Unraveling plant-pollinator interactions from a south-west Andean forest in Colombia.

Background: Loss of biological connectivity increases the vulnerability of ecological dynamics, thereby affecting processes such as pollination. Therefore, it is important to understand the roles of the actors that participate in these interaction networks. Nonetheless, there is a significant oversight regarding the main actors in the pollination networks within the highly biodiverse forests of Colombia. Hence, the present study aims to evaluate the interaction patterns of a network of potential pollinators that inhabit an Andean Forest in Totoró, Cauca, Colombia. Methods: The interactions between plants and potential pollinators were recorded through direct observation in 10 transects during six field trips conducted over the course of one year. Subsequently, an interaction matrix was developed, and network metrics such as connectance, specialization, nestedness, and asymmetry of interaction strength were evaluated by applying null models. An interpolation/extrapolation curve was calculated in order to assess the representativeness of the sample. Finally, the key species of the network were identified by considering degree (k), centrality, and betweenness centrality. Results: A total of 53 plant species and 52 potential pollinator species (including insects and birds) were recorded, with a sample coverage of 88.5%. Connectance (C = 0.19) and specialization (H2' = 0.19) were low, indicating a generalist network. Freziera canescens, Gaiadendron punctatum, Persea mutisii, Bombus rubicundus, Heliangelus exortis, Chironomus sp., and Metallura tyrianthina were identified as the key species that contribute to a more cohesive network structure. Discussion: The present study characterized the structure of the plant-pollinator network in a highly diverse Andean forest in Colombia. It is evident that insects are the largest group of pollinators; however, it is interesting to note that birds form a different module that specializes in pollinating a specific group of plants. On the other hand, the diversity and generality of the species found suggest that the network may be robust against chains of extinction. Nevertheless, the presence of certain introduced species, such as Apis mellifera, and the rapid changes in vegetation cover may affect the dynamics of this mutualistic network. So, it is imperative to apply restoration and conservation strategies to these ecosystems in order to enhance plant-animal interactions and prevent the loss of taxonomical and functional diversity.

Assessment of the psychometric properties of the Spanish version of EORTC QLQ-MY20 and evaluation of health-related quality of Life outcomes in patients with relapsed and/or refractory multiple myeloma in the real-world setting in Spain: results from the CharisMMa study.

We evaluated the psychometric properties of the Spanish version of the European Organization for Research and Treatment of Multiple Myeloma (MM) specific quality-of-life (QoL) questionnaire module (QLQ-MY20) in relapsed/refractory MM (RRMM) patients. This was an observational, cross-sectional, multicenter study using EORTC QLQ-C30 and QLQ-MY20 in RRMM patients (ClinicalTrials.gov ID NCT03188536). We assessed the non-response rate, ceiling/floor effects, internal consistency, test-retest reliability, and validity. The study included 276 patients (53.3% males, mean [SD] age of 67.4 [10.5] years). The EORTC QLQ-MY20 showed a low non-response rate, very low ceiling and floor effects, and good internal consistency. The test-retest reliability assessment revealed good temporary stability, the construct validity analysis stated four main factors similar to the ones of the original version, and the criterion validity assessment showed no differences between groups. In conclusion, the Spanish version of EORTC QLQ-MY20 is a reliable and valid tool for assessing QoL in RRMM patients.

Hepatitis C Virus: Insights Into Its History, Treatment, Challenges, and Future Directions.

Hepatitis C virus (HCV) is a global public health concern with significant impacts. It primarily spreads through blood-to-blood contact, such as sharing needles among drug users. Given the wide prevalence of risk factors, HCV continues to pose a major threat. Hence, it is crucial to understand its characteristics, structure, and genotypes to prevent, treat, and potentially eradicate it. This narrative review aims to explore the history of HCV treatment, highlight the breakthroughs achieved with direct-acting antiviral (DAA) therapy, address potential barriers to HCV eradication, and discuss future treatment possibilities. For this article, relevant studies were identified using various databases, including PubMed, ClinicalTrials.gov, and Journal Storage. The literature search revealed that after identifying HCV and studying its characteristics, interferon alfa and ribavirin became primary treatment options. However, due to their limited coverage against different HCV genotypes, ethnic variations, and suboptimal sustained virological response, the development of DAAs became essential. Combining various DAAs, such as sofosbuvir and velpatasvir, for a duration of 12 weeks has become the standard HCV treatment, with effectiveness against most genotypes. Additionally, ongoing clinical trials have shown promising results for other drugs such as CDI31244/sofosbuvir/velpatasvir, sofosbuvir/coblopasvir, and daclatasvir/asunaprevir. Despite the success of DAAs and ongoing efforts to discover more effective treatments, the high costs of DAAs pose a significant challenge to eradicating HCV, as not all patients can afford these expensive therapies. Furthermore, the ability of HCV to mutate limits the potential for vaccine development. Therefore, it is crucial to focus on developing more cost-effective strategies to control the spread of HCV and create novel, highly effective, and affordable DAAs.

Effect of high-volume insecticide spraying on sand fly vectors in household gardens in Spain.

This study investigated the efficacy of high-volume spraying with the adulticide α-cypermethrin alone and in combination with the larvicide diflubenzuron on the density of sand flies in gardens of three detached households in periurban areas in southeast Spain. Treatments were applied four times between June and August 2016, and four nearby sites, two households and two non-urbanized sites, were untreated controls. The number of sand flies collected between May and October 2016 using sticky interception and light attraction traps, was 4446 specimens. Species identified morphologically included Sergentomyia minuta (n = 2101; 48%), Phlebotomus perniciosus (n = 1922; 44%), Phlebotomus papatasi (n = 173; 4%), Phlebotomus sergenti (n = 161; 4%) and Phlebotomus ariasi (n = 36; 1%). Sand flies were detected in both treated and untreated sites. The proportion of positive sticky traps and the median (range) density of sand flies in positive traps were 61% traps and 7 (2-172) sand flies/m2 /day in untreated sites, and 43% traps and 4 (1-56) sand flies/m2 /day in treated sites (p < 0.05). Similarly, for light traps, it was 96% traps and 30 (3-168) flies/trap/day, and 83% traps and 3 (1-12) sand flies/trap/day, respectively (p < 0.05). However, sand fly density followed a comparable seasonal pattern in untreated and treated sites and did not consistently decrease following insecticide applications. These results were confirmed with mixed negative binomial modelling of sand fly density adjusted for time since application, month, environmental setting and site. The limited efficacy of the treatments, added to their cost, the impact of insecticides on non-target organisms and human health, and the risk of development of insecticide resistance, should dissuade similar outdoor applications to control sand fly vector populations in residential areas.

Evaluación de la eficiencia de dos estructuras para jardines de coral de la especie Acropora cervicornis (Scleractinia: Acroporidae) en el Parque Nacional Portobelo

Introducción: En Panamá y el mundo, desde hace varios años, los arrecifes se han visto amenazados por acciones como el desarrollo costero, sedimentación, contaminación por entidades terrestres, que son las más frecuentes, dando lugar a la implementación de metodologías de restauración. Objetivo: Comparar la eficiencia de dos estructuras de jardines coralinos en el crecimiento de la especie Acropora cervicornis en Bahía Huertas, Parque Nacional Portobelo, Colón Panamá. Métodos: Se colocaron dos estructuras de tipo fijo, utilizando PVC junto con bases de concreto hidráulico, y dos estructuras tipo tendedero, utilizando PVC e hilo de nylon. Los fragmentos de coral que se colocaron en las estructuras provenían de Bahía Huertas e Isla Buenaventura. El crecimiento de los corales se determinó mediante el método de medición de crecimiento lineal y extensión tisular. Resultados: Se plantaron un total de 80 fragmentos de coral, lo que representa una extensión estimada de 9.71 m de coral en Bahía Huertas. Tras nueve meses de estudio, se estimó un total de 15.3 m de coral totalmente nuevo (1 525.6 cm), donde sólo el 4.6 % pertenecía a los fragmentos de la estructura de tipo fijo, mientras que el 95.4 % pertenecía a la estructura de tipo tendedero. Conclusiones: La estructura tipo tendedero fue la más eficiente y viable dentro del estudio, con mejor tasa de supervivencia y crecimiento en comparación con la estructura tipo fija, donde sí presentó una mortalidad del 45 %.

Introduction: In Panama and the world, for several years, reefs have been threatened by actions such as coastal development, sedimentation, and contamination by terrestrial entities, which are the most frequent, giving rise to the implementation of restoration methodologies. Objective: To compare the efficiency of two coral garden structures in the growth of the species Acropora cervicornis in Huertas Bay, Portobelo National Park, Colon Panama. Methods: Two fixed-type structures were placed, using PVC together with hydraulic concrete bases, and two clothesline-type structures, using PVC and nylon thread. The coral fragments that were placed in the structures came from Bahía Huertas and Buenaventura Island. The growth of the corals was determined using the linear growth and tissue extension measurement method. Results: A total of 80 coral fragments were planted, representing an estimated extension of 9.71 m of coral in Huertas Bay. After nine months of study, a total of 15.3 m of totally new coral (1 525.6 cm) was estimated, where only 4.6 % belonged to the fragments of the fixe-type structure, while 95.4 % belonged to the clothesline-type structure. Conclusions: The clothesline-type structure was the most efficient and viable within the study, with better survival and growth rate compared to the fixed type structure, where it did present a mortality of 45 %.

Techno-functional properties and antioxidant capacity of the concentrate and protein fractions of Leucaena spp. seeds / Propiedades tecno-funcionales y capacidad antioxidante del concentrado y fracciones proteicas de semillas de Leucaena spp

Introduction: The use of vegetable proteins as ingredients in food systems is based on their functional properties. The water and oil holding capacity, foaming, and emulsifying capacity/stability, and antioxidant assay of the protein fractions - albumins, globulins 7S/11S, glutelins and prolamins - isolated from Leucaena seed were evaluated. Objective: The objective of this study was to evaluate the functional properties and antioxidant capacity of the concentrate and protein fractions of ripe Leucaena spp. seeds. Materials and methods: Ripe Leucaena seeds were collected and evaluated in Oaxaca, Mexico (16°59'21''N 96°43'26''O) during the months of February-April 2021.The protein concentrate was isolated by isoelectric precipitation (pH=9, pH=4). The albumins, globulins, glutelins and prolamins were isolated based on their solubility properties in different extracting solutions. Results: Glutelins constituted the main protein fraction (75.88%). Prolamins were not found. The glutelins fractions showed the highest oil holding capacity (0.93±0.08 mL g-1). The albumins fraction had the highest water holding capacity (2.53±0.15 mL g-1), foaming capacity and foam stability (71.83±1.26 % and 70.00±0.00%, respectively) and antioxidant capacity (18.09±0.88%). The globulins exhibited the highest emulsifying capacity and emulsion stability (56.83±1.76% and 55.67±1.20%, respectively). Conclusions: The concentrate and protein fraction of Leucaena seeds showed different techno-functional and antioxidant properties of interest for the food industry, like those showed by other commercial vegetable proteins(AU)

Introducción: El uso de proteínas vegetales como ingredientes en sistemas alimentarios se basa en sus propiedades funcionales. Se evaluó la capacidad de retención de agua y aceite, la capacidad/estabilidad espumante y emulsionante y el ensayo antioxidante de las fracciones proteicas -albúminas, globulinas 7S/11S, glutelinas y prolaminas- aisladas de las semillas de Leucaena. Objetivo: El objetivo de este estudio fue evaluar las propiedades funcionales y la capacidad antioxidante del concentrado y las fracciones proteicas de las semillas maduras de Leucaena spp. Materiales y métodos: Las semillas maduras de Leucaena fueron recolectadas y evaluadas en Oaxaca, México (16°59'21''N 96°43'26''O) durante los meses de febrero-abril del año 2021. Se usó harina de Leucaena desgrasada para la preparación de las fracciones proteicas. El concentrado proteico se aisló por precipitación isoeléctrica (pH=9, pH=4). Las albúminas, globulinas, glutelinas y prolaminas se aislaron en función de sus propiedades de solubilidad en diferentes soluciones de extracción. Resultados: Las glutelinas constituyeron la principal fracción proteica (75,88%). No se encontraron prolaminas. La fracción de glutelinas mostró la mayor capacidad de retención de aceite (0.93±0,08 mL g-1). La fracción de albúminas presentó la mayor capacidad de retención de agua (2,53±0,15 mL g-1), capacidad espumante y estabilidad de la espuma (71,83±1,26% y 70,00±0,00%, respectivamente) y capacidad antioxidante (18,09±0,88%). Las globulinas mostraron la mayor capacidad emulsionante y estabilidad de la emulsión (56,83±1,76 y 55,67±1,20%, respectivamente). Conclusiones: El concentrado y las fracciones proteicas de las semillas de Leucaena mostraron diferentes propiedades tecno-funcionales y antioxidantes de interés para la industria alimentaria, similares a los reportados por diversas proteínas vegetales comerciales(AU)

PytuTester: RaspberryPi open-source ventilator tester.

PytuTester is an open-source ventilator tester developed to help bio-engineers in the design and verification of new ventilator prototypes. A ventilator tester allows measuring the flow, pressure, volume, and oxygen concentration provided to the patient. During the global pandemic COVID-19, several open-source ventilators prototypes were developed; however, due to high cost and demand testers, they were not available. In this context, a low-cost tester was developed using a Raspberry Pi and medical-grade sensors for the test ventilators prototypes. This paper presents the design files, software interface, and validations tests. Our results indicate that the tester has good accuracy to evaluate the efficacy and performance of new prototypes. When tested on two ventilator designs developed in Paraguay, PytuTester reported flow profiles that were concordant with the industry-standard VT650 Gas Flow Analyzer. PytuTester was then field deployed to test several DIY ventilator designs in low-resource areas.

Dispositivo de barrera protectora con presión negativa utilizado en urgencias odontológicas durante la pandemia de COVID-19 / Negative pressure protective barrier device used in dental emergencies during the COVID-19 pandemic

RESUMEN Introducción: el personal de salud involucrado en la atención odontológica tiene alto riesgo de contagio por exposición constante a aerosoles durante la atención de urgencias. Es necesario incrementar sus medidas de protección personal durante la pandemia por SARSCoV-2. Objetivo: describir la experiencia de odontólogos, auxiliares y pacientes respecto al uso de un dispositivo de barrera para contención de aerosoles en la atención de urgencias odontológicas. Materiales y métodos: estudio descriptivo de corte transversal con 21 pacientes adultos atendidos por urgencias odontológicas en una institución privada durante el mes de junio en la ciudad de Bogotá. Los 21 pacientes, cinco odontólogos tratantes y cinco auxiliares de odontología diligenciaron un cuestionario estructurado sobre la experiencia con el dispositivo, medidas de bioseguridad implementadas y percepción de seguridad respecto al posible contagio del virus. Resultados: la edad promedio de los pacientes fue 41 años (DE±14). La duración de los procedimientos fue de 45 minutos (DE±12) en promedio. No hubo eventos adversos durante los procedimientos con el uso del dispositivo. El 95 % de los pacientes se sintieron más seguros durante la atención con el uso del dispositivo de barrera. Conclusiones: a pesar de la muestra limitada, la alta percepción de seguridad respecto al uso del dispositivo en la atención de urgencias odontológicas reportada aquí, sugiere que pueden llegar a complementar las medidas de bioseguridad y equipos de protección personal establecidos por autoridades sanitarias durante la pandemia.

SUMMARY Introduction: Health personnel involved in dental care are at high risk of contagion from constant exposure to aerosols during emergency care. It is necessary to increase its protection measures during the SARS-CoV-2 pandemic. Objective: Describe the experience of dentists, assistants and patients regarding the use of a barrier device to contain aerosols in dental emergency care. Materials and methods: A descriptive cross-sectional study with 21 adult patients attended by dental emergencies in a private institution during June in the city of Bogotá. The 21 patients, five treating dentists and five dental assistants filled out a structured questionnaire about the experience with the device, the biosafety measures implemented and the perception of safety regarding the possible contagion of the virus. Results: The mean age of the patients was 41 years (SD ± 14). The duration of the procedures was 45 minutes (SD ± 12) on average. There were no adverse events during the procedures with the use of the device. 95% of patients felt safer during care with the use of the barrier device. Conclusions: Despite the limited sample, the high perception of safety regarding the use of the device in dental emergency care reported here suggests that they may complement the biosecurity measures and personal protective equipment established by health authorities during the pandemic.

Evaluation of the Effect of a Live Interview in Journalism Students on Salivary Stress Biomarkers and Conventional Stress Scales.

A career in journalism can be very stressful, as journalists frequently have to deal with uncontrolled situations such as conducting live interviews. Therefore, training is essential during their career, both for the development of communication skills and for the improvement of the real and effective capacity to perform the tasks of their professional activity. The aim of this study was to assess the levels of stress in students before and after a practical training in a professional television set using subjective (State-Trait Anxiety Inventory (STAI) and Likert scale) and objective (salivary cortisol and alpha-amylase) methods. The results indicate that a live interview produces stress in the students as revealed by increased concentrations of cortisol and alpha amylase in saliva. Furthermore, students with lower initial concentrations of these biomarkers obtained better grades in evaluation, suggesting that greater control of anticipatory stress could be associated with a better activity performance.

Fractura triplanar del extremo distal del radio: reporte de caso / Triplanar fracture of the distal end of the radius: case report

Triplanar fractures are those that occur in 3 separate planes at the end of an immature long bone. They typically cross the epiphysis in the sagittal plane, physis in the transverse plane, and metaphysis in the coronal plane. Due to their nature, they are considered an intra-articular injury. Radiographically, these fractures often resemble a Salter-Harris type III epiphyseal slip injury on the anterior-posterior view and a Salter-Harris type II on the lateral view and are consequently considered a Salter-Harris type IV. Only 5 cases of triplanar fracture of the distal end of the radius have been reported so far in the literature, the second case managed with open reduction and internal fixation is reported. This is a 12-year-old patient with a history of trauma to the right wrist after a traffic accident, with deformity and functional limitation and diagnostic images that confirm a fracture at the metaphyseal level of the distal end of the right radius with radial displacement and a stroke. fracture that goes through the epiphysis, physis and metaphysis. An open reduction and internal fixation was performed. 2 years after the injury, the patient is asymptomatic, with preserved ranges of joint mobility and diagnostic images with evidence of adequate consolidation of the fracture, absence of alteration of the physis and consequent normal growth of the radius. Level of Evidence: IV.

Changes in salivary proteins can reflect beneficial physiological effects of ejaculation in the dog.

The objective of this study was to study the changes in salivary proteins that occur in the dog after the ejaculation process. Saliva samples from eight dogs before and after induced ejaculation were analyzed by proteomic using Tandem Mass Tag (TMT) labeling and LC-MS/MS analysis. A total of 33 salivary proteins showed significant changes after the ejaculation process. The up-regulated proteins that showed changes of higher magnitude were mucin-7 (MUC-7), peroxiredoxin-4 (PRDX4) and galectin-3 (LEGALS3) whereas proteins such as alpha-1-acid glycoprotein (A1G1) and alpha-1B-glycoprotein (A1BG) were the most down-regulated. MUC-7 and PRDX4 expression in saliva after ejaculation could be associated with the protective "environment" created by the organism to exert pr 3o-fertility activities and antioxidants benefits in spermatozoa. Also LEGALS3 increment could be associated with an improvement of wellbeing and could contribute to a positive global effect in the body. Down-regulations of A1G1 and A1GB proteins found in saliva after ejaculation could be associated with a reduction in systemic inflammation. Overall it can be concluded that, changes in proteins in saliva that are produced after ejaculation can reflect a state of increase immune defenses, improvement of antioxidant status and low inflammation.

Differences in the Exposure of the Lumbar Nerve Root Between Experts and Novices: Results From a Realistic Simulation Pilot Study With Force Sensors.

STUDY DESIGN: Nonrandomized prospective trial. OBJECTIVE: Several studies could demonstrate "learning curves" in almost every single surgical procedure for unexperienced surgeons. This is in sharp contrast to the rising quality requirements in public health care to provide surgical training at patients "expense." The aim of this study was to visualize, measure, and set a baseline of the pressure load on the spinal nerve root during a simulated microdiscectomy on a standardized and validated model (RealSpine) under the influence of the level of surgical experience and individual skills. METHODS: Five highly experienced spine surgeons and 5 trainees without considerable surgical experience were selected to perform a standardized microsurgical discectomy on a validated RealSpine simulator. Force-torque sensors were integrated in this simulator to measure the load on the nerve root. The forces were recorded every 125 ms. RESULTS: We could identify cumulative for the total intervention as well as for defined single surgical steps of this procedure and as well in between the single subjects a significant higher tension and contusion forces on the nerve for the trainee group (Δp contusion 83-765 Ncs and Δp tension 159-1131 Ncs for the trainees. Δp contusion 16-171 Ncs and Δp tension 27-146 Ncs for the experts). CONCLUSION: We could measure a difference between unexperienced and experienced surgeons regarding the manipulations of the nerve root during a standardized simulated microdiscectomy. This possibility could be the starting point for a new and innovative surgical education to improve outcome without negative side effects of "learning curves."

Interdisciplinary Collaboration Between Veterinary and Communication Students to Promote Communication Skills: A Qualitative Pilot Study.

Interdisciplinary collaborations are increasingly gaining popularity, as are active in higher education and innovative learning strategies. However, relatively little research has been performed related to interdisciplinary learning methodologies in higher education. In the present work, a pilot activity between communication and veterinary students was performed, consisting in performance of mock interviews at a professional television studio. Besides some drawbacks such as low participation rates by veterinary students, the activity was associated with a number of benefits, including enhanced acquirement of communication skills, greater topic-related knowledge assimilation, and reinforced practical application of the theoretical concepts.

The Long-Term Effect of Medically Enhancing Melanin Intrinsic Bioenergetics Capacity in Prematurity.

BACKGROUND: The ability of the human body to produce metabolic energy from light modifies fundamental concepts of biochemistry. OBJECTIVE: This review discusses the relationships between the long-accepted concept is that glucose has a unique dual role as an energy source and as the main source of carbon chains that are precursors of all organic matter. The capability of melanin to produce energy challenges this premise. METHODS: The prevalent biochemical concept, therefore, needs to be adjusted to incorporate a newly discovered state of Nature based on melanin's ability to dissociate water to produce energy and to re-form water from molecular hydrogen and oxygen. RESULTS AND DISCUSSION: Our findings regarding the potential implication of QIAPI-1 as a melanin precursor that has bioenergetics capabilities. CONCLUSION: Specifically, we reported its promising application as a means for treating retinopathy of prematurity (ROP). The instant report focuses on the long-term treatment medical effects of melanin in treating ROP.

Teaching the basics of the One Health concept to undergraduate veterinary students.

The increasingly acknowledged "One Word, One Health" (OH) concept studies the collaborative multi-disciplinary approaches for the assessment of human and animal health and the environment. This study provides information about a module of activities created to teach the OH concept to undergraduate veterinary students. The module consisted of three different activities: theoretical classes, teamwork for the preparation of different concepts and practical examples related to OH, and public presentations of the students of these practical cases. This module was evaluated by two questionnaires' consisting of online surveys, which were filled in before (questionnaire 1, Q1) and after (questionnaire 2, Q2) the module about OH. Before the course, 80% of students recognized to have a poor or very poor knowledge about OH, and a 71% failed to include the three main items of the OH concept (humans, animals and environment) in their answers. After the course, the general knowledge about OH was improved and most students evaluated the course positively, although the lack of time and the high number of students per group were pointed out as the main drawbacks. In conclusion, the module of activities described in this report contributed to the increase of OH knowledge by veterinary students and could be a resource for future advances to improve the teaching of the OH concept in the curricula at University level in Veterinary and other Degrees related with OH.

Profile of phenolic acids, antioxidant activity and total phenolic compounds during blue corn tortilla processing and its bioaccessibility.

Varieties of blue maize are used to produce tortillas. The Bolita genotype is frequently underused, despite its rich content of bioactive compounds. The composition of derived products is influenced by maize processing. The impact of processing on the total phenolic content (TPC), phenolic acid profile (PAP), antioxidant activity (AA) and color was evaluated in tortillas produced from blue Bolita landraces. The properties were determined in anatomical fractions, nixtamal, wastewater, masa and tortillas. Vanillic, ferulic, p-coumaric and chlorogenic acids were identified in anatomical fractions. Nejayote had the highest AA. The compounds in the soluble fraction contributed mainly to the AA, whereas the free and bound fractions were largely responsible for the TPC. Syringic and ferulic acids were the most common compounds in the nejayote, and p-coumaric acids were the most common compounds in the masa. Only ferulic acid was detected after simulated gastric digestion, and temperature, pH and milling had an important effect on the PAP.

Detection of Leishmania infantum DNA by real-time PCR in saliva of dogs.

This study developed a real-time quantitative PCR (qPCR) assay to detect L. infantum kinetoplast DNA (kDNA) in canine saliva. The qPCR showed an efficiency of 93.8%, a coefficient of correlation of 0.996 and a detection limit of 0.5 fg/reaction (0.005 parasites), although it detected until 0.25 fg/reaction (0.0025 parasites). When samples from 12 dogs experimentally infected with L. infantum were collected, L. infantum kDNA was detected at 16-weeks post-infection (wpi) in 41.7% and 91.7% of saliva and bone marrow samples, respectively, and at 47-wpi in 75% of both samples. L. infantum kDNA can be detected by qPCR in canine saliva, with lower sensitivity in the early stages of infection and a lower parasite load estimation compared to bone marrow. However, saliva had similar sensitivities to bone marrow in the later stages of the infection and could be used to detect L. infantum kDNA being aware of its limitations.

Evaluation of the circadian rhythm of anti-Leishmania IgG2 and IgA antibodies in serum and saliva of dogs with clinical leishmaniosis.

In this study, the circadian rhythm of IgG2 and IgA specific antibodies in serum and saliva samples of 6 dogs experimentally infected with Leishmania infantum was assessed. Sampling was performed at 8.00, 12.00, 16.00, 20.00, and 00.00 h on two consecutive days. Anti-Leishmania antibody levels in serum were expressed without any correction, whereas in saliva were shown in different ways: without any correction, adjusted by protein concentration and corrected by the salivary flow rate. No significant differences in anti-Leishmania IgG2 antibody levels in serum and saliva samples with or without correction were found. Significant differences were found when anti-Leishmania IgA levels were corrected by the salivary flow rate. In addition, a greater intra-individual variation of antibody levels was observed in saliva than in serum. However, this variation did not modify the serological status of the dogs. Therefore, it could be concluded that there is no circadian rhythm in serum and saliva samples and sampling can be performed at any time of the day.

Serum proteome of dogs at subclinical and clinical onset of canine leishmaniosis.

The objective of this study was to identify changes in serum proteome in dogs that may occur after an experimental infection at subclinical and clinical stages of canine leishmaniosis (CanL). For this purpose, canine pre- and post-infection with Leishmania infantum serum proteomes in the same dogs were analysed by a high-throughput label-based quantitative LC-MS/MS proteomic approach. A total of 169 proteins were identified, and 74 of them including complement C8 alpha chain, adiponectin, transferrin, sphingomyelin phosphodiesterase acid-like 3A and immunoglobulins showed different modulation between the different stages of CanL. These proteins could be considered as potential serum biomarkers of early diagnostic or disease progression in CanL. Additionally, biological pathways modulated during CanL such as blood coagulation or gonadotropin-releasing hormone receptor were revealed, which could help to understand the pathological mechanisms of the disease.

One-year follow-up of anti-Leishmania antibody concentrations in serum and saliva from experimentally infected dogs.

The quantification of anti-Leishmania antibodies in serum and saliva by a time-resolved immunofluorometric assay is useful for the diagnosis and treatment monitoring of dogs with clinical leishmaniasis. We compared the kinetics of anti-Leishmania IgG2 and IgA antibodies in serum and saliva from 11 Beagle dogs experimentally infected with Leishmania infantum. Most dogs showed detectable concentrations of anti-Leishmania IgG2 earlier in serum (between 3 and 4 months p.i.) than in saliva (between 4 and 6 months p.i.). Overall, a high correlation between concentrations of anti-Leishmania IgG2 in serum and saliva (r = 0.853; P < 0.0001) was observed. The quantification of anti-Leishmania IgA showed less diagnostic value than IgG2, since detectable amounts of IgA were not observed in the saliva of four dogs and in the serum of one dog. In addition, a very low correlation between anti-Leishmania IgA in serum and saliva (r = 0.289; P < 0.001) was observed. Our results indicate that the antibodies against L. infantum in saliva appear approximately 1 month later than in serum, and suggest that there is a threshold for the passing of immunoglobulins from serum to saliva in dogs. These facts should be taken into consideration for a proper interpretation of saliva assays for quantification of antibodies.