Rose Bengal Research Articles

A retrospective study of risk factors associated to brucellosis seropositivity and bacterial isolation in bovine carcasses with cervical bursitis from the states of Mato Grosso, Pará and Tocantins - Brazil.

Visible Light-Responsive Green Mediated Synthesized ZnCuO Nanocomposites: Characterization and Photocatalytic Application Toward the Degradation of Rose Bengal Dye

Brucellosis, a contagious zoonotic bacterial infection affecting livestock and wildlife, is primarily caused by Brucella abortus, globally. However, in South Africa, the true prevalence of bovine brucellosis remains unknown because of a lack of epidemiological data. Therefore, this study used diagnostic data to evaluate and determine the seropositivity of bovine brucellosis based on Rose Bengal test (RBT) screening and confirmation with complement fixation test (CFT) in Limpopo and Free State provinces between 2013 and 2022. The use and limitations of this data were also evaluated based on the bovine brucellosis scheme in South Africa. The study revealed the overall seropositivity of 4.2% (n = 8980/212 440) for bovine brucellosis based on RBT and CFT in series. In Limpopo province, the brucellosis seropositivity was slightly higher at 4.3% (n = 7488/173 011) compared to 3.8% (n = 1492/39 429) in Free State province. Analysis of brucellosis distribution over the study period indicated significant variation (p < 0.001) both between and within the provinces. Notably, the highest prevalence in Limpopo occurred during 2013–2017, whereas in Free State, peak prevalence was observed in 2013 and 2016. Challenges preventing an accurate reflection of the brucellosis seropositivity in these provinces for the period include a lack of data on vaccination history and herd status of the samples submitted, as well as the inability to match the CFT results from different laboratories, because some laboratories are only accredited to perform the RBT.ContributionInsights gained from retrospective studies such as this study can play crucial roles in shaping effective control and preventative measures against bovine brucellosis. Given the challenges in obtaining confirmatory test results, we suggest that brucellosis tests be conducted at a single central laboratory or that the government provides a central database where all laboratories can enter their data. Furthermore, information submitted to the laboratories must make herd and vaccine history compulsory for sample submission to ensure more accurate data.

Background This study evaluates the diagnostic value of etiological and serological testing and the clinical characteristics of spine brucellosis. Methods A retrospective analysis was conducted on 200 cases of spine brucellosis diagnosed between 2012 and 2025, alongside 200 non-infected controls. Bacterial cultures, clinical features, serological results (Standard Agglutination Test [SAT] and Rose Bengal Test [RBT]), and imaging findings were analyzed. Results Among 200 patients with spinal involvement, Brucella melitensis was confirmed via bacterial culture in 29 cases (14.5%), with spinal tissue culture demonstrating the highest diagnostic yield (58.6%, 17/29) and a median detection time of 3 days. Spinal brucellosis constituted 25% of all brucellosis cases, exhibiting a male predominance (76.5%) and a mean patient age of 52.2 ± 10.1 years. The median hospital stay was 14.7 ± 7.8 days, while the median duration of chronic spinal brucellosis was 4.4 months (IQR: 1.5–11.5). The lumbar spine was the most frequently involved site (78.3%), followed by the cervical (8.6%) and thoracic (5.1%) regions. CT imaging revealed characteristic lesions, including bone destruction (53.0%), intervertebral space stenosis (38.5%), disc herniation (22.0%), degenerative changes (4.5%), and osteomyelitis (1.0%). Diagnostic sensitivity varied significantly across methods: blood culture (48.0%, 12/25), lumbar tissue culture (65.4%, 17/26), SAT(91.3%, 157/172), and RBT(95.9%, 165/172). Notably, all 15 serologically false-negative cases (7 by RBT and 15 by SAT) occurred in male patients. ROC curve analysis identified C-reactive protein (CRP) as a robust biomarker, with a cutoff of 1.21 mg/L yielding 85.2% sensitivity and 71.7% specificity (Z = 6.167, p &amp;lt; 0.001). Multivariable regression identified female sex (OR = 2.44, 95% CI: 1.20–4.96) and lumbar involvement (OR = 1.61, 95% CI: 0.75–3.46) as independent predictors of chronicity (p &amp;lt; 0.05). All patients received standard 3-month combination therapy with doxycycline and rifampicin, with surgical intervention required in 45.5% (91/200) of cases. Clinical outcomes were favorable across the cohort, with rare reports of severe complications. Conclusions Brucella Spondylitis, typically chronic and lumbar-predominant, presents with bone destruction, gap stenosis, and disc herniation on imaging. Diagnosis relies on lumbar tissue culture, SAT, RBT, and CT. Early diagnosis and combined medical-surgical management improve outcomes.

IntroductionPhotodynamic therapy (PDT) is a promising cancer treatment. However, the efficacy of photosensitizers such as rose bengal (RB) is often limited by poor delivery. Dendrimer-based nanocarriers can enhance PDT efficacy in vitro, but their in vivo safety profile remains largely uncharacterized. This study aimed to assess the systemic safety of three different dendrimer-based RB delivery systems in a healthy mouse model.MethodsBALB/c mice were randomly divided into eight groups and received weekly intraperitoneal injections of either PBS (control), free RB, or one of three carriers (phosphorus dendrimer 1cat, dendrimersome DG2, PPI G3 dendrimer) with or without RB. Body weight was monitored weekly. Blood and urine samples were collected over four weeks for comprehensive biochemical and microscopic analysis, assessing markers for liver, kidney, and muscle function.ResultsNo significant changes in body weight were observed across any of the groups. Analysis of blood biochemical parameters (including ALT, AST, urea, creatinine, and LDH) and urine profiles revealed no statistically significant differences between any treatment group and the PBS control group over the four-week study period. The observed minor fluctuations in some parameters were not dose- or time-dependent and remained within normal physiological ranges.ConclusionThe three tested nanocarrier systems - phosphorus dendrimer 1cat, dendrimersome DG2, and PPI G3 dendrimer - and their respective rose bengal formulations are well tolerated and do not induce systemic toxicity in BALB/c mice at the tested concentrations. These findings support their safety for in vivo applications and provide a basis for future efficacy studies in tumor-bearing animal models.

Introduction Donkeys are vital to livelihoods in Kenya, yet their exclusion from national disease surveillance leaves potential health risks underexplored. Brucellosis, a significant zoonosis, remains poorly characterized in donkeys despite frequent close contact with humans. This study aimed to determine the seroprevalence, molecular detection, and risk factors for Brucella spp. infection in donkeys, and to assess owner knowledge, attitudes, and practices (KAP) across seven Kenyan counties representing diverse production systems. Methods Between October 2024 and February 2025, a cross-sectional survey sampled 392 donkeys. Serum was tested using the Rose Bengal Plate Test (RBPT) and indirect ELISA (iELISA). Donkeys testing seropositive on either test (n = 42) had their corresponding whole blood samples subjected to DNA extraction for PCR analysis, targeting Brucella abortus and Brucella melitensis . Structured interviews with owners were conducted to assess knowledge, attitudes, and practices (KAP). Mixed-effects logistic regression in R was used to identify risk factors. Results Overall seroprevalence was 10.7% by RBPT, 2.0% by iELISA, and 0.0% by PCR. All iELISA-positive cases (n = 8) were from Turkana (4), Narok (3), and Nairobi (1). Young donkeys (&amp;lt;3 years) had significantly higher odds of being seropositive (aOR = 11.8; 95% CI: 1.70–81.99; p = 0.013). Owner knowledge was low—only 25.3% had heard of brucellosis and risky practices were common, with 91.1% assisting foaling without protective equipment and 19.4% consuming donkey products, often raw. Conclusion Donkeys in Kenya may contribute to Brucella transmission within mixed livestock systems and to humans. Inclusion of donkeys in brucellosis surveillance, targeted community education, and improved diagnostics are recommended. These findings provide the first field-based evidence of donkey brucellosis in Kenya and underscore the importance of integrating donkeys into One Health strategies to reduce zoonotic risk.

ABSTRACTProgrammed cell death (PCD) mediates plant development and environmental interactions. Photosynthesis‐derived singlet oxygen (1O2) is one of key reactive oxygen species (ROS) implicated in acclimation and PCD responses to environmental stress conditions. Using Arabidopsis thaliana cell suspension culture system, we characterized the PCD induced by Rose Bengal (RB), a photosensitizer generating 1O₂ upon light exposure. Obtained results reiterated that RB‐induced PCD is light and chloroplast dependent. Further, we demonstrate that PCD induced by RB involves calcium signaling and mitochondria, thus sharing common features with other forms of regulated cell death in plants. The PCD induced by RB was associated with early transcriptional reprogramming, involving switching off the primary metabolism and activation of stress response and cell death related pathways (e.g., oxidative stress, hypoxia, immunity, and salicylic acid). The constructed gene regulatory network featured 1O2‐responsive genes and suggested involvement of transcription factor ANAC102 in retrograde regulation of RB‐induced PCD. Interestingly, treatment with RB also induced light independent toxicity, showing features of uncontrolled, necrotic cell death. Presented findings highlight RB as a valuable tool for studying 1O2‐induced PCD that may advance future work on chloroplast‐mediated oxidative stress responses and enhancing plant resilience to climate change‐related stressors through targeted modulation of ROS pathways.

Precise intraoperative integration of bioelectronic devices with wet tissue surfaces remains a challenge due to the limited spatial control of adhesion sites. Here, an in situ spatially programmable electrical bioadhesive (termed "STICH")is reported that enables site-selective adhesion and functional coupling via light-activated bonding with wet biological tissue. Upon irradiation with patterned green light, Rose Bengal in a chitosan/silver nanowire hydrogel matrix generates singlet oxygen, which oxidizes amino acid residues into carbonyl groups on the tissue surface. The covalent bonding is then formed between the newly formed reactive carbonyl group and amine groups on chitosan. The spatially programmable adhesive allows robust tissue bonding with a lap-shear strength of 160kPa and precise adhesion regions at ≈2 µm resolution. The light-patternable adhesive enables spatially resolved mechanical coupling for directional electromechanical sensing on ex vivo cardiac tissue. The low impedance adhesive interface also provides spatially programmed electrical coupling for in vivo neuromuscular stimulation on intraoperatively selected muscle groups. This platform advances microscale device-tissue integration and paves the way for reconfigurable bioelectronic therapies.

Background and Objectives: Brucellosis, a contagious infection caused by Brucella spp, remains the most widely reported bacterial zoonosis globally. Since the clinical manifestations are often non-specific, reliable laboratory confirmation, in ac- cordance with World Health Organization recommendations, is essential. This study reports human brucellosis cases between 2017 and 2025 based on serological confirmation; it also discusses approaches to improve diagnostic accuracy for better surveillance, timely treatment, and support public health strategies. Materials and Methods: A total of 95 serum samples were obtained from patients presenting with clinical manifestations suggestive of brucellosis. Initial screening was performed using the Rose Bengal test, and positive or equivocal samples were further analyzed by Enzyme-Linked Immunosorbent Assay to detect both IgG and IgM antibodies for serological confirmation. Results: Among the 95 patients investigated, the Rose Bengal test yielded positive results in 69.5% of cases. Serological confirmation by ELISA demonstrated IgM seropositivity in 57.9% of patients and IgG seropositivity in 55.8%. The diagnos- tic performance of ELISA showed a sensitivity of 83.3% for IgM detection and 80.3% for IgG detection. Regarding patient demographics, the mean age was 37.9 ± 16.4 years, with a slight male predominance (54.7%). Conclusion: The study reveals a considerable proportion of brucellosis-positive cases, confirming the value of serological testing in endemic regions such as Morocco. Nonetheless, serology should be complemented with advanced diagnostic methods, including PCR to improve both the accuracy and timeliness of diagnosis. These findings support the adoption of integrated diagnostic approaches and the reinforcement of laboratory capacity in high-risk areas.

Stroke is a leading global health concern, with cerebral infarction accounting for 62% of cases. Despite advances in acute-phase treatments, functional impairments such as motor deficits remain prevalent. This study investigates the potential of human induced pluripotent stem cell (iPSC)-derived cerebral cortical neurons for neural regeneration and motor function recovery in a female mouse model of ischemic stroke. Cerebral infarction was induced using the Rose Bengal photothrombosis method, followed by transplantation of iPSC-derived cortical neurons into the area adjacent to the infarction. Behavioral recovery was assessed using the foot fault and cylinder tests. Histological analysis was performed to evaluate graft integration and neurite extension. Foot fault test demonstrated significant improvements in fine motor function in the transplantation group compared to the vehicle group. However, no recovery was observed in the cylinder test, which assesses gross motor function. Neurite extension from grafted cells was observed along the corticospinal tract, with axonal projections reaching the spinal cord in 68% of transplanted mice. In addition, neurite outgrowth extended to the thalamus, superior colliculus, and vestibular nucleus, suggesting integration into multiple neural circuits. Histological analysis revealed that 16.4% and 47.3% of grafted cells expressed CTIP2 and SATB2, respectively, indicating the presence of both deep- and upper-layer cortical neurons. This study demonstrates that iPSC-derived cortical neurons extend axons along the corticospinal tract and can promote fine motor recovery after stroke. However, further research is needed to validate functional connectivity and long-term safety. These findings offer a promising avenue for developing cell-based therapies for stroke patients.

Abstract Organic dye contamination of aquatic habitats has become a global issue, necessitating the urgent adoption of radical remediation methods using environmentally benign techniques. This study aims to synthesize and characterize a novel NiO@Chitosan (NiO@CS) composite material that can be used as a photocatalyst for wastewater treatment. The successful preparation of nanocomposites by a coprecipitation method is followed by characterization studies using various techniques, including X‐ray diffraction (XRD), scanning electron microscopy (SEM), Fourier‐transform infrared spectroscopy (FTIR), and UV‐visible spectroscopy, to measure structural and optical properties. The NiO nanoparticles are homogeneously distributed within the chitosan matrix and establish an appreciable crystallographic structure, improving photocatalytic activity. Under UV light irradiation, a degradation efficiency of 86.0% for Crystal Violet (CV) and 86.6% for Rose Bengal (RB) is achieved in 60 min for the NiO@CS nanocomposite. Moreover, the catalyst maintained over 85% of its initial activity for three consecutive cycles, exhibiting excellent reusability and stability. These results indicate the promise of NiO@Chitosan nanocomposites as efficient, recyclable, and eco‐friendly photocatalysts for dye‐based water pollution.

ABSTRACTDespite the efficacy of current hepatitis B vaccines, approximately 10% of immunocompetent individuals remain non-responsive, underscoring the urgent need for novel adjuvants to enhance vaccine-induced immunity. In this study, we investigated PV-10, a 10% Rose Bengal solution, for its potential to activate the stimulator of interferon genes (STING) pathway and enhance both innate and adaptive immune responses. Through molecular docking, we demonstrated that Rose Bengal binds to the ligand-binding domain of STING with an affinity of −7.1 kcal/mol (−7.0 to −9.0 kcal/mol for moderate binding), promoting dimer stabilization via hydrophobic and hydrogen-bonding interactions. In the human acute monocytic leukemia cell line THP-1, treatment with PV-10 induced the phosphorylation of key downstream signaling proteins, including TBK1, IRF3, and NF-κB p65, and uniquely generated a high molecular weight STING band indicative of dimer formation. Cytokine profiling revealed a time-dependent increase in pro-inflammatory cytokines and chemokines following PV-10 treatment. Furthermore, in an in vitro model, dendritic cells were pulsed with hepatitis B surface antigen (HBsAg)-derived peptides, HBV-1 (TVELLSFLPSDFFPSV, extended HBsAg epitope) and HBV-2 (FLPSDFFPSV, minimal cytotoxic T lymphocyte epitope) and then the pulsed DCs were used to prime CD8+ T-cells. HBsAg–primed CD8+ T-cells exhibited significantly enhanced IFN-γ secretion when co-cultured with HBsAg-positive hepatoma cells in the presence of PV-10 compared to vehicle-treated controls. These findings indicate that PV-10 functions as a potent STING agonist, stabilizing STING dimerization, and eliciting an immune microenvironment conducive to robust antigen presentation and T-cell activation, thereby demonstrating its potential as a novel adjuvant for improving hepatitis B vaccine efficacy, particularly in vaccine non-responders.

New photoactive nanofiber materials based on an aminolyzed polycaprolactone membrane with demonstrated cytocompatibility were developed. Two photoactive compounds, the photosensitizer Rose Bengal and the nitric oxide photodonor 4-nitro-3-(trifluoromethyl)aniline, were covalently bonded to the nanofiber surface, with or without a glutaraldehyde linker. The surface functionalization was confirmed via X-ray photoelectron spectroscopy, UV-vis absorption, and steady-state and time-resolved luminescence spectroscopy. Upon excitation with green or blue light, these materials efficiently generate antibacterial species, including singlet oxygen, with a slight contribution of hydrogen peroxide and nitric oxide. A potent light-induced antibacterial effect was demonstrated against Escherichia coli. Furthermore, the functionalized photoactive membranes, especially those with a glutaraldehyde linker and photosterilized by light, not only excluded the material toxicity but also demonstrated improved cell adhesion and proliferation when tested with adipose tissue-derived stem cells. These materials, which offer a unique combination of light-controlled surface sterilization and high cellular compatibility, are promising for advanced tissue engineering applications.

Brucellosis remains an underreported zoonotic disease in Ecuador. Its control program in cattle integrates diagnostic testing, vaccination, and eradication incentives, although participation is largely voluntary. Since 2025, vaccination has become compulsory nationwide. Human surveillance remains largely passive, and strain-level data are very limited. This study applied an integrated approach, combining serology (Rose Bengal and SAT-EDTA), microbiological culture, and molecular diagnostics, to assess the presence and diversity of Brucella spp. in cattle and pigs from six slaughterhouses in the northern Andean highlands. A total of 2054 cattle and 1050 pigs from Carchi, Imbabura, and Pichincha were sampled. Among cattle, 133 (6.5%; 95% CI: 5.5–7.6) were seropositive, and viable B. abortus strains were isolated from 17 (12.8%). Genus identification was confirmed by IS711-PCR, while species- and biovar-level differentiation was achieved with AMOS-PCR; additional assays targeting the ery gene and RB51 marker were used to distinguish field from vaccine strains. Biotyping and molecular analysis revealed a predominance of B. abortus biovar 4 (13/17 isolates) over biovar 1, all confirmed as field strains. In pigs, 10 animals (0.95%) tested seropositive, but no isolates were recovered, highlighting limitations of serology in swine. Most livestock, including the positives, originated locally, reinforcing the representativeness of our findings. The successful isolation and molecular characterization of B. abortus demonstrates the value of combining diagnostic strategies beyond serology. These results underscore the utility of active surveillance when supported by traceability systems; this approach may also contribute to guide interventions to reduce infection risk in livestock and humans.

Atopic dermatitis (AD) is a chronic inflammatory skin disorder marked by barrier dysfunction and immune dysregulation. Colonization of lesional skin by Staphylococcus aureus, present in up to 80–100% of cases, exacerbates inflammation, in part through production of superantigenic toxins. While standard treatments such as topical corticosteroids, antibiotics, and antiseptic baths are widely used, their outcomes remain variable and often inadequate, highlighting the need for alternative strategies that minimize adverse effects and resistance development. In this study, we evaluated antimicrobial photodynamic inactivation (aPDI) using rose bengal (RB), a photosensitizer activated by visible light, as a potential approach to reduce S. aureus colonization and virulence. Across in vitro, ex vivo, and murine in vivo models, RB-mediated aPDI significantly decreased S. aureus viability and markedly attenuated the expression and activity of staphylococcal enterotoxins. Transcript and protein analyses confirmed substantial reductions in superantigenic activity post-aPDI. These effects were dependent on the combination of both RB and light, with no significant impact observed with either component alone. Our findings indicate that RB-based aPDI may represent a promising non-antibiotic approach to limit S. aureus viability and toxin activity in the context of AD. Our data contribute to the understanding of how photodynamic inactivation affects S. aureus virulence and highlight a model for studying the impact of microbial factors on skin immune responses in AD.

Seroprevalance of Brucella abortus in humans in Izmir Menemen and comparison of diagnostic tests; an experience between July 2012 and March 2022.

A cross-sectional study was carried out from March 2023 to March 2024 to assess the seroprevalence and risk factors of brucellosis in goats in the Nyangatom district, South Omo Zone, Southern Ethiopia. A total of 768 extensively managed local female goats of different age groups, were bled and data on hypothesized risk factors were collected using a semi-structured questionnaire. The preliminary screening of the sera for brucella antibodies was done using the Rose Bengal plate test (RBPT) and sera positive for RBPT were further examined by the complement fixation test (CFT). The overall seroprevalence of brucellosis was 12.9 . Among the risk factors considered, seroprevalence was associated with parity, litter size, flock size, and history of occurrence of abortion (p

Background/Objectives: The rapid progression of stroke often results in irreversible brain damage and poor outcomes when treatment is delayed. Prophylactic administration of FAD012 (3,5-dimethyl-4-hydroxycinnamic acid), a synthetic derivative of ferulic acid (FA), has demonstrated cerebroprotective effects in ischemic models through antioxidant and endothelial protective mechanisms. This study investigated the effects of FAD012 on cerebral infarction and blood–brain barrier (BBB) integrity using a photothrombotic stroke model in rats. Methods: Male Sprague Dawley rats received a single intraperitoneal injection of FAD012 or FA (100 or 300 mg/kg) 60 min prior to stroke induction. Under isoflurane anesthesia, the middle cerebral artery was exposed, and stroke was induced by intravenous administration of Rose Bengal followed by green laser irradiation. Cerebral blood flow (CBF) was monitored by laser Doppler flowmetry. BBB disruption was evaluated by Evans Blue extravasation and immunohistochemistry for tight junction (TJ) proteins. Results: Control rats exhibited extensive infarction, BBB disruption, and reduced expression of claudin-5, occludin, and ZO-1, along with fragmented collagen IV. In contrast, FAD012 (300 mg/kg) significantly attenuated CBF reduction, reduced infarct size, preserved BBB integrity, and maintained TJ protein expression, with greater efficacy than an equivalent dose of FA. FAD012 also preserved the expression and phosphorylation of endothelial nitric oxide synthase (eNOS), a key marker of vascular integrity. The CBF-preserving effect of FAD012 was completely abolished by NG-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase inhibitor. Conclusions: These findings suggest that FAD012 protects endothelial function, thereby contributing to the maintenance of CBF and BBB integrity, supporting its potential as a prophylactic therapeutic agent for ischemic stroke.

Brucellosis is an important zoonotic disease among livestock and humans worldwide and in Low-and-Middle Countries (LMICs) including Tanzania. The aim of this study was to determine the factors that could influence seroprevalence of brucellosis in commercial cattle farms. A cross-sectional study was conducted in a total of 54 commercial cattle farms randomly selected in Kagera, Mara, and Mwanza regions of the Lake Zone in Tanzania. Serum samples were collected from 1,080 cattle comprising both dairy and beef animals of both sexes and tested for Brucella-specific antibodies using Rose Bengal Plate Test (RBPT). Positive samples were confirmed by using competitive Enzyme-linked Immunosorbent Assay (c-ELISA). Animals in each farm were randomly selected for blood collection whereby a total number of 20 adult animals from both sexes were involved in the study. Descriptive statistics and multivariable regression analysis were conducted to assess the risk factors associated with brucellosis.The overall seroprevalence of brucellosis was 6.9% at the animal level and 51.9% at the farm level. Medium scale farms Odds ratio (OR = 11.304; Confidence Interval 95% CI 1.140 – 112.108;) and small - scale farms (OR = 37.170; 95% CI 1.119 – 1235.006) demonstrated a significantly higher likelihood of seropositivity to brucellosis than large - scale farms. Dairy cattle farms were less likely to be seropositive (OR = 0.046; 95% CI 0.003 – 0.728;) than beef cattle farms. The findings from the present study indicated that more than half of the farms are positive for Brucella antibodies. The study also revealed that the scale of production and functional type of cattle increase the risk of seropositivity. The findings provide baseline information for the development of targeted intervention programme in the control of brucellosis.

Human brucellosis is an acute febrile illness responsible of causing serious threats to pregnant women and their developing fetus. It is a neglected disease having zoonotic potential resulting in variety of complications. The present study aimed to detect sero-prevalence of brucellosis in pregnant women and to find out the role of various demographic and potential risk factors associated with this disease during January to July 2024. In this cross-sectional study (n=300) blood samples were collected from pregnant women ranging from 18 to 45 years of age. The information about personal data, demographics and risk factors was gathered via pre-designed structured questionnaire. The anti-Brucella antibodies were detected using Rose Bengal Plate test (RBPT) followed by enzyme linked immunosorbent assay (ELISA). The seropositive samples were subjected to qRT-PCR for molecular detection of Brucella. The data was subjected to descriptive statistics, Chi square test and Odds ratio using Minitab version 18. The result of study showed the sero-prevalence of brucellosis 9.33% among pregnant women with higher 11.36% in age group (18-28 years). Among the demographic factors, the occupation of farming (P =0.005; OR = 3.51, 95%CI: 1.56-7.85) and absence of education (P = 0.001; OR = 7.20, 95%CI: 2.79-18.59) showed significant association with human brucellosis. Additionally, of the potential risk factors analyzed, keeping animals at home (P =0.001; OR = 4.64, 95%CI: 2.05-10.50), and lack of knowledge about brucellosis (P =0.047; OR = 3.53, 95%CI: 1.03-12.03) were found statistically significant. A comprehensive awareness should be given to females regarding risk factors and spread of brucellosis. Consumption of pasteurized dairy products and adopting personal protection while dealing with animals will prevent pregnant women and their unborn from human brucellosis and its complications.