Protective efficacy of the oral vaccine Tc24:Co1 produced in Schizochytrium sp. against Trypanosoma cruzi infection in a mouse model.

Trypanosoma cruzi parasite - causal Chagas disease agent - affects about 7 million people; no vaccine is available, and current medications have not been entirely effective. Multidisciplinary efforts are necessary for developing clinical vaccine prototypes. Thus, this research study aims to assess the expressed and whole-cell administration protection of the oral vaccine prototype Tc24:Co1 using Schizochytrium sp. microalga. High recombinant protein expression yields (675 µg/L) of algal culture were obtained. Additionally, Schizochytrium sp.-Tc24:Co1 resulted stable at 4 °C for up to six months and at 25 °C for three months. After receiving four oral doses of the vaccine, the mice showed a significant humoral immune response and a parasitemia reduction associated with a lack of heart inflammatory damage compared with the unvaccinated controls. The Schizochytrium sp.-Tc24:Co1 vaccine demonstrates to be promising as a prototype for further development showing protective effects against a T. cruzi challenge in a mouse model.

Trypanosoma cruzi Tc24 Antigen Expressed and Orally Delivered by Schizochytrium sp. Microalga is Immunogenic in Mice.

Chagas disease-caused by the parasite Trypanosoma cruzi-is a neglected tropical disease for which available drugs are not fully effective in the chronic stage and a vaccine is not available yet. Microalgae represent a promising platform for the production and oral delivery of low-cost vaccines. Herein, we report a vaccine prototype against T. cruzi produced in a microalgae platform, based on the candidate antigen Tc24 with a C terminus fusion with the Co1 peptide (Tc24:Co1 vaccine prototype). After modeling the tertiary structure, in silico studies suggested that the chimeric protein is antigenic, not allergenic, and molecular docking indicated binding with Toll-like receptors 2 and 4. Thus, Tc24:Co1 was expressed in the marine microalga Schizochytrium sp., and Western blot confirmed the expression at 48 h after induction, with a yield of 632 µg/L of algal culture (300 µg/g of lyophilized algal cells) as measured by the enzyme-linked immunosorbent assay (ELISA). Upon oral administration of whole-cell Schizochytrium sp. expressing Tc24:Co1 (7.5 µg or 15 µg of Tc24:Co1 doses) in mice, specific serum IgG and intestinal mucosa IgA responses were detected in addition to an increase in serum Th1/Th2 cytokines. In conclusion, Schizochytrium sp.-expressing Tc24:Co1 is a promising oral vaccine prototype to be evaluated in an animal model of Trypanosoma cruzi infection.

Plant-Based Vaccines: Antigen Design, Diversity, and Strategies for High Level Production.

Vaccines for human use have conventionally been developed by the production of (1) microbial pathogens in eggs or mammalian cells that are then inactivated, or (2) by the production of pathogen proteins in mammalian and insect cells that are purified for vaccine formulation, as well as, more recently, (3) by using RNA or DNA fragments from pathogens. Another approach for recombinant antigen production in the last three decades has been the use of plants as biofactories. Only have few plant-produced vaccines been evaluated in clinical trials to fight against diseases, of which COVID-19 vaccines are the most recent to be FDA approved. In silico tools have accelerated vaccine design, which, combined with transitory antigen expression in plants, has led to the testing of promising prototypes in pre-clinical and clinical trials. Therefore, this review deals with a description of immunoinformatic tools and plant genetic engineering technologies used for antigen design (virus-like particles (VLP), subunit vaccines, VLP chimeras) and the main strategies for high antigen production levels. These key topics for plant-made vaccine development are discussed and perspectives are provided.

Plant-made vaccines against parasites: bioinspired perspectives to fight against Chagas disease.

Introduction: Three decades of evidence have demonstrated that plants are an affordable platform for biopharmaceutical production and delivery. For instance, several plant-made recombinant proteins have been approved for commercialization under good manufacturing practice (GMP). Thus far, plant-based vaccine prototypes have been evaluated at pre- and clinical levels. Particularly, plant-made vaccines against parasitic diseases, such as malaria, cysticercosis, and toxoplasmosis have been successfully produced and orally delivered with promising outcomes in terms of immunogenicity and protection. The experience on several approaches and technical strategies over 30 years accounts for their potential low-cost, high scalability, and easy administration.Areas covered: This platform is an open technology to fight against Chagas disease, one of the most important neglected tropical diseases worldwide.Expert opinion: This review provides a perspective for the potential use of plants as a production platform and delivery system of Trypanosoma cruzi recombinant antigens, analyzing the advantages and limitations with respect to plant-made vaccines produced for other parasitic diseases. Plant-made vaccines are envisioned to fight against Chagas disease and other neglected tropical diseases in those countries suffering endemic prevalence.

Characterization and Structural Performance in Bending of CLT Panels Made from Small-Diameter Logs of Loblolly/Slash Pine.

The main objective of this work was to study the structural viability of using small-diameter logs of Uruguayan Loblolly/Slash pine, mainly from thinning operations, to design cross-laminated timber (CLT) panels. A visual grade named "CTH" (coniferous thinning) was proposed, and 45 specimens of sawn timber boards were tested, resulting in 51% lower bending strength than that of the minimum strength class C14. Subsequently, 20 CLT panels were manufactured and experimentally tested, the results showed that the bending strength of the CLT panels was 43% above that of the individual layers. Additionally, the structural performance of the CLT panels for use in floors was calculated, and the thickness-span relationship depending on strength class and imposed load are presented. Results showed than the use of CTH timber to design CLT floors implies a volume (m³/m²) 17% higher than that using C24 timber.

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

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.

Expression of the VP40 antigen from the Zaire ebolavirus in tobacco plants.

MAIN CONCLUSION: The plant cell is able to produce the VP40 antigen from the Zaire ebolavirus , retaining the antigenicity and the ability to induce immune responses in BALB/c mice. The recent Ebola outbreak evidenced the need for having vaccines approved for human use. Herein we report the expression of the VP40 antigen from the Ebola virus as an initial effort in the development of a plant-made vaccine that could offer the advantages of being cheap and scalable, which is proposed to overcome the rapid need for having vaccines to deal with future outbreaks. Tobacco plants were transformed by stable DNA integration into the nuclear genome using the CaMV35S promoter and a signal peptide to access the endoplasmic reticulum, reaching accumulation levels up to 2.6 µg g-1 FW leaf tissues. The antigenicity of the plant-made VP40 antigen was evidenced by Western blot and an initial immunogenicity assessment in test animals that revealed the induction of immune responses in BALB/c mice following three weekly oral or subcutaneous immunizations at very low doses (125 and 25 ng, respectively) without accessory adjuvants. Therefore, this plant-based vaccination prototype is proposed as an attractive platform for the production of vaccines in the fight against Ebola virus disease outbreaks.

[Variations of the nutritional condition of lobsters Panulirus argus (Decapoda: Palinuridae) in Eastern region of the Gulf of Batabanó, Cuba]. / Variaciones en la condición nutricional de langostas Panulirus argus (Decapoda: Palinuridae) en la región este del Golfo de Batabanó, Cuba.

Nutritional condition can affect survival and growth rate of crustaceans, and this is mostly affected by habitat conditions. This study describes the space-temporary nutritional changes in this commercially important species. With this aim, the variations in the nutritional condition (K) of lobsters from four zones (1, 2, 4 and 5) in the Gulf of Batabanó, Cuba, were determined. For this, the weight/length ratio (K=Pt/Lt) was calculated using animals captured in 1981 and 2010. The nutritional condition between areas and sexes, and years and sexes, was contrasted by a bifactorial ANOVA, and the overall length and weight of lobsters were compared using a t-Test for independent samples and unifactorial ANOVA. It was found that the nutritional condition was significantly greater in males than in females. In addition, significant variations between zones were detected for both years. Nutritional condition of lobsters from Zone five was the highest for 1981, while it was Zone two for 2010. Lobsters nutritional state showed significant variations between years, being greater in 1981 (2.34 +/- 0.84g/mm) than in 2010 (1.96 +/- 0.49g/mm). The inter-zones variations as well as the inter-annual ones seem to be related to the reported variations of the bottom type and the vegetation cover. Seasonal variations in the abundance and distribution of benthic organisms, that constitute food for lobsters, could also be influencing. The differences between sexes, however, were assumed as a consequence of the methodology used and the sexual dimorphism of the species. Due to other K estimation methods, that do not include morphometric measurements, these differences were not detected. We suggested that the P. argus nutritional condition is a good estimator of the habitat condition. Besides, according to the applied K estimation methodology, it was found that different groups of lobsters that have resemblant nutritional condition, did not necessarily observe similarities in the overall mean length or weight, so they could exist under different habitat conditions.

Variaciones en la condición nutricional de langostas Panulirus argus (Decapoda: Palinuridae) en la región este del Golfo de Batabanó, Cuba / Variations of the nutritional condition of lobsters Panulirus argus (Decapoda: Palinuridae) in Eastern region of the Gulf of Batabanó, Cuba

Variations of the nutritional condition of lobsters Panulirus argus (Decapoda: Palinuridae) in Eastern region of the Gulf of Batabanó, Cuba. Nutritional condition can affect survival and growth rate of crustaceans, and this is mostly affected by habitat conditions. This study describes the space-temporary nutritional changes in this commercially important species. With this aim, the variations in the nutritional condition (K) of lobsters from four zones (1, 2, 4 and 5) in the Gulf of Batabanó, Cuba, were determined. For this, the weight/length ratio (K=Pt/Lt) was calculated using animals captured in 1981 and 2010. The nutritional condition between areas and sexes, and years and sexes, was contrasted by a bifactorial ANOVA, and the overall length and weight of lobsters were compared using a t-Test for independent samples and unifactorial ANOVA. It was found that the nutritional condition was significantly greater in males than in females. In addition, significant variations between zones were detected for both years. Nutritional condition of lobsters from Zone five was the highest for 1981, while it was Zone two for 2010. Lobsters nutritional state showed significant variations between years, being greater in 1981 (2.34±0.84g/mm) than in 2010 (1.96±0.49g/mm). The inter-zones variations as well as the inter-annual ones seem to be related to the reported variations of the bottom type and the vegetation cover. Seasonal variations in the abundance and distribution of benthic organisms, that constitute food for lobsters, could also be influencing. The differences between sexes, however, were assumed as a consequence of the methodology used and the sexual dimorphism of the species. Due to other K estimation methods, that do not include morphometric measurements, these differences were not detected. We suggested that the P. argus nutritional condition is a good estimator of the habitat condition. Besides, according to the applied K estimation methodology, it was found that different groups of lobsters that have resemblant nutritional condition, did not necessarily observe similarities in the overall mean length or weight, so they could exist under different habitat conditions.

En Cuba, la región del Golfo de Batabanó es la más importante zona de pesca de langosta debido a que de ella se extrae anualmente alrededor del 70% de la captura nacional. Con el objetivo de determinar variaciones espacio-temporales en la condición nutricional (K) de langostas Panulirus argus del Golfo de Batabanó, Cuba, se calculó dicha condición mediante una relación pesolargo (K=Pt/Lt), para lo que se utilizaron ejemplares de cuatro zonas (uno, dos, cuatro y cinco) del este del golfo, capturados en los años 1981 y 2010. Se encontró que en todos los casos analizados la condición nutricional fue significativamente mayor en machos que en hembras. Se detectaron variaciones significativas entre zonas en 1981 y 2010. También entre años el estado nutricional de las langostas presenta variaciones significativas, de manera que en 1981 (2.34±0.84g/mm) fue mayor que en 2010 (1.96±0.49g/mm). Tanto las variaciones interzonales como las interanuales parecen estar relacionadas con variaciones reportadas en el tipo de fondo y la cobertura vegetal. Variaciones estacionales en la abundancia y distribución de organismos bentónicos que constituyen alimento para las langostas también podrían estar influyendo. Las diferencias entre sexos en cambio, se suponen consecuencia de la metodología utilizada y del dimorfismo sexual característico de esta especie, por cuanto otros métodos de estimación de K que no involucran mediciones morfométricas, no detectan estas diferencias.