Evaluación del efecto de las artemisininas provenientes del extracto etanólico de Artemisia cina sobre L3 de Haemonchus contortus en una técnica de explantes abomasales

The new approach of integrated nematode control has driven to look for other alternatives that consider steadily and sustainable the environment. The aim of the present study was to obtain different ethanolic fracctions from Artemisia cina plant, using artemisinins, and to evaluate the nematicide association capacity effect with infective larvae (L3) and abomasal explant. The artemisinins from A. cina were obtained by thin layer chromatography and were evaluated on unsheathed L3 of Haemonchus contortus. The abomasal technique explants were performed for triplicate and compared as follows: three control with levamisole (7.5 mg/ml), commercial artemisinin (1 mg/ml), and distillate water, and six A. cina fracctions at 1 mg/ml each one: Ac3k, Ac3b, Ac3a, Ac3h, Ac3i and Ac4b. No significant differences between fractions of A. cina and commercial artemisinin (p>0.05) were obtained. These findings led us to conclude that there was no association capacity effect of fractions of A. cina on unsheathed L3 of H. contortus, to the abomasal tissue. It is important to continue with more studies to determine the parasitic phase in which the artemisinin viability is susceptible to be affected.El nuevo enfoque de control integrado de parásitos obliga a buscar alternativas que consideren el medioambiente, es decir, sostenibles y sustentables. El objetivo de este trabajo fue fraccionar el extracto etanólico de la planta Artemisia cina, obtener artemisininas y conocer el efecto de estas sobre la capacidad de asociación de las L3 de Haemonchus contortus en explantes abomasales. El extracto etanólico se fraccionó por medio de la metodología establecida para Artemisia japónica, y se identificaron las artemisininas por medio de cromatografía en capa fina, teniendo como referencia artemisininas comerciales. Las artemisininas se utilizaron sobre L3 desenvainada, se realizó la técnica de explantes abomasales por triplicado y se compararon así: levamisol (7.5 mg/ml), artemisinina comercial (1 mg/ml), agua y seis diferentes fracciones de A. cina a dosis de 1 mg/ml: Ac3k, Ac3b, Ac3a, Ac3h, Ac3i, Ac4b. No se obtuvieron diferencias significativas entre las fracciones de A. cina y artemisinina comercial (p&gt,0.05). Se concluye que no existió efecto de las artemisininas obtenidas del extracto etanólico de A. cina sobre la capacidad de asociación de las L3 de H. contortus al tejido abomasal. Es importante continuar con más estudios de la artemisinina para determinar sobre qué fase del parásito afecta su viabilidad

Utilização de bolos ruminais de liberação de sulfametazina sódica para o controle de coccidiose ovina

Avaliou-se o efeito de bolos intrarruminais de liberação de sulfametazina sódica no controle de coccidiose ovina. Foram utilizados 60 cordeiros machos e fêmeas em dois sistemas de alimentação: pastoreio intensivo diurno com recolhimento noturno em aprisco e engorda em confi namento, com grãos. Os animais foram divididos em quatro grupos: Confi namento-Bolo (CB, n=20); Confi namento-Sem Bolo (CSB, n=10); Pastoreio-Bolo (PB, n=20) e Pastoreio-Sem Bolo (PSB, n=10). Avaliou-se quinzenalmente o efeito do tratamento sobre o ganho médio de peso (GP), a eliminação de oocistos e identifaram-se as espécies de Eimeria envolvidas. Não se demostraram diferenças no GP entre os animais dos grupos PB= 0,32kg e PSB= 0,2kg (p 0,65), e sim nos grupos CB= 3,31kg e CSB= 4,28kg (P0,02). A eliminação de oocistos foi menor nos grupos que receberam o bolo: CB: 428 e PB:844 ooc/g que nos grupos CSB:2169 e PSB:1533 ooq/g de fezes (p0,009), indicando que o bolo controlou a parasitose. As espécies de Eimeria presentes nos animais confi nados foram: E. ovina 30%, E. ovinoidalis 24%, E.  ranulosa 16% e E. parva 15%, outras espécies 7%. Nos animais em pastoreio E. ovinoidalis 28%, E. ovina 22%, E. granulosa 15%   E. parva 10%, outras espécies 8%. Ao fi nal do experimento, observaram-se diferenças relativas nas espécies identifi cadas nos diferentes grupos, indicando que a condição de alimentação e o tratamento com o bolo afetou quantitativamente a parasitose e a  proporção de espécies envolvidas

Efecto in vitro de los extractos hidroalcohólico y etanólico de semilla de papaya (Carica papaya) en Haemonchus contortus

Among the main health problems for sheep production are gastrointestinal parasites, of which the most important nematode is Haemonchus contortus. Current control strategies focus on the effectiveness, along with the low impact on the ecosystem. The study of the integral control of parasites allows the investigation of alternatives with probable anthelmintic effect, such as the use of papaya (Carica papaya) seeds. This study aimed at evaluating the in vitro effect of hydroalcoholic and ethanolic extracts of papaya seeds on eggs and larvae of H. contortus. The study was carried out with the equipment and facilities of Laboratory 3 of the Multidisciplinary Research Unit at the Facultad de Estudios Superiores Cuautitlán, UNAM. The extracts were prepared through maceration in solvent. A sheep was inoculated with 5000 L3 of H. contortus, a strain isolated and maintained in the facilities. Fecal samples were collected, and eggs were recovered for the hatch inhibition assay. The eggs and L3 were pretreated with hydroalcoholic and ethanolic extract of papaya seed (2.5 mg/ml), distilled water, ethanol 90 %, and levamisole (7.5 mg/ml). The abomasal explants technique was used to evaluate the effect of the extracts on L3 and its association to the abomasal mucosa. An ANOVA was performed for repeated samples using the Statgraphics program. The ethanolic extract of the papaya seed inhibited 92 % of hatching, and the hydroalcoholic extract 50 % (p <0.05). In the case of the explants, the ethanolic extract showed 10 % of the larvae not associated with the abomasal tissue, followed by the hydroalcoholic extract with 60 % of non-associated larvae (p<0.05). In conclusion, the ethanolic extract has an effect on eggs, while the hydroalcoholic extract inhibits the association to the abomasal tissue.Entre los principales problemas sanitarios para la producción ovina están los parásitos gastroentéricos; de ellos, el más importante es Haemonchus contortus. Las estrategias actuales de control enfocan la eficacia, junto con el bajo o nulo impacto al ecosistema; el estudio del control integral de parásitos permite la investigación de alternativas con probable efecto antihelmíntico, entre ellas el uso de semilla de papaya (Carica papaya). El objetivo de este trabajo fue evaluar el efecto in vitro de los extractos hidroalcohólico y etanólico de la semilla de papaya (C. papaya) sobre huevos y larvas de H. contortus. El trabajo se llevó a cabo con el equipo e instalaciones del Laboratorio 3 de la Unidad de Investigación Multidisciplinaria en la Facultad de Estudios Superiores Cuautitlán, UNAM. Se elaboraron los extractos por medio de maceración en solvente. Se inoculó un ovino con 5000 larvas de tercer estadio (L3) de H. contortus, cepa aislada y mantenida en dichas instalaciones. Se tomaron muestras de materia fecal y se recuperaron huevos para el ensayo de inhibición de la eclosión. Los huevos y las L3 fueron pretratados con extracto hidroalcohólico y etanólico de semilla de papaya (2.5 mg/ml), agua destilada, etanol 90 % y levamisol (7.5 mg/ml). Se efectuó la técnica de explantes abomasales, evaluando el efecto de los extractos sobre las L3 y su asociación a la mucosa abomasal. Se realizó un ANOVA para muestras repetidas por medio del programa Statgraphics. El extracto etanólico de semilla de papaya mostró el 92 % de inhibición de la eclosión; el extracto hidroalcohólico mostró 50 % (p<0.05). En el caso de los explantes, el extracto etanólico mostró el 10 % de larvas no asociadas al tejido abomasal, seguido por el extracto hidroalcohólico, con el 60 % de larvas no asociadas (p<0.05). Se concluye que el extracto etanólico tiene efecto sobre huevos, mientras que el extracto hidroalcohólico inhibe la asociación al tejido abomasal

In vitro effect of the hydroalcoholic and ethanolic extracts of papaya seeds (Carica papaya) against Haemonchus contortus

1 recurso en línea (páginas 53-59).Among the main health problems for sheep production are gastrointestinal parasites, of which the most important nematode is Haemonchus contortus. Current control strategies focus on the effectiveness, along with the low impact on the ecosystem. The study of the integral control of parasites allows the investigation of alternatives with probable anthelmintic effect, such as the use of papaya (Carica papaya) seeds. This study aimed at evaluating the in vitro effect of hydroalcoholic and ethanolic extracts of papaya seeds on eggs and larvae of H. contortus. The study was carried out with the equipment and facilities of Laboratory 3 of the Multidisciplinary Research Unit at the Facultad de Estudios Superiores Cuautitlán, UNAM. The extracts were prepared through maceration in solvent. A sheep was inoculated with 5000 L3 of H. contortus, a strain isolated and maintained in the facilities. Fecal samples were collected, and eggs were recovered for the hatch inhibition assay. The eggs and L3 were pretreated with hydroalcoholic and ethanolic extract of papaya seed (2.5 mg/ml), distilled water, ethanol 90 %, and levamisole (7.5 mg/ml). The abomasal explants technique was used to evaluate the effect of the extracts on L3 and its association to the abomasal mucosa. An ANOVA was performed for repeated samples using the Statgraphics program. The ethanolic extract of the papaya seed inhibited 92 % of hatching, and the hydroalcoholic extract 50 % (p<0.05). In the case of the explants, the ethanolic extract showed 10 % of the larvae not associated with the abomasal tissue, followed by the hydroalcoholic extract with 60 % of non-associated larvae (p<0.05). In conclusion, the ethanolic extract has an effect on eggs, while the hydroalcoholic extract inhibits the association to the abomasal tissue.Entre los principales problemas sanitarios para la producción ovina están los parásitos gastroentéricos; de ellos, el más importante es Haemonchus contortus. Las estrategias actuales de control enfocan la eficacia, junto con el bajo o nulo impacto al ecosistema; el estudio del control integral de parásitos permite la investigación de alternativas con probable efecto antihelmíntico, entre ellas el uso de semilla de papaya (Carica papaya). El objetivo de este trabajo fue evaluar el efecto in vitro de los extractos hidroalcohólico y etanólico de la semilla de papaya (C. papaya) sobre huevos y larvas de H. contortus. El trabajo se llevó a cabo con el equipo e instalaciones del Laboratorio 3 de la Unidad de Investigación Multidisciplinaria en la Facultad de Estudios Superiores Cuautitlán, UNAM. Se elaboraron los extractos por medio de maceración en solvente. Se inoculó un ovino con 5000 larvas de tercer estadio (L3) de H. contortus, cepa aislada y mantenida en dichas instalaciones. Se tomaron muestras de materia fecal y se recuperaron huevos para el ensayo de inhibición de la eclosión. Los huevos y las L3 fueron pretratados con extracto hidroalcohólico y etanólico de semilla de papaya (2.5 mg/ml), agua destilada, etanol 90 % y levamisol (7.5 mg/ml). Se efectuó la técnica de explantes abomasales, evaluando el efecto de los extractos sobre las L3 y su asociación a la mucosa abomasal. Se realizó un ANOVA para muestras repetidas por medio del programa Statgraphics. El extracto etanólico de semilla de papaya mostró el 92 % de inhibición de la eclosión; el extracto hidroalcohólico mostró 50 % (p<0.05). En el caso de los explantes, el extracto etanólico mostró el 10 % de larvas no asociadas al tejido abomasal, seguido por el extracto hidroalcohólico, con el 60 % de larvas no asociadas (p<0.05). Se concluye que el extracto etanólico tiene efecto sobre huevos, mientras que el extracto hidroalcohólico inhibe la asociación al tejido abomasal.Bibliografía: páginas 58-59

Bolos intrarruminales con liberación controlada de minerales traza. Revisión

Los minerales traza son nutrientes esenciales para el mantenimiento de la vida, el crecimiento y la reproducción. Las deficiencias de minerales en rumiantes afectan las funciones fisiológicas y metabólicas que con frecuencia causan enfermedades. El diseño y uso de los bolos intrarruminales de liberación controlada (BILC) es una alternativa para corregir la falta de los microelementos en el organismo. El propósito de esta revisión es evidenciar la información disponible sobre los diferentes tipos de BILC de minerales traza, así como de los métodos de fabricación que incluye: técnicas de extrusión en caliente, granulación por fusión y fusión directa. Además, se describen los efectos de BILC relacionados en la salud, en los parámetros productivos y reproductivos en rumiantes

Molecular method for the characterization of Coxiella burnetii from clinical and environmental samples: variability of genotypes in Spain

BACKGROUND: Coxiella burnetii is a highly clonal microorganism which is difficult to culture, requiring BSL3 conditions for its propagation. This leads to a scarce availability of isolates worldwide. On the other hand, published methods of characterization have delineated up to 8 different genomic groups and 36 genotypes. However, all these methodologies, with the exception of one that exhibited limited discriminatory power (3 genotypes), rely on performing between 10 and 20 PCR amplifications or sequencing long fragments of DNA, which make their direct application to clinical samples impracticable and leads to a scarce accessibility of data on the circulation of C. burnetii genotypes. RESULTS: To assess the variability of this organism in Spain, we have developed a novel method that consists of a multiplex (8 targets) PCR and hybridization with specific probes that reproduce the previous classification of this organism into 8 genomic groups, and up to 16 genotypes. It allows for a direct characterization from clinical and environmental samples in a single run, which will help in the study of the different genotypes circulating in wild and domestic cycles as well as from sporadic human cases and outbreaks. The method has been validated with reference isolates. A high variability of C. burnetii has been found in Spain among 90 samples tested, detecting 10 different genotypes, being those adaA negative associated with acute Q fever cases presenting as fever of intermediate duration with liver involvement and with chronic cases. Genotypes infecting humans are also found in sheep, goats, rats, wild boar and ticks, and the only genotype found in cattle has never been found among our clinical samples. CONCLUSIONS: This newly developed methodology has permitted to demonstrate that C. burnetii is highly variable in Spain. With the data presented here, cattle seem not to participate in the transmission of C. burnetii to humans in the samples studied, while sheep, goats, wild boar, rats and ticks share genotypes with the human population

Designing and evaluation of sodium selenite nanoparticles in vitro to improve selenium absorption in ruminants

Sodium selenite is used to prevent selenium deficiency known as nutritional muscular dystrophy or white muscle disease. In ruminants, selenium supplements are transformed partiality in insoluble form by ruminal microorganisms and its process decrease the selenium absorption in digestive gastrointestinal. However, the objective in this research was focused in encapsulated sodium selenite to be release into of a pH less than four, similarity to an intestinal environment. It was encapsulated by nanoprecipitation and emulsion–evaporation methods, within polymeric nanoparticles. The effect of these methods, polymer proportion (Eudragit RL and RS) and solvent (ethanol and acetone) on the physicochemical (drug entrapment, polidispersity index (PDI) and z potential) and morphological characteristics (particle morphology and particle size) were evaluated. Particle size from each nanoparticles, formulation ranged from 36.64 to 213.86 nm. Particle size, z potential and PDI increased (P ≤ 0.01) when nanoprecipitation and ethanol were used. No significant differences (P > 0.05) were observed when different polymeric proportions were used. Selenium entrapment was 26% when emulsion–evaporation method was used and 78% with nanoprecipitation. Nanoparticles produced by nanoprecipitation were spherical and had a great variation in particle size; on the other hand, nanoparticles produced by emulsion–evaporation were spherical as well as amorphous and presented a homogeneous nanopartcicle size distribution. The release of selenium from nanoparticles was higher in acid pH (less than 4), this condition may represent a better availability of the mineral in the small intestine

Impact of a Bacillus Direct-Fed Microbial on Growth Performance, Intestinal Barrier Integrity, Necrotic Enteritis Lesions, and Ileal Microbiota in Broiler Chickens Using a Laboratory Challenge Model

Decreases in the use of antibiotics and anticoccidials in the poultry industry have risen the appearance of necrotic enteritis (NE). The purpose of this study was to evaluate the effect of a Bacillus direct-fed microbial (DFM) on growth performance, intestinal integrity, NE lesions and ileal microbiota using a previously established NE-challenged model. At day-of-hatch, chicks were randomly assigned to three different groups: Negative control (NC), Positive control (PC) challenged with Salmonella Typhimurium (day 1), Eimeria maxima (EM, day 13) and Clostridium perfringens (CP, day 18–19), and Bacillus-DFM group (DFM) challenged as the PC. Body weight (BW) and body weight gain (BWG) were measured weekly. Total feed intake (FI) and feed conversion ratio (FCR) were evaluated at day 21. Liver samples were collected to assess bacterial translocation and blood samples were used to measure superoxide dismutase (SOD) and fluorescein isothiocyanate-dextran (FITC-d). Intestinal contents were obtained for determination of total IgA and microbiota analysis. NE lesion scores (LS) were performed at day 21. Chickens consuming the DFM significantly improved BW and had a numerically more efficient FCR compared to PC at day 21. Additionally, there were no significant differences in FCR between the DFM group and NC. Furthermore, the DFM group showed significant reductions in LS, IgA and FITC-d levels compared to the PC. However, there were no significant differences in SOD between the groups. The microbiota analysis indicated that the phylum Proteobacteria was significantly reduced in the DFM group in comparison to PC. At the genus level, Clostridium, Turicibacter, Enterococcus, and Streptococcus were reduced, whereas, Lactobacillus and Bacillus were increased in the DFM group as compared to PC (p &lt; 0.05). Likewise, the DFM significantly reduced CP as compared to PC. In contrary, no significant differences were observed in bacterial composition between NC vs. DFM. In addition, beta diversity showed significant differences in the microbial community structure between NC vs. PC, and PC vs. DFM. These results suggest that the dietary inclusion of a selected DFM could mitigate the complex negative impacts caused by NE possibly through mechanism(s) that might involve modulation of the gut microbiota

Risk Of Recurrence After Withdrawal Of Anticoagulation In Patients With Unprovoked Venous Thromboembolism: External Validation Of The Vienna Nomogram And The Dash Prediction Score

[ES] Introducción: Las escalas predictivas de recurrencias de ETV son útiles para decidir la duración del tratamiento anticoagulante. Aunque hay varias escalas, desconocemos la aplicabilidad de las mismas en nuestro medio. Por ello nos planteamos validar el modelo predictivo DASH y el nomograma de Viena a 12 meses.-- Métodos: Estudio retrospectivo de pacientes consecutivos no seleccionados con ETV no provocada desde 2006 hasta 2014. Comparamos la capacidad de predecir recurrencias de ETV de la escala DASH y el nomograma de Viena. La validación se realizó estratificando a los pacientes como de bajo o alto riesgo, según cada escala (discriminación) y comparando las recurrencias observadas frente a las esperadas (calibración).-- Resultados: De 353 pacientes evaluados, se analizaron 195, con una edad media de 53,5+/-19 años. Hubo 21 recurrencias a 1 año (10,8%, IC95%: 6,8-16%). Según la escala DASH, fueron catalogados de bajo riesgo el 42%, observando ETV recurrente en el grupo de bajo fue del 4,9% (IC95%: 1,3-12%) vs. el grupo de alto riesgo en que fue del 15% (IC95%: 9-23%) (p<0,05). Según el nomograma de Viena, fueron catalogados de bajo riesgo el 30%, observando ETV recurrente en el grupo de bajo vs. alto riesgo en el 4,2% (IC95%: 0,5-14%) vs. 16,2% (IC95%: 9,9-24,4%) (p<0,05).-- Conclusiones: Nuestro estudio valida la escala DASH y el nomograma de Viena en nuestra población. El modelo predictivo DASH sería el más aconsejable, tanto por su sencillez como por la capacidad de identificar a más pacientes de bajo riesgo frente al nomograma de Viena (42% vs. 30%).[EN ] Introduction: Scales for predicting venous thromboembolism (VTE) recurrence are useful for deciding the duration of the anticoagulant treatment. Although there are several scales, the most appropriate for our setting has not been identified. For this reason, we aimed to validate the DASH prediction score and the Vienna nomogram at 12 months.-- Methods: This was a retrospective study of unselected consecutive VTE patients seen between 2006 and 2014. We compared the ability of the DASH score and the Vienna nomogram to predict recurrences of VTE. The validation was performed by stratifying patients as low-risk or high-risk, according to each scale (discrimination) and comparing the observed recurrence with the expected rate (calibration).-- Results: Of 353 patients evaluated, 195 were analyzed, with an average age of 53.5 ± 19 years. There were 21 recurrences in 1 year (10.8%, 95% CI: 6.8%-16%). According to the DASH score, 42% were classified as low risk, and the rate of VTE recurrence in this group was 4.9% (95% CI: 1.3%-12%) vs. the high-risk group that was 15% (95% CI: 9%-23%) (p <.05). According to the Vienna nomogram, 30% were classified as low risk, and the rate of VTE recurrence in the low risk group vs. the high risk group was 4.2% (95% CI:0.5%-14%) vs. 16.2% (95% CI: 9.9%-24.4%) (p <.05).-- Conclusions: Our study validates the DASH score and the Vienna nomogram in our population. The DASH prediction score may be the most advisable, both because of its simplicity and its ability to identify more low-risk patients than the Vienna nomogram (42% vs. 30%).Peer reviewe

Nirmatrelvir/ritonavir in COVID-19 patients with haematological malignancies: a report from the EPICOVIDEHA registry

Background: Nirmatrelvir/ritonavir treatment decreases the hospitalisation rate in immunocompetent patients with COVID-19, but data on efficacy in patients with haematological malignancy are scarce. Here, we describe the outcome of nirmatrelvir/ritonavir treatment in a large cohort of the latter patients. Methods: This is a retrospective cohort study from the multicentre EPICOVIDEHA registry (NCT04733729) on patients with haematological malignancy, who were diagnosed with COVID-19 between January and September 2022. Patients receiving nirmatrelvir/ritonavir were compared to those who did not. A logistic regression was run to determine factors associated with nirmatrelvir/ritonavir administration in our sample. Mortality between treatment groups was assessed with Kaplan-Meier survival plots after matching all the patients with a propensity score. Additionally, a Cox regression was modelled to detect factors associated with mortality in patients receiving nirmatrelvir/ritonavir. Findings: A total of 1859 patients were analysed, 117 (6%) were treated with nirmatrelvir/ritonavir, 1742 (94%) were treated otherwise. Of 117 patients receiving nirmatrelvir/ritonavir, 80% had received ≥1 anti-SARS-CoV-2 vaccine dose before COVID-19 onset, 13% of which received a 2nd vaccine booster. 5% were admitted to ICU. Nirmatrelvir/ritonavir treatment was associated with the presence of extrapulmonary symptoms at COVID-19 onset, for example anosmia, fever, rhinitis, or sinusitis (aOR 2.509, 95%CI 1.448-4.347) and 2nd vaccine booster (aOR 3.624, 95%CI 1.619-8.109). Chronic pulmonary disease (aOR 0.261, 95%CI 0.093-0.732) and obesity (aOR 0.105, 95%CI 0.014-0.776) were not associated with nirmatrelvir/ritonavir use. After propensity score matching, day-30 mortality rate in patients treated with nirmatrelvir/ritonavir was 2%, significantly lower than in patients with SARS-CoV-2 directed treatment other than nirmatrelvir/ritonavir (11%, p&nbsp;=&nbsp;0.036). No factor was observed explaining the mortality difference in patients after nirmatrelvir/ritonavir administration. Interpretation: Haematological malignancy patients were more likely to receive nirmatrelvir/ritonavir when reporting extrapulmonary symptoms or 2nd vaccine booster at COVID-19 onset, as opposed to chronic pulmonary disease and obesity. The mortality rate in patients treated with nirmatrelvir/ritonavir was lower than in patients with targeted drugs other than nirmatrelvir/ritonavir. Funding: EPICOVIDEHA has received funds from Optics COMMIT (COVID-19 Unmet Medical Needs and Associated Research Extension) COVID-19 RFP program by GILEAD Science, United States (Project 2020-8223)