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Many powerful methods have been employed to elucidate the global transcriptomic, proteomic, or metabolic responses to pathogen-infected host cells. However, the host glycome responses to bacterial infection remain largely unexplored, and hence, our understanding of the molecular mechanisms by which bacterial pathogens manipulate the host glycome to favor infection remains incomplete. Here, we address this gap by performing a systematic analysis of the host glycome during infection by the bacterial pathogen Brucella spp. that cause brucellosis. We discover, surprisingly, that a Brucella effector protein (EP) Rhg1 induces global reprogramming of the host cell N-glycome by interacting with components of the oligosaccharide transferase complex that controls N-linked protein glycosylation, and Rhg1 regulates Brucella replication and tissue colonization in a mouse model of brucellosis, demonstrating that Brucella exploits the EP Rhg1 to reprogram the host N-glycome and promote bacterial intracellular parasitism, thereby providing a paradigm for bacterial control of host cell infection., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Brucella species are Gram-negative intracellular bacterial pathogens that cause the worldwide zoonotic disease brucellosis. Brucella can infect many mammals, including humans and domestic and wild animals. Brucella manipulates various host cellular processes to invade and multiply in professional and non-professional phagocytic cells. However, the host targets and their modulation by Brucella to facilitate the infection process remain obscure. Here, we report that the host ubiquitin-specific protease, USP8, negatively regulates the invasion of Brucella into macrophages through the plasma membrane receptor, CXCR4. Upon silencing or chemical inhibition of USP8, the membrane localization of the CXCR4 receptor was enriched, which augmented the invasion of Brucella into macrophages. Activation of USP8 through chemical inhibition of 14-3-3 protein affected the invasion of Brucella into macrophages. Brucella suppressed the expression of Usp8 at its early stage of infection in the infected macrophages. Furthermore, we found that only live Brucella could negatively regulate the expression of Usp8 , suggesting the role of secreted effector protein of Brucella in modulating the gene expression. Subsequent studies revealed that the Brucella effector protein, TIR-domain containing protein from Brucella , TcpB, plays a significant role in downregulating the expression of Usp8 by targeting the cyclic-AMP response element-binding protein pathway. Treatment of mice with USP8 inhibitor resulted in enhanced survival of B. melitensis, whereas mice treated with CXCR4 or 14-3-3 antagonists showed a diminished bacterial load. Our experimental data demonstrate a novel role of Usp8 in the host defense against microbial intrusion. The present study provides insights into the microbial subversion of host defenses, and this information may ultimately help to develop novel therapeutic interventions for infectious diseases., Competing Interests: The authors declare no conflict of interest.

Aim: In this study, we aimed to discuss the clinical features, laboratory findings, treatment and outcome of seven cases of neurobrucellosis from a tertiary care center and review the available global literature., Materials and Methods: The diagnosis of neurobrucellosis was established using the following criteria in our setting: (1) signs and symptoms of neurological infection with examination of cerebrospinal fluid (CSF) revealing signs of meningitis, (2) isolation of Brucella spp. from blood and/or CSF and/or antibody titer ≥1:160 in serum using standard agglutination test (SAT) and/or the presence of anti-Brucella antibodies in CSF and/or detection of Brucella spp.-specific DNA from CSF using PCR. A literature search was performed to review previous cases of neurobrucellosis published worldwide during the last 30 years., Results: The proportion of neurobrucellosis was 2.8% in our setting. Fever with headache and altered sensorium were the major presenting complaints. Brucella melitensis was isolated from blood culture in 6 patients. From the literature search, a total of 221 cases of neurobrucellosis were reviewed and analyzed. Meningitis (32.6%), loss of hearing (25.8%) and encephalitis (14.9%) were the most common clinical features. Involvement of cranial nerves, polyradiculopathy and paraplegia were the major complications found in patients with neurobrucellosis., Conclusions: Neurobrucellosis should always be considered in the differential diagnosis of befitting neurological, rheumatological, and neuropsychiatric presentations in endemic regions for brucellosis. To prevent morbidity and mortality associated with neurobrucellosis, a multimodal diagnostic approach is essential for early and accurate diagnosis and effective treatment.

Background: Abortions cause tremendous economic losses in food-producing animals and may lead to food insecurity., Objectives: This study aimed to characterize Brucella spp. and other abortigenic pathogens from aborted tissues of cattle., Methods: For cattle, aborted tissues (n = 19) were cultured, and Brucella spp. were detected using the genus-specific 16S-23S ribosomal DNA interspacer region (ITS) assay and speciated using Brucella abortus, Brucella melitensis, Brucella ovis, and Brucella suis (AMOS) and Bruce-ladder PCR assays. Brucella negative samples were screened using the eight abortigenic pathogens PCR panel. Samples from an abortion outbreak that occurred within a goat tribe were included in this investigation. Sera of females (n = 8) and males (n = 2) were analyzed using the Rose Bengal Test (RBT) and indirect enzyme-linked immunosorbent assay (i-ELISA), while vaginal swabs (n = 3) and aborted tissues (n = 1) were cultured and characterized., Results: The ITS-PCR detected Brucella DNA in cultures from two aborted tissues of cattle (10.5%, [2/19]), which were identified as B. melitensis (n = 1), and B. abortus (n = 1) using AMOS and Bruce-ladder PCR assays. Campylobacter fetus (n = 7) and Leptospira spp. (n = 4) including co-infections (n = 2) of C. fetus and Leptospira spp. were identified from the Brucella negative samples of cattle. Goats (100.0%, 10/10) were brucellosis seropositive on RBT and i-ELISA. Mixed infections caused by B. melitensis and B. abortus were isolated from the vaginal swabs (n = 3) and aborted tissues (n = 1)., Discussion and Conclusions: This is the first identification of abortion-associated pathogens in aborted cattle indicating the enormous financial losses and a threat to public health. It is therefore essential to include these identified pathogens in the surveillance scheme of veterinary and human services., (© 2022 The Authors. Veterinary Medicine and Science published by John Wiley & Sons Ltd.)

Human brucellosis represents a serious public health concern in Hulun Buir and requires a comprehensive epidemiologic analysis to define adapted control measures. The present study describes the case numbers, constituent ratios and incidence rate of human brucellosis. Conventional biotyping, that is abortus, melitensis, ovis and suis (AMOS)-PCR and multi-locus variable-number tandem repeat analysis (MLVA) were used to characterize the Brucella strains. Between 2003 and 2018, a total of 23,897 human brucellosis cases were reported, with an incidence rate of 56.03/100,000, which is 20 times higher than the country's average incidence. This incidence rate increased year after year, culminating in 2005 and decreased between 2011 and 2018. Because Hulun Buir relies on a nomadic livestock rearing system, brucellosis spreads easily among different animal species and humans. In Xin Barag Left Banner and Xin Barag Right Banner, the incidence rates were, respectively, 226.54/100,000 and 199.10/100,000, exceeding those observed in other areas. Most of the cases occurred in the 25- to 45-year-old group, accounting for 65.74% of the cases (15,709/23,897), and among farmers, accounting for 66.71% (15,942/23,897). The male to female incidence ratio was 2.67:1. The higher incidence in younger people and the large gender ratio reflected the unique traditional production and lifestyle of nomads. Most reported cases were observed from April to June, indicating that more than 40% of the cases were related to the delivery of domestic livestock. The biotyping showed that the 44 isolated strains were all B. melitensis, including 12 Brucella melitensis biovar (bv) 1 and 32 B. melitensis bv. 3. The strains displayed a genetic similarity of 80%-100%. Our hypothesis is that human brucellosis outbreak in this region may be originating from a limited source of infection, so further investigation is necessary. The epidemic situation of human brucellosis in Hulun Buir is extremely serious, strengthened surveillance and control in animals' brucellosis should be priority., (© 2021 Wiley-VCH GmbH.)

Neurobrucellosis is a serious central nervous system (CNS) inflammatory disorder caused by Brucella, and outer membrane protein-31 (Omp31) plays an important role in Brucella infection. This study aims to determine whether Omp31 can induce autophagy in BV-2 microglia. Another goal of the study is to further examine the effect of autophagy on the nuclear transcription factor κB (NF-κB) p65 signaling pathway. We observed that Omp31 stimulated autophagy by increasing microtubule-associated protein 1 light chain 3B (LC3B-II) levels and inducing autophagosome formation at 6 h and 12 h. Concomitantly, Omp31 induced tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) expression in a time-dependent manner but reduced the expression of TNF-α at 6 h. We utilized Omp31 with or without rapamycin or 3-methyladenine (3-MA) to treat BV-2 microglia, and it demonstrated further that Omp31 induced autophagy by promoting LC3B-II, Beclin-1 proteins expression and inhibiting the p62 protein levels. Furthermore, we explored the effects of autophagy on the NF-κB p65 pathway through western blot analysis, RT-qPCR assay, enzyme-linked immunosorbent assay (ELISA) and immunofluorescence. The data suggest that Omp31 as well as rapamycin, the autophagy inducer, can decrease TNF-α levels through the inhibition of the NF-κB p65 signaling pathway. Taken together, Omp31 can function as a catalyst in both autophagy induction and NF-κB p65 signal inhibition. Furthermore, Omp31-induced autophagy may inhibit the expression of TNF-α by negatively regulating NF-κB p65 signaling pathway., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: A novel Brucella strain closely related to Brucella (B.) melitensis biovar (bv) 3 was found in Croatian cattle during testing within a brucellosis eradication programme., Case Presentation: Standardised serological, brucellin skin test, bacteriological and molecular diagnostic screening for Brucella infection led to positive detection in one dairy cattle herd. Three isolates from that herd were identified to species level using the Bruce ladder method. Initially, two strains were typed as B. melitensis and one as B. abortus, but multiplex PCR based on IS711 and the Suis ladder showed that all of them to belong to B. melitensis, and the combination of whole-genome and multi-locus sequencing as well as Multi-Locus Variable numbers of tandem repeats Analysis (MLVA) highlighted a strong proximity within the phylogenetic branch of B. melitensis strains previously isolated from Croatia, Albania, Kosovo and Bosnia and Herzegovina. Two isolates were determined to be B. melitensis bv. 3, while the third showed a unique phylogenetic profile, growth profile on dyes and bacteriophage typing results. This isolate contained the 609-bp omp31 sequence, but not the 723-bp omp31 sequence present in the two isolates of B. melitensis bv. 3., Conclusions: Identification of a novel Brucella variant in this geographic region is predictable given the historic endemicity of brucellosis. The emergence of a new variant may reflect a combination of high prevalence among domestic ruminants and humans as well as weak eradication strategies. The zoonotic potential, reservoirs and transmission pathways of this and other Brucella variants should be explored.

Brucellosis is a significant zoonotic infection in Iran impacting both humans and animal health status. A number of reasons such as few clinical signs complicate the diagnosis of this infection in Camelidae. Despite the ubiquitous use of serological tests for the first screening of brucellosis in camel, this approach showed several restrictions because of the intracellular properties of this organism as well as decline antibody titers in chronic stage. This study aimed at identifying the presence of Brucella spp. in blood and lymph node samples collected from slaughtered male camels of Sistan-Baluchistan province by serology, culture and polymerase chain reaction (PCR) assay. For this purpose, 2854 blood camel samples were sampled and analyzed for Brucella detection by serological screening tests. The molecular detection of IS711 gene and Bruce-ladder assay as well as culture were performed using the lymph nodes of all seropositive camel (n = 10) and 30 seronegative samples. Results showed that 0.35% (10/2854), 0.24% (7/2854) and 0.21% (6/2854) of blood samples were positive by RBPT, SAT and 2ME, respectively. However, 0% (0/10) and 70% (7/10) of lymph node samples collected from seropositive camels were positive for Brucella infection by culture and PCR, respectively. Furthermore, 6.6% (2/30) of seronegative lymph node specimens showed the presences of Brucella by PCR and culture assay. The results of the present study indicated the low seroprevalence of Brucella infection in male camels of the Sistan-Baluchistan province and highlighted the complementary role of PCR techniques for a better screening of Brucella infection among seronegative camels. Moreover, the potential shedding of Brucella within undiagnosed camel milk and secretions is a serious problem which may result in further spread and maintenance of Brucella infection among both human and livestock. Thus, for brucellosis detection and control, our results suggested that a first PCR screening supported by a bacteriological isolation on positive samples should be performed along with the serological test in endemic countries to identify the source and prevent the uncontrolled spread of the disease among camels., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background: Brucellosis is one of the most common diseases that afflicts both humans and animals. Bacteria react to stress conditions using different mechanisms one of which is Toxin-Antitoxin (TA) systems. It is believed that the Toxin-Antitoxin (TA) systems have a key role in the chronicity of the disease. This study investigated the expression of TA system genes under acid and antibiotic stresses in Brucella spp., Methods: Fifty Brucella isolates (17 isolated from animals and 31 isolated from human specimens, and two standard strains) were analyzed using PCR (using two pairs of primers). Then, to determine the effects of sub-MIC of gentamicin on bacterial survival and growth, colony forming unit was quantitated and turbidity was assessed following the treatment of Brucella spp, with ½ MIC of gentamicin at different time intervals. Furthermore, the colony forming unit of Brucella spp, was assessed under acid stress (pH = 5.5) compared to the control (pH = 7.6). Moreover, the expression of TA system genes in Brucella spp, was evaluated 1 h after treatment using qRT-PCR method., Results: A total of 50 isolates, including 41 (82%) Brucella melitensis and 7 (14%) Brucella abortus with two standard strains Brucella melitensis (16 M) and Brucella abortus (B19) were investigated. Our results revealed the reduced growth of Brucella spp. in the presence of sub-MIC of gentamicin compared to the control. Furthermore, according to the results of qRT-PCR assay, gentamicin could increase the expression of TA system genes. Also, results of qRT-PCR showed that under acid stress, the expression of TA system gene COGT/COGAT decreased compared to the control., Conclusion: Although the exact role of the TA systems in response to stress is still unclear, our study provided information on the effect of the type II TA systems under the acid and antibiotic stress conditions. However, further studies are still required., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Brucellosis' surveillance and control programs require robust laboratory techniques that can reliably identify and biotype Brucella strains and discriminate between vaccine and field infection. In the recent years, Matrix Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) has revolutionized the routine identification of several microorganisms in clinical microbiology laboratories. Nevertheless, its application on Brucella spp. identification is limited since there are no reference spectra in the commercial databases, due to the microorganism's potential bioterrorist use. In this study, a custom MALDI-TOF MS reference library was constructed and its performance on identification at species level was evaluated using 75 Brucella spp. isolates. Furthermore, distinct peak biomarkers were detected for biovar assignment and discrimination from vaccine strain Rev.1. Analysis of mass peak profiles allowed Brucella accurate identification at genus and species level (100%) with no misidentifications. Despite the high intrageneric similarity, MALDI-TOF MS database succeeded in classifying at biovar level, 47 out of 62 B. melitensis bv. 3 isolates (75.81%), whereas all B. melitensis strains, except for one, were correctly discriminated from vaccine strain Rev.1. MALDI-TOF MS appeared to be a rapid, cost-effective and reliable method for the routine identification of brucellae which reduces time consumption in pathogen identification and could replace in the near future the current conventional and molecular techniques. Its ability to differentiate vaccine from field infection could facilitate brucellosis' monitoring systems contributing in the effective control of the disease., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

In the present study, a total of 1102304 serum samples were collected to detected human brucellosis between the years 2012 and 2016 in Inner Mongolia. Overall, an average of 3.79% anti-Brucella positive in Inner Mongolia was presented but the range of positive rates were among 0.90 to 7.07% in 12 regions. Seroprevalence of human brucellosis increased gradually from 2012 to 2016. However, the incidence rate of human brucellosis showed a declining trend. One hundred and seven Brucella strains were isolated and identified as B. melitensis species, and B. melitensis biovar 3 was the predominant biovar. MLVA-11 genotypes 116 was predominant and had crucial epidemiology to the human population. All 107 strains tested were sorted into 75 MLVA-16 genotypes, with 54 single genotypes representing unique isolates. This result revealed that these Brucellosis cases had epidemiologically unrelated and sporadic characteristics. The remaining 21 shared genotypes among two to four strains, confirming the occurrence of cross-infection and multiple outbreaks. Extensive genotype-events were observed between strains from this study and Kazakhstan, Mongolia, and Turkey, these countries were key members of the grassland silk road. Long-time trade in small ruminants (sheep) in these countries has possibly promoted the spread of Brucella spp . in these regions.

Background: Brucellosis is an infectious and contagious zoonotic bacterial disease of both humans and animals. In developing countries where brucellosis is endemic, baseline data on the prevalence of brucellosis, using abattoir facilities, is important., Objectives: The aim of this study was to determine the seroprevalence of antibodies against Brucella in slaughter cattle at Gauteng province, South Africa and to characterize isolates of Brucella spp., Methods: In this cross-sectional study, un-clotted blood samples with corresponding organ tissue samples were collected from slaughtered cattle. Serological [Rose Bengal test (RBT), complement fixation test (CFT) and indirect ELISA (iELISA)], molecular (PCR) and bacteriological methods were used to detect Brucella antibodies and Brucella spp. from 200 slaughtered cattle in 14 abattoirs., Results: The RBT revealed a seroprevalence of brucellosis as 11.0% (22 of 200) and iELISA confirmed 5.5% (11 of 200). The estimated seroprevalence from RBT and iELISA was 5.5% while RBT and CFT was 2.0% (4 of 200). Brucella melitensis (n = 6) and B. abortus (n = 5) were isolated from 11 cattle tissues (5.5%) as confirmed to species level with AMOS PCR and differentiated from vaccine strains with Bruce-ladder PCR. Seven of the 11 isolates originated from seropositive cattle of which five were biotyped as B. abortus bv 1 (n = 2) and B. melitensis bv 2 (n = 1) and B. melitensis bv 3 (n = 2)., Conclusions: This is the first documentation of B. melitensis in cattle in South Africa. The zoonotic risk of brucellosis posed by Brucella-infected slaughter cattle to abattoir workers and consumers of improperly cooked beef cannot be ignored., (© 2019 The Authors. Veterinary Medicine and Science Published by John Wiley & Sons Ltd.)

Background: The objective of this study was to investigate Brucella infection in farm animals in Saham, Oman, with reference to a survey carried out by the Ministry of Agriculture & Fisheries (MAF) for Brucellosis during the period of May to July 2016 in Saham, following an outbreak of human brucellosis. We wanted to apply different serological, bacteriological and molecular tests in a time frame (phase 1, 2 & 3) with reference to the pivotal time of a human brucellosis outbreak to ascertain the status of the disease in Saham area where the MAF survey was conducted. Blood samples were collected from farm animals and sera were screened in parallel for Brucella antibodies using different serological tests., Results: Using the RBT test, phase 1 sera showed seropositivity in sheep at 2.6%, (95% CI: 0.5-13.5%), in camel (5.9%, 1.1-27.0%), but not in sera from goats and cattle (0%). Using I-ELISA, seropositivity in goat was 3.1% (0.6-15.8%), with no positive sheep and cattle. Using c-ELISA for camel we found a seropositivity of 5.9% (1.1-27.0%). Furthermore, CFT seropositivity in goats was 21.9% (CI: 11.3-38.9), cattle and sheep sera were negative and camel was 5.9% (1.1-27.0%). In phase 2, the seropositivity in goats was 1.9% (1.4-2.6%), sheep 4.5% (3.5-5.8%), cattle 1.1%, (0.5-2.3%) and camels 18.2% (5.1-47.7%), Phase 3 sera were collected 6 months after the human brucellosis outbreak. With RBT, the seropositivity in goats was 3% (1.0-8.5%), sheep 2% (0.6-7.1%) cattle 1% (0.2-5.5%). With I-ELISA, goats & camels were negative, sheep were 3% (1.0-8.5%) and cattle 1% (0.2-5.5%). Moreover, B. melitensis was isolated from a bronchial lymph node of the RBT and I-ELISA seropositive cow and confirmed by Multiplex PCR and biochemical tests., Conclusion: Using a retrospective study analysis of animal sera and following up after a human brucellosis outbreak, the present study showed a slight decrease in seropositivity of infected animals after the MAF implemented test and slaughter policy. The most interesting finding in this study was the isolation, identification and molecular characterization of Brucella melitensis in a cow (spillover), which is not a preferential host for Brucella melitensis.

Brucellosis is a zoonosis caused by bacteria of the Brucella genus. In ruminants, brucellosis causes abortion, followed by chronic infection and secretion of bacteria in milk. In humans, it usually presents as flu-like symptoms, with serious complications if untreated. Epidemiological studies have only recently established that brucellosis can also cause pregnancy complications in women, but the pathogenic mechanisms are unknown. Pioneering studies in ruminants showed that Brucella infect trophoblasts and then colonise the placenta where they grow to high density. A recent study showed that the main zoonotic Brucella species can infect human cytotrophoblasts (CTB) and extravillous trophoblasts (EVT). In this work, we show that Brucella papionis (associated with stillbirth in primates) also infects human trophoblasts. However, it replicates actively in CTB, whereas its replication is very restricted within EVT. We also observed alteration of several trophoblastic functions upon infection by B. papionis or Brucella melitensis (the most prevalent species in human brucellosis). Infection altered the production of hormones, the ability of CTB to form syncytiotrophoblasts, and the invasion capacity of EVT. We also found that infection can spread between different types of trophoblasts. These findings constitute a new step in understanding how Brucella infection causes adverse pregnancy outcomes., (© 2019 John Wiley & Sons Ltd.)

Bacteria in the genus Brucella are the cause of brucellosis in humans and many domestic and wild animals. A rapid and culture-free detection assay to detect Brucella in clinical samples would be highly valuable. Nanomaterial optical fiber biosensors (NOFS) are capable of recognizing DNA hybridization events or other analyte interactions with high specificity and sensitivity. Therefore, a NOFS assay was developed to detect Brucella DNA from cultures and in tissue samples from infected mice. An ionic self-assembled multilayer (ISAM) film was coupled to a long-period grating optical fiber, and a nucleotide probe complementary to the Brucella IS 711 region and modified with biotin was bound to the ISAM by covalent conjugation. When the ISAM/probe duplex was exposed to lysate containing ≥100 killed cells of Brucella , or liver or spleen tissue extracts from Brucella- infected mice, substantial attenuation of light transmission occurred, whereas exposure of the complexed fiber to non- Brucella gram-negative bacteria or control tissue samples resulted in negligible attenuation of light transmission. Oligonucleotide probes specific for B. abortus , B. melitensis , and B. suis could also be used to detect and differentiate these three nomenspecies. In summary, the NOFS biosensor assay detected three nomenspecies of Brucella without the use of polymerase chain reaction within 30 min and could specifically detect low numbers of this bacterium in clinical samples.

In the present study, protective efficacy conferred by a cocktail protein consisted of Brucella L7/L12 ribosomal, truncated outer membrane protein 31 (TOmp31) and SOmp2b recombinant proteins in CpG ODN 1826+ Montanide ISA 70VG or Poly (I:C) adjuvants was evaluated and compared in BALB/c mice. Immunization of mice with both vaccine regimens elicited strong specific IgG responses (higher IgG2a titers over IgG1 titers), provided T helper1 (Th1) oriented immune responses and conferred protection levels compatible to the live vaccines against Brucella challenge. Vaccination of BALB/c mice with the cocktail protein in CpG ODN 1826+ Montanide ISA 70 V G adjuvants induced higher levels of antibody, IFN-γ/IL-2 and conferred more protection levels against B. melitenisis and B. abortus challenge than did the cocktail protein in Poly (I:C) formulation. In conclusion, both vaccine regimens are capable of stimulating specific Th1- biased immune responses and conferring cross protection against B. melitensis and B. abortus infections. Therefore, they could be introduced as new potential candidates for the development of subunit vaccines against Brucella infection., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Allergic asthma is a chronic Th2 inflammatory disease of the lower airways affecting a growing number of people worldwide. The impact of infections and microbiota composition on allergic asthma has been investigated frequently. Until now, however, there have been few attempts to investigate the impact of asthma on the control of infectious microorganisms and the underlying mechanisms. In this work, we characterize the consequences of allergic asthma on intranasal (i.n.) infection by Brucella bacteria in mice. We observed that i.n. sensitization with extracts of the house dust mite Dermatophagoides farinae or the mold Alternaria alternata ( Alt ) significantly increased the number of Brucella melitensis, Brucella suis , and Brucella abortus in the lungs of infected mice. Microscopic analysis showed dense aggregates of infected cells composed mainly of alveolar macrophages (CD11c + F4/80 + MHCII + ) surrounded by neutrophils (Ly-6G + ). Asthma-induced Brucella susceptibility appears to be dependent on CD4 + T cells, the IL-4/STAT6 signaling pathway and IL-10, and is maintained in IL-12- and IFN-γR-deficient mice. The effects of the Alt sensitization protocol were also tested on Streptococcus pneumoniae and Mycobacterium tuberculosis pulmonary infections. Surprisingly, we observed that Alt sensitization strongly increases the survival of S. pneumoniae infected mice by a T cell and STAT6 independent signaling pathway. In contrast, the course of M. tuberculosis infection is not affected in the lungs of sensitized mice. Our work demonstrates that the impact of the same allergic sensitization protocol can be neutral, negative, or positive with regard to the resistance of mice to bacterial infection, depending on the bacterial species.

Brucella spp. are intracellular pathogenic bacteria remarkable in their ability to escape immune surveillance and therefore inflict a state of chronic disease within the host. To enable further immune response studies, Brucella was engineered to express the well-characterized chicken ovalbumin (OVA). Surprisingly, we found that CD8 T cells bearing T cell receptors (TCR) nominally specific for the OVA peptide SIINFEKL (OT-1) reacted to parental Brucella -infected targets as well as OVA-expressing Brucella variants in cytotoxicity assays. Furthermore, splenocytes from Brucella -immunized mice produced gamma interferon (IFN-γ) and exhibited cytotoxicity in response to SIINFEKL-pulsed target cells.To determine if the SIINFEKL-reactive OT-1 TCR could be cross-reacting to Brucella peptides, we searched the Brucella proteome using an algorithm to generate a list of near-neighbor nonamer peptides that would bind to H2K b Selecting five Brucella peptide candidates, along with controls, we verified that several of these peptides mimicked SIINFEKL, resulting in T cell activation through the "SIINFEKL-specific" TCR. Activation was dependent on peptide concentration as well as sequence. Our results underscore the complexity and ubiquity of cross-reactivity in T cell recognition. This cross-reactivity may enable microbes such as Brucella to escape immune surveillance by presenting peptides similar to those of the host and may also lead to the activation of autoreactive T cells., (Copyright © 2018 Harms et al.)

Objective: The aim of this study was to evaluate an ongoing outbreak of brucellosis in southern region of Bosnia and Herzegovina (BIH) on the epidemiological, clinical and molecular level., Patients and Methods: This study included 19 patients affected by brucellosis between 2015 and 2017, in Trebiševo (BIH). Out of 19 patients, 16 were admitted to and treated at the Department of Infectious diseases of the University Clinical Hospital Mostar, while three patients were treated in ambulatory care setting. Epidemiological, clinical and microbiological parameters were investigated. The Rose Bengal test (RBT) positive sera were serologically confirmed by complement fixation test (CFT). We also analyzed blood cultures, and isolates were additionally serotyped. Molecular analyses were performed with Bruce-ladder multiplex polymerase chain reaction (PCR) and multiple locus variable number of tandem repeat analysis of 16 loci (MLVA-16) assay., Results: Fifteen out of 19 patients had been professionally exposed to the bacterium, while four patients acquired brucellosis without prior contact with infected animals. In seven out of eight (87.5%) patients with localized form of brucellosis, we detected significantly higher values of C-reactive protein (CRP) or erythrocyte sedimentation rate (P<0.001). B. melitensis was isolated from 13/16 (81.3%) blood culture samples, and additionally serotyped as biovar 3. Using MLVA16 assay, 11 isolates were genotyped. We observed complete genotype matches among 8/11 B. melitensis isolates, while 3/11 isolates differed in Bruce04 locus., Conclusion: Overall, our study confirms the usefulness of MLVA-16 method in the epidemiological and molecular research of brucellosis during epidemic that, most likely, originated from the same source., (Copyright © 2018 by Academy of Sciences and Arts of Bosnia and Herzegovina.)

Brucellosis is the most common bacterial zoonotic disease worldwide and no vaccine is available for the prevention of human brucellosis. In humans, brucellosis is mostly caused by Brucella melitensis and Brucella abortus. The Outer membrane protein 31 (Omp31) and L7/L12 are immunodominant and protective antigens conserved in human Brucella pathogens. In the present study, we evaluated the humoral and cellular immune responses induced by a fusion protein designed based on the Truncated form of Omp31 (TOmp31) and L7-L12 antigens. Vaccination of BALB/c mice with the recombinant fusion protein (rL7/L12-TOmp31) provided the significant protection level against B. melitensis and B. abortus challenge. Moreover, rL7/L12-TOmp31 elicited a strong specific IgG response (higher IgG2a titers) and significant IFN-γ/IL2 production and T-cell proliferation was also observed. The T helper1 (Th1) oriented response persisted for 12 weeks after the first immunization. The rL7/L12-TOmp31 could be a new potential antigen candidate for the development of a subunit vaccine against B. melitensis and B. abortus., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The author describes a study, carried out in the Province of Córdoba, Spain, to test the efficacy of a live vaccine prepared from the Rev I strain of Brucella melitensis against caprine brucellosis and to determine the extent of natural infection in goats and humans in the Province.It was found that the vaccine significantly increased the resistance of the goats to infection without inducing a carrier state of the vaccine strain and that the immunity persisted for at least 15 months-the period of test.Serum agglutination tests, milk ring tests, and milk culture tests on goats showed that approximately 16-29% of the individual animals examined would be considered infective on the basis of one or other of the tests. Of the 118 herds tested, 111 were discovered to be harbouring infected animals.Serum agglutination tests on humans revealed that 25 of the 880 people tested (2.8%) had titres of 160 International Units (IU) or above and that, on the basis of a diagnostic titre of 80 IU or above, 7% of the population would be regarded as showing evidence of a past or present infection.

The use of gel diffusion precipitin plates in the study of brucella is described.Cultures of Brucella melitensis yielded a diffusible precipitin antigen which produced lines with sera of rabbits, goats, and cattle that had been infected with either Br. abortus or Br. melitensis. The precipitin lines varied in number from one to three.A diffusible precipitin antigen could not be prepared from Br. abortus and Br. suis cultures by the methods which were effective in producing such an antigen from Br. melitensis.No precipitin reactions occurred with sera from cattle in brucella-free herds which had been inoculated with Strain 19 vaccine, although some of these sera were positive to the agglutination test.

Experiments have been carried out on monkeys, goats and guinea-pigs to define as closely as possible the degree of attenuation of the Rev I strain of B. melitensis. Earlier studies had conclusively demonstrated the effectiveness of the strain as an immunizing agent of the three animal species and had suggested that the degree of attenuation was such as to warrant limited study in humans. Results of such a limited study suggested more intensive measurement of the virulence of the strain in other stocks of animals as well as in individual animals rendered increasingly susceptible. A comparison of Rev I with B. abortus, strain 19-BA, and with a fully virulent strain of B. melitensis in guinea-pigs confirmed that the BA strain was more attenuated than Rev I. Cynomolgus monkeys were effectively immunized by Rev I and showed temporary signs of generalized infection. Human isolates of the Rev I strain were striking in the temporary infectivity possessed by rough colony types.

Glenchur, Harry (Minneapolis Veterans Administration Hospital, and University of Minnesota School of Medicine, Minneapolis), Ulysses S. Seal, Horace H. Zinneman, and Wendell H. Hall. Antigenicity of some Brucella melitensis cell fractions. J. Bacteriol. 85:363-368. 1963.-The components of a Brucella melitensis strain were obtained by differential centrifugation and diethylaminoethyl-cellulose ion-exchange chromatography. Rabbit antisera to these Brucella fractions were tested for agglutinins, precipitins, and blocking phenomena. Dermal hypersensitivity was also determined in rabbits. Insoluble (cell-wall) fractions provoked agglutinins, blocking antibodies and skin sensitivity, and precipitins for soluble antigens. Most soluble antigen fractions were capable of producing all the secondary immunological phenomena.

Continuing previous studies, the authors investigated the immunogenicity of low doses of Brucella melitensis strain Rev I in Cynomolgus monkeys challenged by the respiratory route and compared the efficacy of Rev I vaccine with that of vaccine from Br. abortus strain 19-BA.Challenge with the virulent Br. melitensis strain 6015 indicated that 19-BA vaccine was considerably more effective when administered intracutaneously than subcutaneously, whereas with Rev I vaccine the difference was slight, but Rev I conferred immunity in much lower doses. It was found that a dose as low as 260 cells of Rev I led to a high degree of immunity, although there was a very pronounced delay in the bacteriological response. The authors conclude that the results obtained justify trials of the safety of Rev I vaccine for man in the low dosage effective for cynomolgi.

A non-specific element has been demonstrated in the resistance of monocytes derived from immunized rabbits. Vaccination by BCG or by an effective anti-brucellosis reagent induces protection in either case against both Mycobacterium tuberculosis and Brucella melitensis when studied by the monocyte culture method. The activity of the antiserum required to demonstrate the resistance of the monocyte is not affected when the agglutinating action of the anti-Brucella rabbit serum is removed by absorption. The ability of the monocytes from specifically immunized rabbits to retard the growth of virulent Brucella was demonstrated, not as an all-or-none phenomenon, but in the light of the unrestricted bacterial multiplication which occurs in monocytes from normal animals.