Search Results (189)

Background: This study aimed to assess the prevalence of SARS-CoV-2 infection, vaccination, and immune status among a population, both Dentists and University Professors, within a clinical setting at one and at 12 months after COVID-19 vaccination. Methods: A cross-sectional study involving 47 professionals (aged 27–52) was conducted in the University Fernando Pessoa. Participants completed an online survey on SARS-CoV-2 infection status and vaccination, received and provided plasma samples for serological analysis. The protocol was approved by the UFP-Ethics Committee. Anti-S1-RBD SARS-CoV-2 IgM and IgG antibody titration values (AU/mL) were measured, by enzyme-linked-immunosorbent assay (ELISA), with reactive immunoglobulins (Ig) seropositivity for values ≥1 AU/mL. Results: SARS-CoV-2 infection rate increased from 8.5% in July 2021 to 48.9% in June 2022, with 8.5% experiencing reinfection. Vaccination rate was 91.5% by July 2021 and increased slightly to 93.6% by June 2022; 72.3% of the sample received a third dose. IgG seropositivity increased from 91.5% to 95.7% in June 2022. After one-year, significant associations were found between IgG seropositivity and both participant’s age (p = 0.009; <50 years) and vaccine doses (p = 0.003; 1–3 doses) received. Conclusions: SARS-CoV-2 infection rate, vaccination, and IgG seropositivity rates were high and increased over one year. The age and vaccination status were associated with the immunity status at 12th month follow-up. Findings highlight variability in IgG seroprevalence due to multiple influencing factors, which justifies future studies. Full article

Background: Vaccination effectively reduces the risk of infection, including COVID-19 yet older adults often receive insufficient attention despite their increased vulnerability. The study aimed to correlate serological results with underlying conditions, vaccination status, and COVID-19 history. Methods: This non-interventional, multicenter study aimed to assess vaccination coverage and SARS-CoV-2 antibody levels among residents of eight long-term care facilities (LTCFs) in Southern Poland. Data collection took place between January and June 2022, with 429 participants recruited based on their ability to provide informed consent and their residency in LTCFs. Sociodemographic data, medical history, and COVID-19-related information—including infection history and vaccination status—were collected through surveys. Blood samples were obtained for serological testing using enzyme-linked immunosorbent assays (ELISA) to detect anti-SARS-CoV-2 antibodies. Statistical analysis, including Spearman’s correlation, revealed significant associations between antibody levels and vaccination status, as well as between RT-PCR-confirmed COVID-19 infections and higher antibody titers. Results: Among the seven different qualitative serological, only the Anti-SARS-CoV-2 NCP (IgG) and Anti-SARS-CoV-2 (IgA) tests showed a positive correlation with the Anti-SARS-CoV-2 QuantiVac (IgG) test, which was used as a comparator. A weak correlation was noted with the age of the residents. Conclusions: Our findings suggest that vaccination positively influences antibody responses, underscoring the importance of immunization among LTCF residents. Additionally, certain comorbidities—such as degenerative joint disease and diabetes—showed weak correlations with higher antibody levels. This study provides valuable insights into the humoral immune response to COVID-19 in vulnerable populations residing in LTCFs. Full article

(1) Background: The ongoing global health threat posed by SARS-CoV-2 requires reliable and accessible diagnostic tools, especially in resource-limited settings where RT-qPCR may be impractical. This study describes the development and validation of two enzyme-linked immunosorbent assays (ELISA) designed to detect anti-SARS-CoV-2 IgG antibodies employing recombinant S1 and S2 spike protein subunits. (2) Methods: The assays were optimized and validated using serum samples from 354 RT-qPCR-confirmed hospitalized patients and 337 pre-pandemic blood donors. (3) Results: The S1-based ELISA achieved a 52.8% sensitivity and a specificity of 93.5%, with an area under the ROC curve (AUC) of 71.6%. In contrast, the S2-based ELISA demonstrated superior diagnostic performance, with a sensitivity of 63.7%, a specificity of 99.7%, and an AUC of 83.1%. Cross-reactivity analysis using sera from individuals with unrelated infectious diseases confirmed the high specificity of the S2-ELISA. Time-stratified analysis revealed that sensitivity increased with time, peaking between 15 and 21 days post-symptom onset. Compared to commercial serological assays, the S2-ELISA demonstrated comparable or improved performance, particularly in specificity and diagnostic odds ratio. (4) Conclusions: The S2-ELISA offers a robust, highly specific, and operationally simple tool for serological detection of SARS-CoV-2 infection. Its strong diagnostic performance and accessibility make it well-suited for implementation in diverse epidemiological settings, particularly where molecular testing is limited. The development of affordable, validated serological assays such as this is critical for strengthening surveillance, understanding transmission dynamics, and informing public health responses. Full article

Background: The Human Papillomavirus (HPV) vaccine generates high antibody titers against targeted HPV types. This study investigated vaccine-induced anti-HPV18 immunoglobulin (IgG) antibody titers and subsequent HPV18/45 infections. Methods: We performed a nested matched case-control study leveraging a prospective longitudinal cohort of adolescent and young adult women (AYW) vaccinated with the quadrivalent HPV vaccine (4vHPV) attending the Mount Sinai Adolescent Health Center (MSAHC) in Manhattan, NY. The case individuals included AYW who had an incident detection of cervical HPV18 (n = 3) or HPV45 (n = 34) DNA after vaccination and were compared to two vaccinated control individuals (HPV18/45-negative); one random control (RC, n = 37) and one high-risk control (HRC, n = 37) selected from the upper quartile of a sexual risk behavior score. Serological titers against HPV18 were measured by end-point dilution and enzyme-linked immunosorbent assay (ELISA) in serum collected before the incident detection of HPV. Matching was performed based on age at first dose, follow-up time, and sexual risk behavior score. Conditional logistic regression was used to assess the association between case-control status and anti-HPV antibody titers, consistent with the matched-pair design. Results: Antibody titers for HPV18 were most different between AYW who developed an HPV18/45 infection compared to high-risk controls OR = 1.66, 95% CI: 0.96–2.85 (p = 0.1629). Analyses of pooled data from vaccinated recipients including who developed HPV16/31 or HPV18/45 infections demonstrated that the odds of a one-log unit increase in anti-HPV16 or 18 antibody titers, respectively, were 40% higher in the combined control groups (RC + HRC, n = 160) (OR = 1.40, 95% CI: 1.09–1.79, p = 0.0135) and 73% higher in the HRC (n = 80) (OR 1.73, 95% CI: 1.34, 2.52, p = 0.0117) compared to HPV16/18/31/45 cases (n = 80). Conclusions: Overall, these findings suggest that higher IgG antibodies to HPV16/18 after vaccination represent an increased likelihood of protection from homologous and cross-reactive HPV types (HPV16/18/31/45). These results show that differences in antibody titers are associated with breakthrough infection after vaccination, suggesting that further study of long-term antibody titers and infection should be pursued. Full article

Infectious bursal disease virus (IBDV) is one of the most important immunosuppressive viruses in poultry, causing the global spread of infectious bursal disease (IBD). It poses a significant threat to the healthy development of the poultry industry. Vaccination is an effective approach for controlling IBDV infection. Therefore, reliable immune monitoring for IBDV is critical for maintaining poultry health. The enzyme-linked immunosorbent assay (ELISA) is a common technique used to detect specific antibodies in clinical serum testing and for the serological evaluation of IBDV vaccines. Among the currently available and under development IBDV vaccines, IBD VP2 subunit-based vaccines account for a considerable proportion. These vaccines stimulate the production of antibodies that are specific only to VP2. However, most IBDV antibody ELISA kits approved for use have applied the whole virus as the coating antigen, which does not adequately meet the diverse requirements for IBDV detection across different conditions. This study utilized a prokaryotic expression system to express the VP2 protein of the IBDV epidemic strain, assembling it into virus-like particles to be used as coating antigens. This approach enabled the establishment of an indirect ELISA method for detecting IBDV VP2 antibody (VP2-ELISA). The optimal coated antigen concentration was determined to be 2.5 μg/mL, with overnight coating at 4 °C; sealing with 5% skim milk at 37 °C for 4 h; serum dilution at 1:500 with incubation at 37 °C for 30 min; secondary antibody dilution at 1:4000 with incubation at 37 °C for 40 min; and then incubation with the substrate solution 3,3′,5,5′-tetramethylbenzidine at room temperature for 20 min. The criterion for interpreting the detection results was OD_450nm ≥ 0.111 indicates IBDV antibody positivity, while OD_450nm < 0.111 indicates negativity. The established VP2-ELISA can specifically detect IBDV-positive sera at the lowest serum dilution of 1:6400, with intra- and inter-batch coefficients of variation of <2%. This indicates that the VP2-ELISA exhibits good specificity, sensitivity, and stability. Detection experiments using 20 laboratory-immunized chicken serum samples and 273 clinical serum samples demonstrated that the results of VP2-ELISA were consistent with those of commercial ELISA kits coated with whole virus. In summary, the VP2-ELISA developed in this study offers advantages in immune response detection for IBD VP2 subunit-based vaccines and is appropriate for evaluating the efficacy of IBD vaccines and detecting clinical serum samples. Full article

Infections with the Hepatitis B (HBV) and Hepatitis C (HCV) viruses share some transmission routes, which is why coinfection with these viruses becomes common, especially in endemic areas. This study evaluated the immunological response profile, viral load, and liver damage in groups monoinfected with HBV or HCV and in those co-infected with HBV/HCV. The groups were composed of 22 patients monoinfected by HCV, 22 patients monoinfected by HBV, and 34 co-infected by HBV/HCV, according to serological markers and molecular biology tests. The study was carried out from December 2017 to October 2019. Virus detection employed enzyme immunoassay, Enzyme-Linked Immunosorbent Assay (ELISA), and real-time PCR, while liver function and fibrosis were assessed using biochemical tests and Fibroscan. To research the immunological profile, cytokines were quantified using the BIO-Plex Pro Human Cytokine. Comparing the groups, both mono- and co-infected patients exhibited a Th1 immune response profile. HCV monoinfection notably showed significantly elevated serum levels of INF-γ (p < 0.01) and TNF-α (p < 0.01). The viral load was significantly higher in the HCV monoinfected group when compared to the other groups. Regarding liver damage, patients with a high level of fibrosis (F4) presented significant levels of cytokines INF-γ (p < 0.001), IL-17 (p < 0.0001), and TNF-α (p < 0.0001). Full article

The increasing number of owned and stray dogs in large cities is becoming a pressing issue due to rising population densities, urban conditions, and poor control over animal reproduction. This situation poses serious epidemiological risks, as dogs can act as reservoirs and transmitters of infectious and parasitic diseases dangerous to humans. This study aimed to investigate the prevalence and carriage of infectious and parasitic diseases in stray dogs in the city of Uralsk as a factor of epidemiological risk. In 2024, 1213 stray dogs were captured from different city districts and examined at the veterinary clinic and laboratory of Zhangir Khan University. Biological samples (blood, urine, feces) from 10% of the animals were analyzed using molecular (PCR), serological (ELISA), and helminthological methods. Serological and molecular analyses revealed the widespread circulation of bacterial pathogens. Antibodies to additional bacterial agents, including Pasteurella multocida, Mycobacterium spp., Listeria monocytogenes, and Leptospira spp., were detected in the samples, indicating an unfavorable sanitary and epidemiological situation in the urban environment. An enzyme-linked immunosorbent assay (ELISA) identified antibodies against Toxocara canis in 50.9% of the dogs and against Echinococcus granulosus in 76.4%, reflecting both active and past infections. The polymerase chain reaction (PCR) results showed the presence of Brucella canis DNA in blood and urine samples, while antibodies to Brucella spp. were detected in 57.8% of the examined dogs, underscoring the significant zooanthroponotic importance of this pathogen and its potential threat to human health. Additionally, T. canis DNA was found in 39.2% of the samples and E. granulosus DNA in 16.6%. A helminthological examination using the Fülleborn method revealed a high rate of helminth infection: Ancylostoma caninum—35.3%, T. canis—32.3%, and Toxascaris leonina—29.4%. The obtained results highlight the significant role of stray dogs as epizootiological and epidemiological reservoirs of zooanthroponotic infections. This poses a serious threat to public health and necessitates the implementation of effective control and prevention measures for infectious and parasitic diseases within urban fauna. Full article

Background/Objectives: Syphilis remains a significant public health challenge worldwide. Accurate and efficient diagnostic tools are essential to controlling the spread of the disease. Current diagnostic approaches primarily rely on serologic treponemal tests (TTs) and nontreponemal tests (NTTs). The aim of this study was to evaluate the diagnostic properties of various serological methods for syphilis diagnosis. Methods: Samples were collected from participants of the Health, Information, and Sexually Transmitted Infection Monitoring (SIM study) between March 2020 and May 2023, using convenience sampling at a mobile health unit in Porto Alegre, Brazil. A total of 250 individuals were tested using the point-of-care (POC) lateral flow treponemal test, Venereal Disease Research Laboratory (VDRL) test, Rapid Plasma Reagin (RPR) test, Enzyme-Linked Immunosorbent Assay (ELISA), and Treponema pallidum hemagglutination assay (TPHA). Of these, 125 participants tested positive for syphilis in the POC screening. Diagnostic properties such as sensitivity, specificity, and predictive values were assessed for the POC test, ELISA, and VDRL test. The TPHA was used as the reference standard for the TT, and the RPR test as the reference standard for the NTT. Results: Among individuals with positive POC test results, 97.6% (122/125) were also positive by the ELISA, and 85.6% (107/125) were positive by the TPHA. Additionally, 48.0% (60/125) and 42.4% (53/125) tested positive by the VDRL and RPR tests, respectively. Using the TPHA as a reference, TT tests showed sensitivities of 97–98% and specificities of 93–95% for detecting anti-Treponema pallidum antibodies using the ELISA and POC test, respectively. For the NTT, the VDRL test demonstrated a sensitivity of 98% and a specificity of 95% compared to the RPR test. The kappa coefficients were 0.85 for the POC test vs. the TPHA, 0.81 for the ELISA vs. the TPHA, and 0.89 for the VDRL vs. the RPR tests, indicating substantial agreement. Conclusions: This study highlights a good diagnostic performance and high agreement levels among the evaluated serological tests for syphilis, reinforcing their utility in clinical and public health settings, as well as epidemiological studies. Full article

Standard serological tests post-vaccination, such as enzyme-linked immunosorbent assay (ELISA), complement fixation, and virus neutralization, are crucial for monitoring African horse sickness (AHS). However, the availability of commercial test kits such as blocking ELISA varies by regions; while they are commonly used in Africa and Europe, their limited availability and high cost in Thailand present significant challenges. Therefore, this study aimed to evaluate an alternative approach using an in-house indirect ELISA based on cell-based monovalent and polyvalent strains of live attenuated AHS virus. This method addresses the cost and accessibility issues faced in Thailand. This study demonstrated promising results: the in-house indirect ELISA showed analytical sensitivity and specificity values of 88.30% and 67.02% for monovalent strains and 87.23% and 84.04% for polyvalent strains, respectively, compared to the blocking ELISA. These findings underscore the efficacy of the in-house ELISA as a viable serodiagnostic tool for AHS. Furthermore, the polyvalent antigen-based in-house indirect ELISA proved to be a reliable alternative for AHS monitoring, particularly in vaccinated horses, offering enhanced specificity. Additionally, this method is simpler, cheaper, faster, and more convenient than blocking ELISA and serum neutralization tests, making it a practical choice for routine AHS surveillance. Full article

Actinobacillus pleuropneumoniae (APP) is a bacterial pathogen causing porcine pleuropneumonia, causing great economic loss to the global pig industry. Although natural apxIV contributes to the prevention and control of porcine pleuropneumonia, its isolation poses a great challenge, and recombinant soluble apxIV proteins tend to carry large molecular weight tags. The traditional serologic methods tend not to accurately detect the apxIV-partially deleted vaccine (GDV). In this study, we screened the soluble protein apxIVA N2 (756 bp) from six apxIV-truncated proteins and applied it to the enzyme-linked immunosorbent assay (ELISA) and colloidal gold immunochromatographic strip for detecting the samples vaccinated with APP GDV. The results indicate that N2 was close to the natural apxIV protein in terms of structure and function as it only contained a single His (0.86 kDa) tag and a single S (2 kDa) tag. Among the six candidate proteins, N2 exhibited the best performance in distinguishing APP-infected samples from those vaccinated with the APP GDV. Both ELISA and colloidal gold immunochromatographic strips based on this protein exhibited an excellent performance in detecting and distinguishing wild-strain-infected samples from those vaccinated with the subunit vaccine or the GDV. In addition, three monoclonal antibodies against different antigenic epitopes were identified using these truncated proteins. Our studies are of great significance for further research on APP, the differential diagnosis of wild strains and vaccine strains, and pig control breeding, exhibiting a broad application prospect in the on-site diagnosis of APP, particularly in remote areas lacking detection instruments and professionals. Full article

Background/Objectives: Feline coronavirus (FCoV) belongs to the family Coronaviridae and includes two pathotypes, the less virulent feline enteric coronavirus (FECV), which replicates in the enteric epithelial cells, and feline infectious peritonitis virus (FIPV), which is more virulent, replicates efficiently within monocytes/macrophages with systemic involvement and may cause feline infectious peritonitis (FIP), a progressive and often fatal disease. The diagnosis of FIP is complex and requires different examinations. Among serological tests, the indirect immunofluorescent antibody test (IFAT), considered the gold standard, and the enzyme-linked immunosorbent assay (ELISA) are the most widely used to detect FCoV antibodies. The aim of this work was the development of FCoVCHECK Ab ELISA, a new rapid indirect test for the detection of FCoV antibodies in feline serum/plasma samples. Methods: FCoVCHECK Ab ELISA was developed after a meticulous set-up and cut-off analysis through several methods, including the Youden’s index and ROC curve, to achieve the best test performance. It was validated by testing 110 feline sera (62 positives and 48 negatives) against the reference IFAT and compared with two other rapid ELISA tests, INgezim Corona Felino (Gold Standard Diagnostics) and ImmunoComb Feline Coronavirus (FCoV) [FIP] Antibody Test Kit (Biogal). Conclusions: FCoVCHECK Ab ELISA agreed with IFAT at 96.4% (93.5% sensitivity, 95% confidence interval (CI): 83.5–97.9%; 100% specificity, 95% CI: 90.8–100%), with ImmunoComb FCoV at 93.6% and with INgezim Corona Felino at 82.7%. Intra- and inter-assay accuracy and precision gave coefficients of variation lower than 20%. Compared to IFAT, the new assay correctly identifies positive and negative samples with a good correlation, and, in addition, it is simpler, faster and provides a less subjective reading of the results. Full article

During the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, the development of efficient serological tests for monitoring the dynamics of the disease as well as the immune response after illness or vaccination was critical. In this regard, low-cost and fast production of immunogenic antigens is essential for the rapid development of diagnostic serological kits. This study assessed the plant-based production of nucleoprotein (N) of SARS-CoV-2 and chimeric receptor-binding domain (RBD) of SARS-CoV-2 presented by hepatitis E virus capsid (HEV/RBD) and validation of the plant-derived proteins as diagnostic antigens for serological tests. The target proteins were expressed in and purified from Nicotiana benthamiana plants. The resulting yield of chimeric HEV/RBD protein reached 100 mg/kg fresh weight and 30 mg/kg fresh weight for N protein. The purified N protein and HEV/RBD protein were used to develop an indirect enzyme-linked immunosorbent assay (iELISA) for the detection of antibodies to SARS-CoV-2 in human sera. To validate the iELISA tests, a panel of 84 sera from patients diagnosed with COVID-19 was used, and the results were compared to those obtained by another commercially available ELISA kit (Dia.Pro D. B., Sesto San Giovanni, Italy). The performance of an HEV/RBD in-house ELISA showed a sensitivity of 89.58% (95% Cl: 75.23–95.37) and a specificity of 94.44% (95% Cl: 76.94–98.2). Double Recognition iELISA based on HEV/RBD and N protein is characterized by a lower sensitivity of 85.42% (95% Cl: 72.24–93.93) and specificity of 94.44% (95% Cl: 81.34–99.32) at cut-off = 0.154, compared with iELISA based on HEV/RBD. Our study confirms that N and fusion HEV/RBD proteins, which are transiently expressed in plants, can be used to detect responses to SARS-CoV-2 in human sera reliably. Our research validates the commercial potential of using plants as an expression system for recombinant protein production and their application as diagnostic reagents for serological detection of infectious diseases, hence lowering the cost of diagnostic kits. Full article

Crimean–Congo hemorrhagic fever (CCHF) is a serious tick-borne disease with a wide geographical distribution. Classified as a level 4 biosecurity risk pathogen, CCHF can be transmitted cross-species due to its aerosol infectivity and ability to cause severe hemorrhagic fever outbreaks with high morbidity and mortality. However, current methods for detecting anti-CCHFV antibodies are limited. This study aimed to develop a novel luciferase immunosorbent assay (LISA) for the detection of CCHFV-specific IgG antibodies. We designed specific antigenic fragments of the nucleoprotein and evaluated their sensitivity and specificity in detecting IgG in serum samples from mice and horses. In addition, we compared the efficacy of our LISA to a commercial enzyme-linked immunosorbent assay (ELISA). Our results demonstrated that the optimal antigen for detecting anti-CCHFV IgG was located within the stalk cut-off domain of the nucleoprotein. The LISA exhibited high specificity for serum samples from indicated species and significantly higher sensitivity (at least 128 times) compared with the commercial ELISA. The proposed CCHFV-LISA has the potential to facilitate serological diagnosis and epidemiological investigation of CCHFV in natural foci, providing valuable technical support for surveillance and early warning of this disease. Full article

Bacteria from the genus Proteus are facultative human pathogens, primarily attacking the urinary tract and wounds. A total of 85 O serogroups have been identified so far among these bacilli. P. mirabilis Bprz 86 was isolated from the fistula of a patient in Łódź, Poland. Enzyme-Linked Immunosorbent Assay (ELISA) and Western blotting studies involving the P. mirabilis Bprz 86 lipopolysaccharide (LPS) and the strain-specific rabbit antiserum indicated that the strain, which does not belong to any of the O1–O85 serogroups, shares a common epitope with Proteus O17 antigens and is identical to another clinical P. mirabilis strain, Sm 120, isolated from the urine of a patient in the area. The O-specific polysaccharide (O antigen) was obtained from P. mirabilis Bprz 86 LPS through mild acid degradation, and the six-constituent structure of its repeating unit was determined using chemical analyses and 1D and 2D ¹H and ¹³C Nuclear Magnetic Resonance (NMR) spectroscopy. It includes (R)-3-hydroxybutanoyl, which, along with fucosamine and glucose residues, forms a fragment also present in the O17 antigens. Based on the obtained serological and chemical data, the two studied P. mirabilis isolates were proposed as candidates for a new successive O serogroup in the genus Proteus, O86. Full article

Sheep-associated malignant catarrhal fever (SA-MCF) is a severe lymphoproliferative vascular disease of cattle that is caused by ovine gammaherpesvirus 2 (OvGHV2), which is a Macavirus within the Gammaherpesvirinae subfamily. SA-MCF occurs worldwide in several mammalian hosts. Alternatively, alcelaphine gammaherpesvirus 1 (AlGHV1) is a Macavirus that causes wildebeest-associated malignant catarrhal fever (MCF), which principally occurs in cattle from Africa. Previous serological assays to evaluate the presence of MCF in mammals used a competitive inhibition enzyme-linked immunosorbent assay (CI-ELISA). This CI-ELISA is based on the 15A antigenic epitope that is common to all Macavirus associated with the development of MCF in their respective hosts. This study evaluated an indirect MCF-specific ELISA assay based on the AlGHV1 C500 strain to detect antibodies against OvGHV2 in 43 closed dairy cattle farms from Southern Brazil. These farms are located in a region where subclinical infections by OvGHV2 have been detected in free-ranging wild boars (Sus scrofa). Sheep or goats were not reared at these farms or within the proximity of these farms. Risk factors associated with seropositivity to OvGHV2 were evaluated, while the possible participation of subclinically infected wild boars in the dissemination of OvGHV2 was estimated using spatial analysis. Sera from 29 dairy cows from 16 farms demonstrated sample/positive (S/P) values considered positive with this MCF-specific ELISA (cutoff S/P, 0.063). The S/P values for the positive dairy cows varied between 0.0633 and 0.2510 (mean, 0.0998; standard deviation, 0.0476). At least one cow was seropositive in 16/43 (37.2%) of these farms, with seropositivity identified in 29/367 (7.9%) of dairy cows maintained at these farms. Additionally, dairy cows raised within the intensive system had a more than threefold higher chance of being seropositive to OvGHV2 relative to those reared within the semi-intensive system. Furthermore, the spatial evaluation revealed that cows on dairy farms within a 50 km radius of the home range of subclinically infected wild boars had an increased risk of being seropositive to this assay. These findings demonstrated that the AlGHV1 C500-specific MCF ELISA can be efficiently used to monitor the occurrence of OvGHV2 in cattle. In addition, the occurrence of subclinically infected free-ranging wild boars within a radius of 50 km from susceptible cattle may be a possible risk factor for the occurrence of OvGHV2-related infections in these animals from Southern Brazil. These initial results are fundamental to understanding the epidemiology of OvGHV2-associated infections and clinical SA-MCF in mammals in Brazil. Full article