Isolation of wild yeasts from Olympic National Park and Moniliella megachiliensis ONP131 physiological characterization for beer fermentation.

Thousands of yeasts have the potential for industrial application, though many were initially considered contaminants in the beer industry. However, these organisms are currently considered important components in beers because they contribute new flavors. Non-Saccharomyces wild yeasts can be important tools in the development of new products, and the objective of this work was to obtain and characterize novel yeast isolates for their ability to produce beer. Wild yeasts were isolated from environmental samples from Olympic National Park and analyzed for their ability to ferment malt extract medium and beer wort. Six different strains were isolated, of which Moniliella megachiliensis ONP131 displayed the highest levels of attenuation during fermentations. We found that M. megachiliensis could be propagated in common yeast media, tolerated incubation temperatures of 37 °C and a pH of 2.5, and was able to grow in media containing maltose as the sole carbon source. Yeast cultivation was considerably impacted (p < 0.05) by lactic acid, ethanol, and high concentrations of maltose, but ONP131 was tolerant to high salinity and hop acid concentrations. This is one of the first physiological characterizations of M. megachiliensis, which has potential for the production of beer and other fermented beverages.

Effect of COVID-19 on vaccination coverage in Brazil.

During the COVID-19 pandemic, recommendations for maintaining physical distance, restricted mobility measures, as well as fear of mass transmission by going to health centers have significantly contributed to the general vaccination coverage, which by and large is decreasing worldwide; thus, favoring the potential re-emergence of vaccine-preventable diseases. In this study, we have used the existing data on vaccination coverage during the pre-pandemic (2019) as well as the pandemic (2020) period to evaluate the impact of coronavirus outbreaks during the vaccination drive in Brazil. Furthermore, we have accumulated data since 2015 among the different regions of the country to acquire more consistent information. The various vaccines analyzed in our study were meningococcal C conjugate, Triple antigen vaccine, 10-valent pneumococcal conjugate, and BCG; subsequently, the data were obtained from the National Disease Notification System. This study revealed that the ongoing immunization drive saw a steep decline of around 10 to 20% during the (2019-2020) pandemic period in Brazil. These results provide strong evidence towards the decreasing trends following the vaccination programs during the COVID-19 pandemic period in Brazil. Furthermore, our results also highlight the importance of adopting widespread multi-component interventions to improve vaccination uptake rates.

Antigenicity assessment of the Theileria equi merozoite antigen (EMA-2) expressed in Pichia pastoris in mice and horses.

Equine theileriosis is a severe equine disease caused by the protozoan Theileria equi, which is prevalent in tropical and subtropical areas. In this study, a recombinant equi merozoite antigen-2 (rEMA-2) of T. equi was used as an immunogen. Two groups of 10 mice each were divided into control and vaccinated groups. Sixty mares seronegative for theileriosis were divided in two groups, one vaccinated and another group as a control animal. Mice and mares of the vaccinated groups were inoculated with 150 µL of the vaccine containing 50 µg of rEMA-2 and 2 mL of the vaccine containing 200 µg of rEMA-2, respectively, at days 0 and 21. The immunogenicity of rEMA-2 was evaluated by ELISA and fluorescent antibody test (IFAT) using serum from vaccinated mice, mares and antigenicity in naturally infected horse. At every point throughout the ELISA study, there were significant differences between the vaccinated and control groups (p < 0.05). The vaccine induced 3- and 4-fold IgG increases in mice at the 14th and 28th day, respectively, compared to the control group. The horses' IgG dynamics showed a significant (p < 0.05) increase in the total IgG titer as early as day 7, which increased until day 28 at which time a more significant (p < 0.001) IgG titer was observed. In evaluating the isotypes, we observed a trend similar to that of total IgG, where IgG(T) (IgG3-5) were significantly (p < 0.05) more elevated than the other isotypes analyzed, followed by IgGb (IgG4-7) and IgGa (IgG1). Positive fluorescence was detected by IFAT, suggesting that the protein is immunogenic and conserves some epitopes identical to the native T. equi antigens present in the equine blood smear. Thus, our results suggest that rEMA-2 can be a promising vaccinal antigen.

Detection of Toxocara canis DNA in tissues of experimentally infected mice.

The main etiological agent of toxocariasis is the helminth Toxocara canis. Several difficulties are found in the diagnosis of this disease, because of nonspecific clinical signs and possible cross-reactions that may occur in the available test, the indirect ELISA. Therefore, molecular diagnosis has been indicated as an alternative to conventional diagnosis. The purpose of this study was to evaluate the polymerase chain reaction (PCR) technique for the identification of T. canis in tissues of experimentally infected mice. To this end, nine mice were inoculated with 1500 embryonated eggs and were divided into two groups, the first euthanized 48 h (G1) and the other 30 days post inoculation (G2). Lungs, brain, liver and blood were collected from all the animals for DNA Extraction and tissue digestion, also was collected blood samples for DNA extraction and ELISA test (serum). Toxocara canis DNA was identified in all the inoculated animals using the ITS-2 target gene. The PCR test successfully identified the parasite in the brain, lung and liver of the animals euthanized 48 h PI and 30 days PI. This technique yielded good results in the identification of the parasite in the brain, being more sensitive than the method for the recovery of larvae, in the group with acute infection (48 h PI). The infection was confirmed by PCR within 48 h after infection, while the ELISA indicated serological conversion occurred only 14 days after inoculation. This study demonstrates the ability of PCR to identify T. canis in the liver, lungs and brain during acute and chronic infection.

Olive oil as an alternative to boar semen cryopreservation / Aceite de oliva como una alternativa para la criopreservación de semen suino / O azeite de oliva como uma alternativa para a criopreservação do sêmen suíno

The aim of this study was to evaluate different concentrations of olive oil for cryopreservation of boar semen. A total of 21 eyaculates of 18 males with motility ≥70% were used. For freezing, the semen was diluted in treatments: control (Cont-egg yolk), "Koroneiki" oil in egg yolk at 0.25% (A025), 0.50% (A050), 0.75% (A075) and 1.0% (A10). Straws with 5.107 sperm/ml were frozen and stored in liquid nitrogen. Thawing was done at 37ºC for 30 seconds and motility, mitochondrial functionality, DNA integrity, oocite penetration rate and the number of sperm per oocite (in the in vitro penetration trial) were evaluated. The variables were compared using the Kruskal-Wallis test. Even though no statistical difference was found between control and treatments containingolive oil (p>0.05), the treatment with 0.25% oil concentration showed tendences towards sperm protection, outperforming the controls in mitochondrial functionality and preservation of the fertilizing capabilities in the in vitro penetration trial. This is why, olive oil could represent an alternative to help cryopreservation of boar semen.

El objetivo de este estudio fue evaluar diferentes concentraciones de aceite de oliva para la criopreservación de semen de verraco. Se utilizaron 21 eyaculados de 18 machos con motilidad igual o superior al 70%. Para la congelación, el semen se diluyó en los tratamientos: control (Cont) (lactosa yema), 0,25% (A025); 0,50% (A050); 0,75% (A075) y 1,0% (A10) de aceite de la variedad "Koroneiki" en lactosa yema. Pajas con 5.107 espermatozoides / ml, fueron congeladas y almacenadas en nitrógeno líquido. La descongelación se produjo a 37°C durante 30 segundos y se evaluó: la motilidad, la funcionalidad de la mitocondria, la integridad del ADN, la tasa de penetración de ovocitos y el número de espermatozoides por ovocito (en el ensayo de penetración in vitro). Las variables se compararon mediante la prueba de Kruskal-Wallis. Aunque no hubo diferencia estadística entre el control y los tratamientos que contienen aceite de oliva (p> 0,05), la concentración de 0,25% de este aceite mostró tendencias de protección de esperma, superando el control en la funcionalidad de las mitocondrias y en la preservación de la capacidad fertilizante en el ensayo de penetración in vitro. Por lo tanto, el aceite de oliva puede representar una alternativa para ayudar en la criopreservación de semen porcino

O objetivo de este estudo foi avaliar diferentes concentrações de azeite de oliva para a criopreservação de sêmen suíno. Utilizaram-se 21 ejaculados de 18 machos com motilidade igual ou superior ao 70%. Para o congelamento, o sêmen se dilui-o nos tratamentos: controle (Cont) (lactose-gema), 0,25% (A025); 0,50% (A050); 0,75% (A075) e 1,0% (A10) de azeite da variedade "Koroneiki" em lactose-gema. Palhetas com uma dose de 5.107 espermatozoides/ ml foram congeladas e armazenadas em nitrogênio liquido. O descongelamento se produz a 37°C durante 30 segundos e se avaliaram: motilidade, funcionalidade da mitocôndria, integridade do DNA, taxa de penetração de ovócitos e o número de espermatozoides por ovócito (no teste de penetração in vitro). As variáveis se compararam mediante o teste de Kruskal-Wallis. Ainda que não houve diferença estatística entre o controle e os tratamentos que continham azeite de oliva (p> 0,05), a concentração de 0,25% de azeite de oliva teve tendências de proteção do esperma, superando o controle na funcionalidade das mitocôndrias e na preservação da capacidade fertilizante no teste de penetração in vitro. Assim sendo, o azeite de oliva pode representar uma alternativa para ajudar na criopreservação do sêmen suíno.