the nodule [4]. LSD was first diagnosed in Zambia in 1929 and is currently known to be endemic in all of African countries except Morocco, Libya, Algeria, and Tunisia [5]. The first outbreak of LSD in Tanzania was reported in 1981, followed by a second outbreak in 1986 [6]. Currently, LSD is considered endemic in Tanzania and numerous outbreaks occur each year [7]. LSD has also become a disease of global concern due to its continuous expansion of geographical borders, with reports of outbreaks in the Middle East, Asia, and Europe [5, 8].

The spread of LSD between countries and regions has been associated with the illegal movement of cattle due to the porosity of borders and the international trade of cattle and cattle products [5]. Therefore, the success of different control measures requires a regional willingness to harmonize control strategies such as movement restriction, vaccination and biosecurity measures [9]. Vectors such as biting flies (Stomoxys calcitrans), ticks and mosquitoes play an important role in LSDV transmission and epidemiology [10, 11]. However, majority of the farmers in Tanzania are not aware of the role of vectors in LSD transmission, and therefore vector control is not considered as one of the means of limiting LSD spread [12]. Transmission via direct contact between infected and susceptible animals is rare but can occur especially in animals in intensive system [9]. Contaminated water and feed, medical treatment through needles and contaminated semen, are other routes for LSDV infection [3, 9].

LSD can clinically be confused with other skin conditions of cattle including diseases caused by other poxviruses [5, 11]. For instance, among the Chordopoxvirinae subfamily, parapoxvirus pseudocowpox (formerly known as pseudocowpox virus, PCPV) and parapoxvirus bovinestomatitis, previously known as Bovine papular stomatitis virus (BPSV) in the parapoxvirus (PPV) genera, are capable of infecting cattle and causing similar clinical presentation as capripoxviruses, therefore creating a challenge in clinical diagnosis [2, 13, 14]. PPVs are transmitted through broken skin or oral mucosa, followed by virus replication near the port of entry that leads to clinical signs that progresses through the stages of macules, papules, vesicles, pustules, and scabs [15, 16]. Reports of co-infection of LSD with PCPV, as well as with other viral diseases characterized by skin leasions, such as foot-and-mouth disease are available, but information on LSD co-infection with BPSV are limited [13, 17,18,19].

Vaccination is known to be the most cost effective means of controlling LSD especially in endemic areas [9]. However, the availability of effective and affordable vaccine against LSD is a global challenge [9, 10]. LSD control in Tanzania is mainly done through vaccination using a live attenuated homologous vaccine, which is known to confer immunity lasting up to 18 months; however, vaccine manufacturer’s recommend annual booster to be implemented [9, 20]. In most cases, vaccine acquisition and vaccination in Tanzania are coordinated by individual farmers. Therefore, the quality of the vaccines used, efficiency and coverage are not well established. This increases the risk of severe post-vaccination reactions and the development of recombinant virus strains, which can further complicate the epidemiology and control of the disease [21,22,23].

The LSDV genome has been regarded as relatively stable for years, with LSDV isolates grouped into two main clusters (1.1 and 1.2) based on the whole genome sequences [24, 25]. Several genomic regions such as P32, G-protein-coupled receptor (GPCR), EEV glycoprotein, RNA polymerase 30 kDa subunit (RPO30), and CaPV homolog of the variola virus B22R, have been used as reliable diagnostic tools to identify and distinguish isolates from the two clusters, as well as Neethling-derived vaccines from field isolates [26,27,28]. However, the recent outbreak of LSDV recombinant strains highlights the likelihood of genetic diversity and evolution within the LSDV genomes [29, 30]. Nevertheless, sequencing of more than one of the above LSDV marker genes in addition to whole genome sequencing to characterize the virus has led to the discovery of variability in LSDV genome and identification of new recombinant strains [21,22,23, 31]. The GPCR gene has become particularly important due to its great display of polymorphism [32] and has been used to detect LSDV strains with 12 nucleotide insertions in their GPCR gene resembling the vaccine strains [20]. Vaccine-like LSDVs have been reported in East African archived isolates that were isolated before reports of LSD outbreaks in the Middle East and Europe [21]. This indicates that vaccine-like LSDVs have been circulating in East Africa for years, suggesting the need for conducting studies to establish its distribution in the region. The use of multiple marker genes has been recommended by several researchers to increase the chances of detecting more variabilities in LSDV genomes [33]. For example, the application of multiple LSDV marker genes to characterize the vaccine-like LSDV from East Africa revealed its genetic difference from the vaccine-like viruses reported in Russia [21]. Since the severity of the disease is influenced by the virulence of the virus strain [34], there is evidence that recombinant strains discovered in Russia caused more severe diseases than the wild type [35]. It is therefore important to continuously monitor genomic changes and variability of circulating LSDVs to better support control programs.

Tanzania has the second largest cattle population in Africa; however, the contribution of the livestock subsector to the national GDP is still below its potential [36]. Diseases are among the major constraints to the performance and development of the livestock sector and its contribution towards improved livelihood. Despite the long history of LSD in Tanzania, the epidemiology and the molecular characteristics of LSDV and other poxviruses circulating in Tanzania have not been established. Therefore, the current study is the first to characterize the LSDV and PPV circulating in the country by sequencing and phylogenetic analysis of four LSDV genes, —GPCR, RPO30, EEV- and the B22R— as well as the B2L gene, which encodes the major virus envelope protein that is widely used to molecularly characterize the genetic variation of PPVs [37]. These data are important for understanding the molecular epidemiology and vaccine selection for disease control.

Material and method

Ethics approval and consent to participate

Approved by the Research and Publications Committee, Sokoine University of Agriculture, Morogoro, Tanzania with approval number SUA/DPRTC/R/186/031. Sampling carried out with the owners’ consent.

Study area, origin of samples and sample collection

The study was undertaken in four districts of Pangani, Kisarawe & Mkuranga and Sumbawanga (Fig. 1). The study districts are located in Tanga (Pangani) in the northern zone, Pwani (Kisarawe & Mkuranga) region, which is in the eastern zone of the country, and Rukwa (Sumbawanga) region on the southern highland zone. All the regions are characterized by agropastoral systems.

Fig. 1
figure 1

Map of Tanzania showing the study areas

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A total of 24 samples including blood, saliva, and tissues, were collected purposively from 12 herds suspected of LSD infection from four outbreaks between February and May 2023 (Table 1). The criteria for collecting the three types of samples aimed to understand the onset of the disease/clinical signs, since the diagnostic time window for LSDV in different sample matrices varies with time of the first appearance of clinical signs. The virus is detected on early stages of infection in saliva and blood, and in late stages of infection in nodule and scab samples [38]. The total number of cattle where the samples were collected ranged from 5 to 40 cattle per herd. LSD-suspected cattle had signs of generalized skin nodules, enlarged superficial lymph nodes, fever, nasal discharges, lacrimation, and loss in body condition.

Whole blood was collected through the jugular vein in vacutainer tube containing EDTA, and skin tissues were collected following the procedure specified as previously described [34]. Saliva samples were collected using sterile swabs and placed in a tube containing 1mL of phosphate-buffered saline (PBS). Samples were transported in a cool box with ice packs to the Centre for Infectious Diseases and Biotechnology (CIDB) in Dar es salaam, Tanzania, for processing and analysis. At the laboratory, tissues were thawed and washed in PBS. The skin tissues were then chopped into small pieces and mixed with PBS and grounded using a motor and pestle, followed by centrifugation at 3000 rpm for 15 min. The tubes containing the saliva samples were also centrifuged, and the resulting supernatant was transferred into 1 mL screw-caped cryovials and stored at -20 °C. Whole blood samples were stored at 4 °C until processed.

Table 1 Description of LSD outbreaks in Tanzania; the location of the herds and type of samples collected from the LSD suspected cases between February and May 2023
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DNA extraction and confirmation of LSDV and BPSV genomes in samples

DNA was extracted using a Qiagen kit (DNeasy Blood & Tissue Kit) following the instructions of the manufacturers. A total of 24 samples (12 tissue samples, 9 from blood and 3 from saliva) were first screened for LSD genome by real-time PCR following the described protocol of Bowden and co-workers [39]. The real-time PCR run included LSDV positive control and a nuclease-free water negative control. To determine any co-infection of CaPV with other poxviruses, the samples were subjected to a high-resolution multiplex melting (HRM) assay that is capable of detecting and differentiating poxviruses belonging to Orthopoxvirus, Capripoxvirus, and Parapoxvirus genera based on the amplicon melting temperature and profile [40]. The HRM assay was performed following the previously described PCR program using a 20 µl reaction volume containing 1X of SsoFast EvaGreen Supermix (Bio-Rad, USA), 200 nM each of the forward and reverse primers for each of the three genera, and 2 µl of template DNA [40]. The HRM assay included positive controls from capripoxviruses, orthopoxvirus and parapoxviruses, as well as a nuclease-free water negative control. The amplification plots were analyzed on Bio-Rad CFX Maestro™ Software version 1.1, and the HRM profiles analyzed using the Precision Melt Analysis™ Software version 1.2 (Bio-Rad, Hercules, CA, USA).

Targeted gene sequencing and characterization of Tanzanian LSDV and BPSV isolates

All CaPV and PPV-positive samples were further processed by PCR amplification using four CaPV-marker genes —RPO30, GPCR, EEV glycoprotein, and B22R— and the partial B2L gene of PPVs, as previously described [41, 42]. The PCR products of the samples successfully amplified for all targeted genes were purified and sent for Sanger sequencing at LGC Genomics (Germany) using both forward and reverse primers of each gene. Raw sequences of the successfully sequenced samples were assembled and edited using Vector NTI software version 11.5 (Invitrogen) [43]. Sequences for each targeted gene with representative CaPV or PPV sequences retrieved from GenBank, were then aligned on MEGA X software using the MUSCLE algorithm and the codon option [44]. The multiple sequence alignments of the EEV glycoprotein and B22R genes were visualized and edited on BioEdit (v7.2.6) [45]. Neighbor-joining trees based on the complete RPO30 and GPCR gene sequences and a maximum likelihood tree based on the partial B2L gene sequences were built using MEGA X, with the evolutionary distances computed using the Maximum Composite Likelihood method and 1000 bootstrap replicates. The phylogenetic trees were visualized using the Interactive Tree of Life (ITOL) tool [46].

Results

Field observations and confirmation of LSD cases

Four suspected LSD outbreaks from four districts of Tanzania, between February and May 2023, were investigated. The common clinical signs observed in LSD-suspected cattle cases were fever, depression, enlarged superficial lymph nodes, inappetence, skin nodules on different parts of the body, lacrimation, nasal discharges, and loss of body condition (Fig. 2).

Fig. 2
figure 2

Skin nodules (indicated by red arrows) in LSD-suspected cows

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Out of the 24 samples analyzed (12 tissue, 3 saliva, and 9 blood), 11 (46%) tested positive for CaPV by real-time PCR, of which 10 were tissue and one was a saliva sample. Further analyses using the poxvirus HRM assay confirmed the 11 samples as CaPV positive and detected BPSV in 1 tissue sample (Table 2).

Table 2 List of sample result status (CaPV or BPSV positive) by real-time PCR or the HRM assay
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Sequence analysis of the CaPV targeted genes

The four targeted CaPV genes from 10 out of the 11 LSDV-positive samples (except for EEV glycoprotein and GPCR genes in LSD_Tan23_NJ11) and the partial B2L gene of the BPSV-positive sample were successfully amplified and sequenced. The sequences for all Tanzanian samples were edited to 606, 1134, 327, 832 and 876 bp, and deposited to GenBank under the accession numbers PP331450 to PP331459 (complete RPO30 gene), PP331460 to PP331469 (complete GPCR gene), PP331470 to PP331479 (partial EEV glycoprotein), PP331480 to PP331489 (partial B22R), and PP505768 (partial B2L gene) (Table 3).

Table 3 Sequence data of the four targeted CaPV genes and the PPVs partial B2L gene of the Tanzanian samples
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The multiple sequence alignments of all targeted LSDV genes showed 100% similarity between the Tanzania LSDV isolates. The phylogenetic analysis based on the RPO30 (Fig. 3A) and GPCR (Fig. 3B) clustered the Tanzanian isolates in LSDV Cluster II with other commonly circulating LSDV field isolates encountered in Africa, the Middle East and Europe. This includes isolates such as LSDV Sudan/06-Obied (GenBank accession number GU119938, LSDV Egypt/89_Ismalia (GenBank accession number GU119947), and LSDV SERBIA/Buj/2016 (Genbank accession number KY702007) .

Fig. 3
figure 3

Neighbor-joining tree based on the complete gene sequences of CaPVs (A) RPO30 and (B) GPCR, with LSDVs from Tanzania, (in red) and visualized using iTOL. The evolutionary distances were computed using the Maximum Composite Likelihood method with 1,000 bootstrap replicates in MEGA X

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The multiple sequence alignment of the partial EEV gene showed that all Tanzanian LSDV isolates were 100% identical to the commonly circulating LSDV field isolates that have a 27-nucleotide insertion at base pairs 175–201. This deletion differentiates the LSDV field isolates from LSDV Neethling-derived vaccines and the LSDV recombinants from Southeast Asia and Russia. In addition, the single nucleotide polymorphism (SNP) at position 240 (A-> G) differentiates Tanzanian LSDVs (as well as common field LSDV isolates) from the group of ancient Kenyan LSDV isolates which are currently mostly encountered in South Asia (Fig. 4).

Fig. 4
figure 4

Multiple sequence alignments of the partial nucleotide sequences of the EEV glycoprotein gene. The Tanzania isolates (in red) were aligned with representative LSDV sequences retrieved from GenBank database. A 27-nucleotide insertion and the SNP present in Tanzanian LSDV isolates are highlighted in the boxes. The dots indicate the identical nucleotides in the alignment

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The B22R sequences of the Tanzanian LSDV isolates were 100% identical to the commonly circulating LSDV field isolates, such as LSDV SERBIA/Buj/2016 (GenBank accession number KY702007) and LSDV Russia/Dages/2015 (GenBank accession number MH893760) as well as to LSDVs from the group of ancient Kenyan isolates currently found in South Asia. However, they differ from LSDV Neethling and LSDV KSGP-0240 derived vaccines that have a nucleotide insertion at positions 102 and 745, respectively (Fig. 5).

Fig. 5
figure 5

Multiple sequence alignments of the partial nucleotide sequences of the B22R gene. The Tanzanian isolates (in red) are aligned with representative LSDV sequences retrieved from GenBank database. The nucleotide insertions in LSDV Neethling and LSDV KSGP-0240 vaccines, which are absent in the Tanzanian LSDV isolates, are shown in the blocks. The dots indicate the identical nucleotides in the alignment

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The phylogenetic analysis based on the partial B2L gene clustered the single Tanzanian parapoxvirus isolate in this study within the BPSV species, together with isolates from USA (GenBank accession numbers AY386265, KM875471, KJ137717), Mexico (GenBank accession number KJ137716), Germany (GenBank accession numbers PP565898, PP565899, KF478805) and Switzerland, (GenBank accession number PP565903) (Fig. 6A). However, the amino acids multiple sequence alignment of the partial B2L, showed 2 non-synonymous mutations (V > L and T > A) in the Tanzanian BPSV that were specific to BPSV isolates from Japan (GenBank accession number LC523627) and Switzerland (GenBank accession number PP565904) (Fig. 6B).

Fig. 6
figure 6

(A) Maximum likelihood tree based on partial B2L gene sequences of PPVs, using the HKY model and gamma-rate distributions, with 1000 bootstrap replicates in MEGA X. The tree includes BPSV samples from Tanzania in red and visualized using iTOL. (B) Amino acid sequence alignment showing the nonsynonymous mutations in the partial B2L protein of the Tanzanian BPSV in red, compared to those from GenBank database. The dots indicate the identical amino acids in the alignments

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Discussion

Although LSD is considered endemic in Tanzania based on some documented reports of unconfirmed cases, according to National Agriculture Census of 2019/2020, over 16% of the households keeping cattle, reported LSD cases in agricultural year 2019/2020 [7]. However, studies confirming the presence of LSDV in Tanzania and its molecular characterization are scant. Similarly, little is known about parapoxvirus infections in cattle in Tanzania. The present study confirmed and characterized LSDV in three regions and BPSV circulation in one region in Tanzania.

In this study, the mortality rate reported for LSD was low, whereby only two out of 12 herds reported mortality with an overall mortality rate of about 0.8% during the outbreaks in 2023. This finding agrees with the previous documented reports, where the LSD mortality rate in most cases is less than 10%, such as those reported in China (0.9%), Bangladesh (0.002%), and Nepal (0.0%) [5, 33, 41, 47]. The observed low mortality could be attributed to the low virulence of the LSDV strain circulating in the country, or to host factors such as the immunity status of the animals due to frequent exposure in endemic areas, as well as the breed or age of the animals. In this study, most outbreaks occurred in farms with indigenous cattle that are believed to be less susceptible to LSDV [38].

No viruses were detected in blood samples. LSDV was confirmed in saliva and skin tissue samples, while BPSV was detected in one tissue sample. This is consistent with previous studies suggesting that skin tissues, nasal swabs, and saliva are more suitable for LSDV genome detection than blood samples [32, 48, 49], due to the short viraemia period of LSDV (6 to 15 days post-infection) [38]. The detection of BPSV from one LSD suspected sample by HRM, followed by the subsequent characterization through sequencing of the B2L gene, highlights the importance of differential diagnosis of pox diseases. Interestingly, the sequence analysis of the partial B2L gene of the Tanzanian BPSV showed the circulation of a different BPSV strain compared to publicly available sequences. This creates a need to conduct further studies to fully understand the distribution of this disease in other regions of the country and to molecular characterize the circulating viruses. The presence of a unique profile for BPSVs in Tanzania suggests that, although never reported and possibly rarely included among differential diseases, BPSV is already established in the country. BPS can be confused with many other diseases presented by skin and erosive lesions, such as, foot-and-mouth disease (FMD) which is also endemic in Tanzania [50].

Multiple sequence alignment and phylogenetic analysis of the four targeted genes —RPO30, GPCR, B22R and EEV genes—, confirmed LSDV in all sequenced samples from three regions in Tanzania. Furthermore, all Tanzanian LSDV isolates were 100% identical, suggesting that the LSD outbreaks in the country are possibly caused by a common LSDV strain. Unrestricted animal movement between districts and regions could be a possible reason for the spread of the LSDV strain in many regions in the country. The circulating LSDV strain in Tanzania is also similar to the common African isolates reported in Namibia, Burkina Faso, Nigeria, and Egypt, as well as to isolates circulating in the Middle East and Europe, based on the phylogenetic analysis of the RPO30 gene sequences [13, 51].

Additionally, the B22R sequence analysis differentiated the Tanzania LSDV isolates from the LSDV Neethling and KSGP-0240-derived vaccines due to single nucleotide insertions in these vaccine strains. This suggests that only the wild-type LSDV is responsible for the current LSD outbreaks in Tanzania despite the weak vaccine policies in the country. It is necessary to evaluate the efficacy of the currently approved LSDV vaccines in Tanzania under field conditions before mass vaccination can be implemented. In a recent study however, the characterization of East African archived LSDV isolates from Ethiopia, Kenya, and Sudan, revealed the presence of LSDV resembling vaccine strain [21]. Therefore, due to the weak control and regulation of animal movement and their products between East African countries, the likelihood of the reported vaccine-like LSDVs spilling over to the neighboring countries including Tanzania is high. This justifies the need to investigate and characterize other LSDV strains that may be circulating in other regions of Tanzania not included in the current study. The similarity of Tanzania LSDV isolates with isolates such as LSDV Kenya (MN072619) further emphasizes the transboundary nature of LSDV. Animal movement and trade between these two neighboring countries (Kenya and Tanzania) could be the reason for the transmission of LSDV, thereby amplifying the circulation of similar strains between the countries. The involvement of vectors in the transmission of LSDV also contributes to the circulation of different LSDV strains across the borders between Tanzania and Kenya. It is therefore important to strengthen the control of animal movement for effective control of LSDV.

Conclusion

This is the first study of the molecular detection and phylogenetic analysis of LSDV and BPSV in Tanzania. The analysis based on the four targeted CaPV gene sequences (—RPO30, GPCR, EEV and B22R—) showed that the Tanzanian LSDV isolates are identical to the common LSDV field isolates encountered in Africa, the Middle East, and Europe, but different from the LSDV recombinants found in Russia and Southeast Asia. Based on the partial B2L gene analysis, the Tanzanian BPSV isolate clustered with other BPSVs from the USA, Germany, and Switzerland. Further studies need to be conducted to better understand the molecular epidemiology of LSDV and BPSV circulating in different regions of the country.

Data availability

DNA sequences generated and analyzed are available in GenBank under accession numbers PP331450 to PP331489 and PP505768.

Abbreviations

BPSV:

Bovine papular stomatitis virus

CaPV:

Capripox virus

CIDB:

Centre for Infectious Diseases and Biotechnology

DNA:

Deoxyribonucleic acid

EDTA:

Ethylenediaminetetraacetic acid

FMD:

Foot-and-mouth disease

GTPV:

Goatpox virus

GDP:

Gross Domestic Product

HRM:

High-resolution melting

GPCR:

G-protein coupled receptor

LSD:

Lumpy skin disease

LSDV:

Lumpy skin disease virus

PCPV:

Pseudocowpox virus

PCR:

Polymerase chain reaction

PPV:

Parapoxvirus

RP030:

RNA polymerase 30 KDa subunit

SPPV:

Sheeppox virus

TVLA:

Tanzania Veterinary Laboratory Agency

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Acknowledgements

Livestock farmers and animal health field officers at Pangani, Sumbawanga, Kisarawe and Mkuranga districts for their assistance and cooperation during samples collection. Shaban Motto (Biotechnologist, TVLA) for assisting us with the map of Tanzania.

Funding

This study was supported by the Tanzania Veterinary Laboratory Agency (TVLA), the Ministry of Livestock and Fisheries, The United Republic of Tanzania and the Veterinary Diagnostic Laboratory (VETLAB) Network initiative of Joint FAO/IAEA Division.

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Authors and Affiliations

  1. Tanzania Veterinary Laboratory Agency, P.O.BOX 9254, Dar es salaam, Tanzania

    Fredy T. Makoga, Jelly S. Chang’a, Charles Mayenga, Stella Bitanyi & Bishop Magidanga

  2. Animal Production and Health Laboratory, Joint FAO/IAEA Centre of Nuclear Techniques in Food and Agriculture, Department of Nuclear Sciences and Applications, International Atomic Energy Agency, Wagramer Strasse 5, P.O. Box 100, Vienna, A-1400, Austria

    Irene K. Meki, Tirumala B. K. Settypalli, Giovanni Cattoli & Charles E. Lamien

  3. Department of Microbiology, Parasitology and Biotechnology, College of Veterinary Medicine and Biomedical Sciences, Sokoine University of Agriculture, P.O.BOX 3020, Morogoro, Tanzania

    Emma Peter & Augustino Chengula

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  1. Fredy T. Makoga
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Contributions

Conceptualization and study designing: F.T.M, A.C, E.P, C.M, J.S.C, S.B; Investigation, sample collection and processing: F.T.M, B.M, J.S.C, C.M; Data analysis and interpretation: I.K.M, T.B.K.S, G.C, C.E.L, Manuscript writing: F.T.M, J.S.C, I.K.M, C.M, S.B, T.B.K.S, G.C, C.E.L. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Jelly S. Chang’a.

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Ethics approval and consent to participate

Approved by the Research and Publications Committee, Sokoine University of Agriculture, Morogoro, Tanzania with approval number SUA/DPRTC/R/186/031. Sampling carried out with the owners’ consent.

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Not applicable.

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The authors declare no competing interests.

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Makoga, F.T., Chang’a, J.S., Meki, I.K. et al. Detection and molecular characterization of lumpy skin disease and bovine papular stomatitis viruses in lumpy skin disease-suspected outbreaks in Tanzania. Virol J 21, 276 (2024). https://doiorg.publicaciones.saludcastillayleon.es/10.1186/s12985-024-02558-w

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Virology Journal

ISSN: 1743-422X