National Veterinary Institute

Background: Brucella canis is an emerging zoonotic pathogen responsible for canine reproductive disorders and public health concerns. This study assessed the seroprevalence of B. canis in dogs from Campania, Southern Italy (2022-2025). Methods: Serum samples (n = 400) were retrospectively screened using an indirect immunofluorescence assay (IFAT), performed according to the manufacturer's instructions. Screening was conducted at a 1:40 cut-off, followed by serial dilutions to determine endpoint titres. Statistical analysis included chi-square tests for univariable screening, followed by nominal logistic regression models to evaluate the association between IFAT positivity and predictive factors (year, province, and sex of dogs). Additionally, a general linear model (GLM) was applied to the seropositive subset (n = 69) to analyse the magnitude of the antibody response, expressed as geometric mean titres (GMTs). Results: The overall seroprevalence was 17.3% (95% CI: 13.6-21.0%). Dog's sex, year of sampling, and province were not significant independent predictors of infection (p > 0.05), but GLM analysis showed that sampling year (p = 0.0024) and province (p = 0.0490) significantly influenced antibody intensity. A significant temporal increase in antibody intensity was observed towards 2025 (p = 0.037), suggesting an intensification of infection pressure. Conclusions: Our results confirm that Brucella canis is an endemic pathogen in the shelter dog population of southern Italy. The high seroprevalence and significant increase in antibody magnitude (GMT) over the study period indicate rising infection pressure, highlighting the urgent need for mandatory screening and a coordinated One Health surveillance strategy to manage zoonotic risk effectively.

Porridge is a culturally important and nutritionally valuable food, widely consumed across nations and age groups. In Ethiopia, it is traditionally prepared for lactating women to support postpartum recovery, often using barley as the primary ingredient. Lactating mothers and their offspring face significant nutritional challenges due to their heightened dietary requirements. However, nutrient rich and healthy local crops as tef, chickpea, and pumpkin offer valuable fortification, helping to address nutritional issues. This study aimed to enhance the nutrient content and sensory acceptance of traditional porridge for lactating mothers. Formulation incorporated barley (0%–100%), brown tef (0%–70%), chickpea (0%–30%), and pumpkin (10%) to optimize nutritional benefits by using a D–optimal mixture design. A standard porridge from barley flour (BF) (100%) served as the benchmark. The nutrient compositions, vitamin A, viscosity and sensory attributes were meticulously evaluated. The results revealed that increasing the amounts of tef and chickpea flour (CF) significantly boosted the porridge nutritional profile. Porridges enriched with CF exhibited elevated levels of protein, fat, and calories, whereas those with higher tef content had abundant calcium, iron, and fiber. A blend comprising up to 15% chickpea with BF yielded porridge with sensory qualities on par with that of barley alone. The optimal formulation, consisting of 41% barley, 30% tef, and 29% chickpea, was identified as the superior choice for developing a nutritious porridge with favorable sensory attributes. It significantly increased protein (22.85%), fiber (29.6%), calcium (70.4%), and iron (fivefold), while reductions in viscosity (44.45%) and overall acceptability (OAA) (23.7%) in the acceptable limits compared to control. This research offers insightful strategies for augmenting the nutritional value of traditional porridge, a staple for breastfeeding mothers in Ethiopia.

BackgroundAvian infectious bronchitis (IB) is a highly contagious respiratory disease that affects the poultry industry globally. The disease is caused by avian infectious bronchitis virus (IBV), member of the genus Gammacoronavirus. In Ethiopia, IBV has been reported in both commercial and backyard chickens based on clinical observation. The objectives of this study were to isolate the virus, conduct molecular based identification, and phylogenetic analysis of the circulating IBV isolates.Methods and materialsA cross-sectional study was conducted between November 2023 and May 2024 in Mekele and Bishoftu, Ethiopia. A total of 49 clinical samples were collected, comprising 12 tissue samples and 39 pooled swab samples. Of these, 6 samples—specifically, 5 swab samples and 1 tissue sample—tested positive for infectious bronchitis virus (IBV) through virus-specific conventional RT-PCR and real-time PCR. Nested PCR was performed using serotype-specific primers. The purified PCR products, which targeted the spike glycoprotein S1 subunit gene and the 3′ UTR of the IBV, were sequenced, followed by phylogenetic tree analysis.ResultsThe six positive samples propagated into specific pathogen free embryonated eggs and exhibited characteristic IBV lesions and mortality observed over five consecutive passages. IBV isolates from Bishoftu (n = 4) and Mekele (n = 2) were amplified using one-step RT-PCR to target 466 bp of the S1 subunit gene and 433 bp of the 3ʹUTR. A BLAST search on the S1 partial gene and 3ʹUTR sequences, nested PCR, and phylogenetic analysis revealed that the present IBV isolates are genetically similar to the Massachusetts serotype. The S1 gene sequences of the five IBV isolates were deposited in GenBank with accession numbers PQ389500 to PQ389504.ConclusionsThis is the first detailed study on IB virus isolation, molecular detection, sequencing, and phylogenetic analysis in Ethiopia. The findings revealed that the outbreaks were caused by the IB virus, which created a serious health risk and economic losses in the chicken industry. To the author’s knowledge, this is the first comprehensive study on the isolation and genetic analysis of IBV in Ethiopia. Further research on the economic impact of IBV in chicken production, farm biosecurity, serotyping of circulating IB virus, and vaccine development based on the local serotypes is recommended.

Livestock trade significantly contributes to the spread of transboundary animal diseases (TADs), like Peste des Petits Ruminants virus (PPRV), due to the long-distance movement of infected animals. This study investigates the structure and dynamics of the small ruminant trade network across 64 markets and 20 destinations, focusing on market connectivity, trade patterns, and disease risk. Eight key markets were identified due to their significant trading activity and connections. During the study period, 365,261 small ruminants were traded, with 54% directed toward market destinations and the remainder to slaughterhouses or cross-border locations, primarily Somaliland and Kenya. Over 85% of transport to these destinations was by truck, while more than 55% of internal movements were on foot. Trade peaked between July and September, accounting for 42% of the annual trade volume and 46% of cross-border movements. Network analysis revealed 172 distinct trade routes with varying activity levels: 12 persistent, 47 frequent, 42 intermediate, and 69 occasional. The network exhibited low connectivity (density = 0.024) and low reciprocity (0.104), with a negative assortativity (−0.52), indicating smaller markets often connected with larger ones. The network's clustering coefficient was 0.729, and the average path length was 2.216, contrasting with simulated random networks that had a clustering coefficient of 0.047 and an average path length of 3.22. Key markets were identified as “gate-keepers” and “pulse takers,” facilitating connections between isolated and central markets. The epidemic threshold (q) and R0 indicated a high risk of disease spread, especially from July to October. The study found that removing key markets drastically reduced network cohesion, with a 10% removal of central nodes decreasing the giant strongly connected component (GSCC) size by ~80%. Peste des Petits Ruminants (PPR) outbreaks in 2023 correlated strongly with market centrality, highlighting the need for targeted monitoring and control measures. This study underscores the importance of understanding network dynamics to manage trade efficiently and mitigate disease risks in small ruminant populations.

BackgroundFetal calf serum (FCS), an existing cell culture supplement, is effective but has several drawbacks, including being expensive, requiring a lengthy process of production, and requiring a hard currency. With this in mind, we planned to evaluate chick embryo extract and egg yolk extracts in cell culture as alternatives to fetal calf serum (FCS).MethodsSpecific pathogen-free eggs were purchased from the National Veterinary Institute, Bishoftu, Ethiopia, and incubated in a humidified incubator at 37 °C for 11 days. Egg yolk extract (EYE) and chick embryo extract (CEE) were collected after the egg was opened with caution not to destroy the yolk sack or the chick embryo itself. Chick fibroblasts and Vero cells were cultured in minimum essential medium (MEM) supplemented with egg yolk extract or chick embryo extract at ratios of 0:10, 1:9, 2.5:7.5, and 5:5% fetal calf serum.ResultsFibroblast cell attachment was better in media supplemented with 5% CEE and 5% FCS. The confluency was also greater than 50% at this concentration. Vero cells cultured with 5% CEE and 5% FCS also exhibited very good cell attachment and a confluency of up to 70%. Viability and confluency were also observed at 5%:5% ratios of 50 and 70%, respectively.ConclusionThis investigation evaluated these two extracts as cell culture supplements and revealed promising results as alternatives to fetal calf serum. The limitation of this study is that it only used two cell types and additional cell lines, and different ratios should be tested. With the above findings, further research using different cell lines, ratios and conditions is warranted.

Lumpy skin disease (LSD) is one of the most economically significant viral diseases of cattle caused by the Lumpy Skin Disease Virus (LSDV), classified as a member of the genus Capripoxvirus and belongs to the family Poxviridae. Nodular skin samples were collected from clinically sick cattle in the districts of Amuru and Wara Jarso Ethiopia to isolate LSD virus. The virus was isolated using primary lamb testis and kidney cells. The isolated LSDV was infected into a healthy calf while maintaining the necessary biosecurity measures to generate skin lesions and to assess disease progression using postmortem examinations. On the fourth day after virus inoculation, the calf developed typical LSD skin nodules with increased rectal temperature, which lasted until the 12th day, when they began to decrease. Viral shedding was detected in nasal, oral, and conjunctival swabs from 6 to 14 days after infection using real-time PCR. Post-mortem tissue specimens tested positive for LSD virus using real-time PCR and virus isolation. This study showed that LSDV were responsible for the LSD outbreaks, and the appearance of typical skin nodules accompanied by fever (> 39.5 °C) defined the virus’s virulent status. The experimental infection with the isolated infectious LSDV could serve as a platform for future vaccine evaluation study using an LSDV challenge model.

Newcastle disease (ND) is caused by virulent strains of avian paramyxovirus type 1, also known as Newcastle disease virus (NDV). Despite vaccination, the frequency of reported outbreaks in Ethiopia has increased. From January to June 2022, an active outbreak investigation was conducted in six commercial chicken farms across areas of central Ethiopia to identify the circulating NDV strains. Thirty pooled tissue specimens were collected from chickens suspected of being infected with NDV. A questionnaire survey of farm owners and veterinarians was also carried out to collect information on the farms and the outbreak status. NDV was isolated using specific-pathogen-free (SPF)-embryonated chicken eggs and detected using haemagglutination and the reverse transcriptase–polymerase chain reaction (RT–PCR). The genotype and virulence of field NDV isolates were determined using phylogenetic analysis of fusion (F) protein gene sequences and the mean death time (MDT) test in SPF-embryonated chicken eggs. The questionnaire results revealed that ND caused morbidity (23.1%), mortality (16.3%), case fatality (70.8%), and significant economic losses. Eleven of thirty tissue specimens tested positive for NDV using haemagglutination and RT–PCR. The MDT testing and sequence analysis revealed the presence of virulent NDV classified as genotype VII of class II velogenic pathotype and distinct from locally used vaccine strains (genotype II). The amino acid sequences of the current virulent NDV fusion protein cleavage site motif revealed 112RRQKR↓F117, unlike the locally used avirulent vaccine strains (112GRQGR↓L117). The epidemiological data, MDT results, cleavage site sequence, and phylogenetic analysis all indicated that the present NDV isolates were virulent. The four NDV sequences were deposited in GenBank with accession numbers F gene (PP726912-15) and M gene (PP726916-19). The genetic difference between avirulent vaccine strains and circulating virulent NDV could explain the low level of protection provided by locally used vaccines. Further studies are needed to better understand the circulating NDV genotypes in different production systems.

Abstract BACKGROUND ANDMETHODS: Brucellosis is a dreadful zoonotic disease affecting humans and all domestic animals including camels worldwide. Serological evidence for Brucella infection in camels has been reported from all pastoralist and agro-pastoralist regions of Ethiopia. Investigations have shown that antibody concentrations are lower in camels than in cattle. However, serological diagnostic kits have been developed for cattle brucellosis is directly transposed for camels without adequate assay validation and genotyping of Brucella species is still absent in camel population in Ethiopia. This study aimed to evaluate gaps associated with Rose Bengal Plate Test (RBPT) and molecular detection of Brucella species from apparently healthy camels slaughtered at the Akaki abattoir. The study applied Brucella genus specific, Brucella abortus (B.abortus) and Brucella melitensis (B.melitensis) species-specific primers on RBPT-positive and retropharyngeal lymph node samples collected from 100 camels’ heads. RESULTS: RBPT revealed the presence of anti-Brucella antibodies in 5 of 100 (5%) slaughtered camels. All RBPT-positive were also positive for PCR. Among the100 lymph node samples examined, 35 (35%) were Brucella positive by PCR. All were found to be B. abortus;however, B. melitessis was not detected in either the serum or lymph node samples. CONCLUSION: To the authors’ knowledge, this investigation is the first report on the molecular detection of B.abortus from camel in Ethiopia. Sequence data confirmed the presence of B. abortus from apparently healthy camels slaughtered at Akaki abattoir, Ethiopia. B. abortus molecular detection rate on lymph nodes samples was seven times greater than that of RBPT. We recommend that advanced research be conducted on camel milk and meat and those camel herders emphasize Ethiopian pastoral areas in particular to understand Brucella epidemiology and its public health significance.

Factors behind intraspecific variation in sensitivity to pathogens remain poorly understood. We investigated how geographical origin in two North European amphibians affects tolerance to infection by the chytrid fungus Batrachochytrium dendrobatidis (Bd), a generalist pathogen which has caused amphibian population declines worldwide. We exposed newly metamorphosed individuals of moor frog Rana arvalis and common toad Bufo bufo from two latitudinal regions to two different BdGPL strains. We measured survival and growth as infections may cause sub-lethal effects in fitness components even in the absence of mortality. Infection loads were higher in B. bufo than in R. arvalis, and smaller individuals had generally higher infection loads. B. bufo had high mortality in response to Bd infection, whereas there was little mortality in R. arvalis. Bd-mediated mortality was size-dependent and high-latitude individuals were smaller leading to high mortality in the northern B. bufo. Bd exposure led to sub-lethal effects in terms of reduced growth suggesting that individuals surviving the infection may have reduced fitness mediated by smaller body size. In both host species, the Swedish Bd strain caused stronger sublethal effects than the British strain. We suggest that high-latitude populations can be more vulnerable to chytrids than those from lower latitudes and discuss the possible mechanisms how body size and host geographical origin contribute to the present results.

Marburg and Ebola filoviruses are two of the deadliest infectious agents and several outbreaks have occurred in the last decades. Although several receptors and co-receptors have been reported for Ebola virus, key host factors remain to be elucidated. In this study, using a haploid cell screening platform, we identify the guanine nucleotide exchange factor CCZ1 as a key host factor in the early stage of filovirus replication. The critical role of CCZ1 for filovirus infections is validated in 3D primary human hepatocyte cultures and human blood-vessel organoids, both critical target sites for Ebola and Marburg virus tropism. Mechanistically, CCZ1 controls early to late endosomal trafficking of these viruses. In addition, we report that CCZ1 has a role in the endosomal trafficking of endocytosis-dependent SARS-CoV-2 infections, but not in infections by Lassa virus, which enters endo-lysosomal trafficking at the late endosome stage. Thus, we have identified an essential host pathway for filovirus infections in cell lines and engineered human target tissues. Inhibition of CCZ1 nearly completely abolishes Marburg and Ebola infections. Thus, targeting CCZ1 could potentially serve as a promising drug target for controlling infections caused by various viruses, such as SARS-CoV-2, Marburg, and Ebola.