Brucella Abortus Research Articles

Gated nanoprobe utilizing metal-organic frameworks for identifying and distinguishing between the wild strains and the vaccine strains of brucella.

In the assay for Brucella, the identification and differentiation of wild strains and vaccine strains present a significant challenge. Currently, there aren't any commercially available product to address this issue. In this study, we have developed a novel gated nanoprobe by utilizing Metal-Organic Frameworks (MOFs) as a scaffold and hairpin DNA as a "gating switch". Specifically, Probe 1 with hairpin structure (P1h) targets a gene that is present in both wild strains Y3 (B. melitensis biovar 3) and vaccine strains A19 (Brucella abortus strains A19). We successfully applied this probe to screen positive samples of Brucella without any cross-reactivity with other substances. Additionally, we identified another specific gene exclusively found in wild strains, which serves as Probe 2 with hairpin structure (P2h) to confirm the strain type. Simultaneous detachment of both P1h and P2h from the MOFs leads to the release of Rhodamine 6G (Rho 6G) and Fluorescein (Flu), specifically indicating the presence of wild strains. If only P1h detaches and the Flu signal is detected, it suggests the presence of vaccine strains. Importantly, this method offers high accuracy, with a detection rate of 90% and a recovery rate of 94.71% to 107.65%, while avoiding cross-reactions with MO and TB. This one-step experiment provides reliable identification and differentiation of Y3 and A19, addressing concerns related to long periodicity, interference from individual variations, and the complex design of primers in existing laboratory methods. Furthermore, our approach successfully detects target 1 (T1) and target 2 (T2) at concentrations ranging from 10-6 M to 10-9 M, with a detection limit of 6.7 × 10-10 M and 6.4 × 10-10 M, respectively. Importantly, our strategy is cost-effective (around $1) and offers higher detection efficiency compared to traditional laboratory methods.

Characterization of the adaptive cellular and humoral immune responses to persistent colonization of Brucella abortus strain RB51 in a Jersey cow.

Brucella abortus strain RB51 is the commercial cattle vaccine used in the United States (US) and many parts of the world against bovine brucellosis. RB51 was licensed for use in 1996, and it has been shown to be safe and efficacious in cattle, eliciting humoral and cellular responses in calves and adult animals. In 2017, an epidemiological trace-back investigation performed by the Centers for Disease Control and Prevention (CDC) identified human cases of brucellosis caused by infection with RB51. These infections resulted from the consumption of unpasteurized dairy products, which were traced back to otherwise healthy animals that were shedding RB51 in their milk. At the current time, six adult Jersey cows have been identified in the U.S. that are shedding RB51 in milk. One of the RB51 shedding cattle was obtained and housed at the National Animal Disease Center (NADC) for further study. Improved understanding of host cellular and humoral immune responses to RB51 in persistently colonized cattle may be achieved by the characterization of responses in shedding animals. We hypothesized, based on the lack of RB51 clearance, that the RB51 shedder animal has a diminished adaptive cellular immune response to RB51. Our data demonstrate that in the presence of persistent RB51 infection, there is a lack of peripheral anti-RB51 CD4+ T cell responses and a concurrently high anti-RB51 IgG humoral response. By understanding the mechanisms that result in RB51 persistence, the development of improved interventions or vaccinations for brucellosis may be facilitated, which would provide public health benefits, including reducing the risks associated with the consumption of non-pasteurized milk products.

Immunoevasion Mechanism of Brucella abortus in Dairy Cows

Immunoevasion Mechanism of Brucella abortus in Dairy Cows

Detectable immunological response in Brucella abortus s 19 vaccinated herds of cattle in vilachchiya veterinary range, north central province, Sri Lanka

Brucellosis a zoonotic disease caused by Gram-negative, intracellular, coccobacillus. Brucellosis, an economically important disease in livestock with zoonotic implications has been reported worldwide. It is transmitted via contaminated animals and their products. Clinical signs of animal brucellosis are abortions, reduced fertility, retention of fetal membranes, endometritis, and substantial decline in milk production over an animal’s lifespan. Orchitis, epididymitis and chronic hygroma had been reported in bulls. The current study focused on to studying the detectable immunological responses to Brucella S 19 live attenuated vaccine using serological tests in cattle of dry zone farms. A questionnaire-based survey was carried out in selected farms with and without a previous history of abortion respectively. Animals that were in 1st to 5th lactation in the 18 farms were selected for screening by Rose Bengal Plate Test (RBPT) and positives were confirmed by Complement Fixation Test (CFT). Three farms each with and without history of abortion were selected for Brucella S19 vaccination from the RBPT and CFT negative farms. Sixty non pregnant heifers over 6 months were selected, and 1ml each of S19 vaccine was given subcutaneously to 30 animals while 30 animals were kept as controls. The serum samples were collected and screened for Brucellosis by RBPT and CFT at 2, 4, 6, 8 and 10 weeks of post vaccination. The second week after the vaccination, 71.42% of the animals showed over 1:4 CFT levels while 28.57% of them did not show any antibody titres for Brucellosis in the Brucella endemic farms. In Brucella non-endemic farms, 68.75% of animals showed over 1:4 CFT titre at 2 weeks after the vaccination. However, 31.25% of animals did not show any CFT titre. No persistent and detectable antibody responses were found 10 weeks post vaccination in heifers of endemic and non-endemic herds. The titers of CFT indicate IgG or IgM humeral response, cell mediated immune response of live attenuated vaccine has not been monitored in the study. Thus, seropositivity after the 10 weeks post vaccination may indicate an infection of Brucellosis since no antibodies from vaccination were found. Further investigation with highly sensitive tests such as ELISA is recommended to have a better insight into this aspect in the future.

Combination of recombinant Brucella abortus dapB and gpm proteins induces host immunity against B. abortus 544 infection using a mouse model

Combination of recombinant Brucella abortus dapB and gpm proteins induces host immunity against B. abortus 544 infection using a mouse model

Big data analysis descriptively of Brucella abortus cases in Indonesia during 2006-2020

Brucellosis represents a formidable zoonotic challenge, primarily impacting animal breeders and raising substantial health and economic concerns. This study delves into the epidemiological analysis of Brucella abortus infections in Indonesia, leveraging the extensive quantitative big data from the World Animal Health Information System (WAHIS) database managed by the World Organisation for Animal Health (WOAH). Covering the period from 2006 to 2020, the research utilizes advanced clustering techniques to dissect the incidence and distribution of this zoonotic disease across various Indonesian provinces and animal species, including cattle, buffaloes, sheep, goats, and swine. The findings reveal a prominent prevalence of the infection in Sulawesi Selatan Province, accounting for 26.71% of cases, with a fluctuating trend observed nationwide and the peak incidence in 2019, notably in Sulawesi Tenggara Province. The study highlights cattle as the species most susceptible to Brucella abortus. The comprehensive analysis underscores the criticality of targeted interventions and sustained surveillance in particular regions, especially focusing on cattle. This research not only enhances understanding of Brucellosis's spread and impact in Indonesia but also aids in formulating effective disease management strategies, thereby addressing both health and economic concerns associated with this zoonotic challenge.

Immuno-SPR biosensor for the detection of Brucella abortus

A proof of principle biosensor for the Brucella abortus recognition onsite is presented. The system is based on a plasmonic optical fiber probe functionalized with an oriented antibody layer immobilized on a short polyethyleneglycol (PEG) interface through carbodiimide chemistry and protein G as an intermediate layer. The biosensor is inserted in a holder built in 3D printing technology, obtaining a custom holder useful for housing the sample to be measured and the equipment. The removable sensor chip is a low-cost Surface Plasmon Resonance (SPR) platform based on D-shaped plastic optical fibers (POFs), built-in in 3D printed connectors, used here for the first time to detect bacteria via a bio-receptor layer specific for its membrane protein. The performances of the biosensor in Brucella abortus recognition are tested by using two different SPR-POF probes combined with the same bio-receptor layer. The best sensor configuration has presented a sensitivity at low concentrations of one order of magnitude greater than the other. A limit of detection (LoD) of 2.8 bacteria/mL is achieved well competitive with other systems but without the need for amplification or special sample treatments. Specificity has been tested using Salmonella bacteria, and reproducibility, regenerability and stability are moreover evaluated. These experimental results pave the way for building an efficient and specific biosensor system for Brucella abortus detection onsite and in a few minutes. Moreover, the proposed POF-based SPR biosensor device, with respect to the already available technologies, could be a Point-of-care-test (POCT), simple to use, small-size and portable, low-cost, don’t necessary of a microfluidic system, and can be connected to the Internet (IoT).

Post-transcriptional control of the essential enzyme MurF by a small regulatory RNA in Brucella abortus.

Brucella abortus is a facultative, intracellular, zoonotic pathogen that resides inside macrophages during infection. This is a specialized niche where B. abortus encounters various stresses as it navigates through the macrophage. In order to survive this harsh environment, B. abortus utilizes post-transcriptional regulation of gene expression through the use of small regulatory RNAs (sRNAs). Here, we characterize a Brucella sRNAs called MavR (for MurF- and virulence-regulating sRNA), and we demonstrate that MavR is required for the full virulence of B. abortus in macrophages and in a mouse model of chronic infection. Transcriptomic and proteomic studies revealed that a major regulatory target of MavR is MurF. MurF is an essential protein that catalyzes the final cytoplasmic step in peptidoglycan (PG) synthesis; however, we did not detect any differences in the amount or chemical composition of PG in the ΔmavR mutant. A 6-nucleotide regulatory seed region within MavR was identified, and mutation of this seed region resulted in dysregulation of MurF production, as well as significant attenuation of infection in a mouse model. Overall, the present study underscores the importance of sRNA regulation in the physiology and virulence of Brucella.

Intracellular Growth Inhibition and Host Immune Modulation of 3-Amino-1,2,4-triazole in Murine Brucellosis.

Catalase, an antioxidant enzyme widely produced in mammalian cells and bacteria, is crucial to mitigating oxidative stress in hostile environments. This function enhances the intracellular survivability of various intracellular growth pathogens, including Brucella (B.) abortus. In this study, to determine whether the suppression of catalase can inhibit the intracellular growth of B. abortus, we employed 3-amino-1,2,4-triazole (3-AT), a catalase inhibitor, in both RAW 264.7 macrophage cells and an ICR mouse model during Brucella infection. The intracellular growth assay indicated that 3-AT exerts growth-inhibitory effects on B. abortus within macrophages. Moreover, it contributes to the accumulation of reactive oxygen species and the formation of nitric oxide. Notably, 3-AT diminishes the activation of the nucleus transcription factor (NF-κB) and modulates the cytokine secretion within infected cells. In our mouse model, the administration of 3-AT reduced the B. abortus proliferation within the spleens and livers of infected mice. This reduction was accompanied by a diminished immune response to infection, as indicated by the lowered levels of TNF-α, IL-6, and IL-10 and altered CD4+/CD8+ T-cell ratio. These results suggest the protective and immunomodulatory effects of 3-AT treatment against Brucella infection.

Comparative Evaluation of Serological, ELISA and Molecular Tests in Diagnosis of Brucella abortus in Human Serum

Background: Brucellosis is one of the most prevalent and common diseases between humans and animals.Prompt diagnosis and timely treatment of this disease can prevent many complications. In this regard, this study aims to comparatively evaluate ELISA, PCR and serological methods to identify Brucella abortus.
 Methods: In this study, the serum of 100 patients referred to Tonekabon private laboratory from July 2020 to January 2021 was examined by PCR, ELISA and Wright, Coombs Wright, and 2ME methods for the detection of Brucella abortus.
 Results: In this study, the mean age of the sample was 43.3 ± 18.2 of which 21% were infected with Brucella abortus according to the above serological methods. According to ELISA test, 22% of the samples were IgM, 6% of the samples were IgG and 16% were PCR positive. Kappa agreement coefficient in Wright and Coombs Wright test and 2me were significant (P <0.001). Serological diagnostic indices and ELISA sensitivity were 68.75% and 68.75%, respectively. The lowest prediction rate of Brucella abortus among diagnostic methods was related to Elisa (IgM). Based on Fisher's exact test, there was no significant relationship between the percentage of Brucella abortus positive PCR cases and age, sex, previous history of infection with Brucella, fever, body aches and dairy consumption.
 Conclusion: Based on the results of our study, the accuracy of all methods is comparative and the lowest accuracy is related to Elisa (IgM) which has a lower level of predictability than other methods. The highest level of prediction belonged to Wright and 2me tests.

Molecular detection and geospatial distribution of <em>Brucella</em> infections in horses in Kano Metropolis, Kano State, Nigeria

Brucellosis is a major zoonotic disease that affects several domestic, wild and marine mammals, and man. It causes serious economic damage in livestock productivity, especially in developing countries, including Nigeria. The study aimed to determine the species of Brucella that infect horses and establish the spatial distribution of the infection in Kano, Nigeria. A total of 328 blood samples were collected from horses. Sera were collected from the samples and kept at -20 0C until tested. The sera were successively subjected to RBPT, SAT-EDTA and multiplex-PCR. Seventy-nine serum samples (24.0%) were positive for Brucella antibodies on RBPT. Out of the RBPT positive samples, 39 (49.4%) were also positive for SAT-EDTA. There was no difference between the results of the two tests (RBPT: p=0.4420; SAT-EDTA: p=0.124). Of the 39 SAT-EDTA positive sera, 32 (82.1%) were confirmed to be Brucella abortus. Fifteen of the 16 mares tested were positive for B. abortus while 73.9% of the stallion were positive. All horses in the age groups of 1-5 and 11-15 years were 100% positive, while 72.0% of the horses in the 6-10 year age group were also positive. The study showed 81.8 % detection rate of B. abortus using AMOS-PCR and that 15 (93.7%) of the female horses tested (16) had B. abortus while 17 (73.9%) of the 23 male horses tested had B. abortus.

Global phylogenomic diversity of Brucella abortus: spread of a dominant lineage

Brucella abortus is a globally important zoonotic pathogen largely found in cattle hosts and is typically transmitted to humans through contaminated dairy products or contact with diseased animals. Despite the long, shared history of cattle and humans, little is known about how trade in cattle has spread this pathogen throughout the world. Whole genome sequencing provides unparalleled resolution to investigate the global evolutionary history of a bacterium such as B. abortus by providing phylogenetic resolution that has been unobtainable using other methods. We report on large-scale genome sequencing and analysis of B. abortus collected globally from cattle and 16 other hosts from 52 countries. We used single nucleotide polymorphisms (SNPs) to identify genetic variation in 1,074 B. abortus genomes and using maximum parsimony generated a phylogeny that identified four major clades. Two of these clades, clade A (median date 972 CE; 95% HPD, 781–1142 CE) and clade B (median date 150 BCE; 95% HPD, 515 BCE–164 CE), were exceptionally diverse for this species and are exclusively of African origin where provenance is known. The third clade, clade C (median date 949 CE; 95% HPD, 766–1102 CE), had most isolates coming from a broad swath of the Middle East, Europe, and Asia, also had relatively high diversity. Finally, the fourth major clade, clade D (median date 1467 CE; 95% HPD, 1367–1553 CE) comprises the large majority of genomes in a dominant but relatively monomorphic group that predominantly infects cattle in Europe and the Americas. These data are consistent with an African origin for B. abortus and a subsequent spread to the Middle East, Europe, and Asia, probably through the movement of infected cattle. We hypothesize that European arrival to the Americas starting in the 15th century introduced B. abortus from Western Europe through the introduction of a few common cattle breeds infected with strains from clade D. These data provide the foundation of a comprehensive global phylogeny of this important zoonotic pathogen that should be an important resource in human and veterinary epidemiology.

Is Brucella excreted in cattle faeces? – Evidence from Punjab, India

Brucellosis is a neglected zoonosis that affects animals and people in much of the underdeveloped world. The disease is endemic in cattle in Punjab, India and controlling it is a public health challenge. Dairy farmers and farm labour commonly handle cattle faeces with bare hands and personal protective equipments are not used. No studies have been conducted about the shedding of Brucella species in faeces of sero positive cattle in the state. This study aimed to isolate and identify the Brucella species from faeces of sero positive cattle in Punjab, India. Faecal samples were collected from 350 Brucella sero positive cattle in Ludhiana district of Punjab, India. Isolation was performed using a pre-enriched Brucella selective broth medium as well as Brucella selective medium agar plates containing horse serum and Brucella selective supplements. Isolates were identified using Gram staining technique and rapid slide agglutination test, and then confirmed by using bcsp31 and 16s rRNA genus specific PCR. Isolates were further identified up to species level by using Bruce-Ladder multiplex PCR. Fourteen Brucella species were isolated, all of which showed coccobacilli on gram staining, positive rapid slide agglutination test and amplification of bcsp31 and 16s rRNA genes. Of the 14 isolates, 11 were identified as Brucella abortus and 3 were identified as Brucella melitensis. The study demonstrates that animal faeces could pose a potential risk for animal and human health and faeces of seropositive cattle must be handled with care.

Effect of selection genotype on immune response to Brucella abortusRB51 in Holstein cattle.

Genetic selection for milk production traits in US Holsteins has affected numerous genes associated with reproduction and immunity. This study compares the transcriptomic response of peripheral blood mononuclear cells to an invitro Brucella abortus strain RB51 (RB51) bacterial challenge between contemporary Holsteins and Holsteins that have not been selected for milk production traits since the mid-1960s. Total RNA was extracted from peripheral blood mononuclear cells from four contemporary and four unselected lactating, primiparous cows following 24-h incubation with or without stimulation with RB51 bacteria. RNA was sequenced and reads analyzed using tools from galaxy.scinet.usda.gov. A total of 412 differentially expressed genes (false discovery rate p < 0.05, log fold change > |1|) were identified. The upregulated genes (genes with higher expression in contemporary than unselected cattle) were enriched for 19 terms/pathways, including alanine, aspartate, and glutamate metabolism, indicating a cellular stress response. Downregulated genes (genes with higher expression in unselected than contemporary cows) were enriched for 37 terms/pathways, representing diverse immune responses, including natural killer cell-mediated immunity, interferon-γ production, negative regulation of interleukin-10 production, and cytokine receptor activity indicating a broad immune response with an emphasis on immune defense. These results provide evidence that differences exist between the two genotypes in response to invitro bacterial challenge. This suggests that contemporary cows, genetically selected for milk production, may have reduced immune function, including limitations in response to intracellular bacteria.

A sensor histidine kinase from a plant-endosymbiont bacterium restores the virulence of a mammalian intracellular pathogen

Alphaproteobacteria include organisms living in close association with plants or animals. This interaction relies partly on orthologous two-component regulatory systems (TCS), with sensor and regulator proteins modulating the expression of conserved genes related to symbiosis/virulence. We assessed the ability of the exoS+Sm gene, encoding a sensor protein from the plant endosymbiont Sinorhizobium meliloti to substitute its orthologous bvrS in the related animal/human pathogen Brucella abortus. ExoS phosphorylated the B. abortus regulator BvrR in vitro and in cultured bacteria, showing conserved biological function. Production of ExoS in a B. abortus bvrS mutant reestablished replication in host cells and the capacity to infect mice. Bacterial outer membrane properties, the production of the type IV secretion system VirB, and its transcriptional regulators VjbR and BvrR were restored as compared to parental B. abortus. These results indicate that conserved traits of orthologous TCS from bacteria living in and sensing different environments are sufficient to achieve phenotypic plasticity and support bacterial survival. The knowledge of bacterial genetic networks regulating host interactions allows for an understanding of the subtle differences between symbiosis and parasitism. Rewiring these networks could provide new alternatives to control and prevent bacterial infection.

Serological and Molecular Survey of Brucella Species in Owners and Their Dogs Living on Island and Mainland Seashore Areas of Brazil.

Background: Although Brucella abortus, Brucella suis, and Brucella canis may infect humans and dogs worldwide, no study to date has assessed and compared owners and their dogs between island and mainland seashore areas. Materials and Methods: Accordingly, the study herein has applied serological tests, including Microplate Agglutination Test with 2-Mercaptoethanol, immunochromatographic assay, and Rose Bengal Test, and a Brucella genus-specific PCR assay to 195 owners and their 148 dogs living on 1 mainland seashore area and three nearby oceanic islands of southern Brazil. Results: No seropositivity to B. abortus and B. suis was detected in owner or dog sera. Anti-B. canis seropositivity was observed in 3/148 (2.0%) dogs, but no owner sample was seropositive to B. canis. In addition, all blood samples from both owners and dogs were negative on Brucella genus-specific PCR assay. Conclusions: The seropositive dogs were not related and lived on the seashore mainland area of Guaraqueçaba city. The absence of seropositivity on the islands and the low seropositivity on the seashore mainland could be attributed to geographic isolation, and suggest the low impact of the disease in the region. Despite being a zoonotic disease, brucellosis by B. canis is not included in the National Program for Control and Eradication of Brucellosis, and its diagnosis and notification are not mandatory. The presence of seropositive dogs highlights the risk to human health and the importance of epidemiological surveillance actions in the region, as well as the need for the implantation of preventive measures to avoid the transmission of the pathogen.

Zoonotic risks of pathogens from dairy cattle and their milk-borne transmission.

Dairy products are major sources of high-quality protein and bioavailable nutrients and dairy production contributes to local, regional and national-level economies. Consumption of raw milk and raw milk products does, however, carry a zoonotic risk, as does direct contact with cattle by farm husbandry staff and other employees. This review will mainly focus on the latter, and deal with it from the standpoint of a well-developed dairy industry, using the example of the Netherlands. With regard to dairy cattle, the main bacterial pathogens are Salmonella spp., Listeria monocytogenes and Leptospira hardjo as well as Brucella abortus and Chlamydia abortus. The main viral pathogens associated with dairy are Rift Valley fever virus, rabies virus, cowpox virus and vaccinia virus. The main parasitological infections are Echinococcus granulosis, Cryptosporidium parvum and Giardia duodenalis, however, the last mentioned have mainly swimming pools as sources of human infection. Finally ectoparasites such as lice and mites and Trichophyton verrucosum may affect employees. Some pathogens may cause health problems due to contamination. Bacterial pathogens of importance that may contaminate milk are Campylolobacter jejuni, Escherichia coli, Mycobacterium avium subsp. paratuberculosis, Leptospira hardjo and Salmonella typhimurium. Excretion of zoonotic viruses in milk is negligible in the Netherlands, and the endoparasite, Toxocara vitulorum is mainly found in suckling and fattening calves, whilst the risk in dairy cattle is limited. Excretion of transmissible spongiform encephalopathies (TSEs) or mycoses in milk are not expected and are, therefore, not of importance here.Being aware of the risks and working according to hygiene standards can substantially limit zoonotic risks for employees. Additionally, diseased employees are advised to limit their contact with cattle and to indicate that they work with cattle when consulting a physician. To prevent zoonotic risks through excretion of pathogens in milk, standard hygiene measures are necessary. Further, using only pasteurised milk for consumption and/or processing of milk can considerably limit the risks. If these measures are not possible, well-constructed monitoring can be followed. Monitoring programmes already exist for pathogens such as for Salmonella spp., Leptospira hardjo and Mycobacterium avium subsp. paratuberculosis. For others, like Campylobacter jejuni and E. coli, programmes are not available yet as far as we know.

Brucellosis: Why is it eradicated from domestic livestock in the United States but not in the Nile River Basin countries?

Brucellosis is one of the most highly infectious zoonotic diseases worldwide and has substantial health and economic impact. Strenuous efforts are essential to combat and prevent this disease from the one-health perspective. Brucellosis is successfully eradicated from domestic animals in the United States, but control strategies continue to eradicate it from wildlife in the Greater Yellowstone Area (GYA). Brucellosis in the Nile River Basin countries (Egypt, Sudan, Ethiopia, and Tanzania) is highly prevalent and endemic. There are several factors behind the failure of eradication of Brucella in these countries. The lack of cooperation between policymakers, health officials, veterinary sectors, and farmers is the key reason that impedes the control and prevention strategies in brucellosis-endemic countries. This review will focus on the epidemiology, prevention, and control strategies of Brucella abortus and Brucella melitensis in the United States and the Nile Basin countries (Egypt, Sudan, Ethiopia, and Tanzania).

Scent detection of Brucella abortus by African giant pouched rats (Cricetomys ansorgei)

BackgroundBrucellosis is a contagious zoonosis caused by bacteria of the genus Brucella. While the disease has been eradicated in most developed countries, it remains endemic in sub–Saharan Africa where access to reliable diagnostics is limited. African giant pouched rats (Cricetomys ansorgei) have been trained to detect the scent of Mycobacterium tuberculosis to increase case detection in sub-Saharan Africa. Given the similar diagnostic challenges facing brucellosis and tuberculosis, we explored the feasibility of training African giant pouched rats to detect Brucella.ResultsAfter 3 months of training, rats reliably identified cultured Brucella, achieving an average sensitivity of 93.56% (SD = 0.650) and specificity of 97.65% (SD = 0.016). Rats readily generalized to novel, younger Brucella cultures that presumably generated a weaker volatile signal and correctly identified at least one out of three fecal samples spiked with Brucella culture during a final test of feasibility.DiscussionTo our knowledge, these experiments are the first to demonstrate Brucella emits a unique odor profile that scent detection animals can be trained to identify. Importantly, cultured E. coli samples were included throughout training and test to ensure the rats learned to specifically identify Brucella bacteria rather than any bacteria in comparison to bacteria-free culture medium. E. coli controls therefore served a crucial function in determining to what extent Brucella abortus emits a unique odor signature. Further research is needed to determine if a Brucella-specific volatile signature is present within clinical samples. If confirmed, the present results suggest trained rats could serve as a valuable, novel method for the detection of Brucella infection.

Imipramine Administration in Brucella abortus 2308-Infected Mice Restores Hippocampal Serotonin Levels, Muscle Strength, and Mood, and Decreases Spleen CFU Count.

Brucellosis infection causes non-specific symptoms such as fever, chills, sweating, headaches, myalgia, arthralgia, anorexia, fatigue, and mood disorders. In mouse models, it has been associated with increased levels of IL-6, TNF-α, and IFN-γ, a decrease in serotonin and dopamine levels within the hippocampus, induced loss of muscle strength and equilibrium, and increased anxiety and hopelessness. Imipramine (ImiP), a tricyclic antidepressant, is used to alleviate neuropathic pain. This study evaluated the effects of ImiP on Balb/c mice infected with Brucella abortus 2308 (Ba) at 14- and 28-days post-infection. Serum levels of six cytokines (IFN-γ, IL-6, TNF-α, IL-12, MCP-1. and IL-10) were assessed by FACS, while the number of bacteria in the spleen was measured via CFU. Serotonin levels in the hippocampus were analyzed via HPLC, and behavioral tests were conducted to assess strength, equilibrium, and mood. Our results showed that mice infected with Brucella abortus 2308 and treated with ImiP for six days (Im6Ba14) had significantly different outcomes compared to infected mice (Ba14) at day 14 post-infection. The mood was enhanced in the forced swimming test (FST) (p < 0.01), tail suspension test (TST) (p < 0.0001), and open-field test (p < 0.0001). Additionally, there was an increase in serotonin levels in the hippocampus (p < 0.001). Furthermore, there was an improvement in equilibrium (p < 0.0001) and muscle strength (p < 0.01). Lastly, there was a decrease in IL-6 levels (p < 0.05) and CFU count in the spleen (p < 0.0001). At 28 days, infected mice that received ImiP for 20 days (Im20Ba28) showed preservation of positive effects compared to infected mice (Ba28). These effects include the following: (1) improved FST (p < 0.0001) and TST (p < 0.0001); (2) better equilibrium (p < 0.0001) and muscle strength (p < 0.0001); (3) decreased IL-6 levels (p < 0.05); and (4) reduced CFU count in the spleen (p < 0.0001). These findings suggest the potential for ImiP to be used as an adjuvant treatment for the symptoms of brucellosis, which requires future studies.