Genetic and Phenotypic Characterization of the Etiological Agent of Canine Orchiepididymitis Smooth Brucella sp. BCCN84.3

Members of the genus Brucella cluster in two phylogenetic groups: classical and non-classical species. The former group is composed of Brucella species that cause disease in mammals, including humans. A Brucella species, labeled as Brucella sp. BCCN84.3, was isolated from the testes of a Saint Bernard dog suffering orchiepididymitis, in Costa Rica. Following standard microbiological methods, the bacterium was first defined as "Brucella melitensis biovar 2." Further molecular typing, identified the strain as an atypical "Brucella suis." Distinctive Brucella sp. BCCN84.3 markers, absent in other Brucella species and strains, were revealed by fatty acid methyl ester analysis, high resolution melting PCR and omp25 and omp2a/omp2b gene diversity. Analysis of multiple loci variable number of tandem repeats and whole genome sequencing demonstrated that this isolate was different from the currently described Brucella species. The smooth Brucella sp. BCCN84.3 clusters together with the classical Brucella Glade and displays all the genes required for virulence. Brucella sp. BCCN84.3 is a species nova taxonomical entity displaying pathogenicity; therefore, relevant for differential diagnoses in the context of brucellosis. Considering the debate on the Brucella species concept, there is a need to describe the extant taxonomical entities of these pathogens in order to understand the dispersion and evolution

Global phylogenomic diversity of Brucella abortus: spread of a dominant lineage

Brucella abortus is a globally important zoonotic pathogen largely found in cattle hosts and is typically transmitted to humans through contaminated dairy products or contact with diseased animals. Despite the long, shared history of cattle and humans, little is known about how trade in cattle has spread this pathogen throughout the world. Whole genome sequencing provides unparalleled resolution to investigate the global evolutionary history of a bacterium such as B. abortus by providing phylogenetic resolution that has been unobtainable using other methods. We report on large-scale genome sequencing and analysis of B. abortus collected globally from cattle and 16 other hosts from 52 countries. We used single nucleotide polymorphisms (SNPs) to identify genetic variation in 1,074 B. abortus genomes and using maximum parsimony generated a phylogeny that identified four major clades. Two of these clades, clade A (median date 972 CE; 95% HPD, 781–1142 CE) and clade B (median date 150 BCE; 95% HPD, 515 BCE–164 CE), were exceptionally diverse for this species and are exclusively of African origin where provenance is known. The third clade, clade C (median date 949 CE; 95% HPD, 766–1102 CE), had most isolates coming from a broad swath of the Middle East, Europe, and Asia, also had relatively high diversity. Finally, the fourth major clade, clade D (median date 1467 CE; 95% HPD, 1367–1553 CE) comprises the large majority of genomes in a dominant but relatively monomorphic group that predominantly infects cattle in Europe and the Americas. These data are consistent with an African origin for B. abortus and a subsequent spread to the Middle East, Europe, and Asia, probably through the movement of infected cattle. We hypothesize that European arrival to the Americas starting in the 15th century introduced B. abortus from Western Europe through the introduction of a few common cattle breeds infected with strains from clade D. These data provide the foundation of a comprehensive global phylogeny of this important zoonotic pathogen that should be an important resource in human and veterinary epidemiology

Brucella Genetic Variability in Wildlife Marine Mammals Populations Relates to Host Preference and Ocean Distribution

Intracellular bacterial pathogens probably arose when their ancestor adapted from a free-living environment to an intracellular one, leading to clonal bacteria with smaller genomes and less sources of genetic plasticity. Still, this plasticity is needed to respond to the challenges posed by the host. Members of the Brucella genus are facultative-extracellular intracellular bacteria responsible for causing brucellosis in a variety of mammals. The various species keep different host preferences, virulence, and zoonotic potential despite having 97–99% similarity at genome level. Here, we describe elements of genetic variation in Brucella ceti isolated from wildlife dolphins inhabiting the Pacific Ocean and the Mediterranean Sea. Comparison with isolates obtained from marine mammals from the Atlantic Ocean and the broader Brucella genus showed distinctive traits according to oceanic distribution and preferred host. Marine mammal isolates display genetic variability, represented by an important number of IS711 elements as well as specific IS711 and SNPs genomic distribution clustering patterns. Extensive pseudogenization was found among isolates from marine mammals as compared with terrestrial ones, causing degradation in pathways related to energy, transport of metabolites, and regulation/ transcription. Brucella ceti isolates infecting particularly dolphin hosts, showed further degradation of metabolite transport pathways as well as pathways related to cell wall/membrane/envelope biogenesis and motility. Thus, gene loss through pseudogenization is a source of genetic variation in Brucella, whichinturn, relates to adaptation to different hosts.This is relevant to understand the natural history of bacterial diseases, their zoonotic potential, and the impact of human interventions such as domestication.Comisión Nacional para la Gestión de la Biodiversidad/[R-028-203-OT]/CONAGEBIO/Costa RicaMinisterio de Ciencia, Tecnología y Telecomunicaciones/[FV-004-13]/MICITT/Costa RicaWellcome Trust/[098051]/WT/LondresWellcome Trust/[106690/Z/14/Z]/WT/LondresUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias de la Salud::Centro de Investigación en Enfermedades Tropicales (CIET

Perspectivas de innovación en gestión, educación ambiental para la adaptación y la mitigación

Esta publicación del libro-foro sobre ciudad y cambio climático responde al aporte de los diferentes profesionales de las entidades públicas y privadas que participaron en calidad de conferencistas, ponentes, panelistas y expositores y compartieron sus experiencias en la ciudad como una contribución al conocimiento de las comunidades acerca de la creciente importancia y consideración de la adaptación y mitigación. Se consideraron acciones de políticas públicas por parte de las administraciones públicas, los sectores económicos y la sociedad, grupos ecológicos y fundaciones ecológicas y de igual forma las acciones y grandes esfuerzos realizados por el Ministerio del Ambiente, el IDEAM, la CAR, la Secretaría de Ambiente, el Jardín Botánico, la Red RAUS y de los grupos de investigación de las universidades

Brucella Genomics: Macro and Micro Evolution

Brucella organisms are responsible for one of the most widespread bacterial zoonoses, named brucellosis. The disease affects several species of animals, including humans. One of the most intriguing aspects of the brucellae is that the various species show a ~97% similarity at the genome level. Still, the distinct Brucella species display different host preferences, zoonotic risk, and virulence. After 133 years of research, there are many aspects of the Brucella biology that remain poorly understood, such as host adaptation and virulence mechanisms. A strategy to understand these characteristics focuses on the relationship between the genomic diversity and host preference of the various Brucella species. Pseudogenization, genome reduction, single nucleotide polymorphism variation, number of tandem repeats, and mobile genetic elements are unveiled markers for host adaptation and virulence. Understanding the mechanisms of genome variability in the Brucella genus is relevant to comprehend the emergence of pathogens

Animal welfare in biomedical research

El bienestar animal es un tema controversial que ha tomado especial interés en los últimos años, a la vez, lo coloca en la mira de la investigación biomédica. El uso de animales es de gran provecho para el avance científico en el campo de la salud; sin embargo, es muy cuestionado por sus implicaciones éticas. En este trabajo, se mencionan medidas útiles para disminuir el dolor y el estrés, procurar bienestar en los animales de investigación, y dar el mejor manejo a los individuos. Se mencionan algunas definiciones y cuestionamientos, desde el punto de vista ético, para el bienestar animal. Además, se cita y comenta la legislación nacional existente al respecto. A lo largo del trabajo, se enfatiza la importancia de conocer la especie con la cual se trabaja, la monitorización rutinaria, la educación y el respeto hacia los animales por parte de quienes los manipulan.Animal welfare represents a controversial topic that has attracted special interest in recent years, and, consequently, has been brought to the attention of biomedical research. The use of animals has greatly benefited scientific progress in the field of medical research. However, it has been widely questioned because of its ethical implications. Useful techniques are included in this article to decrease pain and stress in research animals, pursue their welfare, and improve their handling. The paper also includes some definitions and ethical issues to insure animal welfare. In addition, the national legislation is quoted and discussed. Emphasis is placed on the importance of knowing the species with which the researcher is working, routinely monitoring, and educating those who handle the animals and promoting respect for them.Universidad Nacional, Costa Rica.Escuela de Medicina Veterinari

Letter to the editor: It is erroneous to directly associate the detection of <i>Helicobacter</i> spp. with gastritis in canines

It has been proposed that Helicobacter may play a significant role in the generation of gastritis and cancer in dogs.  However, despite several independent efforts, there is still controversy regarding the true ability of Helicobacter to cause gastric lesions in these animals. The foregoing is due in part to the fact that gastritis is a multifactorial disease. Studies focused on naturally acquired infections have been unable to establish a positive correlation between Helicobacter colonization and gastritis.  Establishing a clear association between the presence of Helicobacter and gastritis may be difficult due to the following reasons: (i) taxonomy of the genus Helicobacter remains unclear, (ii) virulence factors of Helicobacter species that colonize dogs are not well known, and (iii) simultaneous infections have been reported with several species of Helicobacter. Current knowledge in taxonomy and pathogenesis of Helicobacter species is insufficient to differentiate between pathogenic and non-pathogenic strains.  Therefore, no direct association should be made between the detection of Helicobacter and gastritis

Animal welfare in biomedical research

El bienestar animal es un tema controversial que ha tomado especial interés en los últimos años, a la vez, lo coloca en la mira de la investigación biomédica. El uso de animales es de gran provecho para el avance científico en el campo de la salud; sin embargo, es muy cuestionado por sus implicaciones éticas. En este trabajo, se mencionan medidas útiles para disminuir el dolor y el estrés, procurar bienestar en los animales de investigación, y dar el mejor manejo a los individuos. Se mencionan algunas definiciones y cuestionamientos, desde el punto de vista ético, para el bienestar animal. Además, se cita y comenta la legislación nacional existente al respecto. A lo largo del trabajo, se enfatiza la importancia de conocer la especie con la cual se trabaja, la monitorización rutinaria, la educación y el respeto hacia los animales por parte de quienes los manipulan.Animal welfare represents a controversial topic that has attracted special interest in recent years, and, consequently, has been brought to the attention of biomedical research. The use of animals has greatly benefited scientific progress in the field of medical research.&nbsp; However, it has been widely questioned because of its ethical implications. Useful techniques are included in this article to decrease pain and stress in research animals, pursue their welfare, and improve their handling. The paper also includes some definitions and ethical issues to insure animal welfare.&nbsp; In addition, the national legislation is quoted and discussed. Emphasis is placed on the importance of knowing the species with which the researcher is working, routinely monitoring, and educating those who handle the animals and promoting respect for them

Tetraiodothyronine, triiodothyronine and cortisol serum levels in Costa Rican dogs using an immunoassay analyzer

Los intervalos de referencia hormonales varían según el método de medición empleado y el ambiente geoquímico donde las poblaciones se desarrollan. Por ende, cada laboratorio debe contar con intervalos de referencia propios para interpretar adecuadamente sus resultados. El objetivo de este trabajo fue establecer un intervalo de referencia para T4 total (TT4), T4 libre (T4L), T3 total (TT3) y cortisol en caninos de Costa Rica mediante el analizador automatizado AIA- 360® de Tosoh Bioscience. Se utilizó suero de 122 caninos clínicamente sanos, seleccionados al azar, para establecer los intervalos de referencia. Adicionalmente, se midió TT4, T4L y TT3 en 123 caninos con sintomatología sospechosa de hipotiroidismo, y se realizó la prueba de supresión con dexametasona a dosis baja en 12 caninos sospechosos de hiperadrenocorticismo. Las muestras se procesaron en el equipo AIA-360® luego de realizar las curvas de calibración respectivas según las indicaciones del fabricante. El intervalo de referencia se estableció con base en los percentiles 3 y 97 de los valores de caninos sanos: TT4 11,58 nmol/L - 50,19 nmol/L; T4L 6,18 pmol/L - 41,18 pmol/L; TT3 0,69 nmol/L - 1,64 nmol/L; cortisol 17,38 nmol/L - 174,64 nmol/L. La concentración de T4L fue estadísticamente menor (p<0,05) en los caninos sospechosos de hipotiroidismo. De los caninos sospechosos de hiperadrenocorticismo, 4 escaparon a la supresión y 7 suprimieron a las 4 horas post dexametasona, pero mostraron efecto “rebote” a las 8 horas posteriores. Actualmente se cuenta con un intervalo de referencia para TT4, TT3, T4L y cortisol en caninos mediante el uso del equipo AIA-360®.Hormonal reference intervals vary according the measurement method used and the geochemical environment where populations develop. Consequently, each lab must have its own reference intervals in order to interpret results properly. This paper was aimed at establishing a reference interval for total T4 (TT4), free T4 (FT4), total T3 (TT3) and cortisol in Costa Rican dogs, using the Tosoh Bioscience’s AIA-360® automatic analyzer. Blood serum from 122 randomly selected clinically healthy canines was used to determine reference intervals. Additionally, TT4, FT4 and TT3 were measured in 123 canines with suspected symptoms of hypothyroidism, and a low dose dexamethasone suppression test was performed in 12 canines with suspicious signs of hyperadrenocorticism. Samples were processed in the AIA-360® equipment after calibration curves were performed following manufacturer’s instructions. The reference interval was estimated based on the 3rd. and 97th. percentiles of healthy canines values: TT4 11.58 nmol/L – 50.19 nmol/L; FT4 6.18 pmol/L – 41.18 pmol/L; TT3 0.69 nmol/L – 1.64 nmol/L; cortisol 17.38 nmol/L – 174.64 nmol/L. FT4 concentration was statistically lower (p<0.05) in canines with suspected symptoms of hypothyroidism. Four of the canines with suspicious signs of hyperadrenocorticism did not suppress, while 7 suppressed at 4 hours after dexamethasone, but rebounded at 8 hours. Currently, a reference interval for TT4, FT4, TT3, and cortisol is available for Costa Rican canines using the AIA-360® analyzer.Universidad Nacional, Costa Rica.Escuela de Medicina Veterinari

Medición de hormonas tiroideas y cortisol en caballos mediante un analizador de inmunoensayo automatizado

Even though thyroid gland diseases are unusual in horses, it is important to have a reference interval for thyroid hormones in this species at every veterinary endocrine laboratory. In addition, cortisol and thyroid hormone measurement is useful in horses, not only for disease diagnosis, but as a tool for monitoring and researching performance and metabolic rate.The objectives of this study included (i) to test the capacity of the automated analyzer AIA 360® (TOSOH Bioscience) to measure thyroid hormones and cortisol in equine serum in a reliable way, and (ii) to establish a reference interval for thyroid hormones in Costa Rican horses using the AIA-360®.A total of 68 healthy and treatment free horses (31 males and 37 females) were sampled. Horses were grouped into three different categories according to their ages. Serum concentrations of total thyroxine (TT4), free thyroxine (FT4), and cortisol were quantified with the AIA-360®. Statistic relationships of hormonal values, gender and age were determined using Kruskal-Wallis, Dunn and Wilcoxon (U Man-Whitney) tests. P &lt;0.05 was considered significant.Selected reference intervals were: TT4 14.16 – 46.33 nmol/L, FT4 3.60 – 16.09 pmol/L and cortisol 23.45 – 166.64 nmol/L. No statistically significant relationship was found between gender, age, and hormone values. Besides, significant relationships (p&lt;0.05) were found between hormone levels and some environmental parameters. Animals living at higher altitudes showed higher cortisol levels, while cortisol decreases as the environmental temperature rises. Moreover, the FT4 levels are higher as precipitation increases. Finally, TT4 seems to be lower as day light length increases; however, no statistical relationship was found (p&lt;0.30).  Our results show that the use of the automated analyzer AIA-360® is suitable to measure thyroid hormones and cortisol, and that the values found are comparable with other studies and analysis techniques.A pesar de que las enfermedades tiroideas son poco comunes en equinos, es importante contar con un intervalo de referencia para hormonas tiroideas en cualquier laboratorio de endocrinología veterinaria. Asimismo, la medición de cortisol y hormonas tiroideas es útil en caballos, no solamente en el diagnóstico de enfermedades, sino que, ambos tipos de hormonas, son utilizadas como herramientas para monitorización e investigación del desempeño físico y la tasa metabólica.Los objetivos de este estudio son: (i) probar la capacidad del analizador automatizado AIA 360® (TOSOH Bioscience) para la medición de hormonas tiroideas y cortisol en suero equino de forma confiable y; (ii) establecer un intervalo de referencia para hormonas tiroideas en caballos costarricenses utilizando el AIA -360®.Se muestreó 68 caballos saludables, libres de medicación (31 machos y 37 hembras) y se agruparon en tres categorías, con base en la edad. Las concentraciones séricas de tiroxina total (TT4), tiroxina libre (FT4) y cortisol fueron cuantificadas en el AIA-360®. La relación estadística, entre sexo, edad y valores hormonales, se determinó con las pruebas de Kruskal-Wallis, Dunn y Wilcoxon (U Man-Whitney). Se consideró significativo un valor de p &lt;0.05.Los intervalos de referencia fueron los siguientes: TT4 14.16 – 46.33 nmol/L, FT4 3.60 – 16.09 pmol/L y cortisol 23.45 – 166.64 nmol/L. Estadísticamente, no se encontró relación significativa entre el sexo, la edad y los valores hormonales. Se observó relaciones significativas (p&lt;0.05) entre los niveles hormonales y algunos de los parámetros medio ambientales en los cuales se encontraban los animales.  A mayor altitud, los niveles de cortisol fueron más elevados; mientras que, conforme aumentaba la temperatura ambiental, los valores de cortisol fueron más bajos. Los niveles de FT4 incrementaron a mayor precipitación. La concentración de TT4 tiende a disminuir conforme aumentan las horas-luz diarias. Sin embargo, no se encontró una relación estadística para ello (p&lt;0.30). Los resultados muestran que el AIA-360® es útil para la medición de hormonas tiroideas y cortisol; además, que los valores encontrados son comparables con otros estudios y técnicas de análisis