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Hemophagocytic lymphohistiocytosis (HLH) is a hyper-inflammatory condition triggered by infections, malignancies, or autoimmune conditions. Brucellosis is a zoonotic disease contracted through exposure to infected animals or consumption of unpasteurized dairy products. The complications of both pathologies may be fatal. This report presents a rare instance of HLH induced by Brucellosis, highlighting the need for increased recognition of this life-threatening association.

Introduction: Human brucellosis is a severe public concern in Shaanxi Province, China, and investigating the epidemiological relationship and transmission pattern of B. melitensis is necessary to devise control strategies. Methods: In this study, a conventional bio-typing approach and whole genome sequencing of single-nucleotide polymorphisms (SNPs) were employed to identify 189 strains. Results: Based on the conventional bio-typing, 189 Brucella strains were identified as B. melitensis , of which 14 were in bv. 1, 145 were in bv. 3, and 30 were variant, and the Brucella strains were distributed in all ten cities in Shaanxi Province. SNP analysis was used to identify genetic variation in 189 B. melitensis genomes, and maximum-likelihood was used to generate a phylogeny that identified two clades (A and B) and 19 sequence types (STs). The two clades were highly diverse and exclusively of Eastern Mediterranean origin. Clade B contained 18 STs (2-19), with most isolates originating from a broad swath, implying that multiple B. melitensis lineages circulated in Shaanxi. The 19 STs were composed of 3 to 46 strains isolated from different counties and years, suggesting that multiple cross-county brucellosis outbreak events are driven by multiple B. melitensis lineages. Global phylogenetic analysis revealed that clade A was close to GTIIb, and clade B was placed in the GTIIh lineage, expanding the known diversity of B. melitensis from China. Conclusion: The human brucellosis epidemic in Shaanxi is driven by multiple indigenous circulating B. melitensis lineages, the knowledge of which will contribute to devising a control strategy and providing the foundation for a comprehensive regional phylogeny of this important zoonotic pathogen. [ABSTRACT FROM AUTHOR]

Introduction: Antimicrobial resistance (AMR) in Brucella melitensis, the causative agent of brucellosis, is of growing concern, particularly in low and middle-income countries. This study aimed to explore the genetic basis of AMR in B. melitensis strains from India. Methods: Twenty-four isolates from humans and animals were subjected to antimicrobial susceptibility testing and whole-genome sequencing. Results: Resistance to doxycycline (20.80%), ciprofloxacin (16.67%), cotrimoxazole (4.17%), and rifampicin (16.67%) was observed. Genome analysis revealed efflux-related genes like mprF, bepG, bepF, bepC, bepE, and bepD across all isolates, however, classical AMR genes were not detected. Mutations in key AMR-associated genes such as rpoB, gyrA, and folP were identified, intriguingly present in both resistant and susceptible isolates, suggesting a complex genotype–phenotype relationship in AMR among Brucella spp. Additionally, mutations in efflux genes were noted in resistant and some susceptible isolates, indicating their potential role in resistance mechanisms. However, mutations in AMR-associated genes did not consistently align with phenotypic resistance, suggesting a multifactorial basis for resistance. Discussion: The study underscores the complexity of AMR in B. melitensis and advocates for a holistic multi-omics approach to fully understand resistance mechanisms. These findings offer valuable insights into genetic markers associated with AMR, guiding future research and treatment strategies. [ABSTRACT FROM AUTHOR]

Background: Brucellosis, a zoonotic disease in Türkiye, which has significant direct and indirect impacts on the healthcare system and livestock. This study, which aimed to investigate the differences among Brucella spp. isolates originating from different regions of Türkiye, for implications for public health and veterinary medicine. Method: Twenty-one isolates from ruminants and two isolates from humans obtained from various regions of Türkiye were utilized in the study. The isolates were identified and biotyped using traditional microbiological procedures, and whole-genome sequencing (WGS) was performed. This was followed by single nucleotide polymorphism (SNP)--based phylogenetic analysis and WGS-based analysis of virulence and resistance genes. Additionally, phenotypic antimicrobial resistance and phage susceptibilities were determined. The obtained data were then compared for concordance, ensuring the validity and reliability of the results. Results: Our study, employing culture methods, polymerase chain reaction (PCR), and WGS analyses, identified 11 Brucella melitensis (bv 3 (n = 9), one each bv 1 and bv 2) and 12 B. abortus (bv 3 (n = 11), bv 9 (n = 1)) isolates All B. abortus isolates were of bovine origin, while the B. melitensis isolates were from sheep (n = 7), goat (n = 1), ram (n = 1), and humans (n = 2). In the whole-genome SNP-based phylogenetic tree, all B. melitensis strains were found to be of the IIb subtype of genotype II associated with the Eastern Mediterranean lineage. Ten different genotypes were identified in the SNP analysis of the isolates, with a maximum SNP difference of 278 and a minimum SNP difference of 4 among these genotypes. According to the WGS-SNP-based phylogenetic tree of B. abortus isolates, they were grouped in clade C1. In the SNP analysis, where ten different genotypes were identified, the SNP difference among these genotypes was a maximum of 316 and a minimum of 6. In the in silico MLST analysis performed with WGS data, B. melitensis isolates were identified as ST8 and ST102 genotypes, while B. abortus isolates were identified as ST2 and ST3 genotypes. The dominant genotypes were ST8 for B. melitensis and ST2 for B. abortus, respectively. Virulence gene analysis conducted based on WGS data of the 23 B. abortus and B. melitensis isolates revealed 43 virulence gene-associated regions in all strains, irrespective of species, host, or isolation year. Although classical resistance-related genes were not detected by WGS-based antimicrobial resistance gene analysis, phenotypic resistance analysis revealed resistance to azithromycin, rifampin, and trimethoprim/sulfamethoxazole in B. abortus and B. melitensis isolates. Conclusion: Both B. melitensis and B. abortus were circulating species in animals and human. The dominant genotypes were ST8 for B. melitensis and ST2 for B. abortus, respectively. All B. melitensis strains were found to be of the IIb subtype of genotype II associated with the Eastern Mediterranean lineage, while B. abortus isolates, they were grouped in clade C1. Further, a comprehensive study with a sufficient number of isolates covering all regions of Türkiye would provide more accurate information about the current epidemiological situation in the country. [ABSTRACT FROM AUTHOR]

Brucellosis and Tuberculosis, both of which are contagious diseases, have presented significant challenges to global public health security in recent years. Delayed treatment can exacerbate the conditions, jeopardizing patient lives. Currently, no vaccine has been approved to prevent these two diseases simultaneously. In contrast to traditional vaccines, mRNA vaccines offer advantages such as high efficacy, rapid development, and low cost, and their applications are gradually expanding. This study aims to develop multi-epitope mRNA vaccines argeting Brucella melitensis and Mycobacterium tuberculosis H37Rv (L4 strain) utilizing immunoinformatics approaches. The proteins Omp25, Omp31, MPT70, and MPT83 from the specified bacteria were selected to identify the predominant T- and B-cell epitopes for immunological analysis. Following a comprehensive evaluation, a vaccine was developed using helper T lymphocyte epitopes, cytotoxic T lymphocyte epitopes, linear B-cell epitopes, and conformational B-cell epitopes. It has been demonstrated that multi-epitope mRNA vaccines exhibit increased antigenicity, non-allergenicity, solubility, and high stability. The findings from molecular docking and molecular dynamics simulation revealed a robust and enduring binding affinity between multi-epitope peptides mRNA vaccines and TLR4. Ultimately, Subsequently, following the optimization of the nucleotide sequence, the codon adaptation index was calculated to be 1.0, along with an average GC content of 54.01%. This indicates that the multi-epitope mRNA vaccines exhibit potential for efficient expression within the Escherichia coli(E. coli) host. Analysis through immune modeling indicates that following administration of the vaccine, there may be variation in immunecell populations associated with both innate and adaptive immune reactions. These types encompass helper T lymphocytes (HTL), cytotoxic T lymphocytes (CTL), regulatory T lymphocytes, natural killer cells, dendritic cells and various immune cell subsets. In summary, the results suggest that the newly created multi-epitope mRNA vaccine exhibits favorable attributes, offering novel insights and a conceptual foundation for potential progress in vaccine development. [ABSTRACT FROM AUTHOR]

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

The impact of vaccine-induced immune responses on host metabolite availability has not been well studied. Here we show that prior vaccination alters the metabolic profile of mice challenged with Brucella melitensis. In particular, glucose levels were reduced in vaccinated mice in an antibody-dependent manner. We also found the glucose transporter gene gluP plays a lesser role in B melitensis virulence in vaccinated wild type mice relative to vaccinated mice unable to secrete antibodies. These data indicate that vaccine-elicited antibodies protect the host in part by restricting glucose availability. Moreover, Brucella and other pathogens may need to employ different metabolic strategies in vaccinated hosts. [ABSTRACT FROM AUTHOR]

PurposeCase presentationConclusionTo present a treatment regimen for Brucellosis endophthalmitis that resulted in a good visual outcome. Additionally, we conducted a literature review on the treatment and visual prognosis of related cases.A 49-year-old woman with the chief complaint of decreased vision and redness in the right eye was initially diagnosed with noninfectious uveitis and prescribed high-dose steroids which led to transient improvement followed by a decline in vision. An infectious cause was suspected. Metagenomic next-generation sequencing of vitreous fluid and serological testing confirmed Brucella melitensis infection. The patient underwent vitrectomy combined with six intravitreal injections of ceftazidime in the right eye in addition to systemic antibiotic treatment. The intraocular inflammation was completely resolved, and the visual acuity recovered to 20/25, which is the best-documented recovery in Brucella endophthalmitis cases, as revealed by the literature review.Vitrectomy combined with repeated intravitreal injections of ceftazidime can enhance the treatment for brucellosis endophthalmitis and achieve a better visual prognosis. [ABSTRACT FROM AUTHOR]

Introduction: Brucellosis is an important zoonosis worldwide, affecting humans and animals. There are no specific medicines available to treat brucellosis. Astragalus polysaccharide (APS) is derived from Astragalus membranaceus and exhibits impressive bioactivity, including anti-aging, anti-tumor, and immunomodulatory functions. Methods: Mice were intraperitoneally inoculated with Brucella melitensis M5 and then treated with APS intraperitoneally injection daily for 7 d. Results: Compared to the M5-infected group, the lower bacteria loads in the APS-treated groups were proved, especially at the acute stage of infection. APS treatment relieved splenomegaly, excess expressions of several pro-inflammatory cytokines (including CXCL1, IFN-γ, IL-1β, IL-2, IL-12p70, and TNF-α). The raised level of IL-4 was observed in APS-treated mice. APS contributed to raising the ratio of M1 macrophage and reducing the ratio of M2 macrophage in the blood. Discussion: The present study provides some evidence on the potential application of APS in controlling and treating brucellosis and should be further explored. [ABSTRACT FROM AUTHOR]

A retrospective study was conducted in Abergele and Ziquala districts in Ethiopia to investigate the occurrence, impact, and potential risk factors for abortion in small ruminants linked to a regional abortion storm. Affected (case) and nonaffected (control) villages were compared to assess infectious and noninfectious risk factors causing abortions. A case village was defined as a village with abortion seen in all households enrolled in the study, while a control village is characterized by presence of abortion in two and fewer households. A questionnaire survey, focus group discussions, and serological examinations were used to assess the differences in the abortion rate between the case and control villages. The Rose Bengal Plate Test for Brucellosis, Toxoplasma gondii Antibody Test Kit for Toxoplasma gondii, and ELISA for Chlamydophila abortus and Coxiella burnetii were used to detect antibodies. Per village 15 household flocks were selected. In the case villages, all flocks were affected by abortion (a mean abortion of 13 animals in Abergele and 9 in Ziquala). In contrast, only three households reported abortions in the control villages. A total of 176 blood samples were collected from the case and control villages for further laboratory diagnosis of possible causes of abortion. Of the examined flocks, 17%, 10.2%, and 2.8% were positive for Coxiella burnetii, Toxoplasma gondii, and Chlamydophila abortus, respectively. However, antibodies against Brucella melitensis were not detected. While the seroprevalence was greater for some infectious agents of abortion, there was no significant difference between the case and control villages. It is clear that the abortion problem in the study villages is complex and likely due to a mix of poor husbandry practices and the presence of infectious diseases. To better understand the underlying causes of abortion, there is a need to conduct a longitudinal study involving testing for more pathogens at the household level combined with reliable data on husbandry practices. [ABSTRACT FROM AUTHOR]

Objective: Brucellosis is an important zoonotic infection that causes multi-organ involvement, and the effectiveness of culture and serological tests used in the diagnosis of the disease is limited. In this study, it was aimed to investigate the effectiveness of Polymerase Chain Reaction (PCR) in the diagnosis of brucellosis. Material and Method: Sixty patients with positive Rose-Bengal Agglutination test and clinically suspected brucellosis and 60 control samples without these features were included in the study. The DNA of Brucella spp. was investigated in the samples by real-time PCR method. In the positive samples, the species identification was made, and virulance genes including omp19, wbkA, manA, mviN, urea and perA were studied with PCR. Results: Brucella spp. DNA was found to be positive in 19 of 32 patients in the patient group who were diagnosed with acute disease or relapse by clinical evaluation and serological tests, and no positive result was obtained in any of the control samples. Accordingly, the sensitivity and specificity of PCR were calculated as 59.3% and 100%, respectively. It was determined that Brucella melitensis was the infecting agent in all positive patients and carried all the virulence genes studied. Conclusion: Developing molecular methods and taking samples from the patient during the period of bacteremia may increase the effectiveness of PCR in the diagnosis of brucellosis. The identification of the pathogen as B. melitensis in all positive patients indicated that the transmission brucellosis to humans in our region was mainly from sheeps and goats, and appropriate control measures should be taken accordingly. [ABSTRACT FROM AUTHOR]

Brucellosis is a chronic and debilitating disease in humans, causing great economic losses in the livestock industry. Making an effective vaccine is one of the most important concerns for this disease. The new mRNA vaccine technology due to its accuracy and high efficiency has given promising results in various diseases. The objective of this research was to create a novel mRNA vaccine with multiple epitopes targeting Brucella melitensis. Seventeen antigenic proteins and their appropriate epitopes were selected with immunoinformatic tools and surveyed in terms of toxicity, allergenicity, and homology. Then, their presentation and identification by MHC cells and other immune cells were checked with valid tools such as molecular docking, and a multi-epitope protein was modeled, and after optimization, mRNA was analyzed in terms of structure and stability. Ultimately, the immune system's reaction to this novel vaccine was evaluated and the results disclosed that the designed mRNA construct can be an effective and promising vaccine that requires laboratory and clinical trials. [ABSTRACT FROM AUTHOR]

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. Author summary: Brucellosis is one of the most widespread bacterial zoonoses worldwide. 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 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, which demonstrates that B. melitensis faces different selective pressures depending on the organ. Then, 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. We observed that 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 the effectiveness of functional genome mapping for the design of new safer live attenuated vaccine candidates. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Brucella melitensis is a major livestock bacterial pathogen and zoonosis, causing disease and infection-related abortions in small ruminants and humans. A considerable burden to animal-based economies today, the presence of Brucella in Neolithic pastoral communities has been hypothesised but we lack direct genomic evidence thus far. We report a 3.45X B. melitensis genome preserved in an ~8000 year old sheep specimen from Menteşe Höyük, Northwest Türkiye, demonstrating that the pathogen had evolved and was circulating in Neolithic livestock. The genome is basal with respect to all known B. melitensis and allows the calibration of the B. melitensis speciation time from the primarily cattle-infecting B. abortus to approximately 9800 years Before Present (BP), coinciding with a period of consolidation and dispersal of livestock economies. We use the basal genome to timestamp evolutionary events in B. melitensis, including pseudogenization events linked to erythritol response, the supposed determinant of the pathogen's placental tropism in goats and sheep. Our data suggest that the development of herd management and multi-species livestock economies in the 11th–9th millennium BP drove speciation and host adaptation of this zoonotic pathogen. Brucella melitensis is a zoonotic bacterial pathogen of livestock that can infect humans and causes brucellosis. Here, the authors sequence an ancient specimen of B. melitensis and show that the species emerged in the Neolithic period, around the time of development of animal management practices. [ABSTRACT FROM AUTHOR]

This study was carried out to evaluate Brucella spp. isolated from various tissue samples of aborted sheep and bovine fetuses sent to the laboratory of Department of Microbiology, Faculty of Veterinary Medicine, Kafkas University between 2011 and 2023 years and determine the Brucella species and biotype diversity that carry a higher risk for abortion complications in these animals. In this context, 155 Brucella spp. isolates obtained from aborted fetuses were identified by species-specific Bruceladder PCR and biotyped using conventional biotyping methods. As a result of the study, B. melitensis and B. abortus were identified in 92.5% (n=74) and 7.5% (n=6) of sheep, B. abortus and B. melitensis were identified in 80% (n=60) and 20% (n=15) of cattle, respectively. B. melitensis biotype 2 in sheep and B. abortus biotype 3 in cattle were found as the dominant biotypes in these definitive hosts. In the Kars region, where brucellosis is endemic, while the biotype responsible for cattle brucellosis (B. abortus biotype 3) maintained its dominance over a 20-year period, there is a profile change from B. melitensis biotype 3 to B. melitensis biotype 2 in sheep. Considering the period covered by the study and the sample size analyzed, the data obtained provide up-to-date and important information about Brucella species and biotypes in Kars region and the animal species that host these agents. [ABSTRACT FROM AUTHOR]

Background: Brucellosis is the most important public health problem worldwide, and the annual incidence of the disease in humans is 2.1 million. The Brucella genome is highly conserved, with over 90% similarity among species. The aim of this study was to perform species-level identification of Brucella spp. strains isolated from humans diagnosed with brucellosis and to further investigate the phylogenetic relationships using multiple locus variable number of tandem repeats analysis (MLVA)-16 and 16S rRNA sequencing analysis. Materials and Methods:Brucella spp. was isolated from the blood cultures of 54 patients who tested positive for brucellosis through serological examinations. Real-time PCR was used to identify the isolates in species, and the genus level of Brucella was confirmed with 16S rRNA. All isolates were subjected to phylogenetic analysis using variable number of tandem repeat analysis with multiple loci. Results: Subsequent analysis via real-time PCR confirmed these isolates to be of the Brucella melitensis species. The 16S rRNA sequence analysis showed 100% homogeneity among the isolates. MLVA revealed the formation of five different genotypic groups. While two groups were formed based on the 16S rRNA sequence analysis, five groups were formed in the MLVA. Conclusions: The study concluded that 16S rRNA sequence analysis alone did not provide sufficient discrimination for phylogenetic analysis but served as a supportive method for identification. MLVA exhibited higher phylogenetic power. The widespread isolation of B. melitensis from human brucellosis cases highlights the importance of controlling brucellosis in small ruminants to prevent human infections. [ABSTRACT FROM AUTHOR]

IntroductionHuman brucellosis is a severe public concern in Shaanxi Province, China, and investigating the epidemiological relationship and transmission pattern of B. melitensis is necessary to devise control strategies.MethodsIn this study, a conventional bio-typing approach and whole genome sequencing of single-nucleotide polymorphisms (SNPs) were employed to identify 189 strains.ResultsBased on the conventional bio-typing, 189 Brucella strains were identified as B. melitensis, of which 14 were in bv. 1, 145 were in bv. 3, and 30 were variant, and the Brucella strains were distributed in all ten cities in Shaanxi Province. SNP analysis was used to identify genetic variation in 189 B. melitensis genomes, and maximum-likelihood was used to generate a phylogeny that identified two clades (A and B) and 19 sequence types (STs). The two clades were highly diverse and exclusively of Eastern Mediterranean origin. Clade B contained 18 STs (2-19), with most isolates originating from a broad swath, implying that multiple B. melitensis lineages circulated in Shaanxi. The 19 STs were composed of 3 to 46 strains isolated from different counties and years, suggesting that multiple cross-county brucellosis outbreak events are driven by multiple B. melitensis lineages. Global phylogenetic analysis revealed that clade A was close to GTIIb, and clade B was placed in the GTIIh lineage, expanding the known diversity of B. melitensis from China.ConclusionThe human brucellosis epidemic in Shaanxi is driven by multiple indigenous circulating B. melitensis lineages, the knowledge of which will contribute to devising a control strategy and providing the foundation for a comprehensive regional phylogeny of this important zoonotic pathogen.

IntroductionAntimicrobial resistance (AMR) in Brucella melitensis, the causative agent of brucellosis, is of growing concern, particularly in low and middle-income countries. This study aimed to explore the genetic basis of AMR in B. melitensis strains from India.MethodsTwenty-four isolates from humans and animals were subjected to antimicrobial susceptibility testing and whole-genome sequencing.ResultsResistance to doxycycline (20.80%), ciprofloxacin (16.67%), cotrimoxazole (4.17%), and rifampicin (16.67%) was observed. Genome analysis revealed efflux-related genes like mprF, bepG, bepF, bepC, bepE, and bepD across all isolates, however, classical AMR genes were not detected. Mutations in key AMR-associated genes such as rpoB, gyrA, and folP were identified, intriguingly present in both resistant and susceptible isolates, suggesting a complex genotype–phenotype relationship in AMR among Brucella spp. Additionally, mutations in efflux genes were noted in resistant and some susceptible isolates, indicating their potential role in resistance mechanisms. However, mutations in AMR-associated genes did not consistently align with phenotypic resistance, suggesting a multifactorial basis for resistance.DiscussionThe study underscores the complexity of AMR in B. melitensis and advocates for a holistic multi-omics approach to fully understand resistance mechanisms. These findings offer valuable insights into genetic markers associated with AMR, guiding future research and treatment strategies.

This study was carried out to evaluate Brucella spp. isolated from various tissue samples of aborted sheep and bovine fetuses sent to the laboratory of Department of Microbiology, Faculty of Veterinary Medicine, Kafkas University between 2011 and 2023 years and determine the Brucella species and biotype diversity that carry a higher risk for abortion complications in these animals. In this context, 155 Brucella spp. isolates obtained from aborted fetuses were identified by species-specific Bruceladder PCR and biotyped using conventional biotyping methods. As a result of the study, B. melitensis and B. abortus were identified in 92.5% (n=74) and 7.5% (n=6) of sheep, B. abortus and B. melitensis were identified in 80% (n=60) and 20% (n=15) of cattle, respectively. B. melitensis biotype 2 in sheep and B. abortus biotype 3 in cattle were found as the dominant biotypes in these definitive hosts. In the Kars region, where brucellosis is endemic, while the biotype responsible for cattle brucellosis (B. abortus biotype 3) maintained its dominance over a 20-year period, there is a profile change from B. melitensis biotype 3 to B. melitensis biotype 2 in sheep. Considering the period covered by the study and the sample size analyzed, the data obtained provide up-to-date and important information about Brucella species and biotypes in Kars region and the animal species that host these agents.

Abstract Brucellosis poses a significant challenge in the medical field as a systemic infection with a propensity for relapse. This study presented a novel approach to brucellosis treatment, enhancing the efficacy of doxycycline and rifampicin through the use of poly (lactic-co-glycolic) acid coupled with cadmium-telluride quantum dots (Dox-Rif-PLGA@CdTe). The double emulsion solvent evaporation method was employed to prepare Dox-Rif-PLGA@CdTe. The study scrutinized the physicochemical attributes of these nanoparticles. The impact of antibiotic-loaded nanoparticles on Brucella melitensis was evaluated through well diffusion, minimum inhibitory concentration (MIC), and cell culture. The chemical analysis results demonstrated a possibility of chemical reactions occurring among the constituents of nanoparticles. Assessments using the well diffusion and MIC methods indicated that the impact of free drugs and nanoparticles on bacteria was equivalent. However, the drug-loaded nanoparticles significantly decreased the colony-forming units (CFUs) within the cell lines compared to free drugs. In conclusion, the synthesis of nanoparticles adhered to environmentally friendly practices and demonstrated safety. The sustained drug release over 100 h facilitated drug accumulation at the bacterial site, resulting in a heightened therapeutic effect on B. melitensis and improved outcomes in brucellosis treatment. The application of these synthesized nanodrugs exhibited promising therapeutic potential.

The aim of the work was to conduct a comparative phylogenetic analysis based on wgSNP of complete genomes of Brucella melitensis strains circulating in the territory of the Russian Federation.Materials and methods. wgSNP typing of 412 B. melitensis strains of the main genetic lineages of brucella from different regions of the world, including 64 strains isolated in the regions of the European and Asian parts of the Russian Federation, was performed. DNA sequencing was conducted on the “Ion GeneStudio S5 Plus” platform (“Life Technologies”, USA) using the “Ion Plus Fragment Library Kit library preparation” (“Life Technologies”, USA), according to the “Ion 520™ & Ion 530™ Kit – Chef” protocol (Revision D.0).Results and discussion. It has been established that the strains circulating in Russia belong mainly to genotype II, which has a wide geographical distribution across the territory of Eurasia. At the same time, subgenotype IIh prevails in the regions of Siberia, and Iii – in the European territory of the country. For the first time, sets of specific SNPs have been identified that allow for intraspecific differentiation of B. melitensis strains. The obtained results made it possible to determine the probable routes of introduction of the causative agent of brucellosis into the territory of the Russian Federation from China and the countries of the Middle East. The prospect of using an optimized wgSNP typing scheme to solve urgent problems in the field of molecular epidemiology of brucellosis is demonstrated. It includes determining the genotype and subgenotype of the pathogen associated with the probable geographical region of origin of infection, and identifying the genetic relations between strains with high accuracy.

Introduction: Tuberculosis, caused by Mycobacterium tuberculosis complex (MTBC), remains a global health concern in both human and animals. However, the absence of rapid, accurate, and highly sensitive detection methods to differentiate the major pathogens of MTBC, including M. tuberculosis, M. bovis, and BCG, poses a potential challenge. Methods: In this study, we have established a triplex droplet digital polymerase chain reaction (ddPCR) method employing three types of probe fluorophores, with targets M. tuberculosis (targeting CFP-10-ESAT-6 gene of RD1 and Rv0222 genes of RD4), M. bovis (targeting CFP-10-ESATs-6 gene of RD1), and BCG (targeting Rv3871 and Rv3879c genes of 1RD1), respectively. Results: Based on optimization of annealing temperature, sensitivity and repeatability, this method demonstrates a lower limit of detection (LOD) as 3.08 copies/reaction for M. tuberculosis, 4.47 copies/reaction for M. bovis and 3.59 copies/reaction for BCG, without cross-reaction to Mannheimia haemolytica, Mycoplasma bovis, Haemophilus parasuis, Escherichia coli, Pasteurella multocida, Ochrobactrum anthropi, Salmonella choleraesuis, Brucella melitensis, and Staphylococcus aureus, and showed repeatability with coefficients of variation (CV) lower than 10%. The method exhibits strong milk sample tolerance, the LOD of detecting in spike milk was 5 × 10³ CFU/mL, which sensitivity is ten times higher than the triplex qPCR. 60 clinical DNA samples, including 20 milk, 20 tissue and 20 swab samples, were kept in China Animal Health and Epidemiology Center were tested by the triplex ddPCR and triplex qPCR. The triplex ddPCR presented a higher sensitivity (11.67%, 7/60) than that of the triplex qPCR method (8.33%, 5/60). The positive rates of M. tuberculosis, M. bovis, and BCG were 1.67, 10, and 0% by triplex ddPCR, and 1.67, 6.67, and 0% by triplex qPCR, with coincidence rates of 100, 96.7, and 100%, respectively. Discussion: Our data demonstrate that the established triplex ddPCR method is a sensitive, specific and rapid method for differentiation and identification of M. tuberculosis, M. bovis, and BCG. [ABSTRACT FROM AUTHOR]

One of the main causes of human brucellosis is Brucella melitensis infecting small ruminants. To date, Rev1 is the only vaccine successfully used to control ovine and caprine brucellosis. However, it is pathogenic for pregnant animals, resulting in abortions and vaginal and milk shedding, as well as being infectious for humans. Therefore, there is an urgent need to develop an effective vaccine that is safer than Rev1. In efforts to further attenuate Rev1, we recently used wzm inactivation to generate a rough mutant (Rev1Δ wzm) that retains a complete antigenic O-polysaccharide in the bacterial cytoplasm. The aim of the present study was to evaluate the placental pathogenicity of Rev1Δ wzm in trophoblastic cells, throughout pregnancy in mice, and in ewes inoculated in different trimesters of pregnancy. This mutant was evaluated in comparison with the homologous 16MΔ wzm derived from a virulent strain of B. melitensis and the naturally rough sheep pathogen B. ovis. Our results show that both wzm mutants triggered reduced cytotoxic, pro-apoptotic, and pro-inflammatory signaling in Bewo trophoblasts, as well as reduced relative expression of apoptosis genes. In mice, both wzm mutants produced infection but were rapidly cleared from the placenta, in which only Rev1Δ wzm induced a low relative expression of pro-apoptotic and pro-inflammatory genes. In the 66 inoculated ewes, Rev1Δ wzm was safe and immunogenic, displaying a transient serological interference in standard RBT but not CFT S-LPS tests; this serological response was minimized by conjunctival administration. In conclusion, these results support that B. melitensis Rev1Δ wzm is a promising vaccine candidate for use in pregnant ewes and its efficacy against B. melitensis and B. ovis infections in sheep warrants further study. [ABSTRACT FROM AUTHOR]

Brucellosis is a notifiable disease induced by a facultative intracellular Brucella pathogen. In this study, eight Brucella abortus and eighteen Brucella melitensis strains from Egypt were annotated and compared with RB51 and REV1 vaccines respectively. RAST toolkit in the BV-BRC server was used for annotation, revealing genome length of 3,250,377 bp and 3,285,803 bp, 3289 and 3323 CDS, 48 and 49 tRNA genes, the same number of rRNA (3) genes, 583 and 586 hypothetical proteins, 2697 and 2726 functional proteins for B. abortus and B. melitensis respectively. B. abortus strains exhibit a similar number of candidate genes, while B. melitensis strains showed some differences, especially in the SRR19520422 Faiyum strain. Also, B. melitensis clarified differences in antimicrobial resistance genes (KatG, FabL, MtrA, MtrB, OxyR, and VanO-type) in SRR19520319 Faiyum and (Erm C and Tet K) in SRR19520422 Faiyum strain. Additionally, the whole genome phylogeny analysis proved that all B. abortus strains were related to vaccinated animals and all B. melitensis strains of Menoufia clustered together and closely related to Gharbia, Dameitta, and Kafr Elshiek. The Bowtie2 tool identified 338 (eight B. abortus) and 4271 (eighteen B. melitensis) single nucleotide polymorphisms (SNPs) along the genomes. These variants had been annotated according to type and impact. Moreover, thirty candidate genes were predicted and submitted at GenBank (24 in B. abortus) and (6 in B. melitensis). This study contributes significant insights into genetic variation, virulence factors, and vaccine-related associations of Brucella pathogens, enhancing our knowledge of brucellosis epidemiology and evolution in Egypt. [ABSTRACT FROM AUTHOR]

Brucellosis is an important bacterial disease of livestock and the most common zoonotic disease. The current vaccines are effective but unsafe, as they result in animal abortions and are pathogenic to humans. Virus-like particles are being investigated as molecular scaffolds for foreign antigen presentation to the immune system. Here, we sought to develop a new-generation vaccine by presenting selected Brucella melitensis T cell epitopes on the surface of Orbivirus core-like particles (CLPs) and transiently expressing these chimeric particles in Nicotiana benthamiana plants. We successfully demonstrated the assembly of five chimeric CLPs in N. benthamiana plants, with each CLP presenting a different T cell epitope. The safety and protective efficacy of three of the highest-yielding CLPs was investigated in a mouse model of brucellosis. All three plant-expressed chimeric CLPs were safe when inoculated into BALB/c mice at specific antigen doses. However, only one chimeric CLP induced protection against the virulent Brucella strain challenge equivalent to the protection induced by the commercial Rev1 vaccine. Here, we have successfully shown the assembly, safety and protective efficacy of plant-expressed chimeric CLPs presenting B. melitensis T cell epitopes. This is the first step in the development of a safe and efficacious subunit vaccine against brucellosis. [ABSTRACT FROM AUTHOR]

Equine brucellosis significantly impacts the health and functionality of horses, leading to complications such as bursitis infection, septic tenosynovitis, septic arthritis, and non-specific lameness resulting from joint infections. In the present study, we used the Rose Bengal plate agglutination test (RBPT), serum agglutination test (SAT), and the 2-mercaptoethanol (2-ME) assays to find equine brucellosis. From June 2018 to September 2022, 876 blood samples were randomly taken from apparently healthy racing horses in certain parts of Iran, such as Kerman, Isfahan, Tehran, Qom, and Kurdistan. DNA extraction was carried out directly on all 63 serum samples identified as seropositive through RBPT. An additional 30 seronegative serum samples were also randomly chosen for study. Bacterial culture was also done on milk, blood, and vaginal swabs taken from seropositive horses.The bacteria that were found in the samples were then put through Bruce-ladder PCR. Our results indicated that 63 (7.1%), 21 (2.3%), and 2 (0.2%) of horses were seropositive using RBPT, SAT, and 2-ME, respectively. Also, none of the 30 DNA-extracted serum samples from seronegative horses tested positive for Brucella DNA, while 44.5% (28/63) of the DNA samples from seropositive horses yielded positive results for Brucella DNA. Out of the seropositive samples, 26 had DNA from Brucella abortus and 2 had DNA from Brucella melitensis. Also, B. melitensis biovar 1 was found in two milk samples from mares in the provinces of Kerman and Isfahan. It was identified using classical biotyping, and molecular assays. It was seen that some of healthy racing horses in some parts of Iran had antibodies against Brucella. The bacteriology and PCR methodologies provide a more comprehensive and reliable means of identifying Brucella spp. infections in horse, especially when the RBPT test came back positive. This underscores the imperative for employing molecular, bacterial, and serological methods in the diagnosis and monitoring of this zoonotic infection. Additionally, this finding suggests that Brucella is being transmitted to equine hosts as a result of its presence in ruminants. The mechanism of transmission may involve interactions between infected ruminants and susceptible equines. This discovery is significant as it underscores the potential cross-species transmission of Brucella and highlights the importance of understanding and managing the spread of the pathogen in both ruminant and equine populations. [ABSTRACT FROM AUTHOR]

Brucellosis poses a significant challenge in the medical field as a systemic infection with a propensity for relapse. This study presented a novel approach to brucellosis treatment, enhancing the efficacy of doxycycline and rifampicin through the use of poly (lactic-co-glycolic) acid coupled with cadmium-telluride quantum dots (Dox-Rif-PLGA@CdTe). The double emulsion solvent evaporation method was employed to prepare Dox-Rif-PLGA@CdTe. The study scrutinized the physicochemical attributes of these nanoparticles. The impact of antibiotic-loaded nanoparticles on Brucella melitensis was evaluated through well diffusion, minimum inhibitory concentration (MIC), and cell culture. The chemical analysis results demonstrated a possibility of chemical reactions occurring among the constituents of nanoparticles. Assessments using the well diffusion and MIC methods indicated that the impact of free drugs and nanoparticles on bacteria was equivalent. However, the drug-loaded nanoparticles significantly decreased the colony-forming units (CFUs) within the cell lines compared to free drugs. In conclusion, the synthesis of nanoparticles adhered to environmentally friendly practices and demonstrated safety. The sustained drug release over 100 h facilitated drug accumulation at the bacterial site, resulting in a heightened therapeutic effect on B. melitensis and improved outcomes in brucellosis treatment. The application of these synthesized nanodrugs exhibited promising therapeutic potential. [ABSTRACT FROM AUTHOR]

Abstract Background Brucellosis, developing complications including arthritis, spondylitis, sacroiliitis, and osteomyelitis, is one of the most common zoonotic diseases in the current world which causes economic losses to the livestock industry and is a great public health concern. Brucella melitensis are the main pathogen of brucellosis epidemics in China, most of which are located in northern China. However, there is limited knowledge about the epidemiology of osteoarthritis-associated brucellosis. This study was aimed to reveal the prevalence of osteoarthritis-associated brucellosis in Inner Mongolia and also to investigate the molecular characteristics of B. melitensis isolates. Methods and results In 2018, the osteoarthritis symptoms of brucellosis in the Brucellosis department of a hospital in Inner Mongolia were investigated. Twenty osteoarthritis-associated B. melitensis strains, isolated from the inpatients in Inner Mongolia during 2013–2017, were subjected to whole genome sequencing. The multilocus sequence type (MLST) and core genome SNP (cgSNP) analysis were conducted to detect molecular epidemiological characteristics. The incidence of brucellosis osteoarthritis symptoms in males (85/120, 70.8%) was significantly higher than that in females (35/120, 29.2%), and the age of patients was concentrated between 41 and 60 years old. In silico analyses indicated ST8 was the prevalent sequence type and the transmission of osteoarthritis-associated B. melitensis among different geographical areas. All strains carry virulence genes, including cgs, lpsA, manCoAg, pgm, pmm, virB4, wbdA and wboA. Conclusion Our study showed the close epidemiologically connection of osteoarthritis-associated B. melitensis strains in northern China. And ST8 was the prevalent sequence type which need our attention.

The control and eradication of brucellosis represents a critical objective for Veterinary and Health Authorities across several countries globally. Efficient surveillance programs play a pivotal role in detecting and managing outbreaks. Epidemiological investigations significantly benefit from standardized and efficient molecular typing techniques and analytical tools, enabling public health laboratories to identify the origin of outbreaks. This study aimed to sequence Brucella spp. strains isolated in Iraq from different ruminant species to verify their molecular epidemiological correlations and, above all, to shed a light on how these Iraqi isolates are positioned in the phylogenetic context of Brucella spp. The 35 isolates under study were from abortion, milk, placenta, and the fetal membranes of sheep, cattle, and buffalo. Genotyping involved various techniques: MLVA-16, Whole Genome Sequencing, MLST, and cgMLST. All the Iraqi isolates from our study clustered in MLVA-16 within the East Mediterranean clade, and all but one grouped together in the same branch of the MST tree. MST analysis showed the minimum distance of one allele between the studied isolates, except for one strain from buffalo, which was positioned farther away from the rest of the isolates. In cgMLST, the majority of strains grouped within a large cluster predominantly comprising genotypes from the Middle East. The application of different control measures in different territories based on molecular epidemiological studies would increase the chances of maximizing public health benefits and minimizing the spread of infection to disease-free or lower prevalence areas. [ABSTRACT FROM AUTHOR]

Background: Spontaneous miscarriage, a leading health concern globally, often occurs due to various factors, including infections. Among these, Coxiella burnetii and Brucella spp. may have adverse effects on pregnancy outcomes. While previous research has established a link between infections and spontaneous miscarriage, our study aimed specifically to investigate the presence of these two pathogens in abortion samples from women who experienced spontaneous miscarriages in Iran. Our study can add to the existing knowledge by focusing on Iran, a region with a high prevalence of C. burnetii and Brucella spp. As a result, it could provide a better understanding and unique insights into the relationship of these pathogens with spontaneous miscarriages in endemic regions. Methods: From March 2021 to March 2022, a total of 728 abortion samples (including placenta and cotyledon) were collected from 409 women who had experienced spontaneous miscarriages in the provinces of Tehran, Fars, and West Azerbaijan in Iran. The specimens included 467 Formalin-Fixed Paraffin-Embedded (FFPE) and 261 fresh frozen samples. After DNA extraction from abortion samples, the quantitative real-time PCR (qPCR) assay targeted a specific fragment of the IS1111 and IS711 elements for molecular identification of C. burnetii and Brucella spp., respectively. Furthermore, the qPCR assay employing specific primers for different species was used to determine the species of Brucella. Results: Among the studied women, 1 out of 409 (0.24%) samples tested positive for Brucella spp., specifically Brucella melitensis. There were no positive specimens for C. burnetii. Conclusions: Our study contributes to understanding the potential involvement of Brucella species in spontaneous infectious abortion within endemic regions. The identification of B. melitensis in this study highlights the need for further research in this area. However, while our results suggest a relatively low or zero identification of these pathogens in our sample population, this does not rule out the possibility of undetected infections. Therefore, it is critical to acknowledge the limitations of the molecular techniques used (qPCR), which may have potential limitations such as sensitivity and specificity. Moreover, because 64.15% of our samples were FFPE, the sensitivity of the qPCR test may be reduced. These raise concerns about the accuracy of the reported prevalence rates and the potential for false positives or negatives. [ABSTRACT FROM AUTHOR]

Background and Aim: Brucella melitensis is responsible for brucellosis, a highly contagious, life-threatening disease that has a high impact in low- and middle-income countries. This study aimed to compare silica nanoparticles (SiO-NPs) loaded with ciprofloxacin with silver nanoparticles (AgNPs) loaded with ciprofloxacin to evaluate the possible replacement of silver by silica to enhance biological activity and reduce cytotoxicity. Materials and Methods: SiO-NPs and AgNPs loaded with ciprofloxacin were characterized using ultraviolet spectroscopy, scanning electron microscopy, and dynamic light scattering microscopy for size demonstration and loading efficiency. Both nanoparticles were treated with B. melitensis Rev 1 to evaluate their biological activity. Nanoparticle toxicity was also evaluated using cytotoxicity and hemolysis assays. Results: SiO-NP was found to have a smaller size (80 nm) and higher loading efficiency with polydispersity index and zeta potential of 0.43 and 30.7 mV, respectively, compared to Ag-NP (180 nm and 0.62 and 28.3 mV, respectively). SiO-NP was potent with a minimum inhibitory concentration of 0.043 µg/mL compared to Ag-NP (0.049 µg/mL), with a lower cytotoxicity and hemolysis activity. Conclusion: SiO-NP, as a drug delivery system for ciprofloxacin, has better antimicrobial activity against B. melitensis with lower cytotoxicity and hemolysis activity. These results can be attributed to the enhanced physical characterization and better loading efficiency when compared to Ag-NP. [ABSTRACT FROM AUTHOR]

Contaguous animal diseases that cause huge losses in livestock production and affect national economies are regulated at international level by the World Organisation for Animal Health (WOAH) through the Terrestrial Animal Health Code. The Code lays down the rules for the listed animal diseases that must be reported to the WOAH and among these important diseases is Brucellosis. This is a contagious bacterial disease caused by several representatives from the Brucella family which are infectious for specific animal species, including the most common livestock animals like cattle, sheep and goats and swine. Moreover, Brucellosis has a high zoonotic potential and could infect humans as well. In order to trace and analyse the distribution of Brucella spp. in Europe over the span of a twenty-years period (2002-2022), we derived data from the ADIS (Animal Disease Information System) managed by the European Commission which showed that five European counties were infected with variations in the prevalence of Brucellosis among the reporting member states. Regarding the identified species, it can be noted that the following were found: the predominant species is Brucella melitensis, followed by Brucella suis, and lastly, Brucella abortus. For the purpose of explainig the measures for disease management and control of Brucellosis we examined the main documents from the relevant secondary legislation of the EU. For the studied period, it can be noted that the most outbreaks of brucellosis were registered on the territory of Italy - 21 outbreaks (Brucella melitensis) from March 2014 in sheep to December 2022 in goats, but in the other years it was also found in sheep and goats. For the same period with Brucella abortus, there were 18 outbreaks from March 2015 in cattle to June 2022 in cattle, but in 2020, they were found in sheep and goats. Brucella suis was not registered. [ABSTRACT FROM AUTHOR]

In this study, the antibacterial effects of postbiotics obtained from Pediococcus acidilactici, Lactiplantibacillus plantarum and Latilacto-bacillus sakei, which were grown in sterile cow's milk and de man rogosa and sharpe (mrs) broth, against some food pathogens (Salmonella spp., Listeria monocytogenes, Escherichia coli o157:h7, and Brucella melitensis) were investigated. It was observed that lactic acid bacteria postbiotics produced in mrs broth formed larger inhibition zones than those developed in cow's milk against pathogenic bacteria. In order to investigate the antimicrobial effect of the postbiotics on chicken breast meat and to compare this effect with lactic acid decontamination, samples contaminated with Salmonella spp. and L. monocytogenes were immersed into the postbiotics of L. plan-tarum and L. sakei, 2.1% lactic acid solution, and distilled water for 10 minutes. Microbial changes in the groups were investigated during the storage at 4°c for 17 days. On the 8th day of storage, it was determined that the number of Salmonella spp. In the groups treated with postbiotics decreased by 0.9 log10 CFU/g compared to the control and distilled water groups. While the number of L. monocytogenes increased during storage in the control and distilled water groups, the postbiotics and 2.1% lactic acid exhibited a bacteriostatic effect on L. monocytogenes during storage period. Compared to the postbiotics, 2.1% lactic acid had higher reduction (1.8 log10 cfU/g) rates against Salmonella spp. (P<0.05), also a significant difference was observed against L. monocytogenes in the first and last days of storage (P<0.05). While the shelf life of chicken breast meat was determined to be 5 days in the control and distilled water groups, postbiotic treatments extended the shelf life of chicken breast meat by an extra 9 days, and 2.1% lactic acid treatment extended an extra 12 days compared to the control and distilled water treatments. [ABSTRACT FROM AUTHOR]

Brucellosis is one of the most prevalent bacterial zoonosis worldwide. Brucella melitensis (B. melitensis), Brucella abortus (B. abortus), Brucella canis (B. canis), Brucella suis (B. suis) are common to cause disease in humans and B. melitensis is the most pathogenic causative agent of brucellosis in humans and animals. Fast, efficient and accurate identification of Brucella reference strains at the strain-level is indispensable for microbiological method quality assurance and downstream applications. B. melitensis 63/9 is recognized as an important reference strain for the microbiological culture collection organizations worldwide, and the identification of B. melitensis strain 63/9 is still lacking. The genomic sequences of B. melitensis 63/9 and nine other Brucella strains were compared. Two specific genes were selected for the multiplex PCR method. Gene BMEA_B0162 with unknown function is the key target to identify B. melitensis 63/9, and gene BMEA_A1238 annotated as TRAP transporter solute receptor is included as a control gene for the Brucella genus. A multiplex PCR was established in this study to differentiate B. melitensis reference strain 63/9 from 39 B. melitensis strains, 13 B. abortus strains, 5 B. suis strains, 6 B. canis strains, 3 E. coli, and 4 Salmonella strains by targeting the BMEA_B0162 and the BMEA_A1238 in the genome. This method allows at least 100 pg of B. melitensis 63/9 genomic DNA to be detected. We established a fast, and a cost-effective method to distinguish B. melitensis 63/9 from other Brucella strains and some non-Brucella bacteria strains with high sensitivity and specificity, making the first report about the identification of Brucella reference strain recognized by World Organization for Animal Health at the strain-level. [ABSTRACT FROM AUTHOR]

Background: Brucellosis impact both animals and humans worldwide. However, using antibiotics for brucellosis remains controversial despite decades of research. Relapse can complicate treatment in this area. Since the mid-1980s, microbiologists, and physicians have studied fluoroquinolones' use for treating human brucellosis. The principal advantages of fluoroquinolones are their intracellular antimicrobial activity, low nephrotoxicity, good pharmacokinetics, and the lack of drug-level monitoring. Fluoroquinolones inhibit disease recurrence. In vitro and clinical data were used to study the prevalence of Brucella melitensis and Brucella abortus fluoroquinolone-resistant isolates. Methods: The PubMed, Scopus, Embase, and Web of Science databases were carefully searched until August 6, 2022, for relevant papers. The number of resistant isolates and sample size were used to estimate the proportion of resistant isolates, fitting a model with random effects, and DerSimonian–Laird estimated heterogeneity. Furthermore, meta-regression and subgroup analyses were used to assess the moderators to identify the sources of heterogeneity. Meta-analysis was performed using R software. Results: Forty-seven studies evaluated fluoroquinolone resistance in Brucella spp. Isolates. Fluoroquinolones have shown high in vitro efficacy against Brucella spp. The resistance rates to ofloxacin, sparfloxacin, fleroxacin, pefloxacin, and lomefloxacin were 2%, 1.6%, and 4.6%, respectively. Conclusion: Clinical in vitro tests demonstrated that fluoroquinolones can eradicate Brucella spp. Owing to first-line medication resistance, recurrence, and toxicity, it is essential to standardize the Brucella antimicrobial susceptibility test method for a more precise screening of resistance status. Fluoroquinolones are less resistant to fluoroquinolone-based treatments in modern clinical practice as alternatives to standard therapy for patients with brucellosis relapse after treatment with another regimen and in patients who have developed toxicity from older agents. [ABSTRACT FROM AUTHOR]

Copyright of Al-Anbar Journal of Veterinary Sciences is the property of Republic of Iraq Ministry of Higher Education & Scientific Research (MOHESR) and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Susceptibility to brucellosis remains prevalent, even in herds vaccinated with conventional vaccines. Efforts are underway to develop an improved brucellosis vaccine, and possibly a universal vaccine, given that Brucella species are highly homologous. To this end, two B. melitensis mutants were developed, znBM-lacZ (znBMZ) and znBM-mCherry (znBM-mC), and were tested for their ability to confer systemic immunity against virulent B. melitensis challenge. To assess the extent of their attenuation, bone-marrow-derived macrophages and human TF-1 myeloid cells were infected with both mutants, and the inability to replicate within these cells was noted. Mice infected with varying doses of znBM-mC cleared the brucellae within 6–10 weeks. To test for efficacy against systemic disease, groups of mice were vaccinated once by the intraperitoneal route with either znBMZ or B. abortus S19 vaccine. Relative to the PBS-dosed mice, znBMZ vaccination greatly reduced splenic brucellae colonization by ~25,000-fold compared to 700-fold for S19-vaccinated mice. Not surprisingly, both znBMZ and S19 strains induced IFN-γ+ CD4+ T cells, yet only znBMZ induced IFN-γ+ CD8+ T cells. While both strains induced CD4+ effector memory T cells (Tems), only znBMZ induced CD8+ Tems. Thus, these results show that the described znBM mutants are safe, able to elicit CD4+ and CD8+ T cell immunity without a boost, and highly effective, rendering them promising vaccine candidates for livestock. [ABSTRACT FROM AUTHOR]

Copyright of Iranian Veterinary Journal is the property of Shahid Chamran University of Ahvaz and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Introduction and aim: Brucellosis is a zoonotic disease. Experimental clinical and laboratory diagnosis is still facing problems in identifying the organism. The present study will diagnose a Brucella infection in camel blood in Qatar using serological assays. Isolation and identification were performed on a camel blood sample. Brucella in bacterial isolates was determined by real-time polymerase chain reaction (RT-PCR) as a gold standard test. Material and methods: A total of 220 samples, 200 random serum samples, and 20 EDTA blood samples were selected among the above-mentioned random samples, and 20 serum samples from camel handlers were collected from Al Shahaniya province, Qatar. The Rose Bengal test (RBT), buffered antigen plate agglutination test (BAPAT), and enzyme linked immunosorbent assay (cELISA) for the monoclonal antibody in serum samples were performed using commercially available kits. For the molecular detection of Brucella, conventional PCR and real-time PCR (GPS kit) were used for the genus-specific insertion sequence IS711. Brucella melitensis (MICROBOSS Hightech GmbH kit) was used to identify subspecies. Results: The results identified by vitek2 compact (30%) showed B. melitensis in 6 samples out of 20 isolates. Both conventional (66.67%) and RT-PCR (83.33%) analyses supported this, demonstrating the presence of Brucella. These tests also showed that Brucella species were present in Rose Bengal 182/200 (91%), BAPAT 182/200 (91%), and cELISA (90%) 180/200 in camel serum. Concusion: To conclude, the prevalence of brucellosis in dromedary camels is higher in this region, and as a matter of urgency, measures should be taken to control the disease. [ABSTRACT FROM AUTHOR]

This study assesses the diagnostic efficacy of a Polymerase Chain Reaction (PCR) assay targeting a 1412 bp segment of the 16S rRNA sequence of Brucella melitensis in suspected cases of brucellosis in goats. The research investigates the assay's performance with both serum and whole blood samples in 42 suspected brucellosis cases. Results indicate a significant difference in sensitivity between serum and whole blood samples, with the PCR assay demonstrating higher sensitivity (90%) in serum compared to whole blood (80%). The specificity of the serum PCR assay is determined to be 100%, emphasizing its precision in identifying negative cases. The study strongly recommends the preferential use of serum samples over whole blood for brucellosis diagnosis in goats using the 16S rRNA-based PCR assay, emphasizing the enhanced sensitivity observed in serum samples. These findings contribute to optimizing diagnostic protocols for brucellosis detection in goat populations, with potential implications for veterinary practice and research endeavors. [ABSTRACT FROM AUTHOR]

ABSTRACT Background Human brucellosis, otherwise known as undulant fever, is one of the most widespread zoonotic diseases in the world. Even though 9%–15% of stillbirths are known to be caused by infectious diseases, the study of the link between human Brucella melitensis and the termination of births in humans is a topic that has received little attention. This study examines if there was an association between infection of undulant fever, an endemic zoonotic disease in the Maltese Islands from 1919 until 1954, and reproductive loss through stillbirths. Methods A univariate descriptive analysis was used to show the temporal trend of undulant fever time, as well as the age and sex distribution. Time series analysis was used to assess the relationship between time (months) and undulant fever cases with stillbirth proportions. Results On the island of Gozo, the majority of undulant fever cases for both males and females occurred in their reproductive period between 15 and 45 years of age. Based on regression analysis, undulant fever had a statistically significant effect on the stillbirth rate for males (t = 2.8986, p = 0.0039). The effect of undulant fever on stillbirths was not significant for females (p = 0.9103). Conclusion This paper highlights the importance of undulant fever as having implications for the health burden in pregnant women and potential fetal loss through stillbirths in the contemporary context.

ObjectiveBrucellosis, a significant zoonotic disease, not only impacts animal health but also profoundly influences the host immune responses through gut microbiome. Our research focuses on whole genome sequencing and comparative genomic analysis of these Brucella strains to understand the mechanisms of their virulence changes that may deepen our comprehension of the host immune dysregulation.MethodsThe Brucella melitensis strain CMCC55210 and its naturally attenuated variant CMCC55210a were used as models. Biochemical identification tests and in vivo experiments in mice verified the characteristics of the strain. To understand the mechanism of attenuation, we then performed de novo sequencing of these two strains.ResultsWe discovered notable genomic differences between the two strains, with a key single nucleotide polymorphism (SNP) mutation in the manB gene potentially altering lipopolysaccharide (LPS) structure and influencing host immunity to the pathogen. This mutation might contribute to the attenuated strain's altered impact on the host's macrophage immune response, overing insights into the mechanisms of immune dysregulation linked to intracellular survival. Furthermore, we explore that manipulating the Type I restriction-modification system in Brucella can significantly impact its genome stability with the DNA damage response, consequently affecting the host's immune system.ConclusionThis study not only contributes to understanding the complex relationship between pathogens, and the immune system but also opens avenues for innovative therapeutic interventions in inflammatory diseases driven by microbial and immune dysregulation.

Co-infection with Brucella melitensis and Coxiella burnetii has been rarely reported. To date, there are only two co-infection case reports from Croatia and China which diagnosed the infections mainly through the use of serological tests. In this report, we present the first case of molecularly confirmed B. melitensis bacteremia and C. burnetii spondylodiscitis co-infection in a goat dairy farmer who presented with lumbosacral spondylodiscitis and bilateral psoas abscesses. From the blood culture, B. melitensis was identified by using 16S rRNA gene sequencing and specific PCR. Lumbar bone tissue was found to be positive for C. burnetii using multiplex real-time PCR and was confirmed with a positive result from conventional PCR which detected the infection through the identification of the IS1111 gene. The patient's condition improved after decompressive laminectomy was performed and administration of antibiotics regimen: intravenous gentamicin, oral rifampicin, and oral doxycycline. From our case, it is important to raise awareness of this underreported co-infection with multiple zoonotic diseases, especially Q fever and brucellosis, which share the same exposure risk. Moreover, we also emphasize the use of advanced molecular techniques to improve the diagnostic efficiency and reduce the use of time-consuming procedures among patients who are continuously exposed to such risk factors in areas with high seroprevalence of these zoonotic diseases.

Jingjing Du,1 Yiqing Tao,2 Jiaxing Yang,1 Jiachang Cai,1 Hongwei Zhou,1 Rong Zhang,1 Yanyan Hu1 1Department of Clinical Laboratory, Second Affiliated Hospital of Zhejiang University, School of Medicine, Hangzhou, People’s Republic of China; 2Department of Orthopedics, Second Affiliated Hospital of Zhejiang University, School of Medicine, Hangzhou, People’s Republic of ChinaCorrespondence: Rong Zhang; Yanyan Hu, Department of Clinical Laboratory, Second Affiliated Hospital of Zhejiang University, School of Medicine, Hangzhou, 310009, People’s Republic of China, Email zhang-rong@zju.edu.cn; yanyanhu@zju.edu.cnAbstract: Brucellosis is a zoonotic disease caused by Brucella spp., with the highest prevalence found in the northern cities of China. In this case report, we present an occurrence of spinal infection caused by B. melitensis in a 67-year-old man residing in a non-endemic area of southern China. The patient initially presented with chest and back pain, which was not accurately diagnosed and treated at a local hospital. Subsequently, due to worsening pain, he was admitted to our hospital. To determine the cause of the infection, we performed CT-guided aspiration biopsy and collected biopsy tissue for metagenomic next-generation sequencing (mNGS) on the second day of hospitalization. Imaging investigations revealed involvement of the thoracic vertebrae, specifically thoracic 4– 7 with the main focus on 5– 6, accompanied by stenosis of the intervertebral space. The mNGS results indicated that the spine infection was caused by B. melitensis. The patient’s history as a shepherd and a positive Rose Bengal plate test (RBPT) further supported the diagnosis of brucella spondylitis. In order to alleviate pain and restore spinal function, the patient underwent posterior internal fixation of the thoracic spine. Treatment was initiated with cefoperazone/sulbactam, followed by doxycycline. Subsequently, the patient was switched to a combination therapy of rifampicin and doxycycline for a duration of six weeks. The patient responded well to treatment, and his condition remained stable. In conclusion, brucellosis is a common disease that can be easily misdiagnosed. This case report highlights the potential value of mNGS in early and rapid diagnosis. We believe that mNGS can serve as an effective tool to improve the diagnosis of spine infections caused by this pathogen.Keywords: spine infection, Brucella melitensis, metagenomic next-generation sequencing, Rose Bengal plate test

Brucellosis is one of the most serious zoonotic diseases in human and farm animals caused by Brucella species. This study aims to: (i) estimate the current prevalence of Brucella among camels in Erbil Governorate; (ii) evaluate the milk ring test as a diagnostic tool for screening of brucellosis in camels; (iii) study the association between months and percentage of positive samples to Brucella. During the period, January - June 2021, a total of 250 raw camel milk samples (130 samples from farms and 120 from sale points) were randomly collected. The brucellosis is diagnosed using the Milk Ring Test (MRT), indirect ELISA, and bacteriological isolation of Brucella species. The prevalence of Brucella antibodies in camel milk samples is 11.6% and 10.4% according to MRT and ELISA, respectively. The overall isolation percentage of Brucella species is 8.4%. The detection rate of isolates in sale points is higher 10.0% than the isolation rate from farm 6.9%. The results also reveal that 4.6% and 5.8% of isolates are Brucella abortus; while, 5.8% and 4.2% are Brucella melitensis from the milk of farm and sale points, respectively. The highest rate of brucellosis according to MRT is observed in February 18.6%, while the lowest rate is documented in May 7.5%. We recommend using MRT for the diagnosis of Brucella spp. in routine screening of brucellosis in milk collection centers, dairy factories, and farm. Customers are also recommended to heat the milk adequately to eliminate this milk-borne pathogen before drinking milk or manufacturing processes.

Abstract Background Brucellosis is a zoonotic disease whose causative agent, Brucella spp., is endemic in many countries of the Mediterranean basin, including Greece. Although the occurrence of brucellosis must be reported to the authorities, it is believed that the disease is under-reported in Greece, and knowledge about the genomic diversity of brucellae is lacking. Methods Thus, 44 Brucella isolates, primarily B. melitensis, collected between 1999 and 2009 from humans and small ruminants in Greece were subjected to whole genome sequencing using short-read technology. The raw reads and assembled genomes were used for in silico genotyping based on single nucleotide substitutions and alleles. Further, specific genomic regions encoding putative virulence genes were screened for characteristic nucleotide changes, which arose in different genotype lineages. Results In silico genotyping revealed that the isolates belonged to three of the known sublineages of the East Mediterranean genotype. In addition, a novel subgenotype was identified that was basal to the other East Mediterranean sublineages, comprising two Greek strains. The majority of the isolates can be assumed to be of endemic origin, as they were clustered with strains from the Western Balkans or Turkey, whereas one strain of human origin could be associated with travel to another endemic region, e.g. Portugal. Further, nucleotide substitutions in the housekeeping gene rpoB and virulence-associated genes were detected, which were characteristic of the different subgenotypes. One of the isolates originating from an aborted bovine foetus was identified as B. abortus vaccine strain RB51. Conclusion The results demonstrate the existence of several distinct persistent Brucella sp. foci in Greece. To detect these and for tracing infection chains, extensive sampling initiatives are required.

Brucellosis is a zoonotic infection that is widely spread across the world. It is becoming more common in Middle Eastern countries such as Qatar, Saudi Arabia, and the Mediterranean region. Despite this, we need to remain vigilant as it is still prevalent in many parts of the world. The most common presentation is musculoskeletal, but it can also present as septic arthritis in the sacroiliac, hip, or knee joints. Brucella melitensis was only found in one extended culture of synovial fluid. Treatment involved a combination of antimicrobial therapy using gentamycin, doxycycline, and rifampin. A high level of suspicion for brucellosis is necessary for any patient coming from an endemic region with non-specific and chronic arthritis to ensure early diagnosis and treatment. In this case, we present a 28-year-old male who was diagnosed with Brucellosis after developing acute septic arthritis.