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1.
Rev. cuba. med. trop ; 75(1)abr. 2023.
Article in Spanish | LILACS, CUMED | ID: biblio-1550874

ABSTRACT

La endocarditis bacteriana secundaria a la infección por Brucella spp., en este caso B. melitensis, como complicación de la brucelosis humana tiene una incidencia baja y, aunque es la presentación clínica con la que se asocia más frecuentemente la mortalidad, no todos los casos son letales, si son tratados oportunamente. Se describe el caso clínico de una endocarditis bacteriana por B. melitensis, diagnosticada en un adulto por el aislamiento del microorganismo en el hemocultivo. Paciente del sexo masculino, de 40 años, con antecedentes de realizar partos en el ganado bovino y consumir leche no pasteurizada. Acudió al médico por presentar durante siete días de evolución de las siguientes manifestaciones clínicas: fiebre, mialgias, artralgias, tos seca y pérdida de peso (15 kg). El hemograma informa: leucopenia, trombocitopenia y anemia; mientras que en un ecocardiograma transesofágico se observó vegetación en la válvula aórtica con una disminución de la función sistólica y en el hemocultivo se aisló B. melitensis. Debido a estos antecedentes, se inició el tratamiento antibacteriano con rifampicina, doxiciclina y gentamicina. El paciente se recuperó y tuvo una evolución clínica satisfactoria. La brucelosis es una enfermedad infrecuente. Debe considerarse en toda persona con fiebre de foco desconocido que resida en zonas endémicas o esté expuesto al cuidado de animales de granja. En esta enfermedad se impone un diagnóstico y tratamiento preciso, por ser una complicación con alta letalidad.


Bacterial endocarditis, secondary to Brucella spp. infection, in this case by B. melitensis, as a complication of human brucellosis has a low incidence. Although it is the clinical presentation most frequently associated with mortality, not all cases are lethal if timely treatment is provided. We describe a clinical case of bacterial endocarditis due to B. melitensis in a 40-year-old male patient with a history of conducting cattle deliveries and consuming unpasteurized milk, diagnosed after isolating the microorganism in blood culture. He presented with the following clinical manifestations after seven days of evolution: fever, myalgias, arthralgias, dry cough and weight loss (15 kg). The hemogram revealed leukopenia, thrombocytopenia, and anemia; while a transesophageal echocardiogram showed vegetation on the aortic valve with decreased systolic function, and B. melitensis was isolated in a blood culture. Considering this medical history, antibacterial treatment was initiated with rifampicin, doxycycline and gentamicin. The patient recovered and had satisfactory clinical evolution. Brucellosis is a rare disease. It should be considered in any person with a fever of unknown origin who lives in endemic areas or is exposed to the care of farm animals. Endocarditis is a highly lethal complication of human brucellosis; therefore, it requires a precise diagnosis and treatment.


Subject(s)
Humans , Male , Adult , Gentamicins/therapeutic use , Brucella melitensis/pathogenicity , Endocarditis, Bacterial/complications
2.
Braz. j. med. biol. res ; 56: e12938, 2023. tab, graf
Article in English | LILACS-Express | LILACS | ID: biblio-1447687

ABSTRACT

Brucellosis has become a global zoonotic disease, seriously endangering the health of people all over the world. Vaccination is an effective strategy for protection against Brucella infection in livestock in developed countries. However, current vaccines are pathogenic to humans and pregnant animals, which limits their use. Therefore, it is very important to improve the safety and immune protection of Brucella vaccine. In this study, different bioinformatics approaches were carried out to predict the physicochemical properties, T/B epitope, and tertiary structure of Omp2b and Omp31. Then, these two proteins were sequentially linked, and the Cytotoxic T lymphocyte associated antigen-4 (CTLA-4) variable region was fused to the N-terminal of the epitope sequence. In addition, molecular docking was performed to show that the structure of the fusion protein vaccine had strong affinity with B7 (B7-1, B7-2). This study showed that the designed vaccine containing CTLA-4 had high potency against Brucella, which could provide a reference for the future development of efficient brucellosis vaccines.

3.
China Tropical Medicine ; (12): 319-2023.
Article in Chinese | WPRIM | ID: wpr-979638

ABSTRACT

@#Abstract: Objective To investigate the clinical characteristics and incidence of Brucella encephalitis and meningitis in children. Methods We report the clinical data of a child with Brucella melitensis meningitis in children, and summarize the incidence, diagnosis methods and treatment of Brucella encephalitis or meningitis in children, taking into account the relevant domestic and foreign literature from January 2014 to December 2020. Results A 4-year-old girl was admitted to the hospital with status epilepticus on March 15, 2021 because of interrupted right limb numbness for 16 hours and convulsions for 2 hours. She had 2 non-febrile convulsions three months before admission and was diagnosed with epilepsy. This incident was acute, accompanied by low fever, with epilepsy as the main manifestation. Cerebrospinal fluid test suggested central nervous system infection, but the nature of infection could not be determined by routine and biochemistry of cerebrospinal fluid.The cerebrospinal fluid next generation sequencing confirmed that the pathogen of the infection was B. melitensis, which was further verified by the peripheral blood antibody test. After effective antibiotics combined with a full course of treatment, the patient recovered after six months of treatment. A total of 60 articles were retrieved in the database, including 29 in Chinese. During this period, a total of 7 cases of brucellosis in children with nervous system involvement were reported, one of which was a case report, and the other 6 cases were mentioned in the comprehensive analysis of children with brucellosis. Conclusions Brucella encephalitis or meningitis in children has a low incidence and various clinical features, which are easy to be misdiagnosed or missed.

4.
Chinese Journal of Endemiology ; (12): 409-413, 2023.
Article in Chinese | WPRIM | ID: wpr-991646

ABSTRACT

Objective:To establish a real-time quantitative PCR assay to detect the copy number of IS711 transposase gene (orfA) in Brucella genome, and the assay is applied to identify the species and biovars of Brucella. Methods:To establish an orfA gene copy number detection system based on Taqman real-time quantitative PCR technique. Primers and probes of bcsp31 and orfA genes were designed, the contents of bcsp31 gene and orfA gene in the same strain with the same DNA concentration were simultaneously detected by real-time quantitative PCR assay, and cycle number (CT value) of the two genes were obtained. According to the differences of CT values of bcsp31 gene and orfA gene, the copy number of orfA gene in Brucella genome was calculated. At the same time, the DNA of Brucella 16M strain was double decreasing dilution to verify the stability of the detection system. Results:A real-time quantitative PCR assay was used to detect bcsp31 gene and orfA gene simultaneously, when the DNA concentration difference of 16M strain was 2 times, the mean difference of CT values measured was 1.00, 95% confidence interval was 0.95-1.05, standard deviation was 0.17, and coefficient of variation was 0.17. The orfA gene copy number of 30 Brucella strains was detected by this detection system. It was found that there were 6, 9, and 7 copy numbers in the biovars 1-3 of Brucella melitensis, respectively. The strain of Brucella suis biovar 2 had 10 copy numbers, which were different from those of the other 4 strains of biovars 1, 3-5. There were 37 copy numbers in Brucella ovis strain. The copy numbers were stable at 5-6 copies in 8 biovars (1-7, 9) of Brucella abortus strains. Conclusions:A real-time quantitative PCR assay for detection of orfA gene copy number in Brucella DNA has been established. This method could identify some Brucella species and biovars strains.

5.
Chinese Journal of Endemiology ; (12): 351-355, 2023.
Article in Chinese | WPRIM | ID: wpr-991635

ABSTRACT

Objective:To study the multi-locus sequence typing (MLST) gene characteristics of Brucella isolates in Guizhou Province. Methods:Brucella strains, which were isolated from 2017 to 2021 in Guizhou Province (preserved in the Bacterial and Viral Seed Bank of Guizhou Center for Disease Control and Prevention) were identified Brucella and species/types by BCSP31-PCR and AMOS-PCR methods, respectively. MLST method was used for genotyping, and Biometrics 8.0 software was used for cluster analysis of the typing results. Results:A total of 32 strains of Brucella were isolated in Guizhou Province and identified as Brucella melitensis ( B.melitensis) by BCSP31-PCR and AMOS-PCR methods. These strains were classified into 2 ST types (ST8 and ST39) by MLST method, with 28 strains of ST8 type(87.5%) and 4 strains of ST39 type (12.5%). The 28 strains of ST8 type were distributed in 7 cities (prefectures) of Guizhou Province, while the 4 strains of ST39 type were only found in Qianxinan Buyi and Miao Autonomous Prefecture. The cluster analysis results showed that ST8 and ST39 types strains were clustered in a group with the reference strain of B.melitensis, and there was only one nucleotide site difference between ST39 and ST8 types in the glk gene, indicating a close genetic relationship. Conclusions:B.melitensis is the main pathogen of the brucellosis epidemic in Guizhou Province in recent years. ST8 is the dominant MLST genotype in Guizhou Province.

6.
Chinese Journal of Endemiology ; (12): 345-350, 2023.
Article in Chinese | WPRIM | ID: wpr-991634

ABSTRACT

Objective:To investigate the effect of recombinant lipoproteins of Brucella outer membrane protein 16, 19 (L16 and L19) on the expression of immune regulatory factors in human monocytic leukemia cell line (THP-1 cells). Methods:THP-1 cells activated with phorbol ester (PMA) were used as an in vitro experimental cell model, and a group design was used to co-culture L16, L19 and THP-1 cells (L16 stimulated group, L19 stimulated group), respectively. THP-1 cells activated with PMA were used as the control group. When co-cultured for 4 hours, immunofluorescence staining (IFS) and Western blotting were used to detect whether L16 and L19 entered the cells, respectively; when co-cultured for 12, 24 hours, real-time fluorescent quantitative PCR was used to measure the mRNA expression levels of interferon regulatory factor 1 (IRF-1) and trans activator protein of major histocompatibility complex class Ⅱ (CⅡTA); Western blotting was used to detect the protein expression levels of T cell immunoglobulin mucin-3 (Tim-3) and γ interferon receptor 1 (IFNGR1). Results:When co-cultured for 4 hours, L16 and L19 were observed entering THP-1 cells in the L16 stimulated group and L19 stimulated group, respectively. When co-cultured for 12 hours, the expression level of IRF-1 mRNA in the L16 stimulated group (0.16 ± 0.15) was significantly lower than that in the control group (1.00 ± 0.00, P < 0.05). When co-cultured for 24 hours, the expression level of CⅡTA mRNA in the L16 stimulated group (0.17 ± 0.10) was significantly lower than that in the control group (1.00 ± 0.00, P < 0.05). When co-cultured for 12 and 24 hours, there were no statistically significant differences in the expression levels of IRF-1 and CⅡTA mRNA between the L19 stimulated group and the control group ( P > 0.05). Western blotting results showed that there were statistically significant differences in the expression levels of INFGR1 and Tim-3 protein among the control group, L16 stimulated group, and L19 stimulated group after co-cultured for 12 and 24 hours ( F = 50.92, 6.80, 148.73, 156.57, P < 0.05). Among them, when co-cultured for 12 hours, the expression level of INFGR1 protein in the L16 and L19 stimulated groups were significantly lower than that in the control group, and the L19 stimulated group was higher than the L16 stimulated group ( P < 0.05), and the expression level of Tim-3 protein in the L19 stimulation group was higher than that in the control group ( P < 0.05). When co-cultured for 24 hours, the expression level of INFGR1 protein in the L16 and L19 stimulated groups were lower than that in the control group, and the L19 stimulated group was higher than that in the L16 stimulated group ( P < 0.05); and the expression level of Tim-3 protein in the L16 stimulated group was higher than that in the control group and L19 stimulated group ( P < 0.05). Conclusions:Brucella L16 can downregulate the expression levels of IRF-1 and CⅡTA mRNA in THP-1 cells. Both L16 and L19 can downregulate IFNGR1 and upregulate Tim-3 protein expression levels.

7.
Chinese Journal of Endemiology ; (12): 328-331, 2023.
Article in Chinese | WPRIM | ID: wpr-991630

ABSTRACT

Objective:To establish a quantitative real-time PCR detection system for Brucella S2 vaccine strain. Methods:Based on the differences in the entire genome sequence between Brucella S2 vaccine strain and other reference strains of Brucella, primers and probes were designed to establish a quantitative real-time PCR detection system for Brucella S2 vaccine strain. The DNA of 22 reference strains of Brucella and 8 non- Brucella control strains were obtained from the National Institute for Infectious Disease Control and Prevention of the Chinese Center for Disease Control and Prevention. At the same time, environmental samples were obtained from the brucellosis vaccine manufacturers, and bacterial DNA from environmental samples was extracted using a blood/tissue genomic DNA extraction kit. The obtained DNA was pre-amplified by conventional PCR, and then subjected to quantitative real-time PCR secondary amplification (nested fluorescence quantitative PCR) using the amplified PCR product as a template. The specific fluorescence curve and corresponding number of cycles (Ct value) were observed, and the sensitivity was tested. Results:The quantitative real-time PCR detection system established did not detect specific fluorescence curves (without Ct values) for 21 reference strains of Brucella and 8 non- Brucella control strains, except for S2 vaccine strains. The established detection system had a minimum detection limit of 4.34 fg (genomic DNA) for detecting the DNA of Brucella S2 vaccine strain; DNA of Brucella S2 vaccine strain was detected in 3 of the 14 environmental samples collected. Conclusion:The quantitative real-time PCR detection system established can detect Brucella S2 vaccine strain in samples, with good sensitivity and specificity.

8.
Chinese Journal of Endemiology ; (12): 269-273, 2023.
Article in Chinese | WPRIM | ID: wpr-991618

ABSTRACT

Objective:To observe multiple locus variable-number tandem repeat analysis (MLVA) typing of Brucella isolated from Himalayan marmot in Qinghai-Tibet Plateau of Qinghai Province, and to explore the relationship between the strains and strains previous isolated from Qinghai Province. Methods:Blood samples of Himalayan marmot were collected in Qinghai-Tibet Plateau of Qinghai Province from March 2019 to October 2020. Pathogens were isolated and cultured from Brucella antibody positive samples identified by using the rose bengal test (RBT). Conventional biological methods and molecular biological methods (BCSP31-PCR and AMOS-PCR) were used for strain identification. At the same time, MLVA method was used to genotype the isolated strains, and cluster analysis was used to analyze the genetic relationships between the strains based on the genotype of 70 Brucella isolated from different hosts in Qinghai Province. Results:A total of 1 466 blood samples of Himalayan marmot were collected from Qinghai-Tibet Plateau. Two strains of Brucella were isolated and cultured from 64 RBT-positive samples, named QH2013054 and QH2013062, respectively. They were identified as Brucella ovis biotype Ⅲ by conventional and molecular biological methods. The MLVA genotyping results showed that QH2013054 and QH2013062 were different at the Bru16 locus, indicating different MLVA genotypes. Cluster analysis showed that strain QH2013054 had the same MLVA genotype as 7 strains, among which 6 strains were from 3 farmers and 3 sheep from the same family in Gonghe County, and 1 strain was from a farmer in Menyuan Hui Autonomous County. The strain QH2013062 had the same MLVA genotype as 4 strains, including 3 strains from 3 farmers in Menyuan Hui Autonomous County and 1 strain from a farmer in Tu Autonomous County of Huzhu. Conclusions:The strains of Brucella isolated from Himalayan marmot in Qinghai-Tibet Plateau of Qinghai Province have the same MLVA genotype as some strains of Brucella isolated from humans and sheep in Qinghai Province. It is speculated that the host humans, sheep and Himalayan marmot in Qinghai-Tibet Plateau may have a common source of infection.

9.
Chinese Journal of Endemiology ; (12): 196-199, 2023.
Article in Chinese | WPRIM | ID: wpr-991604

ABSTRACT

Objective:A nested-PCR assay is developed to detect and identify the genomic DNA of Brucella vaccine A19 strain. Methods:The whole genomic sequences of Brucella vaccine A19 strain and other Brucella spp. strains were compared and analyzed. The primers were designed by nucleotide difference sites. The nested-PCR assay was established to detect and identify Brucella vaccine A19 strain. The genomic DNA of Brucella vaccine A19 strain was extracted and diluted. The diluted template DNA was tested for sensitivity of using nested-PCR assay. And the specificity of nested-PCR assay was tested for the genomic DNA of other Brucella spp. strains and non- Brucella spp. strains. Results:The minimum detection limit of the nested-PCR assay was 3.43 fg. The nested-PCR assay established for amplification of Brucella vaccine A19 strain showed 246 bp electrophoresis bands, while other Brucella spp. strains showed 314 bp electrophoresis bands, and non- Brucella spp. strains did not produce electrophoresis bands. Conclusions:The nested-PCR assay established has the characteristics of high sensitivity and specificity. It can be detected when there is one copy of Brucella vaccine A19 strain genomic DNA in the reaction system. This method is particularly suitable for the detection and identification of trace genomic DNA of Brucella vaccine A19 strain in sample.

10.
Chinese Journal of Endemiology ; (12): 24-29, 2023.
Article in Chinese | WPRIM | ID: wpr-991572

ABSTRACT

Objective:To learn about the genotyping of human Brucella isolated from Sichuan Province. Methods:BCSP31-PCR and AMOS-PCR were used to identify the genus and biotype of the 66 strains isolated from confirmed human brucellosis cases in Sichuan Province from 2014 to 2020, respectively. The isolated strains were genotyped by multi-locus sequence typing (MLST)-9. The sequence type (ST) was compared trough the online MLST database. A minimum spanning tree (MST) was constructed to cluster the newly discovered and known ST using the BioNumerics software version 7.6.Results:The 66 strains isolated from human cases of brucellosis in Sichuan Province from 2014 to 2020 were Brucella, and 65 of them were Brucella melitensis while one strain was Brucella abortus. The MLST method identified three known STs (ST-8, ST-39 and ST-2) and one newly type (ST-101). Among them, ST-8 was the main ST in Sichuan Province (90.91%, 60/66), another 4 strains of Brucella melitensis were ST-39, and 1 strain of Brucella abortus was ST-2. The newly type ST-101 was isolated from Leshan City in 2019, belonging to the Brucella melitensis and closely related to the evolution of ST-8. Conclusion:Brucella melitensis is the main epidemic Brucella strain in Sichuan Province, ST-8 is predominant genotype, with a small amount of ST-39, ST-101 and ST-2.

11.
Article in Spanish | LILACS-Express | LILACS | ID: biblio-1537055

ABSTRACT

En la producción zootécnica, la brucelosis y la leptospirosis ocasionan problemas reproductivos y son una limitante en salud y en producción animal. El objetivo fue determinar la presencia de anticuerpos contra Leptospira spp. y Brucella abortus, en una población bufalera, en el municipio de Tierralta, Córdoba. Se realizó un estudio descriptivo de corte transversal, que incluyó un total de 144 búfalos de la raza Murrah, destinados al doble propósito. Para el diagnóstico de Leptospira spp., se implementó la prueba de aglutinación microscópica, con 13 serogrupos, 19 serovares pertenecientes a 5 especies de Leptospira patógenas y para brucelosis Rosa de Bengala y C-Elisa. La seroprevalencia para Leptospira spp. fue del 87,5 %, el serogrupo Mini fue el de mayor frecuencia, pero Grippotyphosa presentó el mayor título. El 16,67 % de los búfalos evaluados presentaron títulos iguales o superiores a 1:800, asociados con infección actual o reciente. La alta seroprevalencia, se puede deber a las características ambientales de la zona, que brinda las condiciones favorables para el crecimiento y el mantenimiento de este patógeno que, sumado al comportamiento de los búfalos de revolcarse, los hace propensos a las infecciones con bacterias del género Leptospira sp., porque a menudo, las fuentes de agua están contaminadas por este patógeno. La seroprevalencia contra B. abortus por Rosa de Bengala y Elisa-C fue del 2,08 %. Todos los títulos determinados corresponden a procesos infecciosos. No hubo signos clínicos de enfermedad y la carencia de registros productivos no permitió determinar el efecto sobre los parámetros reproductivos.


In animal production, brucellosis and leptospirosis cause reproductive problems and limit animal health and production. The objective was to determine the presence of antibodies against Leptospira spp. and Brucella abortus in a buffalo population in the municipality of Tierralta, Cordoba. A descriptive, transversal study was carried out including a population of 144 Murrah´s breed buffalos destined for beef and milk production. For the Leptospira spp. diagnostic, was used a rapid slide agglutination test with 13 serogrups and 19 serovars belonging to 5 pathogenic Leptospira species was implemented and for Brucella abortus Rose Bengal and C-Elisa was used. Seroprevalence of Leptospira spp. was 87,5 %, serogrup Mini was the most frequent, but Grippotyphosa showed the higher titer. The 16,67 % of the buffaloes evaluated presented titles equal to or above than 1:800 associated with current or recent infection. High seroprevalence may be due to environmental characteristics of the zone, which gives favorable conditions for the growth and maintenance of this pathogen, these factors in conjunction with the habit of wallowing makes them prone to suffering infections caused by bacteria of the genera Leptospira sp. since water sources are often contaminated. Seroprevalence against B. abortus by Rose-Bengal and C-Elisa was 2,08 %, the determined titers correspond to infectious processes. There were no clinical manifestations of disease and the effects on reproductive parameters were not determined because of the lack of productive registries.

12.
Chinese Journal of Infectious Diseases ; (12): 545-550, 2022.
Article in Chinese | WPRIM | ID: wpr-956452

ABSTRACT

Objective:To analyze the clinical characteristics of Brucella infection in Shenzhen City, and to provide reference for clinical diagnosis and treatment of patients with Brucella infection. Methods:The clinical characteristics of 57 patients with Brucella infection from January 1, 2018 to December 31, 2020 in The Third People′s Hospital of Shenzhen were retrospectively analyzed. The clinical characteristics of patients with brucellosis and latent Brucella infection, patients with or without comorbidities were compared respectively, and magnetic resonance imaging (MRI) and lumbar puncture examination findings of 57 patients were also analyzed. Statistical analysis was performed using Wilcoxon rank sum test and chi-square test. Results:Among the 57 patients with Brucella infection, 10 cases (17.5%) were latent infections and 47 cases (82.5%) were brucellosis patients. Among brucellosis patients, 91.5%(43/47) had fever and 74.4%(32/43) had maximum body temperature ≥38.1 ℃, 40.4%(19/47) had chills orshivering, 25.5%(12/47) had hyperhidrosis, 17.0%(8/47) had fatigue, 21.3%(10/47) had headache, 23.4%(11/47) had neck/back/low back pain, and 31.9%(15/47) had joint pain. A total of 18 cases (38.3%) had comorbidities. Cases with positive blood cultures in latent infection and brucellosis were seven and 39, respectively. The time from symptom onset to diagnosis was 30.0 (15.0, 67.5) days in 18 patients of brucellosis with comorbidity, which was longer than 20.0 (13.0, 30.0) days in 29 patients without comorbidity. Neck/back/low back pain and joint pain occurred in patients with brucellosis with comorbidity were seven and nine, respectively, and those without comorbidity were four and six, respectively, with statistically significant differences ( Z=-2.00, χ2=3.90 and 4.39, respectively, all P<0.050). Of the 11 brucellosis patients with neck/back/low back pain, six had spondylitis. Of the 15 brucellosis patients with joint pain, six had arthritis. Lumbar puncture examination did not indicate meningitis in six cases of latent infection, while revealed six cases of brucellosis meningitis in 32 brucellosis patients. Fifty-four patients had good outcomes, and three patients were cured after an extended course of treatment. Conclusions:Although patients with latent Brucella infection have no comorbidities, they have a high positive blood culture rate. Active standardized anti- Brucella treatment is recommended. MRI examination of relevant sites is recommended in brucellosis patients with joint, neck/back/low back pain, and lumbar puncture is recommended in brucellosis patients regardless of headache.

13.
International Journal of Biomedical Engineering ; (6): 287-293, 2022.
Article in Chinese | WPRIM | ID: wpr-989260

ABSTRACT

Objective:To prepare camel-derived nanoantibodies that can bind to the recombinant protein VirB12 antigen with high affinity and lay the foundation for further research.Methods:Xinjiang Bactrian camels were immunized six times with VirB12 recombinant protein, total RNA was extracted from lymphocytes isolated from peripheral blood, and the VHH gene fragment was amplified by nested PCR to construct a phage VHH display library. ELISA solid-phase affinity and enrichment methods were used for screening. After three rounds of affinity screening, the clones enriched in the second and third rounds were randomly picked out, and the binding of a nanoantibody with soluble expression to VirB12 was analyzed by ELISA. After sequence determination and multiple alignment, repetitive sequences were removed, and finally five non-redundant sequences were obtained, which were named D1, E6, H8, H9, and H10. The five identified nanoantibody genes were transformed into the WK6 strain, and the soluble expression of an intercellular substance was carried out at 16 °C. After purified expression of Ni-NTA, the binding ability and thermal stability of nanoantibodies and the antigen VirB12 protein were detected by Western Blot and ELISA.Results:Five strains of nanoantibodies were expressed in WK6 bacteria in soluble form. SDS-PAGE showed that the purity of five anti-VirB12 nanoantibodies was close to 90%, and they had high antigen-binding activity and obvious antigen-antibody concentration dependence. All four strains of nanoantibodies showed high thermal stability, and after being treated at 90 ℃, they could still retain more than 60% binding activity.Conclusions:In the study, a VHH phage display library with a capacity of 2.8×10 8 cfu/ml was constructed from Xinjiang Bactrian camel lymphocytes immunized with VirB12 recombinant protein. Five anti-VirB12 nanoantibodies with high affinity and thermal stability were obtained through solid-phase screening and enrichment and soluble monoclonal ELISA detection. These results laid the foundation for further development of VirB12 nanoantibodies.

14.
NOVA publ. cient ; 20(3): [17], 2022 enero-junio. gráficos, mapas, tablas e ilustraciones
Article in Spanish | LILACS | ID: biblio-1397025

ABSTRACT

Resumen Introducción. La Brucelosis bovina, es una enfermedad bacteriana e infecto contagiosa, causada por Brucella abortus. Se transmite a través de la ingestión de forrajes y aguas contaminadas con descargas vaginales infectadas, que conlleva a una patología del sistema reproductor en bovinos, que impactan la sanidad pecuaria y la economía de la agroindustria. Objetivo. Evaluar el comportamiento de la brucelosis bovina en el municipio de Aguazul, Casanare (Colombia) y los factores asociados al desarrollo de esta enfermedad. Metodología. Estudio descritptivo de 26.187 muestras de suero sanguíneo de ganado bovino evaluadas, que corresponden a 260 predios del municipio de Aguazul. Se empleó técnicas serológicascomo rosa de bengala, fluorescencia polarizada y ELISA competitiva para la evaluación de positividad a Brucella abortus y se evaluaron las pérdidas económicas asociadas a positividad en los ensayos de laboratorio. Resultados y conclusiones. La positividad a brucelosis bovina correspondió al 1%, que corresponde a hembras menores de 24 meses de edad y entre 37 a 48 meses y, machos entre los 57 a 68 meses de edad. Se sugiere consolidar esfuerzos en investigación para evaluar los factores que contribuyen a la seropositividad en el ganado y el riesgo para la propagación y mantenimiento de la enfermedad.


Abstract Introduction. Bovine brucellosis is a contagious bacterial and infectious disease caused by Brucella abortus. It is transmitted through the ingestion of forage and water contaminatedwith infected vaginal discharges, which leads to a pathology of the reproductive system inbovines, which impacts livestock health and the economy of agribusiness. Aim. To evaluatethe behavior of bovine brucellosis in the municipality of Aguazul, Casanare (Colombia) and the factors associated with the development of this disease. Methodology. Descriptive study of 26,187 bovine blood serum samples evaluated, corresponding to 260 farms in the municipality of Aguazul. Serological techniques such as rose bengal, polarized fluorescence, and competitive ELISA were used to evaluate positivity to Brucella abortus and the economic losses associated with positivity in laboratory tests were evaluated. Results and conclusions. The positivity to bovine brucellosis corresponded to 1%, which corresponds to females under 24 months of age and between 37 to 48 months and males between 57 to 68 months of age. It is suggested to consolidate research efforts to evaluate the factors that contribute to seropositivity in cattle and the risk for the spread and maintenance of the disease


Subject(s)
Humans , Animals , Cattle , Brucellosis, Bovine , Brucella abortus , Communicable Diseases , Agribusiness
15.
Braz. j. infect. dis ; 26(2): 102351, 2022. tab
Article in English | LILACS-Express | LILACS | ID: biblio-1384115

ABSTRACT

Abstract Purpose Brucellosis is a zoonotic disease of great public health importance. In wild animals, Brucella abortus is one of the most diagnosed species, mainly in enzootic environments where domestic animals share the same environment. B. abortus is common in environments shared by cattle, wild, and domestic animals. This study aimed to detect the presence of B. abortus DNA in free-ranging and captivity felids at Mato Grosso State, Brazil. Method Polymerase chain reaction, based on the genetic element IS711, was performed in blood samples collected from 23 free-ranging and captive felids. The species represented include Leopardus colocolo, Leopardus pardalis, Leopardus wiedii, Panthera onca, Puma concolor, and Puma yagouaroundi. Results DNA amplification of B. abortus was observed in only one captive P. concolor (4.34%). Conclusion The detection of this pathogen in captive animals using molecular tools demonstrates the importance of monitoring, as it raises concerns about the possibility of transmission between humans and wild and domestic animals, especially in regions of vast biodiversity, such as in the State of Mato Grosso, Brazil.

16.
Chinese Journal of Endemiology ; (12): 246-252, 2022.
Article in Chinese | WPRIM | ID: wpr-931530

ABSTRACT

Brucellosis is an infectious zoonosis caused by Brucella infection. It is widely prevalent all over the world and brings great losses to the development of public health, animal husbandry and social economy in China. However, the understanding of the specific clinical indications of brucellosis, the virulence factors of Brucella and its pathogenesis is very limited, which leads some limitations in the clinical diagnosis and treatment of brucellosis. Starting from the current epidemic situation of brucellosis, this article focuses on the virulence factors of Brucella and pathogenesis of brucellosis, and comprehensively summarizes the activity track and state of Brucella in the infected organism during the pathogenesis of brucellosis, so as to provide new ideas for the in-depth study of the pathogenesis of brucellosis, and a comprehensive reference for vaccine development and clinical diagnosis and treatment.

17.
Chinese Journal of Endemiology ; (12): 180-185, 2022.
Article in Chinese | WPRIM | ID: wpr-931518

ABSTRACT

Objective:To analyze the incidence of brucellosis and the genotypes of Brucella isolates or nucleic acids in Shaanxi Province, to get the epidemiological and molecular genetic characteristics, and to provide scientific basis for precise prevention and control of human brucellosis. Methods:Log into the Chinese Disease Control and Prevention Information System, collect the incidence data of human brucellosis of Shaanxi Province in 2020, and analyze the epidemiological characteristics. Bacteriology and PCR methods were used to identify the isolates or nucleic acids, and multiple locus variable-number tandem repeat analysis (MLVA) was used for molecular typing. BioNumerics (Version 7.6) software was used to analyze the results of MLVA.Results:In 2020, 1 086 cases of human brucellosis were reported in Shaanxi Province, the incidence rate was 2.80/100 000, involving 86 counties (districts), the epidemic peak was from March to September (865 cases), male-to-female ratio was 2.68 ∶ 1.00 (791 ∶ 295), 79.74% (866 cases) in the age group of 30 to 69 years old, and 83.43% (906 cases) of the cases were farmers. Biotype identification of 36 isolates showed that 4 isolates were mutant Brucella melitensis, 3 isolates were Brucella melitensis 1 and 29 isolates were Brucella melitensis 3. The 36 isolates and 7 nucleic acids were identified as Brucella by BCSP31-PCR and Brucella melitensis by AMOS-PCR. MLVA-16 genotyping, panel1 showed two genotypes: type 42 (1-5-3-13-2-2-3-2) and type 63 (1-5-3-13-2-3-3-2), panel2A showed 4-41-8 and panel2B showed high variability. Thirty-six isolates and 7 nucleic acids were divided into 33 genotypes, of which 27 genotypes were single isolates and 6 genotypes were shared. Conclusions:The situation of human brucellosis prevention and control in Shaanxi Province is grim. MLVA-16 is a mature genotyping method, which determines the existence of multiple genotypes of Brucella isolates or nucleic acids in Shaanxi Province, which can provide scientific information for precise prevention and control of human brucellosis, outbreak analysis and epidemiological traceability.

18.
Chinese Journal of Endemiology ; (12): 1013-1018, 2022.
Article in Chinese | WPRIM | ID: wpr-991565

ABSTRACT

Objective:To identify the biotypes of blood culture positive Brucella in patients with acute brucellosis in Xinjiang Uygur Autonomous Region (hereinafter referred to as Xinjiang), and to learn about the clinical characteristics of these patients. Methods:Among the 30 patients diagnosed with acute brucellosis in First Affiliated Hospital, School of Medicine, Shihezi University from January 2019 to July 2021, the positive strains of blood culture Brucella of 12 patients with acute brucellosis were used to identify the biotype by AMOS-PCR. Then the general and clinical data of the 12 patients were collected, including demographic characteristics, epidemiological characteristics, clinical characteristics, laboratory tests, treatment and prognosis. Results:AMOS-PCR results showed that all 12 strains were Brucella melitensis. Among the 12 patients, 9 were males and 3 were females, all of whom had a clear history of livestock contact, and the onset time was concentrated in April to September. All 12 patients showed fever, followed by headache and dizziness (9/12), weakness (8/12), digestive symptoms (8/12), hyperhidrosis (7/12), lumbago and backache (6/12), and arthralgia (6/12). One patient was complicated with pleural effusion and pericardial effusion. One patient was complicated with pleural effusion, pelvic effusion and ascites. Only ascites occurred in 2 patients. Liver function was abnormal in 3 patients, lactate dehydrogenase increased in 6 patients, and blood creatinine decreased in 9 patients. Among the 12 patients, 10 patients were treated with rifampicin combined with doxycycline. The other 2 patients were treated with levofloxacin and doxycycline, supplemented by symptomatic treatment with hepatoprotective drugs due to impaired liver function. After discharge, the patients received out-of-hospital oral anti-infectious drugs for 1-2 courses (6 weeks each course), and all indicators returned to normal at 6 months and 12 months after discharge. Conclusions:All the 12 patients with acute brucellosis in Xinjiang are infected with Brucella melitensis. In addition to the common clinical features, patients are characterized by decreased creatinine and increased lactate dehydrogenase. The standardized anti- Brucella treatment is still the main treatment strategy.

19.
Arq. bras. med. vet. zootec. (Online) ; 73(6): 1294-1300, Nov.-Dec. 2021. tab
Article in English | LILACS, VETINDEX | ID: biblio-1355682

ABSTRACT

Brucella ovis, a non-zoonotic species, is the etiological agent of ovine brucellosis, an infectious disease of clinical or subclinical occurrence in sheep flocks. Until then, there is no serological study of anti-Brucella ovis antibodies in purebred sheep herds. This study aimed to determine the presence of anti-Brucella ovis antibodies in purebred sheep flocks with breeding purposes from Parana State. Blood samples from 728 animals, of which 563 were females and 165 males, between 8 and 56 months of age from the six major sheep producing mesoregions of Parana, were submitted to detection of anti-Brucella ovis antibodies by the Agar Gel Immunodiffusion technique using an antigen from the bacteria Brucella ovis (Reo 198). The results indicate the presence of this disease in purebred sheep from Parana State in a low occurrence of 0.27% (2/728). The only two positive animals were rams, Santa Inês breed, from the same flock in the East Center region of Parana, without clinical disease. In conclusion, Brucella ovis is present in purebred sheep in Parana State, Brazil, and this low occurrence may have occurred due to rigorous breeding systems that may contribute to reduce the transmission of this disease.(AU)


Brucella ovis, espécie não zoonótica, é o agente etiológico da brucelose ovina, doença infecciosa de ocorrência clínica ou subclínica. Atualmente, não existe estudo sorológico de anticorpos anti-Brucella ovis em rebanhos de ovinos puros de origem. Este estudo teve como objetivo determinar a presença de anticorpos anti-Brucella ovis em rebanhos ovinos de raça pura de origem, com fins reprodutivos do estado do Paraná. Amostras de sangue de 728 animais, sendo 563 fêmeas e 165 machos, entre oito e 56 meses de idade, pertencentes a seis principais mesorregiões produtoras de ovinos no Paraná, foram submetidas à detecção de anticorpos anti-Brucella ovis pela técnica de imunodifusão em ágar gel usando-se um antígeno da bactéria Brucella ovis (Reo 198). Os resultados indicam a presença da doença em ovinos puros de origem do estado do Paraná em baixa ocorrência de 0,27% (2/728). Os dois únicos animais positivos foram reprodutores da raça Santa Inês, do mesmo rebanho da região Centro Leste do Paraná, sem manifestação clínica. Em conclusão, Brucella ovis está presente em ovinos puros de origem no estado do Paraná, e essa baixa ocorrência pode ter ocorrido devido a sistemas rigorosos de criação, que podem contribuir para a redução da transmissão dessa doença.(AU)


Subject(s)
Animals , Brucellosis/epidemiology , Sheep/immunology , Brucella ovis/immunology , Sheep Diseases/immunology , Brazil , Immunodiffusion/veterinary
20.
Rev. med. Risaralda ; 27(2): 153-160, jul.-dic. 2021. tab, graf
Article in Spanish | LILACS-Express | LILACS | ID: biblio-1365901

ABSTRACT

Resumen Introducción: La fiebre de malta (brucelosis) es una infección zoonótica producida por cocobacilos gramnegativos, intracelulares facultativos, los cuales se transmiten por el consumo de productos animales no pasteurizados infectados, el contacto de la piel o las membranas mucosas con tejido animal infectado, fluidos animales infectados e inhalación de partículas aerosolizadas infectadas. Caso: Hombre de 34 años residente en zona rural, quien se dedicaba a la ganadería, ingreso a urgencias por presentar cuadro clínico de 15 días de evolución consistente en picos febriles no cuantificados asociados a escalofríos, astenia, adinamia y mialgias. Mediante la correlación clínico-patológica se llegó al diagnóstico de infección por Brucella Abortus. Conclusión : Esta patología es más frecuente en varones adultos. Dentro del cuadro clínico, Los estudios de serológicos (anticuerpos, aglutinación y ensayo inmunocromatográfico) tienen la mayor sensibilidad y especificidad diagnostica. El tratamiento se da con medicamentos que actúen en entornos intracelulares ácidos (tetraciclinas, aminoglucosidos, fluoroquinolonas), esto con el fin de controlar la enfermedad, prevenir las complicaciones y evitar las recaídas.


Abstract Introduction: Malta fever (brucellosis) is a zoonotic infection produced by intracellular gram-negative coccobacilli, which is transmitted by the consumption of infected unpasteurized animal products, skin contact or mucous membranes with infected animal tissues and fluids, and inhalation of infected aerosolized particles. Case: A 34-year-old man living in a rural area, who works in livestock, was admitted to the emergency department for presenting a clinical picture of 15 days of evolution of unquantified febrile peaks associated with symptoms such as chills, asthenia, adynamia and myalgia. The diagnosis of infection with Brucella Abortus was given through clinical-pathological correlation. Conclusion: This pathology is more frequent in adult males. Serological studies (antibodies, agglutination and immunochromatographic assay) prove to have the highest sensitivity and diagnostic specificity in the clinical picture. The treatment is given with medication that acts on intracellular acidic environment (tetracyclines, aminoglycosides, fluoroquinolones), this in order to control the disease, and prevent complications and relapses.

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