Spermatozoa Obtained From Alpaca vas deferens. Effects of Seminal Plasma Added at Post-thawing

The viscous seminal plasma (SP) is currently a major impediment to the handling of ejaculate and the development of some biotechnologies in South American camelids. The vas deferens-collected spermatozoa of alpacas is a useful technique to avoid this problem. On the other hand, SP contains a large protein component that has been implicated in the function of spermatozoa within the female reproductive tract. In this sense, the low fertility achieved using transcervical insemination with frozen-thawed spermatozoa in alpacas could be improved by adding SP. This study aimed to evaluate the effect of the whole SP on some in vitro parameters of alpaca spermatozoa after the freezing-thawing-process and the fertility after artificial insemination. It would contribute to a better understanding of the interaction between thawed sperm cells and SP. Spermatozoa were obtained by surgically diverted vas deferens. The samples were diluted with a Tris-based extender, packaged in straws, and frozen. At thawing, each straw was divided into two post-thawing conditions: with the addition of 10% of PBS (control) or with 10% SP (treatment). The sperm cells were evaluated using dynamic parameters, sperm cell morphology, and morphometry. Fertility was assessed by an artificial insemination trial. All in vitro parameters were analyzed by ANOVA. A heterogeneity test was scheduled for the fertility trial. After the freezing-thawing process, motility and plasma membrane functionality was improved when SP was added. No differences were found for post-thaw viability between the control and treatment samples. The percentage of normal cells was higher with SP at post-thawing, and a decrease of the presence of bent tailed spermatozoa with a droplet in the SP group was observed. The length of the head spermatozoa was 3.4% higher in the samples with PBS compared to those in which SP was added. Females pregnant at day 25 post-insemination were 0/12 (with SP inside the straw) and 1/10 (without SP inside the straw). In conclusion, the presence of 10% SP at post-thawing improves sperm cells' motility, functionality, and morphology, indicating that it would be beneficial to improve the frozen-thawed alpaca's physiology spermatozoa. More fertility trials must be developed to increase this knowledge

Caracterización ultrasonográfica modo-B y Doppler del cuerpo lúteo en llamas

Mode B and Doppler ultrasonography in ruminants, as a technique, has allowed establishing new concepts on the reproductive physiology of females, through the study of follicular dynamics and morphometry of the corpus lu-teum, therefore, the objective was to characterize using mode B ultrasound and Doppler the corpus luteum in recipients’ llamas. 37 llamas were used as recipients (27 for mode B and 10 Doppler ultrasound) that were syn-chronized by applying buserelin acetate 0.0096 mg, nine days later 0.048 mg of Prostaglandin F2α analog was applied, two days later the second dose of buserelin acetate in the same dose to a ovulation and subsequent for-mation of the corpus luteum, The B-mode evaluation of the morphometry consisted of observing the ecotexture, area, diameter and volume of the corpus luteum with the use of a SonoStar SS-8® ultrasound machine at 6.5 MHz frequency and 6 cm depth equipped with a transrectal linear transducer, with the same technique with a Draminski 4Vet® Doppler device, the percentage of luteal area of vascularization (% AVL) was determined. The data were subjected to a descriptive analysis, being 0.119±0.032 cm2 in area, 12.7±1.7 mm in diameter and 0.84±0.32 cm3 in volumen of CL, to determine the relationship between metric measurements the Pearson correlation was used observing a positive relationship between area/diameter of 0.6602, observing a high positive correlation between area and volume, finally, the characteristics of % AVL was 34.97 for the CLs of recipients llamas. In conclusion, the morphometric characteristics evaluated in mode B (area, diameter, and volume) have a positive correlation and the % AVL could be applied as a tool in efficient reproduc-tive management for the selection of receptors in camelids, understanding that it is one of the first reports of these characteristics in llamas. 0.7506, area/volume of 0.9289 and diameter/volume of La ultrasonografía modo B y Doppler en los rumiantes, como técnica ha permitido establecer nuevos conceptos sobre la fisiología reproductiva de las hembras, a través del estudio de la dinámica folicular y morfometría del cuerpo lúteo, por tanto, el objetivo fue caracterizar mediante ultrasonografía modo B y Doppler el cuerpo lúteo en llamas receptoras. Se utilizaron 37 llamas como receptoras (27 para ultrasonografía modo B y 10 Doppler) que fueron sincronizadas aplicando acetato de buserelina 0.0096 mg, nueve días posteriores se aplicó 0.048 mg de análogo de prostaglandina F2α, a los dos días posteriores se aplicó la segunda dosis de acetato de buserelina en la misma dosis para garantizar la ovulación y posterior formación del cuerpo lúteo, la evaluación en modo B de la morfometría consistió en observar la ecotextura, área, diámetro y volumen del cuerpo lúteo con el uso de un equipo ultrasonógrafo SonoStar SS-8® a 6.5 MHz de frecuencia y 6 cm de profundidad equipado con un transductor lineal transrectal; con la misma técnica con un equipo Draminski 4Vet® Doppler se determinó el porcentaje de área luteal de vascularización (% AVL). Los datos fueron sometidos a un análisis descriptivo siendo de 0.119±0.032 cm2 de área, 12.7±1.7 mm de diámetro y 0.84±0.32 cm3 de volumen de CL, para determinar la relación entre medidas métricas se utilizó la correlación de Pearson observando una relación positiva entre área/diámetro de 0.7506, área/volumen de 0.9289 y diámetro/volumen de 0.6602, observando una correlación alta positiva entre área y volumen, finalmente las características de % AVL fue de 34.97 para los CL de llamas receptoras. En conclusión, las características morfométricas evaluadas en modo B (área, diámetro y volumen) tienen una correlación positiva y el % AVL podría aplicarse como herramienta en el manejo reproductivo eficiente para la selección de receptoras en camélidos, entendiendo que es uno de los primeros reportes de estas características en llamas

Efecto de la yema de huevo sobre la calidad de semen descongelado de paco (Piaractus brachypomus)

The objective of the study was to evaluate the effect of the type of egg yolk (quail-YC vs hen-YG) on the quality of frozen paco (Piaractus brachypomus) semen (volume, colour, concentration, motility, vitality and fertility). Eighteen breeders (15 males and 3 females) were used, whose spermiation and ovulation, respectively, were induced with carp pituitary extract. Semen was collected by massaging the abdomen of the fish. Fresh semen was evaluated and then diluted with a cryoprotective solution that included YC or YG. Diluted semen was frozen and thawed at 30 days for post-thaw evaluation and fertility testing. A notable decrease in seminal parameters was found due to the effect of freezing. Motility was lower for YC compared to YG (14.8 vs. 18.8%; p<0.05), while vitality (31 vs. 28.8%), activation time (1.4 vs. 1.3 min) and fertility (18.4 vs. 20.4%) were similar between groups. In conclusion, the motility was improved with the addition of chicken egg yolk, while the vitality, activation time and fertility were not influenced by the type of yolk.El objetivo del estudio fue evaluar el efecto del tipo de yema de huevo (codorniz-YC vs gallina-YG) sobre la calidad del semen de paco (Piaractus brachypomus) (volumen, color, concentración, motilidad, vitalidad y fertilidad) congelado. Se utilizaron 18 reproductores (15 machos y 3 hembras) cuya espermiación y ovulación, respectivamente, fue inducida con extracto de hipófisis de carpa. El semen fue colectado mediante masaje del abdomen del pez. El semen fresco fue evaluado y luego diluido con una solución crioprotetora que incluyó YC o YG. El semen diluido fue congelado y descongelado a los 30 días para la evaluación pos-descongelación y prueba de fertilidad. Se encontró una notoria disminución de los parámetros seminales por efecto de la congelación. La motilidad fue menor para la YC comparado a la YG (14.8 vs. 18.8%; p<0.05), mientras que la vitalidad (31 vs. 28.8%), tiempo de activación (1.4 vs. 1.3 min) y la fertilidad (18.4 vs. 20.4%) fueron similares entre grupos. En conclusión, la motilidad fue mejorada con la adición de yema de huevo de gallina, mientras que la vitalidad, tiempo de activación y fertilidad no fueron influenciados por el tipo de yema

Sustentabilidad productiva de la instalación de sistemas silvopastoriles: una revisión sistemática basada en la realidad de Perú y Colombia

Most of the world's livestock is developed in open field or monoculture systems, which generates economic and environmental losses, such as soil erosion, loss of biodiversity of flora and fauna, low meat and milk production, and less monetary income. Faced with this situation, there are sustainable alternatives for livestock production, which are friendly to the environment, being silvopastoral systems. That is why, in this review, it is proposed to analyze the productive sustainability of the installation of silvopastoral systems, placing greater emphasis on the floristic composition of plots with silvopastoral systems in Peru and Colombia, as a case study. The information review was based on the PRISMA statement considering inclusion and exclusion criteria. Comparisons were made of pasture production and quality, productivity of dairy and fattening cattle, social and environmental opportunities, and the floristic composition of such silvopastoral systems. The installation of silvopastoral systems increases the productive yields of pastures (dry matter, digestibility and protein), of animals such as greater milk and meat production, and greater biodiversity of forage species. In both countries it is observed that the implementation of these systems has improved opportunities for producers dedicated to livestock due to the beneficial economic, environmental and social impacts that they provide. In this sense, it is a challenge for scientists, governments and entities involved in sustainable systems to continue looking for new viable alternatives and validate them for livestock production.La mayor parte de la ganadería a nivel mundial se viene desarrollando en sistemas a campo abierto (sca) o monocultivos, la cual genera pérdidas económicas y ambientales, así como erosión de suelos, pérdida de biodiversidad de flora y fauna, baja producción de carne y leche, y menos ingresos monetarios. Frente a esta situación, existen alternativas sostenibles y sustentables de producción pecuaria que son amigables con el medioambiente, como los sistemas silvopastoriles (ssp). Por lo que, en esta revisión, se plantea analizar la sustentabilidad productiva de la instalación de ssp, al hacer un mayor énfasis en la composición florística de parcelas con ssp en Perú y Colombia, como estudio de caso. La revisión de información fue basada en la declaración Prisma, considerando criterios de inclusión y exclusión, se realizaron comparaciones de producción y calidad del pasto, productividad del ganado lechero y de engorde, oportunidades sociales y ambientales y la composición florística de tales ssp. La instalación de ssp incrementa los rendimientos productivos de los pastos (materia seca, digestibilidad y proteína) y de los animales, como mayor producción de leche y carne, y mayor biodiversidad de especies forrajeras. Además, en ambos países se observa que la implementación de estos sistemas ha mejorado las oportunidades para los productores dedicados a la ganadería, por los beneficiosos impactos económicos, ambientales y sociales que estos brindan. En ese sentido, es un reto para los científicos, gobiernos y entidades involucradas en sistemas sostenibles para seguir buscando nuevas alternativas viables y validarlas para la producción ganadera

Genomic investigations of unexplained acute hepatitis in children

Since its first identification in Scotland, over 1,000 cases of unexplained paediatric hepatitis in children have been reported worldwide, including 278 cases in the UK1. Here we report an investigation of 38 cases, 66 age-matched immunocompetent controls and 21 immunocompromised comparator participants, using a combination of genomic, transcriptomic, proteomic and immunohistochemical methods. We detected high levels of adeno-associated virus 2 (AAV2) DNA in the liver, blood, plasma or stool from 27 of 28 cases. We found low levels of adenovirus (HAdV) and human herpesvirus 6B (HHV-6B) in 23 of 31 and 16 of 23, respectively, of the cases tested. By contrast, AAV2 was infrequently detected and at low titre in the blood or the liver from control children with HAdV, even when profoundly immunosuppressed. AAV2, HAdV and HHV-6 phylogeny excluded the emergence of novel strains in cases. Histological analyses of explanted livers showed enrichment for T cells and B lineage cells. Proteomic comparison of liver tissue from cases and healthy controls identified increased expression of HLA class 2, immunoglobulin variable regions and complement proteins. HAdV and AAV2 proteins were not detected in the livers. Instead, we identified AAV2 DNA complexes reflecting both HAdV-mediated and HHV-6B-mediated replication. We hypothesize that high levels of abnormal AAV2 replication products aided by HAdV and, in severe cases, HHV-6B may have triggered immune-mediated hepatic disease in genetically and immunologically predisposed children

Metodología para la formación de competencia investigativa en los estudiantes de la Universidad de Guantánamo

Teniendo en cuenta las insuficiencias detectadas en la actividad práctico investigativa de los estudiantes de las facultades Ciencias Sociales y Humanísticas, y Agroforestal de la Universidad de Guantánamo,se elaboró una metodología para la formación de la competencia investigativa de estudiantes universitarios. En esta metodología se establecen cuatro fases que, de manera progresiva, a partir de niveles de complejización (iniciación, potenciación y transformación), favorecen el desempeño investigativo de los educandos para enfrentar su objeto social una vez graduados, y contempla la preparación más adecuada del claustro de profesores de la facultad para guiar este proceso formativo

Factores que afectan la tasa de preñez en receptoras de embriones producidos in vitro bajo condiciones de altura

The aim of this study was to determine the factors that affect pregnancy in recipient females of embryos produced in vitro under high altitude conditions. The study was carried out at the Universidad Nacional del Altiplano (UNA), located in Puno, Peru, at an altitude of 3970 m. 50 recipient females (nulliparous, primiparous and multiparous) Brown Swiss were used. Ovaries were collected from cows at a local slaughterhouse and taken to the UNA animal reproduction laboratory, where the oocytes were collected, and in vitro maturation and fertilization, culture and embryo evaluation were carried out. The transport time to the laboratory was 2 to 3 h. Quantitative factors such as body condition score (CC), diameter of the corpus luteum (CL) and number of parities, and qualitative factors such as the place of deposit of the embryo (right or left horn) and embryo quality, were evaluated using the Wilcoxon and Chi test. square, respectively. Likewise, a binary logistic regression was applied to study all the factors together. The highest pregnancy rate occurred when the recipients had a 2.65 CC, 19.54 mm CL diameter and in primiparous as quantitative factors and when transferred to the left ipsilateral horn (9/12) with grade 1 embryos (G1: 11/ 17) as qualitative factors. However, the binary logistic regression model did not determine significance as a whole.El objetivo del estudio fue determinar los factores que afectan la preñez en hembras receptoras de embriones producidos in vitro bajo condiciones de altura. El estudio se llevó a cabo en la Universidad Nacional del Altiplano (UNA), ubicada en Puno, Perú, a una altitud de 3970 m. Se utilizaron 50 hembras receptoras (nulíparas, primíparas y multíparas) Brown Swiss. Se colectaron ovarios de vacas sacrificadas en un centro de beneficio local y se llevaron al laboratorio de reproducción animal de la UNA, donde se colectaron los ovocitos, y se procedió con la maduración y fertilización in vitro, cultivo y evaluación de embriones. El tiempo de transporte hasta el laboratorio fue de 2 a 3 h. Se evaluaron factores cuantitativos como condición corporal (CC), diámetro de cuerpo lúteo (CL) y número de partos, y factores cualitativos como lugar de depósito del embrión (cuerno derecho o izquierdo) y calidad de embrión, mediante la prueba de Wilcoxon y Chi cuadrado, respectivamente. Asimismo, se aplicó una regresión logística binaria para el estudio de todos los factores en conjunto. La mayor tasa de preñez se presentó cuando las receptoras tenían 2.65 de CC, 19.54 mm de diámetro de CL y en primíparas como factores cuantitativos y cuando se transfirió al cuerno ipsilateral izquierdo (9/12) con embriones de grado 1 (G1: 11/17) como factores cualitativos. No obstante, el modelo de regresión logística binaria no determinó significancia en conjunto

Ovarian Follicular Dynamics and Its Functional Significance in Relation with Follicle Deviation, Vaginal Cytology, and Hormone Profiles in Llamas (<i>Lama glama</i>)

The reproductive physiology in camelid species has its particularities. The present study aimed to characterize the ovarian follicular dynamics and its functional significance in relation to follicular deviation, vaginal cytological characteristics, and sexual hormone profiles in llamas as the first report in South American camelids. Non-pregnant, multiparous llamas (Lama glama; n = 10; age: 48–72 mo.; BCS: 2.5–3.0) were enrolled in the study. The ultrasonographic assessment was carried out transvaginally and follicular ablation was performed (day 0) when follicles were larger than 7 mm. The follicle number and diameter were scored daily throughout the process for a proper evaluation of the deviated follicles and to monitor the presence of new follicle pools (1.5 to 2.5 mm diameter). Vaginal cytological evaluation (parabasal, intermediate, and superficial cells) was performed every other day until day 6. Endocrine profiles (17β estradiol, anti-Mullerian hormone, testosterone, and progesterone) during pre- and post-follicular deviation were determined by using the ELISA assay. Differential follicular dynamics both in the presence of a single dominant follicle (DF) and in codominance during the follicular deviation process were detected in llamas (p p > 0.05). Differential patterns among the different hormone concentration levels regarding the 17β estradiol, anti-Mullerian hormone, progesterone, and testosterone during follicular deviation were observed, with the latter being significantly different along the deviation process (p < 0.05). In conclusion, the use of vaginal cytology assessment would not be sufficient to determine the follicular phases in llamas. Therefore, complementary analyses, such as ultrasonography and endocrine assessment, are strongly recommended to determine follicular dynamics during the follicular deviation