The Goat Cytotoxic T Lymphocyte-Associated Antigen-4 Gene: mRNA Expression and Association Analysis of Insertion/Deletion Variants with the Risk of Brucellosis.

The cytotoxic T lymphocyte-associated antigen-4 (CTLA4) gene, a member of the immunoglobulin superfamily, is crucial for maintaining immune homeostasis and preventing autoimmune diseases. Studies have shown that polymorphisms in the CTLA4 gene are linked to an increased risk of brucellosis in humans, but its association with brucellosis in goats remains unexplored. In this study, the tissue expression profile of CTLA4 in goats was investigated, and the correlation between InDel polymorphisms in the CTLA4 gene and susceptibility to brucellosis in goats was examined. The findings reveal the widespread expression of CTLA4 in goat tissues, particularly in the spleen and testes. The tested goat populations presented genotypes insertion/insertion (II), insertion/deletion (ID), and deletion/deletion (DD) at both the P1 and P2 loci, and an association analysis revealed significant differences in the distribution of genotypes and allele frequencies at the P1 and P2 loci of the CTLA4 gene between the Brucella goat case and the control groups (p < 0.05). Specifically, compared with the II genotype, the P1 and P2 loci were significantly associated with an elevated risk of brucellosis development in goats under both the codominant (ID/II) and dominant (ID + DD/II) models (P1, p = 0.042, p = 0.016; P2, p = 0.011, p = 0.014). Additionally, haplotype analysis indicated that haplotypes IP1DP2, DP1IP2, and DP1DP2 were significantly associated with an increased risk of brucellosis in goats compared to the reference haplotype IP1IP2 (p = 0.029, p = 0.012, p = 0.034). Importantly, the Lipopolysaccharide (LPS) stimulation of peripheral blood monocytes and/or macrophages from goats with the II, ID, and DD genotypes resulted in increased CTLA4 expression levels in the II genotype, leading to a robust LPS-induced inflammatory response. Through bioinformatic analysis, the observed effect of the InDel locus on Brucella pathogenesis risk in goats could be attributed to the differential binding of the transcription factors nuclear factor kappaB (NF-κB) and CCAAT/enhancer-binding protein α (C/EBPα). These findings offer potential insights for breeding strategies against brucellosis.

Disruption of Erythritol Catabolism via the Deletion of Fructose-Bisphosphate Aldolase (Fba) and Transaldolase (Tal) as a Strategy to Improve the Brucella Rev1 Vaccine.

Brucellosis is a bacterial zoonosis caused by the genus Brucella, which mainly affects domestic animals. In these natural hosts, brucellae display a tropism towards the reproductive organs, such as the placenta, replicating in high numbers and leading to placentitis and abortion, an ability also exerted by the B. melitensis live-attenuated Rev1 strain, the only vaccine available for ovine brucellosis. It is broadly accepted that this tropism is mediated, at least in part, by the presence of certain preferred nutrients in the placenta, particularly erythritol, a polyol that is ultimately incorporated into the Brucella central carbon metabolism via two reactions dependent on transaldolase (Tal) or fructose-bisphosphate aldolase (Fba). In the light of these remarks, we propose that blocking the incorporation of erythritol into the central carbon metabolism of Rev1 by deleting the genes encoding Tal and Fba may impair the ability of the vaccine to proliferate massively in the placenta. Therefore, a Rev1ΔfbaΔtal double mutant was generated and confirmed to be unable to use erythritol. This mutant exhibited a reduced intracellular fitness both in BeWo trophoblasts and THP-1 macrophages. In the murine model, Rev1ΔfbaΔtal provided comparable protection to the Rev1 reference vaccine while inducing fewer adverse reproductive events in pregnant animals. Altogether, these results postulate the Rev1ΔfbaΔtal mutant as a reproductively safer Rev1-derived vaccine candidate to be studied in the natural host.

Development of a colloidal gold immunochromatographic test strip for detecting the smooth Brucella.

Brucellosis, caused by Gram-negative Brucella, spreads in human and animal populations through contact with infected animals and products. Developing a rapid and sensitive detection technology for pathogen is crucial to reduce the risk of this disease transmitting between animal populations and to humans. We produced a monoclonal antibody LPS-6B5, which shows high affinity to LPS and limited cross-reactivity with other bacteria. Based on LPS-6B5, a colloidal gold immunochromatographic assay (GICA) was developed which demonstrates high sensitivity and specificity in detecting cultured B. melitensis, B. abortus and B. suis. The Gold Immunochromatographic Assay (GICA) strips exhibited the most sensitive detection limits, with a value of 7.8125 × 105 CFU/mL for Brucella melitensis, surpassing the sensitivity levels observed for Brucella abortus and Brucella suis. It is also suitable for clinical and field samples, providing a cost-effective and user-friendly alternative to traditional methods.

Spatiotemporal risk of human brucellosis under intensification of livestock keeping based on machine learning techniques in Shaanxi, China.

As one of the most neglected zoonotic diseases, brucellosis has posed a serious threat to public health worldwide. This study is purposed to apply different machine learning models to improve the prediction accuracy of human brucellosis (HB) in Shaanxi, China from 2008 to 2020, under livestock husbandry intensification from a spatiotemporal perspective. We quantitatively evaluated the performance and suitability of ConvLSTM, RF, and LSTM models in epidemic forecasting, and investigated the spatial heterogeneity of how different factors drive the occurrence and transmission of HB in distinct sub-regions by using Kernel Density Analysis and Shapley Additional Explanations. Our findings demonstrated that ConvLSTM network yielded the best predictive performance with the lowest average RMSE of 13.875 and MAE values of 18.393. RF model generated an underestimated outcome while LSTM model had an overestimated one. In addition, climatic conditions, intensification of livestock keeping and socioeconomic status were identified as the dominant factors that drive the occurrence of HB in Shaanbei Plateau, Guanzhong Plain, and Shaannan Region, respectively. This work provided a comprehensive understanding of the potential risk of HB epidemics in Northwest China driven by both anthropogenic activities and natural environment, which can support further practice in disease control and prevention.

The challenge of etiologic diagnosis of subacute and chronic meningitis: an analysis of 183 patients.

Subacute and chronic meningitis (SCM) presents significant diagnostic challenges, with numerous infectious and non-infectious inflammatory causes. This study examined patients aged 16 and older with SCM admitted to referral centers for neuroinfections and neuroinflammations in Mashhad, Iran, between March 2015 and October 2022. Among 183 episodes, tuberculous meningitis was the most common infectious cause (46.5%), followed by Brucella meningitis (24.6%). The cause of SCM was definitively proven in 40.4%, presumptive in 35.0%, and unknown in 24.6% of cases. In-hospital mortality was 14.4%, and 30.5% of survivors experienced unfavorable outcomes (Glasgow Outcome Scale 2-4). Patients with unknown causes had a significantly higher risk of death compared to those with presumptive or proven diagnoses (risk ratio 4.18). This study emphasizes the diagnostic difficulties of SCM, with one-quarter of cases remaining undiagnosed and over one-third having only a presumptive diagnosis. Improving diagnostic methods could potentially enhance prognosis and reduce mortality.

Spatiotemporal distribution and ecological factors of brucellosis among children from 2016 to 2020 in Inner Mongolia, China.

OBJECTIVE: In recent years, the increasing incidence of brucellosis in children has become more serious. However, relatively few studies have been conducted to characterize the spatialtemporal distribution of brucellosis in children. This study aimed to analyze the spatiotemporal distribution characteristics and ecological influencing factors of brucellosis incidence among children in Inner Mongolia. METHODS: This study used data on brucellosis incidence in children aged 0-14 years reported in Inner Mongolia from 2016 to 2020. A Bayesian model was used to analyze the spatial and temporal distribution of brucellosis in children from 2016 to 2020 in Inner Mongolia. Geographical weighted regression model was used to analyze the ecological factors related to the incidence of brucellosis in children. RESULT: Bayesian spatiotemporal analysis indicated that the highest brucellosis risk and increased disease incidence were observed in Hinggan, Inner Mongolia, in children aged 0-14 years. Alxa had the lowest risk but the incidence rate increased rapidly. The incidence of childhood brucellosis was positively associated with the number of sheep at the year-end (ß: 2.5909 ~ 2.5926, P < 0.01), average temperature (ß: 2.8978 ~ 2.9030, P < 0.05), and precipitation level (ß: 3.3261 ~ 3.3268, P < 0.01). CONCLUSION: From 2016 to 2020, the overall incidence of brucellosis in children in Inner Mongolia showed an upward trend, with cases exhibiting spatial aggregation. We should focus on areas where the incidence of brucellosis in children is rising rapidly. The incidence of childhood brucellosis was associated with the number of sheep at the year-end, average temperature and precipitation level. IMPLICATIONS AND CONTRIBUTION: The findings suggest that brucellosis in children is not to be taken lightly. For children should also focus on protection, take corresponding protective measures. While we focus on high-risk areas, we must also monitor areas where the risk of disease is low, but the incidence is rising fast, to prevent outbreaks in low-risk areas from becoming high-risk areas.

Detection and molecular typing of epidemic Brucella strains among camels, sheep, and cattle in Xinjiang, China.

Brucellosis is a zoonotic disease that can result in symptoms including infertility, abortion, testicular inflammation, and arthritis in affected sheep and cattle. The disease can spread through a range of mechanisms, making outbreaks difficult to control such that affected farms often suffer severe economic losses. In addition, humans can be affected by brucellosis, and the number of cases continues to rise annually. As rates of brucellosis in Xinjiang, China have been increasing substantially in recent years, this study was developed to assess brucellosis seroprevalence among herds of cattle, sheep, and camels in Xinjiang through the use of common diagnostic procedures like the Rose Bengal test (RBT) and PCR. A cross-sectional approach was used to analyze the seroprevalence of brucellosis across 720 total farm animals (320 camels, 250 sheep, and 150 cattle). RBT incidence rate for brucellosis was detected in 60 serum samples, 14 (4.38%) from camels, 45 (18%) from sheep, and 1 (0.67%) from cattle. PCR was performed as a confirmatory approach for these RBT-positive samples, with 55 ultimately being confirmed to be positive 13 (4.06%), 41 (16.4%), and 1 (0.67%) samples from camels, sheep, and cattle, respectively. In this survey, sheep exhibited the highest brucellosis seroprevalence using these two analytical techniques, while cattle exhibited the lowest seroprevalence and camels exhibited an intermediate rate. AMOS-PCR analyses identified Brucella melitensis as the unknown bacterium in some of these samples. These results offer new insight regarding brucellosis seroprevalence among farm animals that can be used to formulate more appropriate prevention and control policies, while also improving awareness of epidemic prevention efforts, the need for routine quarantine and disinfection, the benefits of scientific breeding and management, and approaches to improving breeding efficiency for the camel, cattle, and sheep industries.

Seronegative brucella meningitis diagnosed by CSF PCR: report on seven cases.

INTRODUCTION: Neurobrucellosis (NB) can be associated with meningitis and present as a headache with or without meningeal signs. Pseudotumor presentation of NB has been reported to be accompanied by lymphocytic predominant cerebrospinal fluid(CSF) pleocytosis. NB is diagnosed by means of isolation of Brucella from blood or CSF and/or the presence of anti-Brucella antibodies in the CSF. Molecular techniques have been used in chronic or challenging cases of NB. CLINICAL FINDINGS: We report on seven cases of NB presenting with different types of headache and signs of meningeal involvement. In five cases, signs of intracranial hypertension were evident in the form of papilledema, sixth nerve palsy and blurred vision. DIAGNOSIS: MRIs of the brain revealed signs of intracranial hypertension in three patients, basal meningeal enhancement in one patient and white matter lesions in one patient. Brucella serology in the blood and CSF was negative in all patients. It was interesting that four patients had normocellular CSF analysis with normal glucose and protein results. The diagnosis was made by Brucella PCR in all patients. CONCLUSION: NB should be considered in the differential diagnoses of pseudotumor cerebri syndrome in endemic areas. It is important to employ molecular techniques using sterile CSF samples in the investigation of Brucella.

Copper sensing transcription factor ArsR2 regulates VjbR to sustain virulence in Brucella abortus.

Brucellosis, caused by the intracellular pathogen Brucella, is a major zoonotic infection that promotes reproductive disease in domestic animals and chronic debilitating conditions in humans. The ArsR family of transcriptional regulators plays key roles in diverse cellular processes, including metal ion homeostasis, responding to adverse conditions, and virulence. However, little is known about the function of ArsR family members in Brucella. Here, we identified ArsR2 as a nonclassical member of the family that lacks autoregulatory function, but which nevertheless plays a vital role in maintaining copper homeostasis in B. abortus. ArsR2 is a global regulator of 241 genes, including those involved in the VirB type IV secretion system (T4SS). Significantly, ArsR2 regulates T4SS production in B. abortus by targeting VjbR which encodes a LuxR-type family transcriptional regulator. Moreover, copper modulates transcriptional activity of ArsR2, but not of VjbR. Furthermore, deletion of arsR2 attenuated virulence in a mouse model. Collectively, these findings enhance understanding of the mechanism by which ArsR proteins regulate virulence gene expression in pathogenic Brucella species.

Design a novel of Brucellosis preventive vaccine based on IgV_CTLA-4 and multiple epitopes via immunoinformatics approach.

Brucellosis is a zoonotic disease caused by Brucella, which is difficult to eliminate by conventional drugs. Therefore, a novel multi-epitope vaccine (MEV) was designed to prevent human Brucella infection. Based on the method of "reverse vaccinology", cytotoxic T lymphocyte epitopes (CTLEs), helper T lymphocyte epitopes (HTLEs), linear B-cell epitopes (LBEs) and conformational B-cell epitopes (CBEs) of four Brucella proteins (VirB9, VirB10, Omp 19 and Omp 25) were obtained. In order to keep the correct protein folding, the multiple epitopes was constructed by connecting epitopes through linkers. In view of the significant connection between human leukocyte antigen CTLA-4 and B7 molecules found on antigen presenting cells (APCs), a new vaccine (V_C4MEV) for preventing brucellosis was created by combining CTLA-4 immunoglobulin variable region (IgV_CTLA-4) with MEV protein. Immunoinformatics analysis showed that V_C4MEV has a good secondary and tertiary structure. Additionally, molecular docking and molecular dynamics simulation (MD) revealed a robust binding affinity between IgV_ CTLA-4 and the B7 molecule. Notably, the vaccine V_C4MEV was demonstrated favorable immunogenicity and antigenicity in both in vitro and in vivo experiments. V_C4MEV had the potential to activate defensive cells and immune responses, offering a hopeful approach for developing vaccines against Brucella in the upcoming years.

MRI findings and classification of brucella spondylitis: a China multicenter study.

OBJECTIVES: To propose a magnetic resonance imaging (MRI) classification of brucellosis spondylitis (BS) to determine a standard treatment method and a standard of post-treatment MRI follow-up. METHODS: This is a multicenter retrospective cohort study. All patients diagnosed with BS at six hospitals for 12 years, were included in the study. Data were collected retrospectively from the medical records of the patients. The classification of the cases was based on MRI. We analyzed MRI features of BS was categorized them into three types. RESULTS: Six hundred fifteen patients were included in the study, 78.9% of whom were males. The mean age was 53.05 ± 11.06 years. Involvement of single vertebrae occurred in 6.2% of patients, while 72.7% involved two vertebrae and 21.1% involvement of more than two vertebrae. The most common of the lumbar spine was 64.6%. The main findings of MRI were affected vertebral endplate 2/3 above (60.7%), intervertebral space narrow (77.9%), intervertebral disc involvement (83.6%), paravertebral abscess (50.9%), disc abscess (49.6%), epidural abscess (43.3%), vertebra height change (22.1%), psoas abscess (22.0%), and abnormal vertebral accessory (10.2%). MR imaging of BS was categorized into three types. Type I: 57 (9.3%) patients; Type II: 526 (85.5%) patients, including IIa (26.5%), IIb (36.9%), and IIc (22.1%); Type IV: vertebral appendage type, 63 (10.2%) cases; Type III: 32 (5.2%) patients. CONCLUSIONS: MRI features of multicenter data can provide theoretical support for diagnosing BS. MRI classification of BS can afford better help for clinical treatment and follow-up after clinical treatment.

Clinical characterization of brucellosis in children from non-pastoral areas: a report of five cases.

BACKGROUND: Brucellosis is a global public health concern and occurs mainly in young adults and the elderly, with children having a lower incidence, thus often leading to delayed treatment. This study aimed to describe the epidemiologic features and clinical characteristics of brucellosis in children. METHODS: In this retrospective study, the clinical data of five children diagnosed with brucellosis in Anhui Provincial Children's Hospital between January 1, 2021 and December 30, 2022 were analyzed. RESULTS: All five cases were from non-pastoral areas, among which three have a history of livestock exposure and originated from the countryside. All patients had medium-high grade fever, mostly accompanied by night sweats and malaise, and three had joint pains. Laboratory tests showed that their white blood cell count was normal or mildly raised, with lymphocytes as the predominant cell population. Four patients had anemia, four had aspartate aminotransferase and alanine aminotransferase abnormality, and two had elevated ferritin levels. All blood samples were positive for Brucella culture, one of which had positive bone marrow culture, and all had positive serology test results. All patients were treated with rifampicin, in combination with sulfamethoxazole or doxycycline for 6 weeks following diagnosis. Four children had a good prognosis, but one child had recurrent joint pain. CONCLUSIONS: The epidemiologic history of children from non-pastoral areas with brucellosis is often unclear; clinical manifestations and laboratory tests lack specificity; and they are easily delayed diagnosis. Clinicians should remain vigilant regarding the possibility of this disease in children with fever of unknown origin. The epidemiological history should be investigated in detail to improve the diagnostic ability of brucellosis. We recommend emphasizing serological testing. Children with brucellosis who receive timely diagnosis and standardized treatment can expect a favorable prognosis.

Diagnosis of brucellosis: Combining tests to improve performance.

INTRODUCTION: Brucellosis, a zoonotic infectious disease caused by bacteria of the genus Brucella, remains a significant global health concern in many parts of the world. Traditional diagnostic methods, including serological tests, suffer from limitations, including low sensibility and high false-positive rates, emphasizing the need for improved diagnostic strategies. In this study, we aimed to optimize diagnostic accuracy by reevaluating serological tests and exploring novel diagnostic algorithms. METHODS: A retrospective observational study was conducted using sera collected between June 2012 and June 2023 at the French National Reference Center for Brucella. Various serological tests, including Rose Bengal plate test (RBT), standard agglutination test (SAT), Brucellacapt, and ELISA for IgM and IgG, were performed. Different diagnostic algorithms were evaluated, combining RBT with SAT, Brucellacapt, and ELISA to enhance the performance of diagnostic tests. RESULTS: Among 3587 sera analyzed, 148 were confirmed cases of human brucellosis. Individual serological tests exhibited good sensitivity and specificity but lacked diagnostic accuracy. However, combining RBT with SAT or Brucellacapt significantly improved diagnostic performance, with reduced false positives. The most promising results were observed when an algorithm was built combining RBT, Brucellacapt, and ELISA for IgM and IgG (a score value of 0.5 with 90.5% for sensitivity, 99.7% for specificity, 92.4% for PPV, and 99.6% for NPV). CONCLUSIONS: Serological tests remain crucial for brucellosis diagnosis, but their limitations necessitate innovative diagnostic approaches. Combining multiple serological tests in diagnostic algorithms shows promise in improving diagnostic accuracy. Efforts to refine diagnostic, strengthen surveillance, and raise awareness are essential for effective brucellosis control, particularly in resource-limited settings.

Diagnosis of an atypical fever in a 55-year-old man. / Diagnostyka nietypowego stanu goraczkowego u 55-letniego mezczyzny.

BACKGROUND: The paper describes a diagnostically challenging case of recurrent fever with an atypical course in a 55-year-old man. The authors highlight the significant difficulty in reaching a correct diagnosis, as well as the doubts and problems associated with the diagnostics. They suggest considering brucellosis as a diagnosis based on the patient's history and the course of the disease characterized by fluctuating, intermittent, and nocturnal fever. OBJECTIVE: The aim of the study was to present an atypical and diagnostically challenging case of recurrent febrile episodes in a 55-year-old man. MATERIAL AND METHODS: A comprehensive diagnostic workup, including laboratory tests, imaging, and specialist consultations, was conducted. The patient's medical history, physical examination, and various diagnostic tests were analyzed. RESULTS: In a patient with recurrent episodes of fever, an infection with Brucella spp. was detected, which was effectively treated with long-term antibiotic therapy. Imaging and laboratory diagnostics, along with specialist consultations, helped rule out other potential causes of the symptoms. The patient achieved lasting improvement following the treatment. CONCLUSIONS: There is a high probability that the patient was infected with Brucella bacteria. Diagnostic challenges and the rarity of the disease, considered non-existent in Poland, hinder prompt confirmation or exclusion of infection. The authors aimed to highlight the possibility of a higher frequency of brucellosis in Poland. They suggest that Brucella bacteria may still be present, causing chronic, unrecognized, recurrent fevers. Based on the case analysis, the authors indicate that the frequent occurrence of this disease in Poland might be higher than reported by statistics.

The effect of Brucella abortus on glial activation and cell death in adult male rat's hippocampus.

A zoonotic disease called brucellosis can cause flu-like symptoms and heart inflammation. The bacteria responsible for this disease can also enter the brain, causing a condition called neurobrucellosis that can result in long-term neurological problems. In this study, researchers aimed to determine the changes in the hippocampal cells of rats infected with Brucella. For the study, 24 adult male albino rats were inoculated with 1 × 106 CFU Brucella abortus 544. The rats were then deeply anesthetized, and their hippocampus samples were taken for stereological, histological, and molecular studies. The results showed that the infected rats had increased microgliosis and astrogliosis. Furthermore, a high level of caspase-3 in their hippocampal tissue indicated their susceptibility to apoptosis. Additionally, there was a decrease in expression of Ki67, which further supported this. Sholl's analysis confirmed a significant failure in glial morphology. The study demonstrated that the pathogen has the ability to destroy the hippocampus and potentially affect its normal physiology. However, more research is needed to clarify various aspects of neurobrucellosis.

Protein disulfide isomerase A4 binds to Brucella BtpB and mediates intracellular NAD+/NADH metabolism in RAW264.7 cells.

The Toll/interleukin-1 receptor (TIR) signaling domain is distributed widely in mammalian Toll-like receptors and adaptors, plant nucleotide-binding leucine-rich repeat receptors, and specific bacterial virulence proteins. Proteins that possess TIR domain exhibit NADase activity which is distinct from the canonical signaling function of these domains. However, the effects of bacterial TIR domain proteins on host metabolic switches and the underlying mechanism of NADase activity in these proteins remain unclear. Here, we utilized Brucella TIR domain-containing type IV secretion system effector protein, BtpB, to explore the mechanism of NADase activity in host cells. We showed that using ectopic expression BtpB not only generates depletion of NAD+ but also loss of NADH and ATP in RAW264.7 macrophage cells. Moreover, immunoprecipitation-mass spectrometry, co-immunoprecipitation, and confocal microscope assays revealed that BtpB interacted with host protein disulfide isomerase A4 (PDIA4). The Brucella mutant strain deleted the gene for BtpB, significantly decreased PDIA4 expression. Furthermore, our data revealed that PDIA4 played an important role in regulating intracellular NAD+/NADH levels in macrophages, and PDIA4 overexpression restored the decline of intracellular NAD+ and NADH levels induced by Brucella BtpB. The results provide new insights into the metabolic regulatory activity of TIR domain proteins in the critical human and animal pathogen Brucella.

Stability of a stochastic brucellosis model with semi-Markovian switching and diffusion.

To explore the influence of state changes on brucellosis, a stochastic brucellosis model with semi-Markovian switchings and diffusion is proposed in this paper. When there is no switching, we introduce a critical value R s and obtain the exponential stability in mean square when R s < 1 by using the stochastic Lyapunov function method. Sudden climate changes can drive changes in transmission rate of brucellosis, which can be modelled by a semi-Markov process. We study the influence of stationary distribution of semi-Markov process on extinction of brucellosis in switching environment including both stable states, during which brucellosis dies out, and unstable states, during which brucellosis persists. The results show that increasing the frequencies and average dwell times in stable states to certain extent can ensure the extinction of brucellosis. Finally, numerical simulations are given to illustrate the analytical results. We also suggest that herdsmen should reduce the densities of animal habitation to decrease the contact rate, increase slaughter rate of animals and apply disinfection measures to kill brucella.

Tracking sero-molecular trends of swine brucellosis in Hawai'i and the central Pacific.

Introduction: Brucellosis is a zoonotic disease of mammals caused by bacterial species of the Brucella genus. The reservoir for disease is typically mammals, with species of Brucella found infecting amphibians, bats, and marine mammals. Brucella spp. can pass directly to humans through contact with infected animals or their products. Brucella spp. can cause chronic debilitating infections in mammals, including humans, and is associated with spontaneous abortions in infected animals, causing reduced fecundity. In Hawai'i, terrestrial species that could harbor Brucella spp. include swine, cattle, horses, and axis deer among others. The numerous feral swine in Hawai'i are known to carry Brucella suis, with evidence supporting infections in cattle. Brucella suis also poses infection risk to humans, dogs, and potentially horses across the state. Methods: In this study, 3,274 feral swine serum samples collected from 5 of the 8 main islands over a 15-year span were analyzed for exposure to B. suis. Of the 558 watersheds in the state, 77 were sampled as part of this effort. Spatial analysis was used to identify watersheds of concern. MLVA and whole genome SNP analysis was used for molecular epidemiological analysis. Results: Statewide seropositivity rates were triple that of feral swine found in the conterminous United States. Smoothed positivity rates were highest on Maui, followed by O'ahu, and the island of Hawai'i. Island-by-island analysis found high brucellosis positivity levels associated with specific watersheds and agricultural areas. Local spatial autocorrelation identified hot spots on O'ahu and Hawai'i. MLVA analysis of available B. suis from Hawai'i found molecular epidemiological connections with B. suis found in French Polynesia and the mainland US while differing from those in Tonga, Western Polynesia. Strains from Hawai'i are phylogenetically closest to strains from the United States. MLVA and SNP analysis found B. suis strains from Hawai'i fell into the genetic group that contains biovar 1 B. suis. Discussion: This work identified islands and watersheds of high brucellosis seropositivity in feral swine of Hawai'i, highlighting the magnitude of the zoonotic risk. Introduction of strains in recent history is unlikely due to modern animal trade and disease control practices. Genomic analysis of strains in Hawai'i and the Pacific area can provide hidden historical and local clues to brucellosis epidemiology in the state.