Sectional evaluation of anatomic structures in cat (Felis catus) thoracic cavity by computed tomography imaging and silicone plastination methods / Evaluación seccional de estructuras anatómicas de la cavidad torácica en gato (Felis catus) con imágenes de tomografía computada y métodos de plastinación de silicona

It was aimed to determine the anatomical structures in thoracic cavity by computed tomography imaging (CT) and compare the cross sectional images in the same specimens which were plastinated after CT imaging. It was also aimed to obtain 3 dimensional (3D) reconstructions of thoracic anatomical structures. Thoracic organs of 3 adult cats were CT imaged and then plastinated in this study. Specimens were plastinated in the same body position in the CT imaging process. CT images and corresponding plastinated cross sections were compared to each other. Anatomical structures of the thoracic cavity in plastinates were in accordance with CT images. Beside the bony structures, other organs such as esophagus, trachea, heart with related vessels, lungs and thoracic muscles were well defined in CT images and plastinates. Moreover, 3D reconstructed images of anatomical structures of thoracic cavity were acquired well. This study is thought to be beneficial for veterinary surgery and radiology fields as well as veterinary anatomy educations.

El objetivo de este trabajo consistió en determinar las estructuras anatómicas en la cavidad torácica mediante tomografía computarizada (TC) y comparar las imágenes transversales en las mismas muestras, que fueron plastinadas después de la TC. También se pretendía obtener reconstrucciones tridimensionales (3D) de estructuras anatómicas torácicas. Se tomaron imágenes de los órganos torácicos de 3 gatos adultos por TC y luego se plastinaron en este estudio. Las muestras se plastinaron en la misma posición corporal en el proceso de obtención de imágenes TC. Las imágenes de TC y las secciones transversales plastinadas correspondientes se compararon entre sí. Las estructuras anatómicas de la cavidad torácica en los preparados plastinados estaban de acuerdo con las imágenes de CT. Además de las estructuras óseas, otros órganos como el esófago, la tráquea, el corazón con vasos relacionados, los pulmones y los músculos torácicos estaban bien definidos en las imágenes de TC y los plastinados. Por otra parte, se captaron bien las imágenes reconstruidas en 3D de las estructuras anatómicas de la cavidad torácica. Pensamos que este estudio es beneficioso para la cirugía veterinaria y los campos de radiología, así como también para la educación de anatomía veterinaria.

The use of singleplex and nested PCR to detect Batrachochytrium dendrobatidis in free-living frogs

Many microorganisms are able to cause diseases in amphibians, and in the past few years one of the most reported has been Batrachochytrium dendrobatidis. This fungus was first reported in Brazil in 2005; following this, other reports were made in specimens deposited in museum collections, captive and free-living frogs. The aim of this study was to compare singleplex and nested-PCR techniques to detect B. dendrobatidis in free-living and apparently healthy adult frogs from the Brazilian Atlantic Forest. The sample collection area was a protected government park, with no general entrance permitted and no management of the animals there. Swabs were taken from the skin of 107 animals without macroscopic lesions and they were maintained in ethanol p.a. Fungal DNA was extracted and identification of B. dendrobatidis was performed using singleplex and nested-PCR techniques, employing specific primers sequences. B. dendrobatidis was detected in 61/107 (57%) and 18/107 (17%) animals, respectively by nested and singleplex-PCR. Nested-PCR was statistically more sensible than the conventional for the detection of B. dendrobatidis (Chi-square = 37.1; α = 1%) and the agreement between both techniques was considered just fair (Kappa = 0.27). The high prevalence obtained confirms that these fungi occur in free-living frogs from the Brazilian Atlantic Forest with no macroscopic lesions, characterizing the state of asymptomatic carrier. We concluded that the nested-PCR technique, due to its ease of execution and reproducibility, can be recommended as one of the alternatives in epidemiological surveys to detect B. dendrobatidis in healthy free-living frog populations.

Perspectivas sobre salud pública veterinaria, seguridad alimentaria y la iniciativa conjunta "Una Salud" / Perspectives on veterinary public health, food security, and the "One Health" joint initiative

Los médicos veterinarios juegan un papel fundamental en la seguridad alimentaria. De su accionar eficiente y transparente depende la salud de millones de personas, la dinamización de las economías de los países, el desarrollo de una actividad pecuaria sustentable en torno los alimentos de este origen y los distintos sistemas agroproductivos que componen las cadenas de valor; y el acceso a mercados internacionales más rentables. Durante las próximas décadas, el cambio en los patrones nutricionales a nivel global, el aumento poblacional esperado, junto con el incremento de las rutas y los volúmenes comercializables de bienes alimenticios, hacen prever que la demanda de alimentos de origen animal se intensificará de forma constante. Para poder enfrentar con éxito estos retos, la profesión veterinaria deberá establecer marcos conceptuales y metodológicos de los perfiles académicos y profesionales más prácticos y actualizados, enfocando la profesión hacia los distintos campos temáticos que componen la salud pública, a nivel de los cursos de grado y posgrado. Por otra parte, deberá también elaborar alianzas interdisciplinarias como la propuesta bajo el enfoque "Una Salud" de la Organización de las Naciones Unidas para la Agricultura y la Alimentación (FAO), la Organización Mundial de Sanidad Animal (OIE) y la Organización Mundial de la Salud (OMS), para establecer marcos de trabajo conjuntos y políticas públicas más acordes con las realidades internas de los países latinoamericanos mediante el abordaje colaborativo, sostenible e integral de la sanidad animal, la seguridad alimentaria y las políticas sanitarias de salud pública.

Veterinarians play a key role in food security. The health of millions of people, stimulation of national economies, development of sustainable livestock production related to this food source, and the different agricultural production systems that compose value chains, and access to more profitable international markets all depend on their efficient and transparent work. Shifting nutritional patterns globally, along with expected population growth, and the increase in marketable food commodity routes and volumes, forecast that demand for animal source food will steadily intensify over the coming decades. To successfully address these challenges, the veterinary profession should establish more practical and up-to-date conceptual and methodological frameworks for academic and professional profiles, focusing the profession on the different public health subject areas, in undergraduate and graduate courses. Furthermore, interdisciplinary alliances should also be developed-such as the "One Health" approach proposed by the United Nations Food and Agriculture Organization (FAO), the World Organization for Animal Health (OIE), and the World Health Organization (WHO)-to establish frameworks for joint work and public policies more in line with the domestic conditions of Latin American countries, using a collaborative, sustainable, and comprehensive approach to animal health, food security, and public health policy.

Medios de contraste en ultrasonografía: agentes de contraste, características de las microburbujas, técnicas instrumentales, artefactos y usos en la clínica veterinaria / Contrast enhanced ultrasound: contrast agents, microbubble features, instrumental techniques, artefacts and usefulness in clinic veterinary

Un Agente de Contraste Ultrasonográfico es una sustancia compuesta por microburbujas de un gas de alto peso molecular encapsulado que tras la administración por vía endovenosa y ser incidida por los ultrasonidos empiezan a resonar generando señales retrodispersas que aumentan la señal acústica de la sangre. Los primeros agentes de contraste de ultrasonografía a base de solución salina agitada utilizados en un examen ecocardiográfico fueron el punto de inicio en el desarrollo de esta técnica de diagnóstico...

Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR

Post-mortem bacterial culture and specific biochemical tests are currently performed to characterize the etiologic agent of bovine tuberculosis. Cultures take up to 90 days to develop. A diagnosis by molecular tests such as PCR can provide fast and reliable results while significantly decreasing the time of confirmation. In the present study, a nested-PCR system, targeting rv2807, with conventional PCR followed by real-time PCR, was developed to detect Mycobacterium tuberculosis complex (MTC) organisms directly from bovine and bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other Actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. Regarding the analytical sensitivity, DNA of the M. bovis AN5 strain was detected up to 1.5 pg by nested-PCR, whereas DNA of M. tuberculosis H37Rv strain was detected up to 6.1 pg. The nested-PCR system showed 100% analytical specificity for MTC when tested with DNA of reference strains of non-tuberculous mycobacteria and closely-related Actinomycetales. A clinical sensitivity level of 76.7% was detected with tissues samples positive for MTC by means of the culture and conventional PCR. A clinical specificity of 100% was detected with DNA from tissue samples of cattle with negative results in the comparative intradermal tuberculin test. These cattle exhibited no visible lesions and were negative in the culture for MTC. The use of the nested-PCR assay to detect M. tuberculosis complex in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis.

Development and assessment of a latex agglutination test based on recombinant MSP5 to detect antibodies against Anaplasma marginale in cattle

The recombinant protein MSP5 has been established as an important antigen for serological diagnosis of Anaplasma marginale by enzyme-linked immunosorbent assay (ELISA). However, due to the high cost of specialized equipment, this technique is not accessible to all laboratories, especially in developing countries in areas where the disease is endemic. The present study describes the standardization of a latex agglutination test (LAT) to detect antibodies against A. marginale based on recombinant MSP5. Compared with indirect enzyme-linked immunosorbent assay (iELISA), the relative sensitivity and specificity of the LAT were 95.21% and 91.86% respectively, with an almost perfect agreement between tests (kappa index = 0.863). These results can be considered important for the serological diagnosis of A. marginale, as they indicate that the test represents a rapid and low cost alternative to ELISA.

Comparative analysis of conventional PCR and real-time PCR to diagnose shrimp WSD

The aims of this study were to standard and optimize a qPCR protocol with FAM-BHQ1 probe, and to compare its sensitivity against TaqMan qPCR and PCR methods to diagnose shrimp WSD. The FAM-BHQ1 qPCR presented higher clinical sensitivity and showed to be a robust alternative to detect WSSV in clinical samples.

Manejo nutricional de cães e gatos domiciliados no estado do Espírito Santo - Brasil / Feeding patterns of dogs and cats in Espírito Santo - Brazil

Obtiveram-se informações sobre o manejo nutricional adotado para cães e gatos domiciliados, avaliou-se o grau de conhecimento dos proprietários e correlacionaram-se esses dados com fatores socioeconômicos dos entrevistados. Um questionário foi respondido por 496 proprietários de cães e/ou gatos. Verificou-se que os proprietários reconhecem que a alimentação está diretamente relacionada com a saúde animal e se preocupam com a qualidade do alimento fornecido. A ração seca é considerada a melhor opção para a alimentação de cães e gatos e é o tipo de alimento mais utilizado pelos proprietários. Os cães são alimentados preferencialmente duas vezes ao dia, e os gatos recebem alimento à vontade. Os proprietários buscam informações sobre a alimentação dos animais principalmente com veterinários ou estudantes de veterinária e lojas agropecuárias ou pet shops. Em geral, os proprietários de maior escolaridade têm preferência pela ração seca. O custo com a alimentação dos animais é considerado alto, especialmente para os proprietários que fornecem rações terapêuticas. Dessa forma, não são todos que podem arcar com esse custo. Observou-se que os proprietários que fornecem esse tipo de ração são os que apresentam maior poder aquisitivo. O tipo de alimento fornecido ao animal, ração ou comida caseira, não é influenciado pelo ambiente no qual o proprietário reside (rural ou urbano).

In this study, further information on nutritional management performed by dog and cat owners was obtained, the level of owners' knowledge about pet nutrition was assessed, and some aspects with socioeconomic background of respondents were correlated. A survey was conducted with 496 dog and cat owners. According to the results, owners recognize that nutrition is closely related to animal health and they care about the quality of food provided. Commercial dry foods are considered the best option for feeding dogs and cats and it is the type of food used by most owners. Dogs are fed preferably twice a day and cats ad libitum. Owners seek information on pet nutrition especially with veterinarians or veterinary students and agricultural shops or pet shops. In general, owners with higher education have a preference for dry foods. The cost with pet food is considered high, especially for owners who provide therapeutic diets. Few owners can afford this cost. We found that owners who provide this type of diet are those with higher purchasing power. The type of diet given to pets, commercial or homemade food, is not influenced by the environment in which the owner resides (rural or urban).

Development of a Disperse Dye Immunoassay Technique for Detection of Antibodies against Neospora caninum in Cattle

In this study a disperse dye immunoassay method was standardized and evaluated for detection of antibodies against Neospora caninum in cattle. Sera from 150 cattle with a recent history of abortion were collected and tested by commercial ELISA kit and a standardized in-house dye immunoassay system. The positivity rate for the sera used in this study was 34.6% for the disperse dye immunoassay (DDIA) compared to 32% obtained by ELISA kit. This study showed no significant difference between DDIA and ELISA. The results indicated that the DDIA provide an economic, simple, rapid and robust test for detection of N. caninum infection in cattle.

A Multiplex real-time PCR for detection of Mycoplasma gallisepticum and Mycoplasma synoviae in clinical samples from Brazilian commercial poultry flocks

Mycoplasma gallisepticum (MS) and Mycoplasma synoviae (MS) are important avian pathogens and cause economic losses to the poultry industry. Molecular biology techniques are currently used for a rapid detection of these pathogens and the adoption of control measures of the diseases. The aim of this study was to develop and validate a technique for simultaneous detection of MG and MS by multiplex real time polymerase chain reaction (PCR). The complete assay (Multiplex MGMS) was designed with primers and probes specific for each pathogen and developed to be carried out in a single tube reaction. Vaccines, MG and MS isolates and DNA from other Mycoplasma species were used for the development and validation of the method. Further, 78 pooled clinical samples from different poultry flocks in Brazil were obtained and used to determine the sensitivity and specificity of the technique in comparison to 2 real time PCR assays specific for MG (MG PCR) and MS (MS PCR). The results demonstrated an agreement of 100% (23 positive and 44 negative samples) between Multiplex MGMS and MG PCR in the analysis of 67 samples from MG positive and negative poultry flocks, and an agreement of 96.9% between Multiplex MGMS and MS PCR in the analysis of 64 samples from MS positive and negative poultry flocks. Considering the single amplification tests as the gold standard, the Multiplex MGMS showed 100% of specificity and sensitivity in the MG analysis and 94.7% sensitivity and 100% specificity in the MS analysis. This new assay could be used for rapid analysis of MG and MS in the poultry industry laboratories.

Sensitive and Rapid Detection of Giardia lamblia Infection in Pet Dogs using Loop-Mediated Isothermal Amplification

Giardia lamblia is recognized as one of the most prevalent parasites in dogs. The present study aimed to establish a loop-mediated isothermal amplification (LAMP) assay for rapid and specific detection of G. lamblia from dogs. The fecal samples were collected and prepared for microscopic analysis, and then the genomic DNA was extracted directly from purified cysts. The concentration of DNA samples of G. lamblia were diluted by 10-fold serially ranging from 10(-1) to 10(-5) ng/microl for LAMP and PCR assays. The LAMP assay allows the amplification to be finished within 60 min under isothermal conditions of 63degrees C by employing 6 oligonucleotide primers designed based on G. lamblia elongation factor 1 alpha (EF1alpha) gene sequence. Our tests showed that the specific amplification products were obtained only with G. lamblia, while no amplification products were detected with DNA of other related protozoans. Sensitivity evaluation indicated that the LAMP assay was sensitive 10 times more than PCR. It is concluded that LAMP is a rapid, highly sensitive and specific DNA amplification technique for detection of G. lamblia, which has implications for effective control and prevention of giardiasis.

Estudio preliminar de patrones de mordedura según forma del cráneo, mediante el análisis morfológico y morfométrico de semiarcadas dentarias de perro doméstico (Canis familiaris) con fines de identificación / Preliminary study of bite patterns according to skull shape, by morphological and morphometric analysis of dental semi arcs of domestic dog (Canis familiaris) for identification

La Odontología forense es una rama de las Ciencias Forenses que se utiliza para establecer la identidad de una víctima o victimario mediante el estudio de las huellas de mordeduras, estimación de edad cronológica, entre otros. Dada la inexistencia de este tipo de estudios en Chile y la magnitud que alcanza el problema de mordidas de perros en Chile, se compararon los patrones de huellas de mordedura según forma del cráneo, mediante el análisis morfológico y morfométrico de semiarcadas dentarias de perro doméstico (Canis familiaris) con fines médico legal. Para ello se obtuvieron modelos dentarios en yeso e impresiones de huellas de mordedura en cera de tres razas puras: Bóxer, Dálmata y Ovejero Alemán. Las mediciones contemplaron la distancia máxima externa entre caninos superiores e inferiores y la distancia máxima externa entre incisivos superiores e inferiores. Los principales resultados muestran que sólo la distancia máxima entre caninos superiores permitiría identificar la raza en forma estadísticamente significativa (p0,05) en los modelos de yeso. En el estudio ciego y desde el punto de vista morfométrico se identificaron especialmente los Dálmata con un alto grado de sensibilidad y especificidad. Un poco menor y en forma variable se observó en Bóxer y Ovejero Alemán. El estudio morfológico permitió identificar e individualizar al 100 por ciento de los perros. Por tanto los métodos morfométricos y morfológicos son herramientas complementarias para identificar e individualizar a un posible perro agresor.

Forensic dentistry is a branch of Forensic Sciences used to establish the identity of a crime victim or perpetrator, among these studies are bite marks and dental chronological age studies. In the absence of such studies in Chile and considering the widespread problem of dog bites in Chile, bite patterns were compared according to skull shape, by morphological and morphometric analysis of dental semi arcs of domestic dog (Canis Familiaris) for medico legal purposes. Plaster dental models and impressions of bite marks were obtained of three pure breeds: Boxer, Dalmatian and German shepherd. Measures included he maximum external distance between upper and lower canine, and maximum external distance between upper and lower incisors. Main results show that only the maximum distance between upper canine would allow statistically significant race identification, (p0.05) in cast models. In a blind study and from a morphometric point of view, Dalmatians were identified with a high rate of sensitivity and specificity. A somewhat lesser and variable form was observed in Boxer and German Shepherd. The morphological study allowed identification and individualization of 100 percent of dogs. Therefore, both morphological and morphometric methods are useful and complementary tools to identify and individualize a potentially aggressive dog.

Molecular Identification and Real-time Quantitative PCR (qPCR) for Rapid Detection of Thelohanellus kitauei, a Myxozoan Parasite Causing Intestinal Giant Cystic Disease in the Israel Carp

Intestinal giant-cystic disease (IGCD) of the Israel carp (Cyprinus carpio nudus) has been recognized as one of the most serious diseases afflicting inland farmed fish in the Republic of Korea, and Thelohanellus kitauei has been identified as the causative agent of the disease. Until now, studies concerning IGCD caused by T. kitauei in the Israel carp have been limited to morphological and histopathological examinations. However, these types of diagnostic examinations are relatively time-consuming, and the infection frequently cannot be detected in its early stages. In this study, we cloned the full-length 18S rRNA gene of T. kitauei isolated from diseased Israel carps, and carried out molecular identification by comparing the sequence with those of other myxosporeans. Moreover, conventional PCR and real-time quantitative PCR (qPCR) using oligonucleotide primers for the amplification of 18S rRNA gene fragment were established for further use as methods for rapid diagnosis of IGCD. Our results demonstrated that both the conventional PCR and real-time quantitative PCR systems applied herein are effective for rapid detection of T. kitauei spores in fish tissues and environmental water.

Development and Clinical Evaluation of a Rapid Serodiagnostic Test for Toxoplasmosis of Cats Using Recombinant SAG1 Antigen

Rapid serodiagnostic methods for Toxoplasma gondii infection in cats are urgently needed for effective control of transmission routes toward human infections. In this work, 4 recombinant T. gondii antigens (SAG1, SAG2, GRA3, and GRA6) were produced and tested for the development of rapid diagnostic test (RDT). The proteins were expressed in Escherichia coli, affinity-purified, and applied onto the nitrocellulose membrane of the test strip. The recombinant SAG1 (rSAG1) showed the strongest antigenic activity and highest specificity among them. We also performed clinical evaluation of the rSAG1-loaded RDT in 182 cat sera (55 household and 127 stray cats). The kit showed 0.88 of kappa value comparing with a commercialized ELISA kit, which indicated a significant correlation between rSAG1-loaded RDT and the ELISA kit. The overall sensitivity and specificity of the RDT were 100% (23/23) and 99.4% (158/159), respectively. The rSAG1-loaded RDT is rapid, easy to use, and highly accurate. Thus, it would be a suitable diagnostic tool for rapid detection of antibodies in T. gondii-infected cats under field conditions.