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Adult horses are susceptible to equine coronavirus (ECoV) and severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), although, only ECoV has been linked to clinical disease. Little information is available regarding the seroprevalence against ECoV and SARS-CoV-2 in adult healthy horses. The goal of the present study was to determine the seroprevalence against two coronaviruses known to infect horses using convenience samples collected from horses recently imported from Europe to the United States from 2019 to 2023. A total of 385 banked serum samples were tested against ECoV and SARS-CoV-2 using previously validated ELISA assays. Prevalence factors including date of arrival in the United States, signalment and country of origin were available for the majority of the horses. A total of 9/385 (2.3%) and 4/385 (1.0%) horses tested seropositive for ECoV and SARS-CoV-2, respectively. The ECoV seropositive horses were all mares, ages 4 to 26 years (median 9 years) and originated from Germany, the Netherlands, Ireland, Belgium and Italy. These mares were predominantly imported during the summer and fall months. All SARS-CoV-2 seropositive horses were mares ages 5 to 10 years (median 7.5 years) imported from the Netherlands and the United Kingdom. The majority of the SARS-CoV-2 seropositive horses were imported during the colder months of the year. The study results support the presence of ECoV in Europe and report on the first SARS-CoV-2 seropositive healthy adult horses outside the United States. Commingling for movements by air and close contact to humans may predispose transmission with ECoV and SARS-CoV-2, respectively.

The rapid spread of SARS-CoV-2 caused the COVID-19 pandemic and accelerated vaccine development to prevent the spread of the virus and control the disease. Given the sustained high infectivity and evolution of SARS-CoV-2, there is an ongoing interest in developing COVID-19 serology tests to monitor population-level immunity. To address this critical need, we designed a paper-based multiplexed vertical flow assay (xVFA) using five structural proteins of SARS-CoV-2, detecting IgG and IgM antibodies to monitor changes in COVID-19 immunity levels. Our platform not only tracked longitudinal immunity levels but also categorized COVID-19 immunity into three groups: protected, unprotected, and infected, based on the levels of IgG and IgM antibodies. We operated two xVFAs in parallel to detect IgG and IgM antibodies using a total of 40 μL of human serum sample in

Toxoplasma gondii is an important zoonotic parasite worldwide, but it has received limited attention in Ukraine. A seroepidemiological study was conducted and samples from 452 wild boars that had been hunted in 2006-2011 in 23 of the 25 regions of Ukraine were tested to estimate T. gondii seroprevalence. A locally available commercial enzyme-linked immunosorbent assay (ELISA) was used for the investigation. Additionally, we tested 92 of the sera using a widely used commercial multi-species ELISA and an indirect immunofluorescence antibody test (IFAT). With the locally available ELISA, 35 of the 452 wild boars tested positive, yielding a seroprevalence estimate of 7.7% (95% confidence interval 5.5-10.5). The seropositive wild boars originated from eight of the regions. Using the majority criteria, 10/92 samples tested using both ELISAs and the IFAT were considered positive, yielding an estimated seroprevalence of 10.9% within the subset of samples. The highest seroprevalence was observed in wild boars hunted in Luhansk (30.0%), Odesa (17.7%) and Kharkiv (12.7%). Seroprevalence was higher in older animals (13.3% for age group >12 months and 7.7% for age group ≤12 months). This is the first seroepidemiological study of T. gondii in wild boars in Ukraine. Assuming that seropositivity indicates presence of infectious parasites in the tissues, eating undercooked meat of wild boars hunted in Ukraine could be a potential source of infection to other hosts, including humans.

Toxoplasma gondii (T. gondii) affects around 30% of humans worldwide. Recently, it has emerged as a significant opportunistic pathogen to immunocompromised patients. Data available is still lacking about toxoplasmosis in cancer patients in Egypt. This study aimed to reveal the current trend of T. gondii in cancer patients in Sohag, Egypt. Sera from 50 cancer patients and 50 healthy controls were screened for Toxoplasma IgG and IgM. Further, buffy coats from both groups were used for detection of T. gondii B1 and RE genes via conventional and nested PCR, respectively. The overall seroprevalence of T. gondii IgG was high (58%). IgG and IgM were detected in 30% and 9% cancer patients, respectively. Patients with solid cancers exhibited a greater IgG seropositivity compared to those with hematologic tumors (77.27% and 46.43%, respectively) (P = 0.03). Concerning the molecular results, only 4 (9%) were positive regarding both PCR assays. In conclusion, T. gondii is highly prevalent in cancer patients in Sohag, Egypt. PCR is strongly recommended to complement serology to diagnose acute or reactivated toxoplasmosis in cancer patients. B1 PCR was found to be equivalent to RE PCR. Nevertheless, thorough large-scale research must be implemented. [ABSTRACT FROM AUTHOR]

Introduction: The first vaccines approved against SARS-CoV-2, mRNA-1273 and BNT162b2, utilized mRNA platforms. However, little is known about the proteomic markers and pathways associated with host immune responses to mRNA vaccination. In this proof-of-concept study, sera from male and female vaccine recipients were evaluated for proteomic and immunologic responses 1-month and 6-months following homologous third vaccination. Methods: An aptamer-based (7,289 marker) proteomic assay coupled with traditional serology was leveraged to generate a comprehensive evaluation of systemic responsiveness in 64 and 68 healthy recipients of mRNA-1273 and BNT162b2 vaccines, respectively. Results: Sera from female recipients of mRNA-1273 showed upregulated indicators of inflammatory and immunological responses at 1-month post-third vaccination, and sera from female recipients of BNT162b2 demonstrated upregulated negative regulators of RNA sensors at 1-month. Sera from male recipients of mRNA-1273 showed no significant upregulation of pathways at 1-month post-third vaccination, though there were multiple significantly upregulated proteomic markers. Sera from male recipients of BNT162b2 demonstrated upregulated markers of immune response to doublestranded RNA and cell-cycle G(2)/M transition at 1-month. Random Forest analysis of proteomic data from pre-third-dose sera identified 85 markers used to develop a model predictive of robust or weaker IgG responses and antibody levels to SARS-CoV-2 spike protein at 6-months following boost; no specific markers were individually predictive of 6-month IgG response. Thirty markers that contributed most to the model were associated with complement cascade and activation; IL-17, TNFR pro-apoptotic, and PI3K signaling; and cell cycle progression. Discussion: These results demonstrate the utility of proteomics to evaluate correlates or predictors of serological responses to SARS-CoV-2 vaccination. [ABSTRACT FROM AUTHOR]

Introduction: Risk stratification for CMV infection in lung transplantation (LT) currently relies on determining donor and recipient CMV IgG before transplantation. However, it has been observed that some patients who test positive for CMV-specific humoral response before kidney transplantation (KT) exhibit a weak or absent CMV-specific cellular response. The significance of this observation in LT is still unknown. Methods: This prospective, multicenter, observational study evaluated the agreement between CMV IgG serology and specific cell-mediated response (specific T cell Enzyme-Linked ImmunoSpot Assay, ELISPOT, against CMV pp65 and IE-1 antigens) in 121 patients on the waiting list for LT. Results: One hundred and four (86%) patients were seropositive for CMV. Discordant humoral and cellular immunologic responses were observed, 29% of seropositive patients had a weak ELISPOT response to IE-1 and 39% to pp65. In 22% of seropositive patients, there was a weak or no response to both antigens. All seronegative patients did not respond to either antigen. Conclusions: Therefore, over 20% of CMV seropositive LT candidates showed weak CMV-specific cellular immune responses despite detectable serological memory against CMV. This may be important in assessing the risk of developing a CMV infection after transplantation. [ABSTRACT FROM AUTHOR]

Canine leishmaniosis (CanL), caused by Leishmania infantum, is a widespread vector-borne disease. In Italy, an endemic region for CanL, overlapping transmission of L. infantum and tick-borne pathogens (TBPs) like Anaplasma phagocytophilum and Ehrlichia canis is increasingly reported. Dogs with clinical leishmaniosis often show higher co-infection rates and pronounced clinicopathological abnormalities. This study presents a systematic literature review and new findings from southern Italy, focusing on co-infections with E. canis and A. phagocytophilum in clinically healthy L. infantum seropositive and seronegative dogs. The systematic review identified two eligible studies. The first reported 34/488 (7%) dogs L. infantum seropositive, with 11.8% also seropositive for A. phagocytophilum. Among 454 seronegative dogs, 3% were seropositive for A. phagocytophilum and 2.4% for E. canis. The second study identified 154/1260 (12.2%) dogs L. infantum seropositive, with co-infection rates of 0.6% and 1.9% for A. phagocytophilum and E. canis, respectively. Among 1106 seronegative dogs, 1.3% were seropositive for A. phagocytophilum and 2.3% for E. canis. In the retrospective study from southern Italy, 90/154 (58.4%) dogs were L. infantum seropositive, with co-infection rates of 4.4% for A. phagocytophilum and 2.2% for A. phagocytophilum and E. canis. Among 64 seronegative dogs, 1.6% showed similar co-infections. This is the first systematic review in Italy, documenting low and comparable co-infection rates with A. phagocytophilum and E. canis in clinically healthy dogs, regardless of L. infantum serostatus. These findings suggest that co-infections may occur independently, offering insights into vector-borne disease dynamics in endemic areas. [ABSTRACT FROM AUTHOR]

Purpose and methods: This prospective study aims to diagnose the etiology of non-focalized fever lasting between 5 and 28 days in the islands of La Palma and El Hierro (Canary Islands, Spain) during 2021, using serology and PCR. Results: The etiological profile described in this study aligns with that of fever of intermediate duration (FID), with zoonoses being the primary cause. Murine typhus (MT) is identified as the leading cause, followed by Q fever (QF). The incidence of MT is the highest reported nationally and comparable to the highest in Europe, with 39.6 cases per 100,000 inhabitants in La Palma and 79.7 cases per 100,000 inhabitants in El Hierro. Q fever, known to be endemic to the Canary Islands, presents incidences of 26.5 cases per 100,000 inhabitants in La Palma and 15.6 cases per 100,000 inhabitants in El Hierro. MT shows no gender differences and has a homogeneous geographical distribution. In contrast, QF is more prevalent in men and has a heterogeneous geographical distribution. Conclusions: The high incidence of MT found in both urban and peri-urban areas is particularly noteworthy. Its potential connection with climate change and/or the growth of the reservoir population in the Canary Islands remains unknown. MT's similarity to QF in terms of clinical signs and treatment, coupled with the absence of a specific protocol for early diagnosis, may have contributed to its underdiagnosis. MT can lead to significant health concerns, including risk of hospitalization, complications, and even death. Therefore, the registration of cases for epidemiological control is deemed essential. [ABSTRACT FROM AUTHOR]

Purpose: The study evaluated the effects of elexacaftor/tezacaftor/ivacaftor (ETI) therapy in people with cystic fibrosis (pwCF) and a clinical history of Aspergillus fumigatus (AF) infection. Methods: This prospective cohort study included pwCF who initiated ETI therapy and had received antifungal treatment in the preceding five years due to allergic bronchopulmonary aspergillosis (ABPA group) or other AF-related clinical manifestations (AF group). A control group of pwCF with no prior respiratory cultures positive for AF was also included. Changes from baseline to 12 months in spirometry measures and lung clearance index (LCI2.5), as well as respiratory colonization by AF, were compared across groups. Annual fold changes in the geometric mean of immunological markers were estimated using generalized estimating equations with a piecewise linear spline model, fitted to data collected from three years before to one-year post-ETI. Results: The study included 16 patients in the ABPA group, 47 in the AF group, and 45 controls. Spirometry and LCI2.5 improvements were comparable across groups. Positive respiratory cultures decreased from 43.8 to 18.8% in the ABPA group (p = 0.30), and from 78.7 to 23.4% in the AF group (p < 0.001). Total IgE and IgG anti-AF decreased in both the ABPA and the AF groups, with annual reductions of 20–42%. No ABPA episodes occurred during ETI therapy. Conclusion: During ETI therapy, pulmonary outcomes improved, AF colonization and sensitization decreased, and no episodes of ABPA were observed in pwCF with a clinical history of AF infection. [ABSTRACT FROM AUTHOR]

Background: Dirofilaria immitis and D. repens are mosquito-borne filaroids that primarily infect dogs but also cats. Diagnosing feline dirofilariosis is challenging because of the low parasitic burdens and transient or absent microfilaremia. To improve detection of antibodies against Dirofilaria spp. in cats, an indirect enzyme-linked immunosorbent assay (ELISA) using somatic antigens of D. immitis was standardized. Methods: Serum samples from cats positive for D. immitis, D. repens, bronchopulmonary metastrongylids and gastrointestinal helminths as well as negative sera were tested to evaluate the sensitivity (Se) and specificity (Sp). Three different antigen concentrations (2, 3 and 4 μg/ml) and two concentrations of horseradish peroxidase (HRP) IgG anti-cat conjugate (1:10,000 and 1:20,000) were used to screen the optimal titration of the test. Once the best conditions were established, Se and Sp were assessed by testing 151 serum samples positive for D. immitis, D. repens and other parasites (i.e. feline lungworms, gastrointestinal helminths) or negative. Results: The optimized ELISA showed 89% Se and 98% Sp in detecting D. immitis infections, presenting a cross-reactivity with D. repens. A cut-off point of 1.150 was established to classify positive and negative samples. Conclusions: The developed ELISA may improve the detection of antibodies against Dirofilaria spp. in cats, also in epidemiological contexts characterized by multiple helminth infections. Future efforts will focus on reducing the cross-reactivity with D. repens, therefore increasing ELISA Sp. [ABSTRACT FROM AUTHOR]

Simple Summary: Hepatitis E virus (HEV) is widespread in humans and pigs worldwide. HEV-1 and HEV-2 genotypes are restricted only to humans, while HEV-3 and HEV-4 have zoonotic potential and are common in humans and animals. The aim of our study was to determine the presence of anti-HEV antibodies in three hundred naturally infected domestic pigs from three different commercial farrow-to-finish farms in South Bačka on the territory of Vojvodina Province, Serbia. The presence of specific anti-HEV IgG was examined by an in-house ELISA, and doubtful results were retested by western blotting (WB). The overall detected seroprevalence was 40.66%, indicating that the Hepatitis E virus is circulating in the intensive farming pig population in Serbian farms. Hepatitis E virus (HEV) is the causative agent of acute hepatitis E infection in humans. Two epidemiological patterns of the disease exist—endemic and sporadic. Genotypes 1 (HEV-1) and 2 (HEV-2) are transmitted through contaminated water and are responsible for the outbreaks of many large-scale epidemics in developing countries of Asia and Africa. Genotypes 3 (HEV-3) and 4 (HEV-4) have zoonotic potential and cause sporadic cases and small outbreaks in high-income countries. The first case of HEV infection in swine was confirmed in 1997 and later detected in other animal species. The aim of this study was to investigate the seroprevalence of HEV infection in pig farms in Vojvodina province, Serbia. Three hundred blood samples were collected from five different categories of pigs from 3 different farms on the territory of the South Bačka district in Vojvodina (Serbia). The analyses were conducted by in-house ELISA, while the western blot method was used as a confirmatory test for doubtful results. The presence of HEV IgG was detected on all three examined farms. The established seroprevalence in Farm A was 37%, 31% in Farm B, and 54% in Farm C. The mean seroprevalence for all farms was 40.66%. A higher seroprevalence was found in fatteners compared to younger categories of pigs. We concluded that HEV is widespread on pig farms with intensive management. Further analyses should be conducted with the aim of implementing a surveillance program to prevent possible human infection. [ABSTRACT FROM AUTHOR]