Detection of mecC-Methicillin-resistant Staphylococcus aureus isolates in river water : a potential role for water in the environmental dissemination

Methicillin-resistant Staphylococcus aureus (MRSA) is a public health concern due to limited treatment options. The recent description of a mecA homologue, mecC in human and cattle, led to studies to detect this new variant in human and other animal species. Detection of mecC in wild boar and fallow deer in a Spanish game estate led us to further investigate the presence of mecC-MRSA at this location. Samples from cattle, wild animals, workers and river water were tested. A further three mecC-MRSA isolates were obtained from river water. Molecular characterization (multilocus sequence typing and spa typing) and antimicrobial susceptibility testing (broth microdilution) showed that isolates were similar to those detected in wild animals. Whole genome sequencing confirmed that the isolates from the river water and wild animals in the same geographic area were all closely related isolates of ST425 mecC-MRSA. The presence of mecC-MRSA in the river water highlights the potential role of water in the dissemination of mecC-MRSA

Oral Vaccination with Heat-Inactivated Mycobacterium bovis Does Not Interfere with the Antemortem Diagnostic Techniques for Tuberculosis in Goats

Vaccination against tuberculosis (TB) is prohibited in cattle or other species subjected to specific TB eradication campaigns, due to the interference that it may cause with the official diagnostic tests. However, immunization with a heat-inactivated (HI) Mycobacterium bovis vaccine via the oral route has been suggested to overcome this issue. In this study, the main goal was to assess the interference of the HI vaccine by different routes of administration using a previous vaccination and re-vaccination (boosting) protocol. TB-free kid goats were divided into three groups: oral (n = 16), intramuscular (IM; n = 16), and control (n = 16). Results showed that there was a significant difference in the percentage of animals positive to the single intradermal test (SIT) and blood based interferon-gamma release assay (IGRA) caused by vaccination when performed in the IM group compared to the oral group (p < 0.001). Nevertheless, no positivity to the SIT or IGRA test was observed in orally vaccinated goats regardless of the different interpretation criteria applied. None of the groups presented positive antibody titers using an in-house ELISA and samples collected 2 months after the boost. These results suggest the potential usefulness of the HI vaccine by the oral route in goats to minimize the interference on diagnostic tests (skin and IGRA tests) and reducing the necessity of defined antigens to replace the traditional purified protein derivatives for diagnosis. Finally, the results pave the way to future efficacy studies in goats using different routes of HI vaccination

Staphylococcus aureus carrying mecC gene in animals and urban wastewater, Spain

Letter to the edito

Hypervitaminosis D has no positive effects on goat tuberculosis and may cause chronic renal lesions

[Background]: There is evidence for a link between vitamin D deficiency and active tuberculosis (TB). In human beings, several trials have evaluated the role of vitamin D supplementation in TB treatment with conflicting results. However, the role of vitamin D supplementation in animal TB control has received less attention. The authors evaluated the benefit of vitamin D supplementation for preventing mycobacterial infection or reducing TB lesions (TBL) in a controlled trial with goats naturally exposed to Mycobacterium caprae. [Methods]: Two groups of goats, a vitamin D-supplemented group and a non-supplemented control group, were housed for 10 months in direct contact with M caprae-infected adult goats. Upon contact with the infected adult goats, all animals were TB-tested every two months. [Results]: No experimental evidence of a protective effect of vitamin D supplementation based on M caprae culture prevalence, TBL prevalence, median TBL score or the proportion of single versus multiple organs presenting TBL was observed. [Conclusion]: The results indicate that, in the conditions used in this study, vitamin D supplementation in goats does not reduce TB infection risk nor the diffusion and severity of TBL. In addition, vitamin D-supplemented goats presented hyperphosphataemia and renal injury with calcifications suggestive of vitamin D intoxication.This work was supported by the Ministerio de Economía, Industria y Competitividad of Spain (RTC-2016-4746-2) and the Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria of Spain (RTA2015-00043-C02-02). AR is the recipient of an Industrial Doctorate contract (DI-15-08110) and JB holds a ‘Juan de la Cierva program’ contract (IJCI-2015-24805), both funded by the Spanish Ministry of Science, Innovation and Universities

Molecular detection of Tritrichomonas foetus in bovine samples: a novel real-time polymerase chain reaction (PCR) assay targeting EF1-alpha-Tf1 and a comparative study of published PCR techniques

The parasite T. foetus causes trichomonosis in cattle but is generally asymptomatic in males. Thus, many bulls carrying the disease go unnoticed, making the detection of T. foetus in bulls an important aspect for its control. Due to drawbacks posed by its cultivation, PCR is a preferred option for diagnostic laboratories. Most published PCR protocols target the genomic region compring the 18S, 5.8S, and 28S rRNA genes and internal transcribed spacers 1 and 2 (rRNA-ITS region), homologous to that of other Tritrichomonas species. There is minimal information on alternative genetic targets and no comparative studies have been published. We compared a protocol based on the microsatellite TfRE (called H94) and five protocols based on the rRNA-ITS region (called M06, M15, G02, G05, and N02). We also designed and evaluated a novel PCR-based assay on the EF1-alpha-Tf1 gene (called V21). The analytical sensitivity and specificity assays for the PCR protocols were performed according to the World Organisation for Animal Health (OIE) directives and the comparative study was performed with a widely used PCR (M06) on clinical samples from 466 breeding bulls. V21 showed a high degree of agreement with our reference M06 (kappa = 0.967), as well as M15 (kappa = 0.958), G05 (kappa = 0.948), and H94 (kappa = 0.986). Protocols H94 and V21 appear to be good approaches for confirming clinical cases in preputial bull samples when genomic regions alternative to rRNA-ITS are required. By contrast, N02 gave false negatives and G02 false positives

Liver Transudate, a Potential Alternative to Detect Anti-Hepatitis E Virus Antibodies in Pigs and Wild Boars (Sus scrofa)

In recent years, cases of hepatitis E virus (HEV) infection have increased in Europe in association with the consumption of contaminated food, mainly from pork products but also from wild boars. The animal’s serum is usually tested for the presence of anti-HEV antibodies and viral RNA but, in many cases such as during hunting, an adequate serum sample cannot be obtained. In the present study, liver transudate was evaluated as an alternative matrix to serum for HEV detection. A total of 125 sera and liver transudates were tested by enzyme-linked immunosorbent assay at different dilutions (1:2, 1:10, 1:20), while 58 samples of serum and liver transudate were checked for the presence of HEV RNA by RT-qPCR. Anti- HEV antibodies were detected by ELISA in 68.0% of the serum samples, and in 61.6% of the undiluted transudate, and in 70.4%, 56.8%, and 44.8% of 1:2, 1:10, or 1:20 diluted transudate, respectively. The best results were obtained for the liver transudate at 1:10 dilution, based on the Kappa statistic (0.630) and intraclass correlation coefficient (0.841). HEV RNA was detected by RT-qPCR in 22.4% of the serum samples and 6.9% of the transudate samples, all samples used for RT-qPCR were positive by ELISA. Our results indicate that liver transudate may be an alternative matrix to serum for the detection of anti-HEV antibodies

Bovine tuberculosis in Spain, is it really the final countdown?

Abstract Bovine tuberculosis (bTB) is a severe zoonotic disease that has major impacts on both health and the economy, and which has been subjected to specific eradication programmes in many countries for decades. This manuscript highlights the relevance of this disease in the context of the European Union (EU) and summarizes the epidemiological situation and the main tools (e.g. antemortem diagnostic tests, slaughterhouse surveillance, laboratories, comprehensive databases, etc.) used to control and eradicate bTB in the various EU countries with a focus on the situation in Spain. A comprehensive description of the specific bTB epidemiological situation in Spain is provided, together with an assessment of the evolution of different epidemiological indicators throughout the last decades. Moreover, the main features of the Spanish bTB eradication programme and its control tools are described, along with the studies carried out in Spain that have allowed the updating of and improvement to the programme over the years with the aim of eradication, which has been established for 2030

Liver Transudate, a Potential Alternative to Detect Anti-Hepatitis E Virus Antibodies in Pigs and Wild Boars (Sus scrofa)

In recent years, cases of hepatitis E virus (HEV) infection have increased in Europe in association with the consumption of contaminated food, mainly from pork products but also from wild boars. The animal’s serum is usually tested for the presence of anti-HEV antibodies and viral RNA but, in many cases such as during hunting, an adequate serum sample cannot be obtained. In the present study, liver transudate was evaluated as an alternative matrix to serum for HEV detection. A total of 125 sera and liver transudates were tested by enzyme-linked immunosorbent assay at different dilutions (1:2, 1:10, 1:20), while 58 samples of serum and liver transudate were checked for the presence of HEV RNA by RT-qPCR. Anti- HEV antibodies were detected by ELISA in 68.0% of the serum samples, and in 61.6% of the undiluted transudate, and in 70.4%, 56.8%, and 44.8% of 1:2, 1:10, or 1:20 diluted transudate, respectively. The best results were obtained for the liver transudate at 1:10 dilution, based on the Kappa statistic (0.630) and intraclass correlation coefficient (0.841). HEV RNA was detected by RT-qPCR in 22.4% of the serum samples and 6.9% of the transudate samples, all samples used for RT-qPCR were positive by ELISA. Our results indicate that liver transudate may be an alternative matrix to serum for the detection of anti-HEV antibodies

"Ludificación" para la enseñanza del método científico en Epidemiología y ciencias afines: una actualización

Siepsa Toolkit es un paquete de herramientas informáticas para la enseñanza de la Epidemiología y ciencias afines en Ciencias de la Salud” desarrollado con financiación obtenida en anteriores convocatorias de los PIMCD. En este proyecto se ha hecho una revisión de las funcionalidades de Siepsa desarrollando nuevos supuestos. Los supuesto se pueden crear tanto en español como en inglés y la aplicación permite la selección entre ambos idiomas. El paquete consta de dos herramientas: Siepsa, la aplicación del alumno/a o “juego”, y Siepsa Profesor que permite crear “juegos” de forma sencilla sin conocimiento de programación. El “juego” Siepsa plantea al alumno/a un escenario en el que se están entrenando para pertenecer al “Centro Superior de Investigaciones Epidemiológicas (C.S.I.E.)” y para alcanzar el objetivo deben resolver una investigación epidemiológica. El programa está estructurado en sucesivos niveles de “juego” que se corresponden con las etapas de cualquier investigación (objetivos, diseño, realización, análisis de resultados, conclusiones y difusión) y que el alumno/a supera contestando correctamente a las preguntas que se le plantean hasta resolver el objetivo principal de la investigación. No permite retroceder y cambiar respuestas, de forma que los errores van contabilizando en un único marcador por partida. El juego se inicia con diez vidas y hay un indicador de vidas en la parte inferior derecha de la pantalla en cada partida. Cada vez que se reinicia se carga una nueva partida en la que cambian de forma aleatoria el orden de las respuestas. El programa Siepsa Profesor permite al profesorado crear tantos supuestos como desee con la temática o el nivel de complejidad adecuado a su alumnado, sin necesidad de tener conocimientos de programación. Por una parte se crea la interfaz gráfica. Se crean las preguntas, las respuestas posibles y las explicaciones que aparecen el feedback tanto a las correctas como a las incorrectas. Una parte fundamental de Siepsa profesor es la creación de variables y las condiciones de que se aplicarán en la creación de la datos de la base de datos de forma aleatoria de modo que reflejen los resultados que se obtendrían en una investigación real. Siepsa Profesor crea las condiciones de la base de datos para Siepsa, pero también puede crear la base de datos de forma independiente para ser utilizada en la docencia. Durante el desarrollo de este proyecto se han diseñado y creado distintas bases de datos con distintos grados de complejidad que se han probado y utilizado especialmente en la enseñanza en másteres