Author: Carlos Eliud Angulo Valadez

Serodiagnóstico e inmunogenicidad producida por proteínas de la glándula salival de Oestrus ovis (Diptera: Oestridae) en ovejas y cabras

Carlos Eliud Angulo Valadez (2009)

En el contenido de la glándula salival larvaria de Oestrus ovis (Díptera:Oestridae) se encuentran antígenos reconocidos por las IgG de ovejas infectadas que pueden ser útiles para el diagnóstico de la estrosis ovina y caprina. Adicionalmente, las larvas parásitas de Hypoderma lineatum (Díptera:Oestridae) secretan proteasas antigénicas usadas para el diagnóstico de la hipodermosis bovina en programas de erradicación. Los objetivo general de esta tesis fue investigar las proteasas-antigénicas y los antígenos en el contenido de la glándula salival de O. ovis y determinar su valor antigénico para el diagnóstico de la estrosis en cabras y ovejas. Para ello se capturaron larvas de tercer estadio (L3) y moscas grávidas para obtener antígenos salivales y larvas de primer estadio (L1) infectivas de O. ovis. Se disectaron las larvas L3 para obtener la glándula salival del parásito y sus productos fueron analizados en pruebas enzimáticas (Apyzim, Azocoll, Zimogramas) e inmunoblots y en el desarrollo de pruebas de ELISA para el serodiagnóstico de la estrosis en ovejas y cabras. Se registró el número y desarrollo de larvas de O. ovis encontradas en cabras y ovejas infectadas. Además, se colectaron muestras de suero sanguíneo de caprinos y ovinos infectados experimental y naturalmente así como moco nasal de ovinos. Las ovejas del rebaño estudiado se organizaron en tres grupos de acuerdo al Progenitor que las engendró. Los resultados encontrados fueron: se detectaron las actividades enzimáticas de fosfatasa ácida, naftol-AS-BI-fosfohidrolasa, esterasa (C4), esterasa lipasa (C8), leucina arilamidasa, α-gludosidasa y N-acetil-β-glucosaminidasa. El pH óptimo de actividad proteolítica fue de 8.0; mientras que la actividad proteolítica se incrementó con la temperatura (10 a 50 °C). Además, las enzimas proteolíticas reaccionaron solamente con inhibidores para serin-proteasas. En los zimogramas, se detectaron bandas de actividad proteolítica en el rango de 20 a 63 kDa; y en los inmunoblots se observaron tres bandas antigénicas, una de ellas relacionada a una banda proteolítica (63 kDa). En ovejas, se encontraron correlaciones negativas entre el establecimiento larvario y/o el desarrollo larvario por un lado y la intensidad de las respuestas sistémicas y locales de IgG por el otro en dos de los tres grupos Progenitor-progenie estudiados [...]

The major antigens recognized by IgG from Oestrus ovis (Díptera:Oestridae) infected sheep have been found in salivary gland content that can be useful for diagnosis of ovine and caprine oestrosis disease. In addition, parasitic larvae of Hypoderma lineatum (Díptera:Oestridae) secrete antigenic proteases used for bovine hypodermosis diagnosis in eradication programs. The general objective of this thesis was to investigate antigenic proteases and proteolytic activity in salivary gland content of O. ovis third instars and to determine their antigenic value for sheep and goat oestrosis diagnosis. Third instars (L3) and gravid flies were collected to obtain salival antigens and infective first instars of O. ovis. L3 larvae were dissected and their contents were analyzed in enzymatic tests (Apyzim, Azocoll, Zymogramas) and immunoblots as well as in ELISA test development to serodiagnosis of sheep and goats oestrosis. The number and development of O. ovis larvae found in sheep and goats were recordered. Aditionally, blood serum samples from sheep and goats infected naturally or experimentally were used as well as nasal mucus of sheep. The sheep of the flock were divided into three groups Ram-families. The results found were: enzymatic activities in salivary gland contents of acid phosphatase, naphthol-AS-BI-phosphohydrolase, esterase (C4), esterase lipase (C8), leucine arylamidase, α-glucosidase and N-acetyl-β-glucosaminidase, were detected. Optimum pH for proteolytic activity was 8.0, while proteolytic activity increased with temperature (10 to 50 °C) then drastically decreased at 60 °C. Proteases in O. ovis salivary gland products belong to the serine subclass. In Zymograms, bands of proteolytic activity were detected in the 20 to 63 kDa range; the immunoblot showed three antigenic bands, one of them related to a protease band (63 kDa). In ovine specie, negative correlations among larval establishment and/or larval development on the one hand and intensity of local or systemic IgG responses on the other hand were found in two out of three studied ram-families [...]

Doctoral thesis

Parásito Oestrus ovis; ovejas y cabras; respuestas inmunes CIENCIAS AGROPECUARIAS Y BIOTECNOLOGÍA CIENCIAS AGRARIAS CIENCIAS VETERINARIAS PARASITOLOGÍA ANIMAL PARASITOLOGÍA ANIMAL

A Perspective on the Development of Plant-Made vaccines in the Fight against ebola virus

SERGIO ROSALES MENDOZA RICARDO NIETO GOMEZ CARLOS ELIUD ANGULO VALADEZ (2017)

"The Ebola virus (EBOV) epidemic indicated a great need for prophylactic and therapeutic strategies. The use of plants for the production of biopharmaceuticals is a concept being adopted by the pharmaceutical industry, with an enzyme for human use currently commercialized since 2012 and some plant-based vaccines close to being commercialized. Although plant-based antibodies against EBOV are under clinical evaluation, the development of plant-based vaccines against EBOV essentially remains an unexplored area. The

current technologies for the production of plant-based vaccines include stable nuclear expression, transient expression mediated by viral vectors, and chloroplast expression. Specific perspectives on how these technologies can be applied for developing anti- EBOV vaccines are provided, including possibilities for the design of immunogens as well as the potential of the distinct expression modalities to produce the most relevant EBOV antigens in plants considering yields, posttranslational modifications, production time, and downstream processing."

Article

Ebola virus, mucosal immunization, low-cost vaccine, global vaccination, molecular pharming, glycoprotein antigen, VP antigen BIOLOGÍA Y QUÍMICA CIENCIAS DE LA VIDA INMUNOLOGÍA VACUNAS

Algevir: An Expression System for Microalgae Based on Viral Vectors

BERNARDO BAÑUELOS HERNANDEZ ELIZABETH MONREAL ESCALANTE OMAR GONZALEZ ORTEGA CARLOS ELIUD ANGULO VALADEZ SERGIO ROSALES MENDOZA (2017)

"The use of recombinant algae for the production of valuable compounds is opening promising biotechnological applications. However, the development of efficient expression approaches is still needed to expand the exploitation of microalgae in biotechnology. Herein, the concept of using viral expression vectors in microalgae was explored for the first time. An inducible geminiviral vector leading to Rep-mediated replication of the expression cassette allowed the production of antigenic proteins at high levels. This system, called Algevir, allows the production of complex viral proteins (GP1 from Zaire ebolavirus) and bacterial toxin subunits (B subunit of the heat-labile Escherichia coli enterotoxin), which retained their antigenic activity. The highest achieved yield was 1.25 mg/g fresh biomass (6 mg/L of culture), which was attained 3 days after transformation. The Algevir system allows for a fast and efficient production of recombinant proteins, overcoming the difficulties imposed by the low yields and unstable expression patterns frequently observed in stably transformed microalgae at the nuclear level; as well as the toxicity of some target proteins."

Article

Schizochytrium sp., marine microalgae, vaccine, replicon, viral elements, geminivirus, recombinant protein yield, transient expression BIOLOGÍA Y QUÍMICA CIENCIAS DE LA VIDA BIOLOGÍA VEGETAL (BOTÁNICA) ALGOLOGÍA (FICOLOGÍA)

Uso de hierbas aromáticas en la producción animal

MARTHA CANDELARIA REYES BECERRIL JOSE ANGEL ARMENTA QUINTANA Carlos Eliud Angulo Valadez (2020)

"El objetivo de este trabajo fue describir el uso de las hierbas aromáticas en la producción animal. Las hierbas aromáticas poseen propiedades nutricionales y medicinales (como antioxidantes e inmunoestimulantes). Algunos estudios han demostrado que la adición de hierbas aromáticas en la dieta para animales puede mejorar el rendimiento productivo y la salud de los sistemas de producción pecuaria. Por ello, la adición de hierbas aromáticas (como el orégano) en las dietas es una alternativa que brinda múltiples beneficios en la producción animal, incluidos la mejora del sabor de las dietas, el consumo del alimento, la nutrición y la salud animal."

"The aim of this study was describing the aromatic herbs use on animal production. The aromatic herbs have nutritional and medicinal properties (such as antioxidants and immunostimulants). Several studies have demonstrated that aromatic herbs addition into animal diets can improve productive performance and health in livestock production systems. Therefore, the addition of aromatic herbs (such as oregano) into diets is an alternative with multiple benefi ts in animal production, including the improvement of diets fl avor, feed consumption, animal’s nutrition and health."

Article

plantas medicinales, aditivos alimenticios, producción de alimentos, salud animal medicinal plants, feed additives, food production, animal health CIENCIAS AGROPECUARIAS Y BIOTECNOLOGÍA CIENCIAS AGRARIAS PRODUCCIÓN ANIMAL NUTRICIÓN NUTRICIÓN

Prospects on the Use of Schizochytrium sp. to Develop Oral Vaccines

Abel Antonio Ramos Vega SERGIO ROSALES MENDOZA BERNARDO BAÑUELOS HERNANDEZ Carlos Eliud Angulo Valadez (2018)

"Although oral subunit vaccines are highly relevant in the fight against widespread diseases, their high cost, safety and proper immunogenicity are attributes that have yet to be addressed in many cases and thus these limitations should be considered in the development of new oral vaccines. Prominent examples of new platforms proposed to address these limitations are plant cells and microalgae. Schizochytrium sp. constitutes an attractive expression host for vaccine production because of its high biosynthetic capacity, fast growth in low cost culture media, and the availability of processes for industrial scale production. In addition, whole Schizochytrium sp. cells may serve as delivery vectors; especially for oral vaccines since Schizochytrium sp. is safe for oral consumption, produces immunomodulatory compounds, and may provide bioencapsulation to the antigen, thus increasing its bioavailability. Remarkably, Schizochytrium sp. was recently used for the production of a highly immunoprotective influenza vaccine. Moreover, an efficient method for transient expression of antigens based on viral vectors and Schizochytrium sp. as host has been recently developed. In this review, the potential of Schizochytrium sp. in vaccinology is placed in perspective, with emphasis on its use as an attractive oral vaccination vehicle."

Article

adjuvant, bioencapsulation, microalgae, oral vaccine, thermostable vaccine, Algevir system BIOLOGÍA Y QUÍMICA CIENCIAS DE LA VIDA INMUNOLOGÍA VACUNAS VACUNAS

Prospects on the Use of Schizochytrium sp. to Develop Oral Vaccines

Abel Antonio Ramos Vega SERGIO ROSALES MENDOZA BERNARDO BAÑUELOS HERNANDEZ Carlos Eliud Angulo Valadez (2018)

"Although oral subunit vaccines are highly relevant in the fight against widespread diseases, their high cost, safety and proper immunogenicity are attributes that have yet to be addressed in many cases and thus these limitations should be considered in the development of new oral vaccines. Prominent examples of new platforms proposed to address these limitations are plant cells and microalgae. Schizochytrium sp. constitutes an attractive expression host for vaccine production because of its high biosynthetic capacity, fast growth in low cost culture media, and the availability of processes for industrial scale production. In addition, whole Schizochytrium sp. cells may serve as delivery vectors; especially for oral vaccines since Schizochytrium sp. is safe for oral consumption, produces immunomodulatory compounds, and may provide bioencapsulation to the antigen, thus increasing its bioavailability. Remarkably, Schizochytrium sp. was recently used for the production of a highly immunoprotective influenza vaccine. Moreover, an efficient method for transient expression of antigens based on viral vectors and Schizochytrium sp. as host has been recently developed. In this review, the potential of Schizochytrium sp. in vaccinology is placed in perspective, with emphasis on its use as an attractive oral vaccination vehicle."

Article

adjuvant, bioencapsulation, microalgae, oral vaccine, thermostable vaccine, Algevir system BIOLOGÍA Y QUÍMICA CIENCIAS DE LA VIDA INMUNOLOGÍA VACUNAS VACUNAS

Chitinolytic bacteria associated to Oestrus ovis L. larvae (Diptera: Oestridae)

GREGORIO RODRIGUEZ VALDEZ Carlos Eliud Angulo Valadez Reyna de Jesús Romero Geraldo RAMON CEPEDA PALACIOS (2018)

"The objectives of this study were: I. To culture, isolate, identify and characterize bacterial species obtained from O. ovis L3 larval tissues and the nasosinusal mucosa of goat hosts and, II. To test the in vitro larvicidal effects of selected chitinolytic bacteria against O. ovis L2 and L3 larvae. A total of thirty three bacteria species were isolated and identified by molecular (16S RNA sequence) and biochemical techniques, and phylogenetic trees were constructed. Aeromonas veronii was the most frequent (30%) species isolated from larvae and mucosa tissues. Four bacteria: Pantoea calida, Pantoea gaviniae, Serratia marcescens and A. veronii showed high in vitro chitinolyitic activity. These bacteria were experimentally inoculated in the rearing media of groups of L2 and L3 larvae. Mortality rates in L2 larvae exposed to isolates of Pantoea gaviniae (100%), Aeromonas veronii (100%), and Serratia marcescens (100%) were statistically higher (P < 0.05) than in the control group (12%) at 48 h post-incubation. In L3 larvae, mortality rate at 48 h post-incubation was found higher (P < 0.05) in larvae exposed to Pantoea calida (35%), than in larvae exposed to the other bacteria and control group (12.5%). In conclusion, this is the first report dealing with the identification of bacterial species associated to O. ovis larvae, showing potential effects to cause damage and mortality in larval instars. The use of these bacteria as biological control agents against O. ovis deserves further investigation."

"Les objectifs de cette étude étaient les suivants : I. Culture, isolement, identification et caractérisation des espèces bactériennes obtenues à partir des larves O. ovis L3 et de la muqueuse nasosinusale des hôtes (caprins). II. Tester les effets larvicides in vitro de certaines bactéries chitinolytiques contre les larves O. ovis L2 et L3. Au total, trente-trois espèces de bactéries ont été isolées et identifiées par voie moléculaire (séquence d’ARN 16S) et par techniques biochimiques. Des arbres phylogénétiques ont été construits. Aeromonas veronii était l’espèce la plus fréquente (30%) isolée à partir de larves et des muqueuses. Quatre bactéries : Pantoea calida, Pantoea gaviniae, Serratia marcescens et A. veronii ont montré une forte activité chitinolytique in vitro. Ces bactéries ont été inoculées expérimentalement dans les milieux d’élevage de larves L2 et L3. Les taux de mortalité des larves de L2 exposées aux isolats de Pantoea gaviniae (100%), Aeromonas veronii (100%) et Serratia marcescens (100%) étaient statistiquement plus élevés (P <0,05) que dans le groupe témoin (12%) à 48 h d’incubation. Pour les larves L3, le taux de mortalité à 48 heures après l’incubation a été plus élevé (P <0,05) chez les larves exposées à Pantoea calida (35%) que chez celles exposées aux autres groupes de bactéries et aux témoins (12,5%). En conclusion, il s’agit de la première identification d’espèces bactériennes associées aux larves d’O. ovis présentant des effets potentiels létaux sur les stades larvaires. L’utilisation de ces bactéries comme agents de lutte biologique contre O. ovis mériterait des recherches approfondies."

Article

Myiasis, larvicidal effect, parasite, goat, sheep nose bots, bacteria, Oestridae effet larvicide, parasite, chèvres, bactéries, Oestrus ovis BIOLOGÍA Y QUÍMICA CIENCIAS DE LA VIDA MICROBIOLOGÍA BACTERIOLOGÍA BACTERIOLOGÍA

Rumbo al desarrollo de una vacuna de bajo costo contra la enfermedad de Chagas

Abel Antonio Ramos Vega Elizabeth Monreal Escalante Carlos Eliud Angulo Valadez (2020)

"La enfermedad de Chagas, causada por el parásito Trypanosoma cruzi, es una de las enfermedades tropicales desatendidas más importantes ya que afecta a 10 millones de personas, principalmente en América Latina. Actualmente no existe una vacuna contra esta enfermedad y los medicamentos disponibles no son del todo eficaces. Al presentarse mayoritariamente en países en desarrollo, la obtención de una vacuna resulta difícil de costear y poco redituable para las compañías farmacéuticas. Es por ello que se propone el desarrollo de vacunas de bajo costo, que sean de entrega oral, permanezcan a temperatura ambiente y que se produzcan rápidamente. Las vacunas recombinantes producidas en plantas y microalgas representan una opción relevante que se discute en este artículo."

"Chagas disease is one of the most important neglected tropical diseases, caused by Trypanosoma cruzi parasite and infects about 10 million people, mainly in Latin America. Current medications are not entirely effective and there is not available vaccine against this disease. Because is presented mostly in developing countries, obtaining a vaccine is diffi cult to pay and not very profi table for pharmaceutical companies. We propose the development of low-cost vaccines: orally delivered, remain at room temperature and produced quickly. For this, recombinant vaccines produced in plants and microalgae represent a relevant option that is discussed in this article."

Article

Vacunas de bajo costo, enfermedad de Chagas, vacunas recombinantes Low-cost vaccines, Chagas disease, recombinant vaccines BIOLOGÍA Y QUÍMICA CIENCIAS DE LA VIDA INMUNOLOGÍA VACUNAS VACUNAS

IDENTIFICATION AND CHARACTERIZATION OF Eimeria spp. DURING EARLY NATURAL INFECTION IN GOAT KIDS IN BAJA CALIFORNIA SUR, MEXICO

IDENTIFICACIÓN Y CARACTERIZACIÓN DE Eimeria spp. DURANTE LA INFECCIÓN NATURAL TEMPRANA EN CABRITOS EN BAJA CALIFORNIA SUR, MÉXICO

RAMON CEPEDA PALACIOS JUAN MANUEL RAMIREZ ORDUÑA RAFAEL RAMIREZ ORDUÑA FELIPE DE JESUS ASCENCIO VALLE CARLOS ELIUD ANGULO VALADEZ (2015)

"The aim of this study was to characterize the natural infection by Eimeria spp. in goat kids, and to describe some pathophysiological responses to eimerosis in kids under intensive rearing conditions in B.C.S, Mexico. Nineteen adult crossbred does naturally infected with mixed Eimeria spp. and 20 Anglo Nubian x Creole crossbred kids were used. Oocyst per gram of feces (OPG) and identification of Eimeria species were determined in does (during the prekidding and post-kidding periods) and kids. Clinical signs, hematocrit, hemoglobin and alkaline phosphatase activity in blood serum were evaluated. OPG (mean±SD) was significantly higher (P<0.05) in pre-kidding (9,478±7,599) than in post-kidding (5,313±2,909) period. Oocyst elimination in feces began at age 59±9 days in kids. Eimerian species identified were E. arloingi, E. jolchijevi, E. ninakohlyakimovae, E. hirci, E. christenseni and E. alijevi. Kids were humanely sacrificed to evaluate pathological lesions. Intestinal lesions and lesion severity showed differences in duodenum, jejunum, ileum, cecum and colon, being more severe in duodenum. In conclusion, OPG increased during the late pregnancy in does which favored a doe-kid transmission mechanism. Our results support the notion of Eimeria reproduction rhythms during the late pregnancy period in goats, and this reproduction contribute to vertical transmission of Eimeria to the newborn. However, coccidian outbreaks are developed and clinically observed only when stressing factors such as when weaning occur. Coccidia had devastating effects on the intestine of kids, which might cause long-term permanent malabsortion consequences."

"El objetivo del estudio fue caracterizar la infección natural temprana por Eimeria spp. en cabritos, y describir respuestas fisiopatológicas en cabritos mantenidos bajo estabulación en B.C.S., México. Se utilizaron 19 cabras adultas cruzadas infectadas y 20 cabritos cruzados de Anglo-NubioxCriollo. Se determinó la cuenta de ooquistes por gramo de heces (OPG) y las especies de Eimeria en cabras (antes y después del parto) y en sus crías. Se registraron los signos clínicos, hematocrito, hemoglobina y actividad de fosfatasa alcalina en suero. Las cuentas de OPG (media±DE) fueron más altos (P<0.05) durante el periodo pre-parto (9,478±7,599) con parado con el post-parto (5,313±2,909). La eliminación de ooquistes en cabritos inició a la edad de 59±9 d. Las especies de Eimeria identificadas en las madres y sus crías fueron E. arloingi, E. jolchijevi, E. ninakohlyakimovae, E. hirci, E. christenseni y E. alijevi. Las lesiones intestinales y su severidad mostraron diferencias en duodeno, yeyuno, ileon, ciego y colon, siendo más severas en duodeno. En conclusión, nuestros resultados soportan la idea que los ritmos de reproducción de Eimeria durante el periodo de lactancia tardía en cabras, y que esta reproducción contribuye a la transmisión vertical de Eimera al cabrito recién nacido. Sin embargo, los brotes de coccidia se desarrollan y se observan clínicamente solo cuando factores estresantes ocurren como el destete ocurren. La coccidiosis tiene efectos serios sobre el intestino de los cabritos, que quizá tiene consecuencias de mala absorción a largo plazo."

Article

Eimeria, infection, goat, kids, pathology lesions infección, cabras, cabritos, lesiones BIOLOGÍA Y QUÍMICA CIENCIAS DE LA VIDA BIOLOGÍA ANIMAL (ZOOLOGÍA) PATOLOGÍA ANIMAL

Plantas como biofábricas de vacunas orales para animales

CARLOS ELIUD ANGULO VALADEZ PERLA CARLOS GARCIA BEATRIZ MEZA MARQUEZ CRISTHIAN FRANCISCO SANDEZ ROBLEDO CRYSTAL ALEJANDRA GULUARTE VELEZ RAZIEL ALEJANDRO SOSA SALAZAR ABEL ANTONIO RAMOS VEGA (2016)

" La producción de vacunas recombinantes en plantas inició hace casi 25 años con la producción de una vacuna proteica contra el virus de Newcastle para la industria avícola (Curtis y Cardineau, 1991).La patente se consiguió en 2006 en los Estados Unidos de Norteamérica. Inicialmente, el concepto se probó en plantas pero posteriormente se extendió a las microalgas hace poco más de 10 años. En la actualidad, el CIBNOR desarrolla vacunas experimentales en plantas y microalgas marinas que puedan ser administradas vía oral y con el objetivo de contribuir a la salud de los animales de importancia en la producción de alimentos para el consumo humano. Durante 5 años de investigación se ha logrado producir la primera vacuna oral contra la enfermedad paratuberculosis del ganado en plantas de alfalfa, aunque aún es necesario evaluarla en vacas, ovejas o cabras. El mismo concepto se ha explorado en las microalgas de nombre Chlamydomonas (dulceacuícola) y Schizochytrium (marina), y recientemente en la microalga marina Phaeodactylum. Sin embargo, en la presente revisión se expondrán los avances sobre el desarrollo de las vacunas comestibles producidas en plantas, lo que se le llama en el lenguaje científico “la frontera del conocimiento”, la contribución que está realizando el CIBNOR en el tema, y se ofrecerá una perspectiva de lo que probablemente contribuiremos en los próximos 5 a 10 años."

"The fi rst recombinant vaccine production began 25 years ago starting with a protein vaccine against the Newcastle virus affecting poultry industry (Curtis y Cardineau, 1991). In 2006, a patent was obtained in the United States of America. At the beginning, this concept was tested in plants, but it was extended to microalgae 10 years ago. Currently, CIBNOR develops experimental vaccines in plants and marine microalgae that can be administered orally with the objective of contributing to health of farming animals relevant in food production for human consumption. During the last 5 years of research, we have produced the fi rst vaccine in alfalfa plants against paratuberculosis (Johne’s disease) in livestock although it still needs to be assessed in cattle, sheep or goats. The same concept has been explored in the microalgae Chlamydomonas (freshwater) and Schizochytrium (marine), and recently in the marine microalgae Phaeodactylum. In this review, however, we will set out recent advances on edible plant-based vaccine, which in scientifi c language is known as border line of knowledge, the contribution of CIBNOR in this research fi eld, and a perspective of our potential contribution in the following 5 to 10 years."

Article

Vacunas en plantas, inmunización de las mucosas, platas como vehículos de administración oral, salud animal, seguridad alimentaria Plant-based vaccines, mucosal immunization, plants as oral delivery vehicles, animal health, food security BIOLOGÍA Y QUÍMICA CIENCIAS DE LA VIDA INMUNOLOGÍA VACUNAS