Marcadores moleculares para la taxonomía e identificación del género Brucella (Alphaproteobacteria) / Molecular markers for the taxonomy and identification of the genus Brucella (Alphaproteobacteria)

Introducción: El género Brucella está incluido en la familia Brucellaceae que pertenece al orden Rhizobiales y es reconocido por su alto grado de patogenicidad. Las bacterias de este género son responsables de la brucelosis, enfermedad que ha sido reportada como una de las zoonosis más importantes a nivel mundial por su incidencia en el ganado y el hombre. Los estudios previos para la clasificación taxonómica del género, se han basado fundamentalmente en el análisis del gen 16S ARNr. Sin embargo, pocas investigaciones se han dirigido a la identificación de marcadores moleculares que distingan a sus miembros de otros grupos de bacterias. Objetivo: Identificar inserciones en secuencias de proteínas conservadas, que pudieran ser utilizados como marcadores moleculares para la taxonomía y diagnóstico de especies del género Brucella. Métodos: Las secuencias homólogas de las proteínas analizadas fueron obtenidas de bases de datos internacionales y, posteriormente, alineadas con el programa ClustalX2, para ello fueron considerados los parámetros sugeridos en la literatura. Resultados: Se identificaron inserciones en las proteínas oxoglutarato deshidrogenasa (componente E1) y ADN ligasa A específicas del género Brucella. Conclusiones: Las inserciones halladas pueden ser empleadas como complemento a los métodos tradicionales de clasificación taxonómica y para el diagnóstico molecular de bacterias incluidas en el género Brucella(AU)

Introduction: Brucella is a genus from the Brucellaceae family, Rhizobiales order. This genus is recognized for its high pathogenicity. Brucella bacteria cause brucellosis, a disease reported as one of the most important zoonoses worldwide due to its incidence in cattle and people. Previous studies on taxonomic classification of the genus have been mainly based on the analysis of gene 16S rDNA. However, few studies have been aimed at identification of molecular markers distinguishing its members from other groups of bacteria. Objective: Identify insertions in preserved protein sequences which could be used as molecular markers for the taxonomy and diagnosis of species from the Brucella genus. Methods: The homologous sequences for the proteins analyzed were obtained from international databases and aligned with the software ClustalX2, considering the parameters suggested in the literature. Results: Insertions were identified in the proteins oxoglutarate dehydrogenase (component E1) and DNA ligase A, specific of the genus Brucella. Conclusions: The insertions found may be used as complements to the traditional methods for taxonomic classification and for the molecular diagnosis of bacteria from the genus Brucella(AU)

Evidência molecular de brucella sp. em ozotoceros bezoarticus (veado campeiro) do Pantanal sul-mato-grossense / Molecular evidence of brucella sp. in deer (ozotoceros bezoarticus) of the southern Pantanal

A presença de Brucella spp. entre animais silvestres pode influenciar a taxa de reprodução destes hospedeiros, além de atuarem como fonte de infecção natural para os animais domésticos e humanos. O objetivo deste estudo foi identificar a presença de Brucella spp. em 44 amostras de sangue de veado campeiro (Ozotoceros bezoarticus) do Pantanal do Sul-Mato-Grossense, utilizando a técnica de PCR. Observou-se que 20,4 por cento (9/44) das amostras foram positivas. A sequência consenso de nucleotídeo obtida no sequenciamento do isolado de veado campeiro apresentou 514 pb e 95 por cento de identidade com virB5 de B. abortus (best hits acesso nr AF226278, e-value 0.0), já na análise filogenética a amostra de Brucella isolada de veado campeiro apresentou-se muito próximo de B. suis. A alta porcentagem de amostras positivas sugere que a brucelose pode ser um problema entre os veados campeiros na área estudada e que estes animais podem representar riscos para outros animais domésticos e silvestres.

The presence of Brucella spp. in wild animals can influence their reproduction rate and may be a source of infection for domestic animals and humans. The objective of this study was to identify the presence of Brucella spp. in 44 blood samples from the deer Ozotoceros bezoarticus in the southern Pantanal of Sul-Mato-Grossense, using the PCR technique. It was seen that 20.4 percent (9/44) of the samples were positive. The consensus sequence was obtained by sequencing these samples, which then showed 514 pb and 95 percent of identity with gene virB5 of B. abortus (best hits accession nr AF226278, e-value 0.0). The phylogenetic analysis of the sample isolated from deer revealed the Brucella to be very close to B. suis. The high percentage of positive samples suggests that brucellosis may be a concern in deer within the studied area, and that these animals may poses a risk for other domestic and wild ones.

Brucellosis / Brucellosis

Brucellosis is a zoonosis commont to mammals and occasionally humans especially people who live, work, have contact with infected animals and/or ingest their products (milk, meat, blood, urine, etc.). Brucellae are intracellular gram negative, aerobic, immobile and sporule-less coccobacilli of which different species exist affecting different mammals: - Brucella melitensis (goats and sheep)- Brucella abortus (cow)- Brucella suis (pigs)Brucellosis, as with typhoid fever, is a cyclical septicaemia acquired by humans through the skin or the digestive tract, either by direct contact with infected animal produce or by its ingestion.Brucellosis is clinically manifested by a period of incubation; one of invasion and a state characterised by undulant fever, together with enlarged spleen and liver; arthralgia and arthritis especially of the cervical and lumbar spine and a mild general malaise. After a varyingperiod of time the condition, the following should be noted:- In humans, 15 to 20 per cent of cases of brucellosis spontaneously recover.- There is a 40 to 50 per cent recurrence rate among non-treated cases.- Only some 8 to 10 per cent of treated cases show recurrence.- Complications occur in about 10 per cent of cases.- Total brucellosis mortality is low, around 2 to 3 per cent.

Liver and kidney dysfunctions among brucella reactant Friesian cows in El-Minya province, Egypt

The present study was carried on 50 Friesian female cows [3-5 years old] in El-Minya Province, Egypt. Serological investigation of cattle for brucellosis revealed that, 68, 56, 60, 48 and 56% of the tested cows were brucella reactant to the Buffered Acidified Plate Antigen Test [BAPAT], Rose Bengal Plate Test [RBPT], Serum Agglutination Test [SAT], Rivanol Test [RT] and Complement Fixation Test [CFT], respectively. Biochemical analysis of the tested cows sera showed that brucella reactant cows had a significant increase in serum Alanine Amino Transferase [ALT], Aspartate Amino Transferase [AST], Alkaline Phosphatase [AP], Urea and Creatinine levels. Significant increase of serum total proteins, globulin, lipids, cholesterol, triacylglycerol, free fatty acids and the high, low and very low density lipoproteins were also detected. On the other hand significant decrease in serum albumin, A/G ratio, phospholipids, calcium, inorganic phosphorus, magnesium, sodium and potassium levels were shown among the bruccella reactant Friesian cows compared to the non-reactant controls. It could be concluded that, in addition to the hazard of distributing brucella infection, brucella reactant cows under study suffered hepatic and renal dysfunction that altered their serum constitution, which in turn implies itself on the animal health and they should be eradicated from the producing cattle herd as soon as possible

Neurobrucellosis [a complication of systemic brucellosis]

This detailed study was performed on six patients presented with diverse neurological features of meningoencephalitis, myelopathy or radiculopathy syndromes. The uncommon possibility of brucellosis as an etiological factor was not born in mind early for most of the patients delaying the proper management to the stage of chronicity and complications. Rapid improvement was recorded after proper diagnosis and adequate antibiotic therapy but long st and ing cases responded to a lesser degree and one patient died. The purpose of this article was to emphasize the importance of brucellosis as a differential diagnostic probability for several neurological syndromes

Estudio de brucelosis a partir de muestras de leche de bovinos en el trópico subhúmedo del estado de Guerrero / Study of brucellosis from bovine milk samples in the subhumid tropics of the state of Guerrero, Mexico

Se estudió la prevalencia de brucelosis por hato y por animal. También se intentó aislar la bacteria a partir de muestras de leche de animales identificados como positivos. Se analizaron muestras de leche (n=323) y sangre (n=323) de bovinos productores de leche de 63 explotaciones pertenecientes a 5 municipios de la región de Tierra Caliente, estado de Guerrero. Mediante la detección de anticuerpos contra Brucella spp, con la prueba de anillo en leche se determinó una prevalencia por hato de 52.38 por ciento con un rango de 42.84 por ciento 75 por ciento, siendo los extremos los municipios de Arcelia y Cutzamala, respectivamente. El mayor número de hatos positivos se encontró en el municipio de Coyuca (11 hatos); sin embargo, la mayor tasa se presentó en Cutzamala (75 por ciento) y Tlalchapa (66.67 por ciento). La prevalencia de la enfermedad por animal, mediante la prueba de aglutinación con antígeno Rosa de Bengala, fue de 16.72 por ciento, con rangos de 5.32 por ciento a 24.79 por ciento, correspondientes a los municipios de Cutzamala y Coyuca respectivamente. Con la prueba de Aglutinación Lenta en Tubo hubo una prevalencia de 17.03 por ciento con rangos de 7.45 por ciento para Cutzamala y de 30.30 por ciento para Altamirano. Las prevelencias encontradas son muy superiores a lo informado para la misma región por otros investigadores. Todas las muestras de leche analizadas resultaron negativas al aislamiento del microorganismo

Brucellosis in AL-Kharj regions of Saudi Arabia

The study determined the sensitivity, specificity and predictive value of serologic response in brucellosis by analysis of 150 consecutive positive serologic tests in Al-Kharj regions of Saudi Arabia. In one year 150 positive patients who had a titer of 1/160 or more were identified. Brucella species was recoved in 21% of blood cultures. The expected greater number of Brucella cultures with high recorded titer was confirmed; 15% of the positive high-titer group had positive Brucella cultures. The lower titers were unuseful as indicators of active disease in this population in which many people have persistent low antibody levels due to endemicity of the disease

Some hematological and haemostatic aspects in Egyptian patients with brucellosis

Haematologic and some haemostatic parameters were investigated in 38 Egyptian patients with brucellosis and 30 normal healthy subjects. The cases were diagnosed by blood culture and the brucella organisms were completely disappeared after treatment by2 weeks. Pretreatment results of haematologic investigations showed very highly significant decrease [P<0.001] in Hb concentration [gm/dl], RBCs count [Cram 3], blood indices [PCV and MCHC, total leucocytic count [Cmm 3], polymorphs% T and eosinophile%. with very highly significant increase [P<0.001] in monocytes% and erythrocyte sedimentation rate [ESR]. Two weeks after therapy, a highly significant decrease [Z> 2.58] was present in the percent of anaemia, leucopenia, thrornbocytopeniu, monocytosis and increase of ESR in brucella group compared to pre-treatment cases. The evaluated haemostatie function tests showed significant changes towards bleeding tendency [platelet count, platelet aggregation and fibrinogen showed a highly significant decrease [P<0.01] while prothrombin time and APTT were Significantly prolonged [p< 0.01] in brucella group. Also, a significant prolongation [P < 0.05] in bleeding time has been found in the same group. The haemostatic ahnormalities were improved after two weeks of treatment [platelet count, platelet aggregation and fibrinogen showed a significant difference [P< 0.05], while prothrombin time and APTT showed a highly significant decrease [P<0.001] compared to pre-treatment values. Also, a significant decrease in bleeding time has been observed in brucella group

study on brucellosis in Al-Anbar governorate

One hundred-twenty three cases of brucellosis were studied. The disease affected both sexes, all age groups, and different occupations. The study revealed that there were more cases from rural than urban areas, and children were less affected than adults and lowest infections were in Winter, The clinical features were similar to that described in the previously published series. All patients showed initial response to treatment with different regimens utillizing one or more of the aritibrucella drugs: tetracyclin, streptomycin. cotrimoxzole and rifampicin. The disease a public health problem which deserves registry and implementing measures of control

Brucellosis in Nineveah province

During the period from August to December 1989, blood and/or synovial specimens of 175 patients from Ninevah Province, were cultured into Brain-Heart Infusion [Difco B37]. The patients comprised 64 males [36.6%] and 111 females [63.4%] with mean age of 30.3 +/- 15.7 years who showed positive agglutinin for brucella. Brucella has been isolated from 124 [70.9%] cases. On the bases of C02 requirement of the isolates, cultural and microscopical characteristics, biochemical tests, dye sensitivity and the growth on a special medium, the majority of the isolates [77.4%] gave the characteristics of Br. abortus. Results of the culture were discussed in relation to that of the serology and other factors

Brucella arthritis in adults: report on forty eight patients

Awareness of the presence of brucellosis and its musculoskeletal manifestations has grown in Kuwait over the past 10 years. In this prospective study, 48 patients with Brucella arthritis were followed up for a period of 2 years or more. Treatment with a combination of tetracycline and streptomycin resulted in prompt recovery. Relapses occurred in 2 patients, but they recovered on being given a second course of treatment

Clinical, biochemical and hematological features of an outbreak of human brucellosis in Benghazi

During the outbreak of human brucellosis in Benghazi, Libya, in June-September 1987, haematological and biochemical parameters were analysed in 18 Libyan patients [age: 16:60 years; sex: 10 male, 8 female]. Since leukopenia with relative lymphocytosis was present in 5/18 [28%] patients only, it was not considered as a general rule for diagnosis of acute brucellosis. The ESR was raised in 14/18 [78%] patients which is generally a feature of acute brucellosis. Although biochemical analyses showed in some cases moderate elevation of serum ALT [8/18], AST [7/18], ALP [2/18] and LDH [5/18], serum ALT, AST and LDH were markedly elevated in only 5/18 [28%], 6/18 [33%] and 3/16 [18%] patients respectively. The implications of these findings and methods of choice for quick confirmed diagnosis of acute brucellosis as a possible cause for pyrexia of unknown origin are discussed