DNA Sensor for the Detection of Brucella spp. Based on Magnetic Nanoparticle Markers.

Due to the limitations of conventional Brucella detection methods, including safety concerns, long incubation times, and limited specificity, the development of a rapid, selective, and accurate technique for the early detection of Brucella in livestock animals is crucial to prevent the spread of the associated disease. In the present study, we introduce a magnetic nanoparticle marker-based biosensor using frequency mixing magnetic detection for point-of-care testing and quantification of Brucella DNA. Superparamagnetic nanoparticles were used as magnetically measured markers to selectively detect the target DNA hybridized with its complementary capture probes immobilized on a porous polyethylene filter. Experimental conditions like density and length of the probes, hybridization time and temperature, and magnetic binding specificity, sensitivity, and detection limit were investigated and optimized. Our sensor demonstrated a relatively fast detection time of approximately 10 min, with a detection limit of 55 copies (0.09 fM) when tested using DNA amplified from Brucella genetic material. In addition, the detection specificity was examined using gDNA from Brucella and other zoonotic bacteria that may coexist in the same niche, confirming the method's selectivity for Brucella DNA. Our proposed biosensor has the potential to be used for the early detection of Brucella bacteria in the field and can contribute to disease control measures.

Intracellular growth of Brucella is mediated by Dps-dependent activation of ferritinophagy.

Bacteria of the genus Brucella cause brucellosis, one of the world's most common zoonotic diseases. A major contributor to Brucella's virulence is the ability to circumvent host immune defense mechanisms. Here, we find that the DNA-binding protein Dps from Brucella is secreted within the macrophage cytosol, modulating host iron homeostasis and mediating intracellular growth of Brucella. In addition to dampening iron-dependent production of reactive oxygen species (ROS), a key immune effector required for immediate bacterial clearance, cytosolic Dps mediates ferritinophagy activation to elevate intracellular free-iron levels, thereby promoting Brucella growth and inducing host cell necrosis. Inactivation of the ferritinophagy pathway by Ncoa4 gene knockout significantly inhibits intracellular growth of Brucella and host cell death. Our study uncovers an unconventional role of bacterial Dps, identifying a crucial virulence mechanism used by Brucella to adapt to the harsh environment inside macrophages.

Biodegradation of monocrotophos by Brucella intermedia Msd2 isolated from cotton plant.

Widespread and inadequate use of Monocrotophos has led to several environmental issues. Biodegradation is an ecofriendly method used for detoxification of toxic monocrotophos. In the present study, Msd2 bacterial strain was isolated from the cotton plant growing in contaminated sites of Sahiwal, Pakistan. Msd2 is capable of utilizing the monocrotophos (MCP) organophosphate pesticide as its sole carbon source for growth. Msd2 was identified as Brucella intermedia on the basis of morphology, biochemical characterization and 16S rRNA sequencing. B. intermedia showed tolerance of MCP up to 100 ppm. The presence of opd candidate gene for pesticide degradation, gives credence to B. intermedia as an effective bacterium to degrade MCP. Screening of the B. intermedia strain Msd2 for plant growth promoting activities revealed its ability to produce ammonia, exopolysaccharides, catalase, amylase and ACC-deaminase, and phosphorus, zinc and potassium solubilization. The optimization of the growth parameters (temperatures, shaking rpm, and pH level) of the MCP-degrading isolate was carried out in minimal salt broth supplemented with MCP. The optimal pH, temperature, and rpm for Msd2 growth were observed as pH 6, 35 °C, and 120 rpm, respectively. Based on optimization results, batch degradation experiment was performed. Biodegradation of MCP by B. intermedia was monitored using HPLC and recorded 78% degradation of MCP at 100 ppm concentration within 7 days of incubation. Degradation of MCP by Msd2 followed the first order reaction kinetics. Plant growth promoting and multi-stress tolerance ability of Msd2 was confirmed by molecular analysis. It is concluded that Brucella intermedia strain Msd2 could be beneficial as potential biological agent for an effective bioremediation for polluted environments.

Investigation of the outbreaks of abortions and orchitis in livestock in Namibia during 2016-2018.

This study investigated outbreaks of seemingly related abortions and orchitis which occurred in the Khomas, Omaheke and Otjozondjupa regions of Namibia from 2016 to 2018, affecting cattle, sheep and goats. Fifty-nine questionnaires were administered, and 48 were completed giving an 81.4% return. The outbreaks were limited to Namibia's east and central regions, mainly on farms rearing cattle, sheep and goats and on farms with a mixture of these species. There was no significant difference between Khomas and other regions on abortion reporting at the farm level [X2 (1, N = 48) = 0.0002, p = 0.987851]. However, there was a significant difference in the abortions at the animal level among the three regions [X2 (2, N = 6246) = 239.8339, p = .00001]. In addition, the proportions of abortions calculated at the animal level at each farm were significantly different when the Khomas region was compared to the other regions. Seventeen cattle sera, 35 sheep sera, 52 caprine sera, 18 bovine liver samples, one caprine liver, five aborted cattle foetuses, two cattle placentas, 18 testes (one bull, eight bucks and nine rams) and ten bull sheath scrapings were collected and tested. Histopathology, microbiology, serology, immunohistochemistry, real-time PCR and mineral analytical techniques were used to establish the aetiology of the abortion and orchitis outbreaks. The gross and histopathological findings on the 18 testicles were characteristic of chronic orchitis. In aborted foetuses, significant histopathological findings included meconium aspiration, funisitis and cardiomyopathy. Placentitis and endometritis were the primary pathologies observed in cows. The bacteria isolated from microbiological samples included Enterococcus spp. (65.5% [19/29]), Enterobacter spp. (6.9% [2/29]) and Streptococcus spp. (10.3% [3/29]), Trueperella pyogenes (3.4% [1/29]), Stenotrophomonas maltophilia (3.4% [1/29]), Staphylococcus epidermidis (3.4% [1/29]), Providencia rettgeri (3.4% [1/29]) and Acinetobacter lwoffii (3.4% [1/29]), mostly opportunistic bacteria. On mineral analysis, 28%, 33%, 83%, 33% and 17% (n = 18) of cattle livers were low in copper, zinc, manganese, selenium and iron, respectively. Twenty-three percent (12/52) of the caprine sera were positive for Brucella melitensis on the Rose Bengal and complement fixation tests. Thirty-five ovine sera were tested for B. melitensis, B. ovis and Coxiella burnetii, and the prevalence for each was 2.9% (1/35). PCR tests on foetuses were all negative for Brucella spp., Coxiella burnetii, Chlamydia spp., Listeria monocytogenes, Salmonella spp., Campylobacter fetus spp., Leptospira pathogenic strains, bovine viral diarrhoea virus, Rift Valley fever virus, Anaplasma phagocytophilum and bovine herpes virus 4 Campylobacter fetus spp. and Trichomonas foetus spp. The authors concluded that Brucella spp., Enterococcus spp., Escherichia coli, Streptococcus spp., Trueperella pyogenes and Coxiella burnetii could have contributed to this outbreak. Micronutrient imbalances and pathogenic abiotic nanoparticles were also identified as possible contributors to the abortion outbreaks.

Proteomics-based screening and antibiotic resistance assessment of clinical and sub-clinical Brucella species: An evolution of brucellosis infection control.

Brucellae are intracellular sneaky bacteria and they can elude the host's defensive mechanisms, resulting in therapeutic failure. Therefore, the goal of this investigation was to rapid identification of Brucella species collected from animals and humans in Saudi Arabia, as well as to evaluate their resistance to antibiotics. On selective media, 364 animal samples as well as 70 human blood samples were cultured. Serological and biochemical approaches were initially used to identify a total of 25 probable cultured isolates. The proteomics of Brucella species were identified using the MALDI Biotyper (MBT) system, which was subsequently verified using real-time polymerase chain reaction (real-time PCR) and microfluidic electrophoresis assays. Both Brucella melitensis (B. melitensis) and Brucella abortus (B. abortus) were tested for antimicrobial susceptibility using Kirby Bauer method and the E-test. In total, 25 samples were positive for Brucella and included 11 B. melitensis and 14 B. abortus isolates. Twenty-two out of 25 (88%) and 24/25 (96%) of Brucella strains were recognized through the Vitek 2 Compact system. While MBT was magnificently identified 100% of the strains at the species level with a score value more than or equal to 2.00. Trimethoprim-sulfamethoxazole, rifampin, ampicillin-sulbactam, and ampicillin resistance in B. melitensis was 36.36%, 31.82%, 27.27%, and 22.70%, respectively. Rifampin, trimethoprim-sulfamethoxazole, ampicillin, and ampicillin-sulbactam resistance was found in 35.71%, 32.14%, 32.14%, and 28.57% of B. abortus isolates, correspondingly. MBT confirmed by microfluidic electrophoresis is a successful approach for identifying Brucella species at the species level. The resistance of B. melitensis and B. abortus to various antibiotics should be investigated in future studies.

A Carbonic Anhydrase Pseudogene Sensitizes Select Brucella Lineages to Low CO2 Tension.

Brucella spp. are intracellular pathogens that cause a disease known as brucellosis. Though the genus is highly monomorphic at the genetic level, species have animal host preferences and some defining physiologic characteristics. Of note is the requirement for CO2 supplementation to cultivate particular species, which confounded early efforts to isolate B. abortus from diseased cattle. Differences in the capacity of Brucella species to assimilate CO2 are determined by mutations in the carbonic anhydrase gene, bcaA Ancestral single-nucleotide insertions in bcaA have resulted in frameshifted pseudogenes in B. abortus and B. ovis lineages, which underlie their inability to grow under the low CO2 tension of a standard atmosphere. Incubation of wild-type B. ovis in air selects for mutations that "rescue" a functional bcaA reading frame, which enables growth under low CO2 and enhances the growth rate under high CO2 Accordingly, we show that heterologous expression of functional Escherichia coli carbonic anhydrases enables B. ovis growth in air. Growth of B. ovis is acutely sensitive to a reduction in CO2 tension, while frame-rescued B. ovis mutants are insensitive to CO2 shifts. B. ovis initiates a gene expression program upon CO2 downshift that resembles the stringent response and results in transcriptional activation of its type IV secretion system. Our study provides evidence that loss-of-function insertion mutations in bcaA sensitize the response of B. ovis and B. abortus to reduced CO2 tension relative to that of other Brucella lineages. CO2-dependent starvation and virulence gene expression programs in these species may influence persistence or transmission in natural hosts.IMPORTANCEBrucella spp. are highly related, but they exhibit differences in animal host preference that must be determined by genome sequence differences. B. ovis and the majority of B. abortus strains require high CO2 tension to be cultivated in vitro and harbor conserved insertional mutations in the carbonic anhydrase gene, bcaA, which underlie this trait. Mutants that grow in a standard atmosphere, first reported nearly a century ago, are easily selected in the laboratory. These mutants harbor varied indel polymorphisms in bcaA that restore its consensus reading frame and rescue its function. Loss of bcaA function has evolved independently in the B. ovis and B. abortus lineages and results in a dramatically increased sensitivity to CO2 limitation.

The Application of a Nanomaterial Optical Fiber Biosensor Assay for Identification of Brucella Nomenspecies.

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 IS711 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.

Antimicrobial Resistance in Leptospira, Brucella, and Other Rarely Investigated Veterinary and Zoonotic Pathogens.

Leptospira, Brucella, and Borrelia are major agents of zoonotic disease, causing high morbidity and, in some cases, significant mortality in humans. For all three genera, prompt diagnosis and appropriate antimicrobial therapy are required to prevent the development of chronic, debilitating illness. Leptospira spp. are intrinsically resistant to several antimicrobial classes; however, there is little evidence in the literature for development of acquired resistance to antimicrobial agents used for clinical treatment of acute leptospirosis. For Brucella infections, there are numerous reports of relapses following therapy, but it is unclear whether this is due to sequestration within infected sites (e.g., bone) or the development of acquired resistance. Brucella have maintained their susceptibility to doxycycline and rifampicin, which in combination remain the most common treatments of brucellosis in humans. In vitro induced point mutations are described as imparting resistance to rifampicin (rpoB) and fluoroquinolones (gyrA). The clinical significance of these mutations is unclear. For Borrelia burgdorferi, although acquired resistance to some antimicrobial agents has been described, resistance due to bacterial persister cells surviving in the presence of antimicrobial, with no apparent increase in the MIC of the organism, have been recently described. Of the remaining veterinary fastidious pathogens, Lawsonia intracellularis is the most interesting from an antimicrobial resistance perspective because it can only be grown in cell culture, making in vitro susceptibility testing challenging. MIC testing has been undertaken on a small number of isolates, and some differences in susceptibility to macrolides have been demonstrated between isolates obtained from different regions.

Brucella - Virulence Factors, Pathogenesis and Treatment.

Brucellae are Gram-negative, small rods infecting mammals and capable of causing disease called brucellosis. The infection results in abortion and sterility in domestic animals (sheeps, pigs, rams etc). Especially dangerous for humans are: Brucella melitensis, Brucella suis, Brucella abortus, and Brucella canis that trigger unspecific symptoms (flu-like manifestation). Brucella rods are introduced via host cells, by inhalation, skin abrasions, ingestion or mucosal membranes. The most important feature of Brucella is the ability to survive and multiply within both phagocytic and non-phagocytic cells. Brucella does not produce classical virulence factors: exotoxin, cytolisins, exoenzymes, plasmids, fimbria, and drug resistant forms. Major virulence factors are: lipopolysaccharide (LPS), T4SS secretion system and BvrR/BvrS system, which allow interaction with host cell surface, formation of an early, late BCV (Brucella Containing Vacuole) and interaction with endoplasmic reticulum (ER) when the bacteria multiply. The treatment of brucellosis is based on two-drug therapy, the most common combinations of antibiotics are: doxycycline with rifampicin or fluoroquinolones with rifampicin. Currently, also other methods are used to disrupt Brucella intracellular replication (tauroursodeoxycholic acid or ginseng saponin fraction A).

DISEASE COMPLEXITY IN A DECLINING ALASKAN MUSKOX (OVIBOS MOSCHATUS) POPULATION.

The muskox ( Ovibos moschatus ) population inhabiting the eastern North Slope (ENS) of Alaska, US declined dramatically during 1999-2006, whereas populations in western Alaska (WA) were stable or increasing. To understand morbidity and mortality factors contributing to the decline, Alaska Department of Fish and Game conducted pathologic investigations of carcasses from 2005 until 2008. Additionally, archived sera from both ENS and WA muskoxen collected during 1984-92, before the documented beginning of the ENS decline; sera collected during 2000, near the beginning of the decline; and contemporary sera (from live capture-release, adult females) collected during 2006, 2007, and 2008 were analyzed to determine whether prevalence of antibody to potential pathogens differed in the two areas or changed over time. The pathogens investigated were those that were believed could cause lameness or poor reproduction or adversely affect general health. Furthermore, trace mineral levels, hemograms, and gastrointestinal parasites were evaluated in live adult females captured 2006-08. Pathologic investigations identified several comorbid conditions, including predation, polyarthritis caused by or consistent with Chlamydophila spp. infection, hoof lesions, copper deficiency, contagious ecthyma, verminous pneumonia, hepatic lipidosis suggestive of negative energy balance, and bacterial bronchopneumonia due to Trueperella pyogenes and Bibersteinia trehalosi . Pathogens suspected to be newly introduced in the ENS muskox population on the basis of serologic detection include bovine viral diarrhea, respiratory syncytial virus, Chlamydophila spp., Brucella spp., Coxiella burnetii , and Leptospira spp., whereas parainfluenza virus-3 antibody prevalence has increased in the WA population. Although multiple disease syndromes were identified that contributed to mortality and, in combination, likely limited the ENS muskox population, further holistic investigations of disease agents, trace mineral status, and nutritional factors in conjunction with intensive demographic and environmental analyses would provide a better understanding of factors that influence Alaskan muskox populations.

Historical perspective of brucellosis: a microbiological and epidemiological overview.

The historical process of brucellosis extends back to humankind's first contact with animals. Although brucellosis is a sporadic disease observed in animals in certain regions of the world, it is an important disease in humans that can affect many organs and systems due to the consumption of contaminated milk or milk products. Studies have shown that the presence of Brucella dates back to 60 million years ago. In 450 BC, Hippocrates described a disease similar to brucellosis. Since Hippocrates' time, brucellosis has been characterized by fever. Our aim is to investigate selfless work undertaken by scientists on the epidemiology, diagnosis and clinical findings of brucellosis until today, and to gain a historical perspective about the disease that is as old as human history, still has importance today, causes economic losses in treated animals and harms human health.

Inhibitory effect of red ginseng acidic polysaccharide from Korean red ginseng on phagocytic activity and intracellular replication of Brucella abortus in RAW 264.7 cells

Korean red ginseng (KRG) has long been used in traditional Korean and Oriental medicine. However, the anti-bacterial mechanism and therapeutic efficiency of KGR for intracellular Brucella infection are still unclear. In this study, the bactericidal activity of Korean red ginseng acidic polysaccharide (RGAP) on Brucella (B.) abortus and its cytotoxic effects on RAW 264.7 cells were evaluated. In addition, B. abortus internalization and intracellular replication in macrophages were investigated after RGAP treatment. RGAP-incubated cells displayed a marked reduction in the adherence, internalization and intracellular growth of B. abortus in macrophages. Furthermore, decreased F-actin fluorescence was observed relative to untreated B. abortus-infected cells. Western blot analysis of intracellular signaling proteins revealed reduced ERK, JNK and p38α phosphorylation levels in B. abortus-infected RGAP-treated cells compared to the control. Moreover, elevated co-localization of B. abortus-containing phagosomes with lysosome-associated membrane protein 1 (LAMP-1) were observed in RGAP-treated cells compared with the control. Overall, the results of this study suggest that RGAP can disrupt phagocytic activity of B. abortus via suppression of mitogen-activated protein kinases (MAPKs) signaling proteins ERK, JNK and p38 levels and inhibit intracellular replication of B. abortus by enhancing phagolysosome fusion, which may provide an alternative control of brucellosis.

Herd-level risk factors for Campylobacter fetus infection, Brucella seropositivity and within-herd seroprevalence of brucellosis in cattle in northern Nigeria.

Brucellosis and campylobacteriosis are economically important diseases affecting bovine reproductive efficiency in Nigeria. A questionnaire-based survey was conducted in 271 cattle herds in Adamawa, Kaduna and Kano states of northern Nigeria using multistage cluster sampling. Serum from 4745 mature animals was tested for Brucella antibodies using the Rose-Bengal plate test and positives were confirmed in series-testing protocol using competitive enzyme-linked immunosorbent assay. Preputial scrapings from 602 bulls were tested using culture and identification for Campylobacter fetus. For each disease, a herd was classified as positive if one or more animals tested positive. For each herd, information on potential managemental and environmental risk factors was collected through a questionnaire administered during an interview with the manager, owner or herdsman. Multiple logistic regression models were used to model the odds of herd infection for each disease. A zero-inflated Poisson model was used to model the count of Brucella-positive animals within herds, with the number tested as an exposure variable. The presence of small ruminants (sheep and/or goats) on the same farm, and buying-in of >3 new animals in the previous year or failure to practice quarantine were associated with increased odds of herd-level campylobacteriosis and brucellosis, as well as increased within-herd counts of Brucella-positive animals. In addition, high rainfall, initial acquisition of animals from markets, practice of gynaecological examination and failure to practice herd prophylactic measures were positively associated with the odds of C. fetus infection in the herd. Herd size of >15, pastoral management system and presence of handling facility on the farm were associated with increased odds, and gynaecological examination with reduced odds of herd-level Brucella seropositivity. Furthermore, the zero-inflated Poisson model showed that borrowing or sharing of bulls was associated with higher counts, and provision of mineral supplement with lower counts of Brucella-positive cattle within herds. Identification of risk factors for bovine campylobacteriosis and brucellosis can help to identify appropriate control measures, and the use of zero-inflated count model can provide more specific information on these risk factors.

Serum levels of copper, selenium and manganese in forestry workers testing IgG positive for Brucella, Borrelia, and Rickettsia.

The aim of this study is to measure the alterations in the trace levels of serum copper (Cu), selenium (Se), and manganese (Mn) in forestry workers testing immunoglobulin G (IgG)-positive for Brucella, Borrelia, and Rickettsia. The study was conducted on a sample of 758 subjects (560 male and 198 female). All the subjects underwent medical examinations, which investigated particularly the presence of clinical signs compatible with zoonoses, and routine blood tests from venous blood sample, which tested previous immunisation versus cited microorganisms and serum concentration of Cu, Se, and Mn. The subjects were divided according to IgG positivity versus the cited microorganisms. The group of subjects with IgG positive versus Brucella showed statistically significant higher Cu levels than controls, while the Mn levels were not; the group of subjects with IgG positive versus Rickettsia showed higher levels of all three tested metals. The concentration of the examined metals did not show statistically significant difference between IgG-positive subjects versus subjects with Borrelia compared to controls. These data could confirm the role of both Cu and Se  in the regulation of immune response.

Prevalencia de brucelosis en columna: Evolución clínica y manejo quirúrgico / Prevalência de brucelose na coluna vertebral: Evolução clínica e conduta cirúrgica / Prevalence of brucellosis in the spine: Clinical course and surgical management

OBJETIVO: Conocer la prevalencia, características clínicas y de tratamiento de la espondilodiscitis por Brucella. MÉTODOS: Un total de 104 pacientes con diagnóstico de espondilitis espinal de los cuales 20 correspondieron a espondilitis por Brucella, tratados de forma conservadora con antibióticos o quirúrgicamente, mediante descompresión y estabilización de la columna en cualquier segmento, en el periodo comprendido entre Enero 2006 a Junio 2011. Se elaboró una hoja de captura para recolección de la información, la cual contenía la identificación del paciente, síntomas referidos, signos físicos, parámetros laboratoriales y tratamiento médico y quirúrgico. RESULTADOS: Trece pacientes del sexo masculino y 7 del sexo femenino, con edad promedio de 59,2 años y desviación estándar de 13,6 años. Tres pacientes tenían afectación del segmento torácico, 13 del segmento lumbar y 4 del segmento lumbo-sacro. Trece pacientes presentaron fiebre, el dolor lumbar se presentó en los 20 pacientes, parestesias en 12, debilidad en 9, artralgias, mialgias y anorexia en 5, radiculopatía en 13, cefalea en 1 y pérdida de peso en 5 pacientes. Veinte casos recibieron tratamiento médico con doxiciclina combinada con rifampicina. En 8 casos se realizó instrumentación posterior combinada con laminectomía y fusión póstero-lateral, en 2 se realizó instrumentación posterior combinada con discectomía del nivel afectado y fusión póstero-lateral y en 1 caso se realizó laminectomía combinada con discectomía y fusión. CONCLUSIÓN: La prevalencia de espondilodiscitis fue del 19,2%, siendo el dolor lumbar y la fiebre síntomas con alto índice de sospecha de espondilitis espinal. La descompresión combinada con estabilización y fusión es el tratamiento quirúrgico de elección.

OBJETIVO: Conhecer a prevalência, as características clínicas e de tratamento da espondilodiscite por Brucella. MÉTODOS: Um total de 104 pacientes com diagnóstico de espondilite espinal, dos quais 20 corresponderam a espondilite por Brucella, tratados de forma conservadora com antibióticos ou cirurgicamente, por descompressão e estabilização da coluna em qualquer segmento, no período de janeiro de 2006 a junho de 2011. Foi elaborado um formulário de dados para coleta das informações, contendo identificação do paciente, sintomas relatados, sinais físicos, parâmetros laboratoriais e tratamento médico e cirúrgico. RESULTADOS: Treze pacientes do sexo masculino e 7 do sexo feminino, com média de idade de 59,2 anos e desvio padrão de 13,6 anos. Três pacientes foram acometidos no segmento torácico, 13 no segmento lombar e 4 no segmento lombossacral. Treze pacientes tiveram febre, a dor lombar estava presente nos 20 pacientes, parestesias em 12, debilidade em 9, artralgias, mialgias e anorexia em 5, radiculopatia em 13, cefaleia em um e perda de peso em 5 pacientes. Vinte casos receberam tratamento clínico com doxiciclina combinada com rifampicina. Em 8 casos, realizou-se instrumentação posterior combinada com laminectomia e fusão póstero-lateral, em 2 realizou-se instrumentação posterior combinada com discectomia do nível afetado e fusão póstero-lateral e em um caso, procedeu-se à laminectomia combinada com discectomia e fusão. CONCLUSÃO: A prevalência de espondilodiscite foi de 19,2%, sendo a dor lombar e a febre sintomas com alto índice de suspeita de espondilite espinal. A descompressão combinada com estabilização e fusão é o tratamento cirúrgico de escolha.

OBJECTIVE: To determine the prevalence, clinical characteristics, and treatment of spondylodiscitis due to Brucella. METHODS: A total of 104 patients diagnosed with spinal spondylitis, of which 20 corresponded to Brucella spondylitis, treated conservatively with antibiotics or surgically by decompression and stabilization of the spine in any segment, from January 2006 to June 2011. We created a data sheet for collecting information containing patient identification, reported symptoms, physical signs, laboratory parameters and medical and surgical treatment. RESULTS: Thirteen patients were males and 7 females, with a mean age of 59.2 years and standard deviation of 13.6 years. Three patients were affected in the thoracic segment, 13 in the lumbar and lumbosacral segment, 4. Thirteen patients had fever, low back pain was present in the 20 patients, in 12 there were paresthesias, weakness in 9, arthralgias, myalgias and anorexia in 5, radiculopathy in 13, headache, and weight loss in 5 patients. Twenty cases received medical treatment with doxycycline combined with rifampicin. In 8 cases, posterior instrumentation combined with laminectomy and posterolateral fusion was performed, on 2, posterior instrumentation combined with discectomy and fusion of the affected level posterolateral was held and in one case, laminectomy and discectomy combined with fusion was performed. CONCLUSION: The prevalence of spondylodiscitis was 19.2%, with lower back pain and fever symptoms with high suspicion of spinal spondylitis. The decompression combined with stabilization and fusion is the treatment of choice.

Efficacy of prolonged antimicrobial chemotherapy for brucellar spondylodiscitis.

The standard treatment of brucellar spondylitis with a combination of two antibiotics for 6-12 weeks is associated with high rates of treatment failure and relapse. The present study aimed to assess the safety and efficacy of a treatment strategy based on the prolonged administration of a triple combination of suitable antibiotics. Eighteen patients with brucellar spondylitis were treated with a combination of at least three suitable antibiotics (doxycycline, rifampin, plus intramuscular streptomycin or cotrimoxazole or ciprofloxacin) until the completion of at least 6 months of treatment, when clinical, radiological and serology re-evaluation was performed. If necessary, the treatment was continued with additional 6-month cycles, until resolution or significant improvement of clinical and radiological findings, or for a maximum of 18 months. At presentation, the median age was 66 years (range, 42-85 years) with male predominance. The median duration of therapy was 48 weeks (range 24-72 weeks). Treatment was discontinued early because of side-effects in only one patient. Surgical intervention was required for three patients. At the end of treatment all patients had a complete response. After completion of treatment, all patients were followed up with regular visits. During the follow-up period (duration 1-96 months, median 36.5 months), no relapses were observed. In conclusion, prolonged (at least 6 months) administration of a triple combination of suitable antibiotics appears to be an effective treatment for brucellar spondylitis.

Neurobrucellosis with thalamic infarction: a case report.

Brucellosis is prevalent in North and East Africa, the Middle East, South and Central Asia, South and Central America and the Mediterranean countries of Europe. In 5% of cases it may lead to central nervous system manifestation presenting most often as a meningitis or meningoencephalitis. Here we report and discuss a case of neurobrucellosis with meningitis with cranial nerves neuritis. A 56-year-old German male was admitted with bilateral abducens nerve palsy, amblyacousia and intractable headaches. An enzyme-linked immunosorbent assay (ELISA) revealed antibodies against Brucella in serum and cerebrospinal fluid (CSF). Additionally, our patient suffered from an infarction of the left thalamus. In conclusion, in cases of unresponsive meningitis or meningoencephalitis and history of travel in endemic regions, neurobrucellosis should be considered. If initial microbiological tests fail, complementary investigations such as ELISA are indicated to detect Brucella species in serum and/or CSF.

Orquiepididimitis por Brucella SPP / Brucella Spp orchiepididymitis

OBJETIVO: Presentar un caso de orquiepididimitis unilateral por Brucella y realizar una pequeña revisión de diagnóstico y tratamiento de la patología. MÉTODO: Presentamos a un varón de 62 años que refiere síndrome constitucional, fiebre y dolor testicular, el diagnóstico se basó en la anamnesis, hemocultivos y estudios de imagen ecográfica. RESULTADOS: El tratamiento se inició con Doxiciclina y Teicoplanina, manteniéndose el primero durante seis semanas y sustituyéndose el segundo por ciprofloxacino (dos semanas en total) al alta, con resolución completa del cuadro. CONCLUSIÓN: Se debe ser exhaustivo en la anamnesis del paciente con orquitis infecciosa, ya que ésta puede revelar datos sugerentes de la etiología del cuadro, siendo de especial relevancia en la afectación por Brucella, ya que se presenta en pacientes de riesgo. Sin embargo se antoja necesaria la realización de hemocultivos que revelen Brucella en sangre para confirmar el diagnóstico. Los tratamientos antibióticos orales actuales son más que suficientes para resolver el cuadro. Siendo el tratamiento quirúrgico excepcional hoy en día (AU)

OBJECTIVE: To report a brucellar orchie-pididymitis case and to review the diagnosis and treatment of this pathology. METHOD: We present the case of a 62 year old man presenting with fever, testicular pain and constitutional syndrome. The diagnosis was made after ultrasound, blood cultures and anamnesis. RESULTS: The treatment was Doxiciclin six weeks and teico-planin, the second one was replaced with ciprofloxacin. CONCLUSION: It is important to make a correct anamnesis to the patient with orchiepididymitis to identify any risk factor for brucellosis. The diagnosis is confirmed with blood cultures. The oral antibiotics are enough to cure patients (AU)

Quinolones for brucellosis: treating old diseases with new drugs.

Although quinolones are theoretically interesting candidates for the treatment of brucellosis, the existing data concerning their efficacy are limited and conflicting. A number of small clinical studies with combination regimens that include quinolones have shown adequacy, but not superiority, although cost-effectiveness, excluding certain disease complications, is an important issue. The emergence of quinolone resistance and its implications is another drawback. Experimental data have yielded contradictory results, although most studies do not indicate a bactericidal effect for quinolones. However, in-vitro studies contrast repeatedly with the clinical response, both in terms of clinical failure, despite in-vitro success, and vice versa.

Susceptibility of Mexican brucella isolates to moxifloxacin, ciprofloxacin and other antimicrobials used in the treatment of human brucellosis.

Brucellosis is a disease of domestic and wild animals that is transmitted to humans and exists worldwide. We assessed the in vitro activity of moxifloxacin, ciprofloxacin, tetracycline, doxicycline, rifampin, streptomycin and trimethoprim-sulfamethoxazole (TMP/SMX) against 97 Brucella strains isolated from clinical samples, animals and dairy products in Mexico. Fluoroquinolones showed an antibacterial activity similar to that of tetracyclines (MIC(90) 0.5). Other drugs commonly used against brucellosis were less active, such as rifampin (MIC(90) 2.0 microg/ml) and streptomycin (MIC(90) 4.0 microg/ml). TMP/SMX showed the poorest activity (MIC(90) 8.0 microg/ml). Fluoroquinolones, either first-generation or the newer 8-methoxi derivatives, might be useful in the therapy of brucellosis, which remains to be assessed in clinical trials.