Background: Theileria spp. are responsible for ovine and caprine theileriosis, leading to significant morbidity and mortality in small ruminants.The present study aims to investigate Theileria spp. infections in small ruminants from Southern Punjab in Pakistan, and genetic characterize revealed Theileria spp. isolates. Methods: A total of 93 sheep and 107 goats were sampled between May and August 2022. Blood smears were examined microscopically, and PCR amplification targeting the 18S rRNA gene was performed to detect Theileria spp. Additionally, specific PCR assays targeting 18S rRNA and ms1 partial sequences were used to identify Theileria ovis and T. lestoquardi, respectively. Results: The prevalence of Theileria spp. was significantly higher using PCR (13.5%) compared to microscopic screening (5%). Sheep showed a higher prevalence rate (19.4%) compared to goats (8.4%) (p = 0.024). Young sheep aged ≤ 1 year were more commonly infected with Theileria spp. (41%) compared to older sheep (p = 0.006). The prevalence of Theileria spp. was higher in sheep-only herds (37.3%) compared to goat-only herds (18%) or mixed-species herds (8.1%) (p = 0.015). The prevalence rates of T. ovis and T. lestoquardi were 9% and 2.5%, respectively, with four animals (2 goats and 2 sheep) showing co-infection. Phylogenetic analysis revealed that our T. ovis 18S rRNA sequence clustered with previously reported sequences from sheep in Turkey, China, Spain, and goats in Tanzania. The obtained T. lestoquardi ms1 partial sequence formed a distinct cluster from other T. lestoquardi isolates in Pakistan and neighboring countries. Conclusion: Theileria spp. co-circulation in Pakistani small ruminants, particularly the presence of T. ovis and T. lestoquardi, highlights the need for attention from animal health decision-makers due to their financial and health impacts.

Background: Rickettsia spp. are vector-borne zoonotic pathogens that cause febrile illness in humans. Rickettsioses is not included in the Colombian national surveillance system and is subsequently expected to be underreported. This cross-sectional study aimed to determine the seroprevalence of Rickettsia spp. and the closely related Orientia tsutsugamushi in two indigenous populations residing in the Sierra Nevada de Santa Marta, Colombia. Materials and Methods: Serum samples (n = 539) were collected from the Wiwa and Koguis people between 2021 and 2022. Serum samples were screened for spotted fever group (SFG) and typhus group (TG) Rickettsia spp. using the Fuller laboratories Rickettsia IgG IFA kit and for O. tsutsugamushi with the Scrub Typhus Detect™ IgG ELISA. Results: We observed an overall seroprevalence of 26.2% (95% confidence interval [CI] 22.5-30.1] for Rickettsia spp. of the SFG, 5.4% (95% CI 3.6-7.6) for Rickettsia spp. of the TG and 4.3% (95% CI 2.7-6.3) for O. tsutsugamushi. Common risk factors for zoonotic disease infections were assessed for 147 of the Wiwa participants. Increased odds of seropositivity for SFG Rickettsia spp. were observed for Wiwa participants who cared for livestock, including assisting with the birth of cattle (odds ratio [OR] = 8.85; 95% CI 1.54-50.90; p = 0.015) and goats (OR = 7.60; 95% CI 1.70-33.90; p = 0.008). Conclusions: These results highlight a notable exposure to Rickettsia spp., especially the SFG, in rural Colombia. Together with recent reports of high mortality for Rocky Mountain Spotted Fever in nearby regions of South America, more detailed investigations focusing on improving knowledge and awareness as well as "One Health" and "causes-of-fever" studies are needed. The characterization of Rickettsia spp. infections in humans, livestock, and tick vectors with their potential transmission routes could make a high impact on these easily treatable diseases.

Background: Aedes aegypti, is the primary vector of dengue, Chikungunya, Zika, and yellow fever viruses. Both natural and human-impacted landscapes have selective pressures on Ae. aegypti, resulting in strong genomic structure even within close geographical distances. Materials and Methods: We assess the genetic structure of this medically important mosquito species at the northern leading edge of their distribution in Southwestern USA. Ae. aegypti were collected during 2017 in the urban communities of El Paso and Sparks, Texas (USA) and in the city of Ciudad Juárez, Mexico. Results: Thousands of nuclear loci were sequenced across 260 captured Ae. aegypti. First, we recovered the genetic structure of Ae. aegypti following geography, with all four major collection communities being genetically distinct. Importantly, we found population structure and genetic diversity that suggest rapid expansion through active-short distance dispersals, with Anapra being the likely source for the others. Next, tests of selection recovered eight functional genes across six outliers: calmodulin with olfactory receptor function; the protein superfamily C-type lectin with function in mosquito immune system and development; and TATA box binding protein with function in gene regulation. Conclusion: Despite these populations being documented in the early 2000s, we find that selective pressures on specific genes have already occurred and likely facilitate Ae. aegypti range expansion.

Background: While the influence of landscape and microclimatic conditions on tick populations is well-documented, there remains a gap in more specific data regarding their relationship to rewilding efforts with large herbivore activity. Objective: This pilot study, spanning from 2019 to 2021, explores the effects of naturalistic grazing by large semi-wild ungulates on tick abundance in the Milovice Reserve, Czechia. Methods: Tick collection was observed using flagging techniques at two distinct sites of rewilding area: one grazed, actively utilized by animals involved in the rewilding project, and one ungrazed, left fallow in neighboring areas utilized only by wild animals. Transects, each measuring 150 m in length and 5 m in width (750 m2), were established at these two sampling locations from March to September between 2019 and 2021. To minimize potential bias resulting from tick movement, a 300 m buffer zone separated the two sites. Data analysis employed a generalized estimating equations (GEE) model with negative binomial regression. The study assessed potential variations in tick abundance between selected transects, considering factors such as plant cover seasonality, temperature, and humidity. Results: During the collection periods, we gathered 586 live ticks, with 20% found in grazed areas and 80% in ungrazed areas. Notably, tick abundance was significantly higher in ungrazed areas. Peaks in tick abundance occurred in both grazed and ungrazed areas during spring, particularly in April. However, tick numbers declined more rapidly in grazed areas. Microclimatic variables like temperature and humidity did not significantly impact tick abundance compared to landscape management and seasonal factors. Conclusion: Rewilding efforts, particularly natural grazing by large ungulates, influence tick abundance and distribution. This study provides empirical data on tick ecology in rewilded areas, highlighting the importance of landscape management and environmental factors in tick management and conservation. Trophic rewilding plays a crucial role in shaping ecosystems and tick population dynamics in transformed landscapes.

Background: Toxoplasmosis is one of the most widespread foodborne parasitic zoonoses caused by the obligate intracellular protozoan Toxoplasma gondii. Although a number of studies have reported on the seroprevalence and risk factors of T. gondii infection in ruminants in China, information about T. gondii infection in cattle in Hunan province of China is not available. Material and Methods: Sera of 985 cattle and 1147 goats were examined for the presence of specific antibodies against T. gondii using the indirect hemagglutination test. Some risk factors related to the presence of cats, herd size, gender, age, and geographical origin were determined using a binary logistic regression. Results: Specific IgG against T. gondii were detected in 8.3% (82/985) and 13.3% (153/1147) of the cattle and goats, respectively. Based on statistical analysis, the presence of cats and gender were considered important risk factors for T. gondii infection in cattle and goats in the farms in this study (P < 0.05). Conclusion: Our results provide a baseline for future prevention and control of T. gondii infection in cattle and goats in Hunan province, subtropical China. This is the first report of T. gondii seroprevalence in cattle in Hunan province, China.

Background: Madariaga virus (MADV), a member of the eastern equine encephalitis virus (EEEV) complex, circulates in Latin America and exhibits distinct evolutionary and ecological features compared to the North American EEEV. While published data have shed light on MADV ecology, several key aspects remain unknown. Methods: In this study, we compiled data on virus isolation, vector competence, and animal serology collected over six decades in Latin America to identify critical knowledge gaps on MADV transmission and ecology. Results: Specific vertebrate animals serving as amplifying hosts and the mosquito species acting as enzootic and epizootic vectors have not yet been identified. Other aspects that remain unclear are the virus current geographic distribution, the role of equines as hosts in epizootic cycles, and the full impact of MADV on human health in endemic regions. Conclusions: The numerous knowledge gaps surrounding MADV, its widespread distribution in Latin America, and its potential to cause severe disease in animals and humans emphasize the urgent need for increased research efforts, heightened awareness, and intensified surveillance towards this potential emerging threat.

Background: Rodent is a reservoir of various zoonotic pathogens. Wanzhou section of the Three Gorges reservoir region (TGRR) is a superior habitat for rodents, and the situation of rodent-borne zoonotic pathogens in this region has not been surveyed in recent years. Materials and Methods: Rodents were night trapped with mousecage or mousetrap in urban and surrounding towns' indoor or outdoor areas of the Wanzhou section of the TGRR, and nucleic acid was extracted from their lung or a mixture of liver, spleen, and kidney. Commercialized qPCR kits for pathogenic Leptospira spp., Rickettsia typhi, Anaplasma phagocytophilum, Bartonella spp., Orientia tsutsugamushi, and Francisella tularensis and qRT-PCR kits for hantavirus (HV), and severe fever with thrombocytopenia syndrome virus (SFTSV) were used for the detection of associated pathogens in collected rodents. Results: From 2021 to 2023, 604 rodents belonging to 10 species were collected. HV and pathogenic L. spp. were detected positive, with infection rates of 0.66% (4/604) and 1.32% (8/604), respectively. B. spp. were detected positive with an infection rate of 4.73% (19/402) in the rodents trapped in 2022 and 2023. Other five pathogens were all detected negative. Conclusion: This study showed that the Wanzhou section of the TGRR had HV, pathogenic L. spp., and B. spp. co-circulation in rodents. Hence, more attention should be paid to the prevention and control of associated rodent-borne diseases.

Background: Chikungunya is a zoonotic disease caused by the Chikungunya virus (CHIKV), primarily transmitted to humans through infected Aedes mosquitoes. The infection is characterized by symptoms such as high fever, musculoskeletal pain, polyarthritis, and a rash, which can lead to severe complications such as encephalitis, meningitis, and even fatalities. While many disease manifestations resemble those of other viral infections, chronic arthritis caused by CHIKV is unique, and its molecular mechanisms remain ill-defined. Materials and Methods: Proteomics data from both cellular and patient levels of CHIKV infection were curated from PubMed and screened using inclusion and exclusion criteria. Patient serum proteomics data obtained from P RIDE underwent reanalysis using Proteome Discoverer 2.2. Enrichment and protein-protein interaction network analysis were conducted on differentially expressed proteins from both serum and cellular datasets. Metabolite data from CHIKV-infected patients were further retrieved, and their protein binding partners were identified using BindingDB. The protein-metabolite interaction pathway was further developed using MetaboAnalyst. Results: The proteomics data analysis revealed differential expression of proteins involved in critical host mechanisms, such as cholesterol metabolism and mRNA splicing, during CHIKV infection. Consistent upregulation of two actin cytoskeleton proteins, TAGLN2 and PFN1, was noted in both serum and cellular datasets, and their upregulations are associated with arthritis. Furthermore, alterations in purine metabolism were observed in the integrative proteome-metabolome analysis, correlating with cytoskeletal remodelling. Conclusion: Collectively, this integrative view sheds light on the involvement of actin cytoskeleton remodeling proteins and purine metabolic pathways in the development of arthritis during CHIKV infection.

Background: Early detection and monitoring of SARS-CoV-2 infections in animal populations living in close proximity to humans is crucial for preventing reverse zoonosis of new viral strains. Evidence accumulated has revealed widespread SARS-CoV-2 infection among white-tailed deer (WTD), (Odocoileus virginianus) populations in the United States except in the southeast region. Therefore, the objective was to conduct surveillance for evidence of SARS-CoV-2 infection among WTD in Mississippi. Materials and Methods: Blood, kidney tissues, and nasal swab samples were collected in 17 counties from hunter-harvested deer during 2021-2022 and 2022-2023.Samples of kidney tissue were collected to evaluate for detecting antibody as a possible alternative to blood that is not always available from dead WTD. Nasal swab samples were tested for SARS-CoV-2 viral RNA by a RT-PCR assay. Sera and kidney tissue samples were tested for SARS-CoV-2 antibody by an enzyme-linked immunoassay (ELISA) and sera by a plaque reduction neutralization test (PRNT80). Results: The results of testing sera and kidney homogenate samples provided the first evidence of SARS-CoV-2 infection among WTD in Mississippi. The infection rate during 2021-2022 was 67% (10/15) based on the detection of neutralizing antibody by the PRNT80 and 26%(16/62) based on the testing of kidney tissue homogenates by an ELISA, and viral RNA was detected in 25% (3/12) of nasal swab samples. In 2022 to 2023, neutralizing antibody was detected in 62% (28/45) of WTD serum samples. In contrast, antibodies were not detected in 220 kidney homogenates by an ELISA nor was viral RNA detected in 220 nasal swab samples. Evidence of WTD activity was common in urban areas during the survey. Conclusion: Overall, the findings documented the first SARS-CoV-2 infection among WTD in Mississippi and showed that WTD commonly inhabited urban areas as a possible source of acquiring infection from humans infected with this virus.

Background: This study conducted in the departments of Oueme and Plateau aims to assess the presence of the dengue virus and its different serotypes in Aedes aegypti and Aedes albopictus, as well as the epidemic risk incurred by the populations. Methods: Collections of adult mosquitoes using human landing catches (HLC) were carried out in six communes, three (Porto-Novo, Adjarra, and Avrankou) in the Oueme department and the rest (Ifangni, Kétou, and Pobè) in the Plateau department. Pools of ten Aedes mosquitoes were formed, and stored at -80°C in RNA later. RT-PCR was used to detect dengue virus, and conventional PCR for the different serotypes. Inspection of water containers and collection of Aedes larvae was performed inside and around each house to calculate the stegomyan indices. Results: In the six communes, the dengue virus was present both in Ae. aegypti and Ae. albopictus. Combined data of the two Aedes species at the communes level revealed infection rates ranging from 80.00% (95% CI: 61.43-92.29) to 96.67% (95% CI: 82.78-99.92). In all the communes, the values of stegomyan indices reached the WHO threshold, which indicates the existence of the risk of an arbovirus epidemic. In addition, the infection rates were similar for Ae. aegypti [88.19% (95% CI: 81.27-93.24)] and Ae. albopictus [86.79% (95% CI: 74.66-94.52)]. The three virus serotypes detected in the pools of Aedes were DENV-1, DENV-3, and DENV-4, with a high prevalence for the first two. Conclusion: This study revealed that three serotypes (DENV-1, DENV-3, and DENV-4) of dengue virus circulate in Ae. aegypti and Ae. albopictus in the departments of Oueme and Plateau. Moreover, the risk of transmission of arboviruses was globally high and variable from commune to commune. This information is essential for informed decision-making in the preventive control of the disease.