New Bruce-ladder multiplex PCR assay for the biovar typing of Brucella suis and the discrimination of Brucella suis and Brucella canis

López-Goñi, Ignacio; García-Yoldi, David; Marín, Clara; Miguel, María Jesús de; Barquero-Calvo, Elías; Guzmán-Verri, Caterina; Albert, David; Garin‐Bastuji, Bruno

doi:10.1016/j.vetmic.2011.06.035

Cited by 118 publications

(75 citation statements)

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“…Seventeen isolates of Gram-negative bacteria compatible with B. canis were isolated from seminal fluid of stud males, from vaginal swabs of bitches, or from aborted fetuses between 2009 and 2013. The bacterial strains were characterized as B. canis by bacteriological analysis (3), molecular Bruce-ladder multiplex PCR assay, and multiplex single nucleotide polymorphism (SNP) detection as reported previously (31,32). One representative B. canis strain named bcanCR12 (here B. canis) isolated from a vaginal swab of a Pomeranian bitch after abortion was chosen for biological studies.…”

Section: Methodsmentioning

confidence: 99%

Brucella canis Is an Intracellular Pathogen That Induces a Lower Proinflammatory Response than Smooth Zoonotic Counterparts

et al. 2015

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e Canine brucellosis caused by Brucella canis is a disease of dogs and a zoonotic risk. B. canis harbors most of the virulence determinants defined for the genus, but its pathogenic strategy remains unclear since it has not been demonstrated that this natural rough bacterium is an intracellular pathogen. Studies of B. canis outbreaks in kennel facilities indicated that infected dogs displaying clinical signs did not present hematological alterations. A virulent B. canis strain isolated from those outbreaks readily replicated in different organs of mice for a protracted period. However, the levels of tumor necrosis factor alpha, interleukin-6 (IL-6), and IL-12 in serum were close to background levels. Furthermore, B. canis induced lower levels of gamma interferon, less inflammation of the spleen, and a reduced number of granulomas in the liver in mice than did B. abortus. When the interaction of B. canis with cells was studied ex vivo, two patterns were observed, a predominant scattered cell-associated pattern of nonviable bacteria and an infrequent intracellular replicative pattern of viable bacteria in a perinuclear location. The second pattern, responsible for the increase in intracellular multiplication, was dependent on the type IV secretion system VirB and was seen only if the inoculum used for cell infections was in early exponential phase. Intracellular replicative B. canis followed an intracellular trafficking route undistinguishable from that of B. abortus. Although B. canis induces a lower proinflammatory response and has a stealthier replication cycle, it still displays the pathogenic properties of the genus and the ability to persist in infected organs based on the ability to multiply intracellularly. Brucellosis is a disease of animals and humans caused by members of the genus Brucella. Zoonotic species such as Brucella melitensis, Brucella abortus, and Brucella suis are facultative extracellular-intracellular stealthy pathogens that are able to overcome innate immunity at early times of infection (1-3) and at specific stages of adaptive immunity (4, 5). In addition to influencing the immune response, these Brucella species are able to circumvent the killing action of professional and nonprofessional phagocytes, transit within phagocytic vacuoles, and replicate extensively within the endoplasmic reticulum of cells (6). These properties allow the bacterium to spread throughout the reticuloendothelial system and promote chronic infection (3).There are other Brucella species that are also relevant pathogens; however, their infective strategies remain unclear and are not in tune with the solid results accepted for the previously mentioned zoonotic brucellae. Among these are Brucella canis, the etiological agent of brucellosis in dogs and a zoonotic pathogen (7). This pathogen induces a subclinical infection that may remain undiagnosed for protracted periods (8-10). B. canis invades the conjunctiva or the oronasal system or penetrates through the venereal route. Then it is distributed to different organs...

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Section: Methodsmentioning

confidence: 99%

Brucella canis Is an Intracellular Pathogen That Induces a Lower Proinflammatory Response than Smooth Zoonotic Counterparts

et al. 2015

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“…Identification of the species within the genus was conducted using multiplex PCR known as the Bruce-ladder method [37]. Other multiplex PCR methods such as Suis-ladder also used in order to identify the correct Brucella species [38], (Figure 7).…”

Section: Brucellosis In Marine Mammals In the Republic Of Croatiamentioning

confidence: 99%

“…With the aim of differentiating the species B.ceti and B. pinnipedialis, additional primers were used [34,38,39]. Correct species could still not be determined with certainty.…”

Section: Brucellosis In Marine Mammals In the Republic Of Croatiamentioning

confidence: 99%

Brucellosis in marine mammals, with special emphasis on the Republic of Croatia

Cvetnić¹,

Duvnjak²,

Đuras³

et al. 2017

Rad Hrvatske akademije znanosti i umjetnosti

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SummaryAfter almost two decades of research and evidence of brucellosis in marine mammals, it has become widely known that this disease is prevalent in marine mammals on a global scale. Positive animals have been found in the northern Atlantic, Mediterranean Sea and in the Arctic, including the Barents Sea. Infected or exposed animals have been detected along the Atlantic and Pacific coasts of North America, off the coasts of Peru, Australia, New Zealand, Hawaii, the Solomon Islands, and in the Antarctic Ocean. Brucellosis most commonly occurs in the harbour porpoise (Phocoena phocoena), followed by the striped dolphin (Stenella coeruleoalba), Atlantic white-sided dolphin (Lagenorhynchus acutus), bottlenose dolphin (Tursiops truncatus), shortbeaked common dolphin (Delphinus delphis) and minke whale (Balaenoptera acutorostrata). A positive serological reaction has been confirmed in 35 species of whales and 14 species of seals, two subspecies of sea otters, one species of freshwater otter and the polar bear. To date, brucellosis has been found in marine mammals in the Mediterranean Sea in Italy and Spain. The first research on brucellosis in dolphins in the Republic of Croatia began in 2015. Brucella sp. was isolated from the lymph node of a bottlenose dolphin found dead in the Poreč region. This was the first evidence of brucellosis in dolphins in the Adriatic Sea. The isolated sample was identified as Brucella ceti strain ST27, making this the first record of this strain in Europe, which may present a significant threat to human health.

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“…Hoy en día es posible la detección simultánea de distintos patógenos en una muestra, utilizando lo que se denomina PCR múltiple. Es muy útil en el estudio y diferenciación de organismos relacionados, ejemplo Brucella (López-Goñi et al, 2011;López-Goñi et al, 2008), o en la diferenciación entre animales vacunados e infectados en planes de erradicación, por ejemplo en los casos de la Enfermedad de Auzjesky (ADV), la rinotraqueitis infecciosa o IBR (Mahajan et al, 2013;Schynts et al, 1999) o la Peste Porcina Clásica. La combinación de la PCR y su aná-lisis posterior con enzimas de restricción (PCR-RFLP) o la amplificación al azar de secuencias repetidas en el genoma (RAPDS) permiten el análisis y diferenciación de aislados virales, herramienta clave a la hora de llevar a cabo estudios epidemiológicos, determinar la virulencia de un virus o para tomar una decisión con respecto al tipo de vacuna que se debe utilizar.…”

Section: Diagnóstico Molecularunclassified

La biotecnología en sanidad animal

García

Casares

Moran

et al. 2014

Arbor

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RESUMEN: El crecimiento de la población mundial en los próxi-mos años, provocará un incremento en la demanda de alimentos. Como consecuencia, el número de animales de producción sufrirá un aumento importante y la Sanidad Animal constituirá un elemento crítico a la hora de tratar y prevenir las enfermedades, garantizando el abastecimiento y la seguridad. La biotecnología contribuye de múltiples formas al desarrollo de la Sanidad Animal. Fundamentalmente, aporta herramientas que ayudan al control y erradicación de las enfermedades. El desarrollo de la ingeniería genética en las últimas décadas ha favorecido el avance de la industria biotecnológica aplicada a la sanidad humana y animal. Existen un gran número de ensayos de diagnóstico basados en desarrollos biotecnológicos, así mismo, cada vez hay más vacunas recombinantes contra enfermedades animales. La prevención y control eficientes de las enfermedades dependerán de mecanismos de detección temprana, que faciliten la toma de decisiones y una respuesta rápida. PALABRAS CLAVE:Vacunación; VLPs; diagnóstico; inmunoensayos; DIVA; ensayos múltiples; ensayos de campo; PCR.ABSTRACT: The growth of the world population in forthcoming years will cause an increase in the demand of food. As a result, the number of working animals is set to rise significantly and Animal Health will become a vital issue when it comes to treating and preventing diseases, and guaranteeing and uninterrupted and safe food supply. Biotechnology contributes to the development of Animal Health in many ways. First and foremost, it provides tools that help control and eradicate diseases. The progress of genetic engineering in recent decades has underpinned the development of the biotechnology industry applied to human and animal health. There are a large number of diagnostic tests based on biotechnological developments, and there is also an increase in the use of recombinant vaccines against animal diseases. An effective prevention and control of diseases will depend on early detection mechanisms that facilitate decisionmaking and enable a rapid response. KEYWORDS:Vaccination; VLPs; diagnostic; immunoassays; DIVA; multiplex assays; field tests; PCR.

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New Bruce-ladder multiplex PCR assay for the biovar typing of Brucella suis and the discrimination of Brucella suis and Brucella canis

Cited by 118 publications

References 16 publications

Brucella canis Is an Intracellular Pathogen That Induces a Lower Proinflammatory Response than Smooth Zoonotic Counterparts

Brucella canis Is an Intracellular Pathogen That Induces a Lower Proinflammatory Response than Smooth Zoonotic Counterparts

Brucellosis in marine mammals, with special emphasis on the Republic of Croatia

La biotecnología en sanidad animal

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