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Con el auge de los grandes modelos del lenguaje neuronales, especialmente aquellos basados en Transformers, la tradicional segmentación en palabras y morfemas que empleaba reglas lingüísticas ha sido reemplazada por algoritmos de segmentación estadísticos. Estos algoritmos son mucho más eficientes y, sin necesidad de intervención humana, son capaces de, a partir de corpus de millones de palabras, construir el vocabulario de palabras y subpalabras que necesitan los grandes modelos del lenguaje monolingües o multilingües. Ocurre, sin embargo, que estas subpalabras no se corresponden siempre con morfemas y esto repercute negativamente en el funcionamiento de los modelos del lenguaje que utilizan estos segmentadores. Cuánto se alejan los vocabularios estadísticos de un vocabulario real de palabras y morfemas de una lengua –lo que denominamos calidad morfológica del vocabulario–, y cuánto repercute esta falta de calidad en la eficacia de los grandes modelos del lenguaje son cuestiones todavía sin resolver. Este artículo aborda la primera cuestión, la calidad morfológica de los vocabularios, aportando un método de evaluación basado en tres medidas de calidad –relevancia, coherencia y corrección morfológica–, y un procedimiento para evaluarlas. El método se aplica para medir la calidad de los vocabularios generados por tres algoritmos de segmentación en subpalabras, BPE, WordPiece y Unigram, utilizados mayoritariamente para la construcción de los grandes modelos del lenguaje. Los resultados que hemos obtenido indican que la calidad morfológica de los mismos es muy baja, por lo que merece la pena buscar nuevas soluciones para mejorar la calidad de los vocabularios de los grandes modelos del lenguaje.

Abstract The World Health Organization has highlighted that cancer was the second-highest cause of death in 2019. This research aims to present the current forecasting techniques found in the literature, applied to predict time-series cancer incidence and then, compare these results with the current methodology adopted by the Instituto Nacional do Câncer (INCA) in Brazil. A set of univariate time-series approaches is proposed to aid decision-makers in monitoring and organizing cancer prevention and control actions. Additionally, this can guide oncological research towards more accurate estimates that align with the expected demand. Forecasting techniques were applied to real data from seven types of cancer in a Brazilian district. Each method was evaluated by comparing its fit with real data using the root mean square error, and we also assessed the quality of noise to identify biased models. Notably, three methods proposed in this research have never been applied to cancer prediction before. The data were collected from the INCA website, and the forecast methods were implemented using the R language. Conducting a literature review, it was possible to draw comparisons previous works worldwide to illustrate that cancer prediction is often focused on breast and lung cancers, typically utilizing a limited number of time-series models to find the best fit for each case. Additionally, in comparison to the current method applied in Brazil, it has been shown that employing more generalized forecast techniques can provide more reliable predictions. By evaluating the noise in the current method, this research shown that the existing prediction model is biased toward two of the studied cancers Comparing error results between the mentioned approaches and the current technique, it has been shown that the current method applied by INCA underperforms in six out of seven types of cancer tested. Moreover, this research identified that the current method can produce a biased prediction for two of the seven cancers evaluated. Therefore, it is suggested that the methods evaluated in this work should be integrated into the INCA cancer forecast methodology to provide reliable predictions for Brazilian healthcare professionals, decision-makers, and oncological researchers.

Ankle osteoarthritis (OA) is characterized by intraarticular, activity-related pain. mainly with weight-bearing activities. Several nonoperative therapies remain as the first choices, but there is still no consensus on which is the most effective. This study aimed to systematically synthesize the evidence regarding the effectiveness of PRP in the management of ankle OA. Searches were conducted on PubMed, Cochrane, CINAHL, and SPORTDiscus to find relevant articles from inception to December 2023. Details regarding study characteristics, PRP procedures, and outcomes were extracted. A quality assessment was developed according to Joanna Briggs Institute’s critical appraisal tool. A total of five studies met the inclusion criteria, with 184 patients receiving PRP therapy (mean age 55.7 years, and 51.1% were female). There were three case series, one prospective cohort, and one randomized controlled trial. Pain and function were the most frequently evaluated outcomes among the selected articles. They showed significant improvements in the short-term follow-ups, mostly in patients with II-III-stage ankle OA. There is currently insufficient evidence regarding the effectiveness and safety of PRPs in the treatment of ankle OA. A paucity of high-level research in the literature was also found.

Fires affect forest dynamics in seasonally dry regions such as the Mediterranean Basin. There, fire impacts on tree growth have been widely characterized in conifers, particularly pine species, but we lack information on broadleaf tree species that sprout after fires. We investigated post-fire radial growth responses in two coexisting Mediterranean hardwood species (the evergreen Quercus ilex, the deciduous Celtis australis) using tree-ring width data. We compared growth data from burnt and unburnt stands of each species subjected to similar climatic, soil and management conditions. We also calculated climate–growth relationships to assess if burnt stands were also negatively impacted by water shortage, which could hinder growth recovery. Tree-ring data of both species allowed us to quantify post-fire growth enhancements of +39.5% and +48.9% in Q. ilex and C. australis, respectively, one year after the fire. Dry spring climate conditions reduced growth, regardless of the fire impact, but high precipitation in the previous winter enhanced growth. High June radiation was negatively related to the growth of unburnt Q. ilex and burnt C. australis stands, respectively. Post-fire growth enhancement lasted for five years after the fire and it was a transitory effect because the growth rates of burnt and unburnt stands were similar afterwards.

Background: Chagas disease (CD), a Neglected Tropical Disease caused by Trypanosoma cruzi, affects millions of people in Latin America and the southern US and spreads worldwide. CD results from close interactions between humans, animals, and vectors, influenced by sociodemographic factors and housing materials. Methods: This study aimed to evaluate how these factors, along with seasonal changes, affect the distribution of CD vectors in an endemic community near Puebla, Mexico, using a cross-sectional survey. A total of 383 people from this area, known for the presence of major vectors such as Triatoma barberi and Triatoma pallidipennis, were surveyed. Results: As a result of the survey, it was found that only 27.4% of respondents knew about CD, and 83.3% owned potential reservoir pets; additionally, the quality of the wall, roof, and floor significantly influenced vector sightings, while the seasonal pattern showed less of an association. Chi-square tests confirmed these associations between vector sightings and housing materials (p < 0.001); vector sightings versus seasonal patterns showed less of an association (p = 0.04), and land use changes did not show an association (p = 0.27). Conclusions: Construction materials play an important role in the sighting of triatomines in homes, so important actions should be taken to improve homes. However, further experimental or longitudinal studies are needed to establish causality.

The purpose of this systematic review was to produce a best evidence synthesis of the effects of Kinesiology Tape (KT) in the treatment of tendinopathies. An electronic search on five databases (PubMed, Cumulative Index to Nursing and Allied Health Literature (CINAHL), Cochrane Library, SportDiscus, and Physiotherapy Evidence Database (PEDro)) was conducted. Studies were included if (1) patients suffered from tendinopathy; (2) isolated KT was applied in at least one group; (3) comparisons between other techniques were developed; (4) outcomes based on pain, function, disability, or quality of life were analysed. Two reviewers independently extracted data and assessed methodological quality using the Physiotherapy Evidence Database (PEDro) scale. A total of 13 articles met the eligibility criteria, involving 454 participants. Nine of these studies presented upper extremity tendinopathies, and four explored lower extremity tendinopathies. Selected papers ranged from low to high quality, with an average score of 5 on the PEDro scale. According to our findings, there is limited evidence to support KT alone for the treatment of tendinopathies beyond the short-term. Due to the mixed methodological quality and the insufficient number of clinical trials, larger, long-term, high-quality studies are needed to support the theory that tendinopathies can benefit from KT applications.

Coastal regions in Southwest Europe have experienced major interventions and transformations of the territory with unprecedented urban development, primarily related to growing tourism activity. The coast is the place where marine and terrestrial processes converge, making it highly vulnerable to the effects of climate change. However, the lack of information on the frequency of these extreme weather events and their impacts on the coast hampers an accurate analysis of the consequences of global change. This paper provides a detailed analysis of the extreme weather events (EWE) that have affected the Atlantic and Mediterranean coasts of Southwest Europe during the period from 1 January 2009 to 28 February 2020, as well as a quantification of their impacts: fatalities, injuries and economic damage. Official sources from France, Portugal and Spain were consulted, along with technical reports, scientific articles, etc., to generate a unified database. A total of 95 significant extreme events have caused 168 fatalities, 137 injuries and almost €4000 M in direct economic losses. Cyclone Xynthia (February 2010) on the French Atlantic coast stands out, having caused 47 fatalities, 79 injuries and substantial economic losses valued at €3000 M. The study shows a slight upward trend in the number of events recorded, especially during the last three years of the analysis, as well as in human losses and damages. The results reveal a higher exposure of the Mediterranean coast of Southwest Europe when compared to the Atlantic, especially the Spanish Mediterranean coast, with 61% of the fatalities recorded there during the study period. This is primarily due to a model of exponential tourism growth on the Mediterranean coast, with an enormous urban and infrastructure development during the last decades. Traditionally, the Mediterranean coast is less prepared to reduce the effects of marine storms, extreme events that are becoming more frequent and virulent in the context of climate and global change. This work highlights the need to create a continuous monitoring system–at the European level–of the impacts of extreme weather events on the coast, where 40% of the European population is concentrated. This observatory should serve as a source of information for risk mitigation policies (predictive, preventive and corrective), as well as for emergency management during disasters.

El presente artículo se propone analizar el par conceptual “privatismo civil / politización de la sociedad”. El objetivo es cuestionar si una de las formas concretas que adoptan hoy los procesos de repolitización de la sociedad civil, a saber, el llamado politainment y la sentimentalización del relato político, constituye o no una superación del privatismo civil. Para ello reconstruiremos primero la aparición del privatismo civil en el capitalismo tardío. En segundo lugar, analizaremos el surgimiento de esa conciencia ciudadana participativa que tiene su germen en los movimientos sociales de protesta postmaterialistas de finales de los sesenta. En tercer lugar, estudiaremos los procesos de sentimentalización y espectacularización del relato político. Sobre la base de este recorrido, en último lugar argumentaremos en contra de la tesis de una superación del privatismo civil.

This paper analyzes a flexible job shop scheduling problem with operators and is motivated by a real-life case study of the aeronautical industry, where each process can be performed on alternative machines (shared by multiple products) and requires not only machines but also some operators to execute the process. One goal of this scheduling is to minimize the number of operators and airframes that are needed in the assembly line. Minimizing the number of airframes in the assembly line, which is particularly important due to the cost of each airframe, is generally ignored in production planning models that are used in prior studies. Moreover, the output of the program refers to a set of schedules that includes different combinations of number of operators and intermediate stock, to enable a better decision making. A mixed integer linear programming model is presented to solve this problem. Experimentation was conducted using real-life examples from the aeronautical industry. The solutions that are presented in this paper outperformed current industrial methods in both quality and calculation time. To the best of our knowledge, this variant has not been addressed in prior studies either in scheduling or in aeronautics contexts. However, minimizing the number of operators and intermediate stock could have significant implications for numerous labor intensive industries, contributing to enhanced and more agile decision making processes.

Students’ acquisition of generic skills is a key to their incorporation into the job world. However, teachers encounter several difficulties when measuring their students’ performance in generic skills. These difficulties increase in online courses based on learning management systems, where there is no direct contact between teachers and their students. To address this problem, this paper adopts a solution based on the use of activity records from learning management systems as indicators of the students’ performance in generic skills. After conducting a survey among teachers of online courses on the suitability of the solution, an artefact was developed and applied within four online courses. Interviews conducted with teachers and the results of a survey for mapping skills and activities provide positive evidence of this solution’s potential.

El desarrollo web está evolucionando muy rápidamente tanto en tecnología como en los servicios que ofrece. La ingeniería de software que subyace, por su parte, no ha sido capaz de adaptarse a los nuevos problemas que presenta esta evolución. La encapsulación y la interoperabilidad han permitido el nacimiento de nuevos paradigmas de desarrollo donde usuarios finales sin conocimientos de programación, intermediarios y proveedores crean, explotan y reutilizan elementos conocidos como «componentes web». Sin embargo, los componentes web todavía carecen de un marco de calidad que sea aceptado universalmente, capaz de cuantificar lo bueno o malo que es un componente.

Research software is currently used by a large number of scientists on a daily basis, and everything indicates that this trend will continue to increase in the future. Most of this scientific software is very often developed by the researchers themselves, who usually make it available to the rest of the scientific community. Although the relationship between science and software is unquestionably useful, it is not always successful. Some of the critical problems that scientists face include a lack of training in software development, a shortage of time and resources, or difficulty in effectively cooperating with other colleagues. Additional challenges arise in the context of increasingly common cross-cutting and multidisciplinary research. This often results in the developed software and code being slow, not reusable, lacks visibility and dissemination, and in the worst cases it is defective and unreliable. Therefore, a multidisciplinary framework is needed to meet the demands of both scientists and software engineers and handle the situation successfully. However, a multidisciplinary team is not always sufficient to solve this problem, and it is necessary to have links between scientists and developers: software engineers with a solid scientific background. This paper presents the approach used in the framework of the PROTOCOL project, and more particularly in the development of its applied software, in which a tool for the characterization of climate agents has been developed. The main guidelines of the development process include, among others, modularity, distributed control version, unit testing, profiling, inline documentation and the use of best practices and tools.

Engineering infrastructures require regular maintenance and/or repair activities that have important social, environmental, and economic impacts. Despite their growing importance, few studies have focused on fully integrated analyses. This work presents a general methodological approach to design databases of engineering maintenance activities for their assessment. This methodology was applied to the case of dredging projects in the ports managed by the Andalusian Regional Government (Spain). The resulting database contains 87 fields of information obtained from the analysis of 70 activities performed between 1993 and 2015. This database is free, public, and available to the scientific community, and it was implemented in PostgreSQL using the PostGIS extension for spatial data; therefore, it can be integrated in a GIS. The assessment of deviations from the initial projects and the comparison between locations enhanced our methodology, which represents a valuable tool not only for scientists and managers to improve the decision-making process when planning future strategies, but also to evaluate the environmental impacts.

Due to the importance of coastline detection in coastal studies, different methods have been developed in recent decades in accordance with the evolution of measuring techniques such as remote sensing. This work proposes an automatic methodology with new water indexes to detect the coastline from different multispectral Landsat images; the methodology is applied to three Spanish deltas in the Mediterranean Sea. The new water indexes use surface reflectance rather than top-of-atmosphere reflectance from blue and shortwave infrared (SWIR 2) Landsat bands. A total of 621 sets of images were analyzed from three different Landsat sensors with a moderate spatial resolution of 30 m. Our proposal, which was compared to the most commonly used water indexes, showed outstanding performance in automatic detection of the coastline in 96% of the data analyzed, which also reached the minimum value of bias of − 0.91 m and a standard deviation ranging from ±4.7 and ±7.29 m in some cases in contrast to the existing values. Bicubic interpolation was evaluated for a simple sub-pixel analysis to assess its capability in improving the accuracy of coastline extraction. Our methodology represents a step forward in automatic coastline detection that can be applied to micro-tidal coastal sites with different land covers using many multi-sensor satellite images.

Non-governmental development organizations (NGDOs) are responsible for ensuring basic human rights for socially marginalised people. In situations of migration, people's opportunity to regularise their migratory status and to secure their fundamental rights to key public services is entirely dependent on communication with the local administration. To support these processes for those who are not fluent in the language(s) of the destination country, NGDO workers must learn how to provide effective interpreter-mediated communication services. In 2019-20, the "Linguistic mediation for development cooperation" (MELINCO) project brought together a multidisciplinary team to explore and respond to the communication needs of third sector organisations serving migrant populations. It included a self-study project involving the development and delivery of a training for NGDO staff, which was aimed at helping them select, prepare, and work with interpreters. The training was delivered online and was based on a needs assessment that involved data collection with NGDO representatives and interpreters. It received highly positive evaluations by participants in terms of course design, contents, and methodology, but our analysis also yielded directions for improvement in terms of extending opportunities for debate, design of practical applications, as well as facilitating participation in virtual learning environments.

The inbound logistic for feeding the workstation inside the factory represents a critical issue in the car manufacturing industry. Nowadays, this issue is even more critical than in the past since more types of car are being produced in the assembly lines. Consequently, as workstations have to install many types of components, they also need to have an inventory of different types of the component in a compact space.The replenishment is a critical issue since a lack of inventory could cause line stoppage or reworking. On the other hand, an excess of inventory could increase the holding cost or even block the replenishment paths. The decision of the replenishment routes cannot be made without taking into consideration the inventory needed by each station during the production time which will depend on the production sequence. This problem deals with medium-sized instances and it is solved using online solvers. The contribution of this paper is a MILP for the replenishment and inventory of the components in a car assembly line.

Large beach cusps (LBC, wavelength of ~ 30 m) are intertidal features that can alternately exist in the swash and in the inner surf zone due to tidal sea level changes. They have a larger cross-shore extent (up to 50 m) than traditional cusps. This extent has been explained by a shift of the swash zone during falling tide. The cusps immerse at rising tide and previous studies indicate that surf zone processes are exclusively destructive. Here, the behaviour of large beach cusps in the inner surf zone is investigated by using a 2DH morphological numerical model applied to Trafalgar Beach (Cádiz, Spain). The model results indicate that the inner surf zone processes do not always destroy the cusps but can in fact reinforce them by considering neither the swash processes nor the tidal changes. More generally, in conditions favouring the presence of the LBC the surf zone of a beach can be unstable, leading to the formation of transverse/oblique sand bars that can have characteristics similar to the LBC. Thus, in principle, the LBC could emerge not only due to swash zone morphodynamics but also due to surf zone morphodynamics or a combination of both.

La actualidad de un mundo en constante cambio invita de una manera especial a los docentes de cualquier ámbito a buscar nuevas maneras de implicar al alumno en su propia formación, para acercarlo a la realidad más profunda del proceso educativo y poder sacar lo mejor de cada uno. El presente estudio nos muestra un programa que busca, a través de diferentes actividades fuera del aula, introducir al universitario, no en los paradigmas y enseñanzas propios de cada asignatura, sino en algo más general que está en la base de toda educación universitaria. Se trata de claves de sentido, como la búsqueda de la verdad, la síntesis de saberes, la formación integral y el servicio a la sociedad.

La construcción de una escala de medida del grado de Responsabilidad Social del universitario y su validación como instrumento de medición del mismo son los objetivos principales de este estudio exploratorio. La aplicación piloto de este cuestionario, elaborado en base a las cinco dimensiones que conforman nuestra definición del concepto de Responsabilidad Social en la Universidad, nos servirá para evaluar el cambio producido en los alumnos que han cursado la asignatura de Responsabilidad Social. De este modo podremos evaluar su incidencia en la formación integral dentro del ámbito universitario.

Abstract Reviewing available information from published literature, museum database, personal communications and from the authors own field data, the conservation status of Cuban bats has been assessed using six qualitative parameters: abundance, distribution, roosting habits, aggregation level, forest dependence, and degree of endemism. The resulting Red List is analogous to that of the IUCN, species having been included in four categories of risk. Four out of the 26 extant bats of Cuba should be considered endangered, four vulnerable to extinction, twelve potentially threatened, and six in a stable situation. Most of the species of bats endemic to Cuba are under some form of threat. The major threats to the survival of Cuban bats are the destruction of forests and the modification of caves, the latter being critical habitats for the mostly cave-dwelling Cuban bat fauna. We argue that its conservation should be the result of a cooperative effort promoting research and habitat management. Riassunto Endemismi minacciati: una valutazione dello stato di conservazione dei chirotteri cubani. Lo stato di conservazione dei chirotteri cubani è stato valutato a partire da sei parametri qualitativi: abbondanza, distribuzione, roost utilizzati, livello di aggregazione, dipendenza da ambienti forestali e grado di endemismo. A questo scopo sono state esaminate le informazioni bibliografiche, i database dei musei e dati non pubblicati, in parte raccolti dagli stessi autori. La Lista Rossa risultante è analoga a quella dell’IUCN, comprendendo quattro categorie di rischio crescente. Delle 26 specie attualmente presenti a Cuba, 4 sono da considerarsi in pericolo di estinzione, 4 "vulnerabili", 12 "potenzialmente minacciate" e 6 "stabili". La maggior parte delle specie endemiche è in qualche misura minacciata. La deforestazione e l’alterazione delle cavità carsiche, che costituiscono un habitat critico per le molte specie troglofile, rappresentano i principali fattori di rischio per la chirotterofauna cubana, la cui conservazione dipende da uno sforzo comune volto a favorire la ricerca e la gestione ambientale.

Background: Long-distance swimmers exert energetic, physiological, and neuromuscular demands that must be matched with adequate body composition to improve their performance in long-distance swimming. Objectives: This review aims to compile all available information on energetic and physiological demands, optimal body composition, nutrition, and ergogenic supplements in long-distance swimming. This will provide an understanding of the specific challenges and needs of this sport and will help swimmers and coaches design more effective training and nutrition plans to optimise performance and achieve their goals. Methods: Databases such as Web of Science, SciELO Citation Index, MEDLINE (PubMed), Current Contents Connect, KCI-Korean Journal Database, and Scopus were searched for publications in English using keywords such as swimming, endurance, energy demands, physiological demands, nutrition, body composition, and ergogenic aids, individually or in combination. Results: There is convincing evidence that several physical indicators, such as propulsive surface area, technical, such as stroke rate, and functional, such as hydration strategies, are related to swimming performance and body composition. Each athlete may have a specific optimal body fat level that is associated with improved sporting performance. The nutritional needs of open water swimmers during competition are quite different from those of pool swimmers. Conclusions: Swimmers with an adequate physique have a high body muscle mass and moderately related anaerobic strength both on land and in the water. These general and specific strength capacities, which are given by certain anthropometric and physiological characteristics, are seen throughout the work, as well as ergogenic and nutritional strategies, which have an important impact on long-distance swimming performance. [ABSTRACT FROM AUTHOR]

The testosterone to cortisol ratio (T:C ratio) is a measure of whether elite athletes are recovering from their training. This study described this hormone balance stress in elite women's basketball. (1) Objectives: to analyse the fluctuation of T:C ratio over a 16-week period and explore itis relation to their athletic performance. The participants characteristics were: (height: 177.6 ± 6.4 cm; body mass: 77.808 ± 12.396 kg age: 26.0 ± 5.9 years; and a playing experience of 14.7 ± 2.9 years with 5.0 ± 1.2 years at the elite level. The T:C ratio at Time 1 is: 4.0 ± 2.4 (n = 12); and at Time 2 is: 5.1 ± 4.3 (n = 12). (2) Methods: during 16 weeks of competition, participants underwent analysis of blood samples to assess various biochemical parameters including hormone levels. In addition, their athletic performance was assessed with the following tests: jumping (SJ, CMJ, ABK, DJ); throwing test with a medicine ball (3 kg); Illinois COD agility test; sprint repeatability with change of direction; 20-m speed test without change of direction; and Yo-yo intermittent endurance test IET (II). (3) Results: The main alterations observed were an increase in T levels (1.687%) and a decrease in C levels (−7.634%) between moments, with an improvement (26.366%) in the T:C ratio. Improvements were also observed in some of the tests developed, such as jumping (SJ: 11.5%, p = 0.029; CMJ: 10.5%, p = 0.03; DJ: 13.0%, p = 0.01), upper body strength (MBT: 5.4%, p = 0.03), translation ability (20 m: −1.7%), repeated sprint ability (RSA: −2.2%), as well as intermittent endurance test (Yy (IET): 63.5%, p = 0.01), with significant changes in some of the performance tests. (4) Conclusions: T:C ratio may differ in a manner unrelated to training volume, showing some variation. These results may be attributed to the accumulation of psychophysiological stress during the season. [ABSTRACT FROM AUTHOR]

As for many other organisms, CRISPR-Cas9 mediated genetic modification has gained increasing importance for the identification of vaccine candidates and drug targets in Neospora caninum, an apicomplexan parasite causing abortion in cattle and neuromuscular disease in dogs. A widely used approach for generating knock-out (KO) strains devoid of virulence factors is the integration of a drug selectable marker such as mutated dihydrofolate reductase-thymidylate synthase (mdhfr-ts) into the target gene, thus preventing the synthesis of respective protein and mediating resistance to pyrimethamine. However, CRISPR-Cas9 mutagenesis is not free of off-target effects, which can lead to integration of multiple mdhfr-ts copies into other sites of the genome. To determine the number of integrated mdhfr-ts in N. caninum, a duplex quantitative TaqMan PCR was developed. For this purpose, primers were designed that amplifies a 106 bp fragment from wild-type (WT) parasites corresponding to the single copy wtdhfrs-ts gene, as well as the mutated mdhfrs-ts present in KO parasites that confers resistance and were used simultaneously with primers amplifying the diagnostic NC5 gene. Thus, the dhfr-ts to NC5 ratio should be approximately 1 in WT parasites, while in KO parasites with a single integrated mdhrf-ts gene this ratio is doubled, and in case of multiple integration events even higher. This approach was applied to the Neospora KO strains NcDGRA7 and NcDROP40. For NcDGRA7, the number of tachyzoites determined by dhfr-ts quantification was twice the number of tachyzoites determined by NC5 quantification, thus indicating that only one mdhfr-ts copy was integrated. The results obtained with the NcDROP40 strain, however, showed that the number of dhfr-ts copies per genome was substantially higher, indicating that at least three copies of the selectable mdhfrts marker were integrated into the genomic DNA during gene editing by CRISPRCas9. This duplex TaqMan-qPCR provides a reliable and easy-to-use tool for assessing CRISPR-Cas9 mediated mutagenesis in WT N. caninum strains. [ABSTRACT FROM AUTHOR]