Evidence of coinfection of pigs with African swine fever virus and porcine circovirus 2.

Archival swine DNA samples from Indonesia and Mongolia, some of which were previously shown to be positive for African swine fever virus, were screened for the presence of porcine circovirus 2 (PCV-2) and porcine circovirus 3 (PCV-3) by PCR. Samples from both countries were positive for PCV-2 (three from Mongolia and two from Indonesia), while none were positive for PCV-3. The PCV-2 amplicons were sequenced, and phylogenetic analysis revealed that the PCV-2 strains belonged to four different genotypes: PCV-2a (Mongolia), PCV-2b (Mongolia and Indonesia), PCV-2d (Indonesia), and PCV-2g (Mongolia). This is the first report of ASFV/PCV-2 coinfection in pigs and the first report of the presence of PCV-2 in Mongolia.

Coinfections of African swine fever virus, porcine circovirus 2 and 3, and porcine parvovirus 1 in swine in Nigeria.

As pig production increases in Africa, it is essential to identify the pathogens that are circulating in the swine population to assess pig welfare and implement targeted control measures. For this reason, DNA samples collected from pigs in Nigeria in the context of African swine fever monitoring were further screened by PCR for porcine circovirus 2 (PCV-2), porcine circovirus 3 (PCV-3), and porcine parvovirus 1 (PPV1). Forty-seven (45%) pigs were positive for two or more pathogens. Sequence analysis identified PCV-2 genotypes a, b, and d, while limited genetic heterogenicity was observed among PCV-3 strains. All except one of the PPV1 sequences were genetically distinct from those previously identified in other countries.

Three different genotypes of porcine circovirus 2 (PCV-2) identified in pigs and warthogs in Namibia.

Porcine circovirus 2 (PCV-2) is the causative agent of porcine circovirus diseases (PCVD). A study was undertaken to determine whether PCV-2 was present in samples collected from commercial pigs (n = 46) and warthogs (n = 42) in Namibia between 2019 and 2020. Twenty-three of the collected samples were positive by PCR (13 from pigs and 10 from warthogs), and a phylogenetic analysis of ORF2 identified three genotypes (PCV-2b and PCV-2d in pigs and PCV-2c in warthogs). This is the first time that PCV-2 has been identified in warthogs and in Namibia. It is also the first report of PCV-2c in Africa.

"SILAB for Africa": An Innovative Information System Supporting the Veterinary African Laboratories.

Introduction: To support African veterinary laboratory services, the Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise puts in place an operational system called "SILAB for Africa" (SILABFA); this is a web application used by a laboratory information management system to support laboratory diagnostic activities and to meet the needs of various African countries. SILABFA was designed to collect and manage all necessary information on samples, tests, and test results.Methods: The system involves the entry of sample data on arrival, the tracking of samples through the various sections of the laboratory, and the collection of test results. It automates the generation of test reports and monitors outbreaks through data interrogation functions and eliminates multiple registrations of the same data on paper records. SILABFA is currently installed in Namibia, Botswana, Zambia, Zimbabwe, Tanzania, Uganda, Kenya, Ethiopia, and Cameroon, and installation in Senegal and Ivory Coast is planned for the next few months. After some years of SILABFA usage, it was natural to want to utilize more and more data collected in a homogeneous and consistent way for epidemiological purposes and to cover informative debts toward ministries and other organizations.Conclusion: To improve the availability of good, detailed, and reliable data, as the epidemiological information, SILABFA has been linked to the local animal identification, registration, and traceability system and other relevant national information systems.

Investigation of infectious laryngotracheitis outbreaks in Namibia in 2018.

Between July and August 2018, two outbreaks of infectious laryngotracheitis caused the death of over 116,000 commercial poultry (layers and broilers) near the city of Windhoek, Namibia. A third outbreak occurred in September 2018 in the north of the country approximately 800 km from the original outbreaks. Sample collection and molecular epidemiological analyses revealed that the outbreaks were most likely caused by poor vaccination practices leading to the reversion to virulence of an ILT vaccine strain. The analyses also indicate that inaccurate declarations were made by one of the farms involved and that illegal movement of animals most likely occurred.

Molecular characterization of lumpy skin disease virus in Namibia, 2017.

Between January and July 2017, lumpy skin disease (LSD) outbreaks were reported in cattle in Namibia. DNA was extracted from skin biopsies taken from 32 cattle, and the RNA polymerase 30 kDa subunit (RPO30) gene of the LSD virus (LSDV) was successfully amplified by PCR. Phylogenetic analysis revealed that the newly sequenced LSDV isolates from Namibia were identical to LSDV isolates identified previously in Burkina Faso, Egypt, Greece, Niger, Serbia and South Africa. Given that only unvaccinated herds were affected by LSD, it is recommended that the current vaccination programmes in Namibia be re-evaluated to allow nationwide coverage.

First genetic characterization of newcastle disease viruses from Namibia: identification of a novel VIIk subgenotype.

The complete sequences of the fusion (F) protein genes of six Newcastle disease virus (NDV) isolates from backyard poultry in Namibia in 2016 have been determined. The F gene cleavage site motif for all of the isolates was 112RRQKRF117, indicating that the viruses are virulent. A phylogenetic analysis using the full F gene sequence revealed that the viruses belong to a novel subgenotype, VIIk. This is the first genetic characterization of NDV isolates from Namibia, and the findings have important implications for Newcastle disease management and control in the region.

Rift Valley Fever Virus among Wild Ruminants, Etosha National Park, Namibia, 2011.

After a May 2011 outbreak of Rift Valley fever among livestock northeast of Etosha National Park, Namibia, wild ruminants in the park were tested for the virus. Antibodies were detected in springbok, wildebeest, and black-faced impala, and viral RNA was detected in springbok. Seroprevalence was high, and immune response was long lasting.

Novel genotype of Tritrichomonas foetus from cattle in Southern Africa.

Bovine trichomonosis caused by Tritrichomonas foetus is a significant reproductive disease of cattle. Preputial samples were collected using sheath washing technique in bulls in Namibia. Thirty-six trichomonad cultures were characterized using the TaqMan-probe commercial real-time polymerase chain reaction (PCR) diagnostic assay (VetMAX™-Gold Trich Detection Kit) and CYBR real-time PCR assay based on TFR3/4 primers. Diagnostic real-time PCRs and DNA sequencing of the internal transcribed region confirmed presence of T. foetus in 35 out of 36 samples. Multilocus genotyping using cysteine proteases (CP1, CP2, CP4, CP5, CP6, CP7, CP8, CP9) and malate dehydrogenase (MDH1) gene sequences demonstrate that the T. foetus in Namibia are genetically distinct from those characterized elsewhere. We report the discovery of a novel genotype of T. foetus in Namibian cattle, distinct from other T. foetus genotypes in Europe, South and North America and Australia. We suggest recognition of a 'Southern African' genotype of T. foetus. Identification of the new genotype of T. foetus demonstrates the need for wider global sampling to fully understand the diversity and origin of T. foetus causing disease in cattle or cats.

Six underreported viral diseases of domesticated and wild swine in Africa: Implications and perspectives.

Pig production is increasing annually in Africa as it is recognized as a significant source of income, livelihood and food security, particularly in rural communities. Understanding the circulating swine pathogens is crucial for the success of this emerging industry. Although there is extensive data available on the African swine fever virus due to its devastating impact on pig production, knowledge about the presence of other viral swine pathogens on the continent is still extremely limited. This review discusses what is currently known about six swine pathogens in Africa: classical swine fever virus, porcine reproductive and respiratory syndrome virus, porcine circovirus-2, porcine circovirus-3, porcine parvovirus-1, and pseudorabies virus. Gaps in our knowledge are identified and topics of future focus discussed.

Molecular survey of feline immunodeficiency virus (FIV) infection in Namibian cats.

Feline Immunodeficiency Virus (FIV) is one of the most important infectious diseases of cats, with potential implications in wildlife conservation. Unfortunately, FIV screening and surveillance in domestic cats remains limited in several African countries, including Namibia. In this study, 279 blood samples from domestic cats in Namibia were analyzed for FIV diagnosis by PCR. The cats represented various regions and were cared for by people largely from rural areas with limited financial means. Only 1.43 % of the samples tested positive, unexpectedly low given their outdoor lifestyles. The infected cats, primarily adult and unsterilized, showed no typical FIV symptoms, suggesting subclinical infections. Genetic analysis of the detected strains indicated a unique FIV strain cluster in Namibia, although with a certain within-country variability, in the absence of consistent geographical clustering. The present study represents the first detection and genetic characterization of FIV in the Namibian domestic cat population. Although the infection frequency was low, also in the rural free-roaming population, the features of the enrolled population could have biased the estimation, suggesting the need for more extensive surveys involving diseased and older cats as well. Additionally, because of the long-lasting subclinical nature of the infection, frequent monitoring activities should be performed that allow prompt isolation of infected animals and the implementation of appropriate control measures if necessary.

High detection frequency and genetic diversity of porcine circovirus 3 (PCV-3) in Namibian backyard farms and warthogs.

Since its first identification in 2015, porcine circovirus 3 (PCV-3) has been reported worldwide with a high frequency and in the presence of several clinical conditions, although its impact on pig health and productivity is still debated. Data on the presence of PCV-3 in Africa are, however, limited. A previous study performed on commercial pigs in Namibia failed to identify the pathogen. In the present study, the viral circulation in backyard farms, characterised by lower biosecurity measures and frequent animal exchange between farms, was assessed. The susceptibility of warthogs to PCV-3 infection and their potential epidemiological role were also evaluated. Tonsils from 77 pigs from backyard piggeries and 55 warthogs were collected in different regions of Namibia and tested by PCR. Positive samples were sequenced and compared to PCV-3 strains circulating globally. Forty-two out of 77 pigs (54.54 %) and 12 out of 55 warthogs (21.82 %) tested positive, demonstrating the presence of PCV-3 in the country and suggesting that the high biosecurity measures implemented in the commercial farms that previously tested negative for PCV-3 probably prevented viral introduction. The partial ORF2 gene was successfully sequenced in samples from 27 pigs and 6 warthogs. Genetically, the identified strains were part of 3 distinct groups which included both backyard pigs and warthogs from different regions of Namibia. There is also evidence for the occurrence of multiple introduction events most likely from Asian countries, either directly into Namibia or through other African countries. Considering the strict Namibian regulations on live animal importation, understanding the source of viral introduction is challenging, although semen importation or the habit of feeding backyard pigs with human food waste might have played a role. Pig exchanges between farms for breeding purposes or wildlife movements could also have been involved in PCV-3 dispersal within Namibia. Despite the significant advances in the field, further studies should be undertaken to properly understand PCV-3 epidemiology in Namibia and its impact on pig productivity and wildlife health.

Rift Valley fever in Namibia, 2010.

During May-July 2010 in Namibia, outbreaks of Rift Valley fever were reported to the National Veterinary Service. Analysis of animal specimens confirmed virus circulation on 7 farms. Molecular characterization showed that all outbreaks were caused by a strain of Rift Valley fever virus closely related to virus strains responsible for outbreaks in South Africa during 2009-2010.

Evidence indicating transmission of porcine parvovirus 1 between warthogs and domestic pigs in Namibia.

Thirty swine samples collected from different regions of Namibia between 2019 and 2020 were screened for the presence of porcine parvovirus 1 (PPV1) by PCR. Eleven samples (37%) were positive. Phylogenetic analysis of a partial sequence of the structural protein gene (VP2) identified two distinct clusters, one which contained sequences that were highly similar to PPV1 previously identified in warthogs in Namibia. These results indicate possible PPV1 transmission between warthogs and domestic pigs and highlight the importance of wildlife as sources of pathogens.

Investigation and sequence analysis of psittacine beak and feather disease virus and avian polyomavirus from companion birds in Windhoek, Namibia.

The commercial farming and trading of parrots and ornamental birds as companion animals are important economic activities in many countries. Some of the bird species farmed/traded are captured from the wild or are closely related to wild birds and therefore represent a risk of pathogen exchange/introduction. Beak and feather disease virus (BFDV) and avian poliomavirus (APV) are among the viruses with the biggest impact on companion bird populations and have been detected in different hosts worldwide. Despite their relevance for both domesticated and wild birds, our knowledge of BFDV and APV epidemiology remains limited in several African countries. In the present study, 143 cloacal swabs were collected from companion birds in Windhoek, Namibia, and tested by polymerase chain reaction for BFDV and APV. Of the samples tested, 35/143 (24.48%) tested positive for BFDV; 11/143 (7.69%) were positive for APV; and 6/143 (4.2%) tested positive for both pathogens. Positive amplicons, consisting of segments of the ORF1 and VP1 genes, were sequenced and compared with sequences from viruses identified in other countries. Four Namibian-only clades of BFDV were identified, loosely related to foreign strains, which suggest the occurrence of multiple introduction events in the past, potentially from South Africa, followed by local, independent evolution. In contrast, the Namibian APV sequences were identical to each other and form a single clade. In both instances, no correlation was observed between the sampling host and the viral phylogeny, suggesting the absence of host-specific adaptation and a remarkable, unconstrained viral circulation within Namibian borders. Therefore, while regulations and control measures developed against foreign strain introduction have proven to be effective over time, the spread of BFDV and APV within Namibia's borders appears undeterred. Additional resources should be dedicated to limit strain circulation in commercial farming facilities, markets and small-scale traders.

Molecular epidemiology of Canine circovirus in domestic dogs and wildlife in Namibia, Africa.

Canine circovirus (CanineCV) is a DNA virus affecting domestic dogs and other wild carnivore species. Despite the potential implications for dogs' health and wildlife conservation, data on CanineCV presence, epidemiology and genetic features from Africa is still poor. In the present study, biological specimens collected between 2020 and 2022 from a total of 32 jackals and 575 domestic dogs were tested for the presence of CanineCV DNA to evaluate its frequency. Furthermore, sequencing was conducted on positive samples to characterize the strains and compare them with publicly available sequences through phylogenetic analysis. A high CanineCV prevalence was observed both in jackals (43.75%; 95 CI: 28.17% - 60.67%) and domestic dogs (27.13%; 95 CI: 23.66% - 30.91%). All aside from one Namibian strain formed an independent clade, suggestive of extremely rare introduction events, followed by local persistence, circulation, and evolution. Remarkably, different recombination events were observed involving strains from both jackals and domestic dogs, which testify to the likely strain exchange between these populations. Distinctive amino acid residues were also observed in jackals. The limitations of the considered host populations however prevent a definitive conclusion on host adaptation, biological, and clinical features. Further studies should be performed to expand our current knowledge of the CanineCV disease scenario in Namibia, other African regions, and associated host species in Africa.

Whole-Genome Sequence and Assembly of Eight Africa Horse Sickness Virus Strains Collected in Namibia and South Africa.

In this report, we describe eight complete genome sequences of African horse sickness virus (AHSV) strains belonging to four different serotypes, namely, AHSV-5, AHSV-6, AHSV-8, and AHSV-9. Samples were collected in Namibia and South Africa from infected horses between 2000 and 2011. As expected, phylogenetic analyses of the variable outer capsid protein VP2 genomic sequences of AHSV-6 and AHSV-8 show higher nucleotide identity between the isolated viruses than that of the relevant reference strains. The full-genome sequence of AHSV will provide useful information on its geographical origin, and it will also be instrumental for comparing the distribution of the Namibian isolate with that of global isolates.

Low Seroprevalence of WNV in Namibian Dogs Suggests a Limited Effectiveness as Sentinels for Infection Monitoring.

West Nile virus (WNV) is an important zoonotic Flavivirus responsible for mild fever to severe neurological disease in humans and horses. Despite the occurrence of major previous outbreaks in Namibia and the likelihood of the current endemicity of the virus, only limited investigations and monitoring activities of WNV have been performed in the country. The use of animal sentinels is a valuable approach toward investigating the infection presence in an area and to predict the potential occurrence of human outbreaks. Serological investigations in dogs hold several advantages, considering their infection susceptibility, the ease of sample handling, and the evaluation of risk factors of pet owners that share the same habit with their pets. To evaluate the usefulness of such a sero-epidemiological investigation in Namibia, a broad serosurvey was performed in 2022 that included 426 archived domestic dog samples from eight Namibian regions. Although the ELISA prevalence, indicative of Flavivirus infection, was relatively high (16.43%; 95 CI: 13.10-20.39%), the virus neutralization test confirmed only a minority of cases, highlighting a prevalence of 2.82% (95 CI: 1.47-4.90%), significantly lower than in Namibian donkeys and reports from other countries. Variables that could explain the recorded differences remain to be explored, including animal exposure, variable vector presence, distribution, and feeding preferences. The study results suggest the limited usefulness of dogs as sentinels for WNV monitoring in Namibia.

First detection and molecular characterization of porcine reproductive and respiratory syndrome virus in Namibia, Africa.

Introduction: The swine sector in Africa plays an important role in local economies, contributing to poverty alleviation and community subsistence. In addition, intensive farming is progressively becoming more important in the region. Therefore, any disease affecting swine populations can have detrimental effects on local communities. Porcine Reproductive and Respiratory Syndrome (PRRS) is among the most important infectious diseases affecting swine worldwide, but information on its epidemiology in Africa is extremely limited. Material and methods: In the present study, 147 healthy butchered pigs, originating from 15 Namibian intensive and rural farms were tested by RT-PCR and the ORF7 genes of positive samples were sequenced for further genetic characterization and phylogenetic analysis. Additionally, 55 warthogs were also evaluated using the same approach. Results: Overall, 7 out of 147 pigs (4.76%) tested positive, all originating from 3 rural farms (with a within-herd detection frequency higher than 14%) characterized by strong epidemiological links. All industrial pig and warthog samples were negative. Sequence analysis revealed that all strains belonged to the Betaarterivirus suid1 species, previously known as PRRSV type I, and were likely imported from Europe at least 6 years ago, evolving independently thereafter. When and how the first introduction occurred could not be determined due to the absence of other African sequences for comparison. Discussion: The present work provides the first detection and characterization of PRRSV molecular epidemiology in Namibia. Based on the present findings, the presence of the PPRSV appears marginal and limited to backyard farms. While biosecurity measures applied in industrial farms appear to be effective in preventing viral introduction, PRRSV circulation in rural settings still represents a potential threat, and considering the socio-economical implication of livestock diseases decreasing animal performances in rural areas, active monitoring should be encouraged to promptly act against emerging menaces and guarantee the welfare of local pig populations.

Highly pathogenic avian influenza H5N1 virus outbreak among Cape cormorants (Phalacrocorax capensis) in Namibia, 2022.

In January 2022, significant mortality was observed among Cape cormorants (Phalacrocorax capensis) on the west coast of Namibia. Samples collected were shown to be positive for H5N1 avian influenza by multiplex RT-qPCR. Full genome analysis and phylogenetic analysis identified the viruses as belonging to clade 2.3.4.4b and that it clustered with similar viruses identified in Lesotho and Botswana in 2021. This is the first genomic characterization of H5N1 viruses in Namibia and has important implications for poultry disease management and wildlife conservation in the region.