Genome-wide analysis of Brucella melitensis growth in spleen of infected mice allows rational selection of new vaccine candidates.

Live attenuated vaccines (LAVs) whose virulence would be controlled at the tissue level could be a crucial tool to effectively fight intracellular bacterial pathogens, because they would optimize the induction of protective immune memory while avoiding the long-term persistence of vaccine strains in the host. Rational development of these new LAVs implies developing an exhaustive map of the bacterial virulence genes according to the host organs implicated. We report here the use of transposon sequencing to compare the bacterial genes involved in the multiplication of Brucella melitensis, a major causative agent of brucellosis, in the lungs and spleens of C57BL/6 infected mice. We found 257 and 135 genes predicted to be essential for B. melitensis multiplication in the spleen and lung, respectively, with 87 genes common to both organs. We selected genes whose deletion is predicted to produce moderate or severe attenuation in the spleen, the main known reservoir of Brucella, and compared deletion mutants for these genes for their ability to protect mice against challenge with a virulent strain of B. melitensis. The protective efficacy of a deletion mutant for the plsC gene, implicated in phospholipid biosynthesis, is similar to that of the reference Rev.1 vaccine but with a shorter persistence in the spleen. Our results demonstrate that B. melitensis faces different selective pressures depending on the organ and underscore the effectiveness of functional genome mapping for the design of new safer LAV candidates.

Detection and quantification of Brucella abortus DNA in water buffaloes (bubalus bubalis) using droplet digital polymerase chain reaction.

Brucellosis represents a major public health concern worldwide. Human transmission is mainly due to the consumption of unpasteurized milk and dairy products of infected animals. The gold standard for the diagnosis of Brucella spp in ruminants is the bacterial isolation, but it is time-consuming. Polymerase Chain Reaction (PCR) is a quicker and more sensitive technique than bacterial culture. Droplet digital PCR (ddPCR) is a novel molecular assay showing high sensitivity in samples with low amount of DNA and lower susceptibility to amplification inhibitors. Present study aimed to develop a ddPCR protocol for the detection of Brucella abortus in buffalo tissue samples. The protocol was validated using proficiency test samples for Brucella spp by real time qPCR. Furthermore, 599 tissue samples were examined. Among reference materials, qPCR and ddPCR demonstrated same performance and were able to detect up to 225 CFU/mL. Among field samples, ddPCR showed higher sensitivity (100%), specificity and accuracy of 93.4% and 94.15%, respectively. ddPCR could be considered a promising technique to detect B. abortus in veterinary specimens, frequently characterized by low amount of bacteria, high diversity in matrices and species and poor storage conditions.

Updated therapeutic options for human brucellosis: A systematic review and network meta-analysis of randomized controlled trials.

BACKGROUND: In clinical practice guidelines, there is no consensus about the medications that should be initially offered to patients with brucellosis. To provide informative evidence, we compared and ranked brucellosis medications based on their efficacy and safety. METHODS: For this systematic review and network meta-analysis, we searched 4 English databases and 3 Chinese databases, from the date of database inception to December 13, 2023. We included randomized controlled trials (RCTs) involving children and adolescents with brucellosis, comparing different antibiotic regimens. We excluded studies explicitly targeting patients with spondylitis brucellosis, endocarditis brucellosis, and neuro-brucellosis. The primary outcomes were overall failure (efficacy) and side effects (safety). Secondary outcomes were relapse and therapeutic failure. Pairwise meta-analysis was first examined. Data were analyzed using random effects network meta-analysis, with subgroup and sensitivity analyses performed. The Confidence in Network Meta-Analysis (CINeMA) framework was used to assess the certainty of evidence. The protocol was preregistered in PROSPERO (CRD42023491331). RESULTS: Of the 11,747 records identified through the database search, 43 RCTs were included in the network meta-analysis. Compared with standard therapy (Doxycycline + Rifampicin), Rifampicin + Tetracyclines (RR 4.96; 95% CI 1.47 to 16.70; very low certainty of evidence), Doxycycline + TMP/SMX (RR 0.18; 95% CI 0.06 to 0.52; low certainty of evidence), Doxycycline + Quinolones (RR 0.27; 95% CI 0.11 to 0.71; low certainty of evidence), Streptomycin + Tetracyclines (RR 0.04; 95% CI 0.01 to 0.16; low certainty of evidence), and Single (RR 0.05; 95% CI 0.02 to 0.16; moderate certainty of evidence) were less efficacious. Doxycycline + Gentamicin ranked the best in efficacy (SUCRA values: 0.94), the second is Triple (SUCRA values: 0.87), and the third is Doxycycline + Streptomycin (SUCRA values: 0.78). CONCLUSIONS: Brucellosis medications differ in efficacy and safety. Doxycycline + Gentamicin, Triple, and Doxycycline + Streptomycin have superior efficacy and safety. Treatment of brucellosis should strike a balance between efficacy, safety, and cost.

A meta-analysis of livestock brucellosis prevalence in Indonesia.

Indonesia has a long history of livestock brucellosis, but the overall pooled prevalence remains unclear. This study aims to determine the pooled estimated prevalence of livestock brucellosis in Indonesia using the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines. Five databases were searched and screened using predefined inclusion and exclusion criteria. Data from included studies were extracted and analyzed using a random effects model in R 4.3.0 for pooled estimated prevalence, subgroup meta-analysis, and meta-regression. Publication bias and sensitivity tests were conducted using Egger's test, funnel plot, trim and fill plot, and leave-one-out. The screening process identified 46 included studies, representing 47,057 samples for brucellosis testing. The pooled estimated prevalence for livestock brucellosis was 3.25% (95% CI, 1.81%-5.78%) with high heterogeneity (Q = 2130.91, p = 0, I2=98%). Subgroup meta-analysis indicated no significant difference in the prevalence of livestock brucellosis across the main islands in Indonesia (p = 0.44) and across provinces in Sulawesi Island (p = 0.83), but significant differences were found among provinces in Java (p < 0.01). The subgroup meta-analysis based on animal type showed no significant difference between cattle, small ruminants, and pig brucellosis estimated prevalence (p = 0.26). Between serological tests, no significant difference was found (p = 0.77). Meta-regression showed no significant difference in brucellosis prevalence from 1988-2023. Egger's test and funnel plot showed publication bias. Trim and fill test indicated 21 studies should be added. As most studies were conducted in Java and Sulawesi Islands, caution should be exercised in interpreting the results, emphasizing the necessity of increasing the study of brucellosis in other regions.

Expression of cytokine and Apoptosis-Associated genes in mice bone Marrow-Derived Macrophages stimulated with Brucella recombinant type IV secretion effectors.

BACKGROUND: Brucellosis is an economically important infectious caused by most commonly by Brucella. Detection of infected animals at the early stage is important for controlling the disease. The diagnostic antigens, usually protein antigens, have attracted much interest. However, the accurate mechanism of immune response is still unknown. The secretory effectors (BPE005, BPE275, and BPE123) of the type IV secretion system (T4SS) were involved in the intracellular circulation process of Brucella and the immune responses of the host. METHODS: Genes encoding three B. abortus effector proteins (BPE005, BPE275, and BPE123) of T4SS were cloned and the recombinant proteins were expressed and purified. The purified recombinant proteins were named rBPE005, rBPE275 and rBPE123. Then, the expressions of Th1- and Th2-related cytokine genes were analyzed in mice bone marrow-derived macrophages (BMDMs) after stimulation with rBPE005, rBPE275, and rBPE123. Furthermore, four apoptosis-associated genes (Caspase-3, Caspase-8, Bax, and Bcl-2) were also detected to explore the damage of the proteins to the cells. RESULTS: Expressions of all Th1- and Th2-related cytokine genes were induced with three proteins, and different cytokine expression patterns induced by each protein depend on the stimulation time and dose of protein. However, expressions of apoptosis-related genes did not change. CONCLUSION: These results showed that the secreted antigens of Brucella induced an immune reaction via the production of Th1- and Th2-type cytokines in BMDMs without exerting any damage on the cells.

Pathogenesis of Brucella suis biovar 1 in the armadillo (Chaetophractus villosus).

Brucella suis biovar 1 has the broadest animal host spectrum. Affects domestic animals and wildlife species. The aim of our study was to investigate the pathogenesis of B. suis biovar 1 infection in the armadillo (Chaetophractus villosus) under experimental conditions. One gravid female and three adult males were inoculated with a suspension containing 1×106 CFU/mL (colony-forming units) of B. suis biovar 1 by oral route. In addition, the gravid female and one male received the same suspension by the conjunctival route. A young male and two females not inoculated were kept in contact with the animals inoculated. The animals that tested seropositive were euthanized. All inoculated armadillos showed positive antibody titres 2 weeks post-inoculation. Of the three uninoculated animals, one female was seropositive for Brucella infection.  Brucella was isolated from the spleen, liver, mesenteric lymph nodes, uterus, testes, and urine. Characteristic histologic lesions were found in the epididymis. These results suggest that armadillos can act as a reservoir for the spread of B. suis infection, and the persistence of Brucella in armadillo tissues constitutes a risk for humans, because of the cultural practice of armadillo meat consumption in rural communities.

Distal femur Brucella osteomyelitis in infancy: A rare case report.

Brucella disease is an infectious disease caused by Brucella bacteria. It is transmitted through the consumption of unpasteurized dairy products and undercooked meat and penetration through the skin of individuals in contact with farm animals. A detailed medical history is of utmost importance in the diagnosis. Headache, cyclical fever, sweating, vomiting, abdominal pain, and wandering arthralgia are among the main clinical symptoms. Brucella infection is usually characterized by inflammation in the musculoskeletal system, and osteomyelitis is rarely seen. In this article, we report a case of osteomyelitis after neglected brucellosis.

Meta-genomic next-generation sequencing in the diagnosis of brucellosis: Five cases from a non-endemic area.

Metagenomic next-generation sequencing (mNGS) in diagnosis of human brucellosis is comparatively unexplored. This report details five human brucellosis cases diagnosed using mNGS based on Illumina sequencing platform, comprising three females and two males, four with epidemiological exposure. In cases 1 and 2, plasma mNGS results showed one positive and one negative for Brucella melitensis, and subsequent blood cultures were both positive. Cases 3, 4 and 5 involved spinal brucellosis, some with paravertebral abscesses. mNGS from infectious tissue samples successfully detected Brucella, with read counts ranging between 30 and 1314, yet cultures were negative in cases 4 and 5. Following antibiotic and surgical treatments, all patients showed clinical improvement. This report shows mNGS testing enhances the detection sensitivity of brucellosis diagnosis.

What is this summary about? Brucella is a type of bacteria that can infect humans and animals. It causes a disease called brucellosis. Symptoms of brucellosis include fever and fatigue, among others. Meta-genomic next-generation sequencing (mNGS) is a tool for sequencing the DNA of bacteria. In this report, we use mNGS to diagnose human brucellosis in five cases.What were the results? Brucella was found in the blood of two infected people, but mNGS found Brucella in only one. Of three people with Brucella infection of the spine, mNGS found Brucella in the infected tissue but Brucella was only cultured in one case. Following antibiotic and surgical treatments, all five patients showed improvement of their symptoms.What do the results of the study mean? mNGS is a relatively rapid and effective diagnostic method that can improve the detection of Brucella in brucellosis.

Understanding brucellosis: knowledge, perceptions, and self-reported prevalence among agro-pastoralists in Nakasongola, Uganda.

BACKGROUND: Brucellosis is an infectious zoonotic disease that poses serious health threats around the world including Uganda. Brucellosis is caused by Brucella spp., the bacteria being transmitted via contact through skin breaks, via inhalation, or orally through the consumption of raw milk and other dairy products. The aim of this study was to investigate self-reported prevalence, knowledge, and perceptions towards brucellosis transmission, within agro-pastoralist communities in the Nakasongola district, central Uganda. METHODS: This study employed a cross-sectional survey design. A semi-structured questionnaire was developed and administered to 398 participants selected through convenience sampling method. The survey gathered information on socio-demographic characteristics, knowledge of brucellosis transmission, symptoms, preventive measures, and self-reported prevalence of brucellosis. Qualitative data involved the use of six focus group discussions, identifying factors for transmission based on their perceived level of risk or impact using ranking by proportional piling. RESULTS: A majority (99.2%, n = 398) had heard about brucellosis and 71.2% were aware of the zoonotic nature of the disease. There were varied responses regarding transmission routes, symptoms, and preventive measures. Self-reported prevalence was relatively high (55.5%). Following adjusted analysis, factors such as subcounty, source of income, knowledge about symptoms of brucellosis, whether brucellosis is treatable, perception, and living close to animals were statistically significant. Participants from Wabinyonyi had 2.7 higher odds of reporting brucellosis than those from Nabiswera, aOR = 2.7, 95%CI 1.4-5.5. Crop farming and livestock had much higher odds of reporting brucellosis than those earning from casual sources, aOR = 8.5, 95%C 1.8-40.1 and aOR = 14.4, 95%CI 3.1-67.6, respectively. Those who had knowledge about symptoms had 6.9 higher odds of reporting brucellosis than who mentioned fever, aOR = 4.5, 95%CI 2.3-18.3. Likewise, living close with animals and handling aborted fetuses (aOR = 0.4, 95%CI: 0.17-0.86), (aOR = 0.2,95% CI: 0.07-0.0.42,) had significantly lower odds for self-reported prevalence compared to those who believed did not cause brucellosis. Risk factors identified included, handling of aborted fetuses and living in close proximity with animals. Overall, there was a moderate statistical agreement in the ranking across the focus groups discussion (Wc = 0.48, p < 0.01; n = 6). CONCLUSION: While awareness of brucellosis is high in the community, understanding of the transmission routes, clinical symptoms, and preventive measures varied. Investigating the understanding and assessing knowledge, perceptions and self-reported prevalence in this region indicated a necessity for risk communication strategies and community-based interventions to better control brucellosis transmission.

Two cases of MPO-ANCA-positive hypertrophic pachymeningitis mimicking as intracranial infection.

Hypertrophic pachymeningitis (HP) is a rare disorder marked by thickening of the dura mater due to diverse etiologies. MPO-ANCA-positive HP represents a variant of AAV confined to the central nervous system, distinguished by the presence of serum MPO antibodies. Distinguishing HP triggered by MPO-ANCA from other causes can be challenging.In this study, we present two cases of MPO-ANCA-positive HP initially misdiagnosed as intracranial infections. Case 1 underwent surgery for chronic suppurative otitis media, with histopathological findings revealing inflammatory changes without definitive suppuration. He was presumed to have a secondary intracranial infection resulting from the surgery. However, his condition deteriorated despite two weeks of antibiotic and antiviral treatment. Case 2 presented with headache and was initially suspected of having intracranial Brucellosis given his serum Brucella positivity. Despite treatment for brucellosis, his symptoms persisted, and he developed visual and hearing impairments. Both patients were ultimately diagnosed with MPO-ANCA-positive HP, exhibiting serum MPO antibody positivity. Their symptoms showed improvement with glucocorticoid and immunosuppressive therapy.Based on these observations, we propose that MPO-ANCA-positive HP may initially present as intracranial infection. For HP patients presenting with headache, mastoiditis, otitis media, and visual loss, it is imperative to conduct ANCA antibody-related tests to enhance diagnostic precision.

A systematic review and meta-analysis of comparative clinical studies on antibiotic treatment of brucellosis.

Brucellosis is a difficult to treat infection that requires antibiotic combinations administered over several weeks for clearance of infection and relapse prevention. This systematic review summarizes current evidence for antibiotic treatment of human brucellosis. PubMed, EMBASE, Scopus, CINAHL, Web of Science, and China Academic Journal databases were searched for prospective studies that had compared different antibiotic regimens for treating human brucellosis in the last 25 years. Thirty-four studies recruiting 4182 participants were eligible. Standard dual therapy with doxycycline + rifampicin had a higher risk of treatment failure compared to triple therapy which added streptomycin (RR: 1.98, 95% CI 1.17-3.35, p = 0.01) or levofloxacin (RR: 2.98, 95% CI 1.67-5.32, p = 0.0002), but a similar or lower risk compared to alternative dual antibiotic combinations (p > 0.05). The same combination had a higher risk of relapses compared to triple therapy which added streptomycin (RR: 22.12, 95% CI 3.48-140.52, p = 0.001), or levofloxacin (RR: 4.61, 95% CI 2.20-9.66, p < 0.0001), but a similar or lower risk compared to other dual antibiotic combinations (p > 0.05). Triple antibiotic therapy is more effective than standard dual therapy with rifampicin and doxycycline. However, the latter is also efficacious and suitable for uncomplicated disease.

[One-stage posterior debridement and spinal internal fixation for the treatment of lumbar Brucellar spondylitis].

OBJECTIVE: To explore the clinical efficacy and safety of one-stage posterior lesion removal and internal spinal fixation in patients with lumbar Brucellosis spondylitis. METHODS: The clinical data of 24 patients admitted from October 2017 to October 2022 were retrospectively analyzed, 2 patients were lost to follow-up at 10 months after surgery, at the final 22 cases were included in the study, including 13 males and 9 females with an average age of (52.00±6.89) years old, were treated with one-stage posterior lesion removal and internal spinal fixation. The operation time, intraoperative bleeding, follow-up time, erythrocyte sedimentation rate(ESR) and C-reactive protein(CRP) before and after operation were recorded. The pain visual analogue scale(VAS), Oswestry disability index(ODI), the Japanese Orthopaedic Association(JOA) score for neurofunction, American Spinal Injury Association(ASIA) spinal cord injury grade and modified MacNab criteria were ussed to evaluate the efficacy. RESULTS: All patients were followed up from 12 to 30 months with an average of (17.41±4.45) months. The operation time was 70 to 155 min with an average of (116.59±24.32) min;the intraoperative bleeding volume was 120 to 520 ml with an average of (275.00±97.53) ml. CRP and ESR levels decreased more significantly at 1 week and at the final follow-up than preoperative levels(P<0.05). VAS, JOA score and ODI at 1 week and at the latest follow-up were more significantly improved than preoperative results(P<0.05). There was no significant difference between ASIA preoperative and 1 week after operation(P>0.05), and a significant difference between preoperative and last follow-up(P<0.05). In the final follow-up, 21 patients had excellent efficacy, 1 patient had fair, and there was no recurrence during the follow-up. CONCLUSION: One-stage transpedicular lesion removal and internal spinal fixation, with few incisions and short operation time, helps the recovery of neurological function, and the prognosis meets the clinical requirements, which can effectively control Brucella spondylitis.

Development and Application of Colloidal Gold Test Strips for the Rapid Detection of Canine Brucellosis.

Brucellosis is a global problem, with the causative agent being the genus Brucella. B. canis can cause undulant fever in dogs, which is a zoonotic disease that can spread not only among dogs but also to humans. This poses a public health threat to society. In this study, a rapid and straightforward immune colloidal gold test strip was developed for the diagnosis of canine brucellosis through the detection of anti-LPS antibodies in serum samples. Rabbit anti-canine IgG conjugated with colloidal gold was employed as the colloidal gold-labeled antibody. The extracted high-purity R-LPS was employed as the capture antigen in the test line (T-line), while goat anti-rabbit IgG was utilized as the capture antibody in the control line (C-line). The colloidal gold strip exhibited high specificity in the detection of brucellosis, with no cross-reaction observed with the common clinical canine diseases caused by Canine coronavirus (CCV), Canine distemper virus (CDV), and Canine parvovirus (CPV). In comparison to the commercial iELISA kit, the sensitivity and specificity of the colloidal gold test strip were found to be 95.23% and 98.76%, respectively. The diagnostic coincidence rate was 98.47%. The findings of this study indicate that colloidal gold test strips may be employed as a straightforward, expeditious, sensitive, and specific diagnostic instrument for the identification of canine brucellosis, particularly in resource-limited regions.

Applying a Fluorescence Polarization Assay for Detection of Brucellosis in Animals Using the Fluorescently Labeled Synthetic Oligosaccharides as Biosensing Tracer.

Brucellosis in animals is an infectious disease caused by bacteria of the genus Brucella. Known methods for diagnosing brucellosis face some challenges, due to the difficulties in isolating and standardizing the natural brucellosis antigen. In this work, we investigated the possibility of using the fluorescence polarization assay (FPA) with synthetic glycoconjugate biosensing tracers to detect antibodies against Brucella as a new methodology for diagnosing brucellosis. Based on the received results, the synthetic fluorescein-labeled trisaccharide tracer is most effective for Brucellosis detection. This tracer is structurally related to the immune determinant fragment of the Brucella LPS buildup of N-formyl-d-perosamine units, connected via α-(1→3)-linkage at the non-reducing end and α-(1→2)-linkage at the reducing end. The sensitivity and specificity in the case of the use of trisaccharide tracer 3b were 71% and 100% (Yuden's method) and 87% and 88% (Euclidean method), respectively, which is comparable with the diagnostic efficiency of traditionally used serological methods, such as the agglutination test (AT), complement fixation test (CFT), and Rose Bengal test (RBT). Given the known advantages of FPA (e.g., speed, compactness of the equipment, and standard reagents) and the increased specificity of the developed test system, it would be appropriate to consider its widespread use for the diagnosis of brucellosis in animals, including rapid testing in the field.

Analysis of the Brucella melitensis epidemic in Xinjiang: genotyping, polymorphism, antibiotic resistance and tracing.

Brucella spp. are facultative intracellular pathogens that cause zoonosis- brucellosis worldwide. There has been a trend of the re-emergence of brucellosis worldwide in recent years. The epidemic situation of brucellosis is serious in Xinjiang. To analyze the epidemic situation of Brucella spp. in Xinjiang among humans and animals, this study identified 144 Brucella isolates from Xinjiang using classical identification and 16 S rRNA sequencing. MLVA, drug resistance testing, and wgSNP detection were also performed. At the same time, analysis was conducted based on the published data of Brucella isolates worldwide. The results showed that the dominant species was B. melitensis biovar 3, which belonged to GT42 (MLVA-8 typing) and the East Mediterranean lineage. The correlation among isolates was high both in humans or animals. The isolates in Xinjiang exhibited higher polymorphism compared to other locations in China, with polymorphism increasing each year since 2010. No amikacin/kanamycin-resistant strains were detected, but six rifampicin-intermediate isolates were identified without rpoB gene variation. The NJ tree of the wgSNP results indicated that there were three main complexes of the B. melitensis epidemic in Xinjiang. Based on the results of this study, the prevention and control of brucellosis in Xinjiang should focus on B. melitensis, particularly strains belonging to B. melitensis bv.3 GT42 (MLVA-8 typing) and East Mediterranean lineage. Additionally, the rifampicin- and trimethoprim-sulfamethoxazole- resistance of isolates in Xinjiang should be closely monitored to avoid compromising the therapeutic efficacy and causing greater losses. These results provide essential data for the prevention and control of brucellosis in Xinjiang and China. Although the isolates from Xinjiang have significant characteristics among Chinese isolates and can reflect the epidemiological situation of brucellosis in China to some extent, this study cannot represent the characteristics of isolates from other regions.

Brucella Prostatitis: A Rare Manifestation of Fever of Unknown Origin, Detected by FDG PET/CT.

ABSTRACT: FDG PET/CT is a well-documented imaging investigation to evaluate fever of unknown origin (FUO). Brucellosis is one of the causes of FUO, which can be missed as it requires a longer incubation period for growth on culture media. Rarely, it can involve the prostate. Here, we present a case of FUO with initial negative blood and urine cultures and no localizing signs or symptoms. 18F-FDG PET/CT revealed hypermetabolism in the prostate and seminal vesicles. A repeat blood and urine culture showed the growth of Brucella species after 5 days of incubation, and the patient responded to Brucella-directed antibiotic therapy.

Differentiation of tuberculous and brucellar spondylitis using conventional MRI-based deep learning algorithms.

PURPOSE: To investigate the feasibility of deep learning (DL) based on conventional MRI to differentiate tuberculous spondylitis (TS) from brucellar spondylitis (BS). METHODS: A total of 383 patients with TS (n = 182) or BS (n = 201) were enrolled from April 2013 to May 2023 and randomly divided into training (n = 307) and validation (n = 76) sets. Sagittal T1WI, T2WI, and fat-suppressed (FS) T2WI images were used to construct single-sequence DL models and combined models based on VGG19, VGG16, ResNet18, and DenseNet121 network. The area under the receiver operating characteristic curve (AUC) was used to assess the classification performance. The AUC of DL models was compared with that of two radiologists with different levels of experience. RESULTS: The AUCs based on VGG19, ResNet18, VGG16, and DenseNet121 ranged from 0.885 to 0.973, 0.873 to 0.944, 0.882 to 0.929, and 0.801 to 0.933, respectively, and VGG19 models performed better. The diagnostic efficiency of combined models outperformed single-sequence DL models. The combined model of T1WI, T2WI, and FS T2WI based on VGG19 achieved optimal performance, with an AUC of 0.973. In addition, the performance of all combined models based on T1WI, T2WI, and FS T2WI was better than that of two radiologists (P<0.05). CONCLUSION: The DL models have potential guiding value in the diagnosis of TS and BS based on conventional MRI and provide a certain reference for clinical work.

Revealing metabolic alterations in brucellosis patients by targeted metabolomics.

Brucellosis, a zoonotic disease caused by brucella infection, presents metabolic profile changes in patients that have not been extensively explored. This study utilized an ultra-high performance liquid chromatography tandem mass spectrometry based targeted metabolomic approach to comprehensively investigated metabolic changes in Brucella patients. Serum samples of brucellosis 50 patients and 50 well-matched healthy controls were analyzed for 228 metabolites, revealing significant alterations in 83 metabolites in brucellosis patients. Notably, disruptions were observed in key metabolite pathways, such as amino acid metabolism, urea cycle, tricarboxylic acid cycle (TCA), and fatty acid metabolism. Patients diagnosed with Brucellosis exhibited distinct differences in the levels of aspartate, glutamate, ß-alanine, and asparagine when compared to controls. Within the urea cycle, a significant downregulation of arginine was observed, whereas ornithine levels were considerably upregulated. In the TCA cycle, concentrations of 2-oxoglutarate, succinate, and malate were significantly elevated, while citrate levels demonstrated a notable decrease. Due to the interruption of the TCA cycle, glycolysis was accelerated to compensate for the resultant energy deficit in Brucella patients. Concurrently, there was a significant increase in the levels of short and medium-chain fatty acids, while long-chain fatty acids showed a marked decrease. The study systematically revealed significant metabolic alterations in Brucellosis patients and further explored the potential correlation between these changes and clinic symptoms, including fatigue, muscle soreness and prolonged fever. The results enhanced our understanding of Brucellosis, offering valuable insights potentially beneficial in formulating more effective treatment strategies and improving prognostic approaches.

Transmission of Brucella canis in a canine kennel following introduction of an infected dog.

Brucella canis is a zoonotic pathogen and the main causative agent of canine brucellosis. In the Netherlands, B. canis had previously only been detected in individual cases of imported dogs. However, an outbreak of B. canis occurred for the first time in a cohort of autochthonous dogs in a breeding kennel in 2019. The outbreak began with a positive serological test result of an imported intact male dog showing clinical symptoms of brucellosis. Consequently, urine and blood samples were collected and tested positive for B. canis by culture, matrix-assisted laser desorption/ionization - time of flight mass spectrometry (MALDI-TOF MS) and whole-genome-sequencing (WGS). Screening of the contact dogs in the kennel where the index case was kept, revealed that antibodies against B. canis could be detected in 23 out of 69 dogs (34 %) by serum agglutination test (SAT). Of the 23 seropositive dogs, B. canis could be cultured from the urine and/or heparin samples of 19 dogs (83 %). This outbreak represents the first documented case of transmission of B. canis to autochthonous contact dogs in the Netherlands. WGS revealed all B. canis isolates belonged to the same cluster, which means the transmission of B. canis in the breeding kennel was most likely caused by the introduction of one infected dog. Comparing this cluster with data from other B. canis isolates, it also appears that characteristic clusters of B. canis are present in several endemic countries. These clusters seem to remain stable over time and may help in locating the origin of new isolates found. This outbreak showed that the international movement of dogs from endemic countries poses a threat to the canine population, while serological screening and WGS proved to be valuable tools for respectively screening and the epidemiological investigation.

A morphological and molecular approach to investigating infectious disease in early medieval Iberia: The necropolis of La Olmeda (Palencia, Spain).

OBJECTIVE: Here we investigate infectious diseases that potentially contribute to osteological lesions in individuals from the early medieval necropolis of La Olmeda (6th-11th c. CE) in North Iberia. MATERIALS AND METHODS: We studied a minimum number of 268 individuals (33 adult females; 38 adult males, 77 unknown/indeterminate sex; and 120 non-adults), including articulated and commingled remains. Individuals with differential diagnoses suggesting chronic systemic infectious diseases were sampled and bioinformatically screened for ancient pathogen DNA. RESULTS: Five non-adults (and no adults) presented skeletal evidence of chronic systemic infectious disease (1.87% of the population; 4.67% of non-adults). The preferred diagnoses for these individuals included tuberculosis, brucellosis, and malaria. Ancient DNA fragments assigned to the malaria-causing pathogen, Plasmodium spp., were identified in three of the five individuals. Observed pathology includes lesions generally consistent with malaria; however, additional lesions in two of the individuals may represent hitherto unknown variation in the skeletal manifestation of this disease or co-infection with tuberculosis or brucellosis. Additionally, spondylolysis was observed in one individual with skeletal lesions suggestive of infectious disease. CONCLUSIONS: This study sheds light on the pathological landscape in Iberia during a time of great social, demographic, and environmental change. Genetic evidence challenges the hypothesis that malaria was absent from early medieval Iberia and demonstrates the value of combining osteological and archaeogenetic methods. Additionally, all of the preferred infectious diagnoses for the individuals included in this study (malaria, tuberculosis, and brucellosis) could have contributed to the febrile cases described in historical sources from this time.