Genome-Based Vaccinology Applied to Bovine Babesiosis

Genomics approaches in veterinary research have been a very useful tool to identify candidates with potential to be used in prevention of animal diseases. In Babesia, genome information analysis has elucidated a wide variety of protein families and some members are described in this chapter. Here, we present some of the most recent studies about B. bovis and B. bigemina genomes where some proteins have been identified with potential to prevent infections by these parasites

An upgraded lab-based method to determine natural γ-ray emitters in NORM samples by using Ge detectors

The regulation existing on NORM activities requires the accurate and quick measurement of natural radionuclides in a wide range of matrices by gamma spectrometry. The aim of this work has been to develop a comprehensive and accurate methodology to determine natural long-lived radionuclides (210Pb, 228,226Ra, 234,228Th and 40K) by gamma spectrometry with Ge detectors in NORM samples. An exhaustive calibration method to obtain the full-energy peak efficiency (FEPE), for each gamma emission energy, as a function of the thickness, apparent density and matrix composition of the cylindrical sample, has been developed. The selected certified calibration standards contain only natural radionuclides belonging to the 238U- and 232Th-series as well as 40K (codes RGU-1, RGTh-1 and RGK-1 from IAEA). Then, the obtained FEPE curves were validated using Genie 2000 simulations and Certified Reference Materials, for which the self-attenuation corrections were considered. Finally, a study on 222Rn losses was done for several samples.Funding for open access charge: Universidad de Huelva / CBU

Occupational, Public and Environmental Radiological Impact Caused by the Phosphoric Acid Industry: The Case of Huelva (Spain)

The production of phosphate fertilizers usually uses as raw material sedimentary phosphate rock, which contains enhanced concentrations from U‐series radionuclides about 10–100 times higher than unperturbed soils. This fact implies the need for evaluating the radiological implications of this activity. In our case, the study has been performed in a large fertilizer industrial complex located at Huelva town (SW of Spain), where sedimentary phosphate rock has been processed since 1965 to 2010, generating annually an average of about 2.5 million tons of a by‐product called phosphogypsum (PG), which has been stored in big stacks 1 km away from Huelva city, covering 1000 ha. The fluxes of the radionuclides of interest along the production process and the effective doses received by the workers have been determined. In addition, the radioecological impact associated to the waste management strategy followed has been evaluated

Value of the toluidine blue test as an aid to determine the biopsy site in actinic prurigo cheilitis

Actinic prurigo (AP) is a type of photodermatosis that primarily affects the Latin American mestizo population. Histologically, AP cheilitis exhibits acanthosis with spongiosis and vacuolation of the basal cell layer overlying a dense lymphocytic inflammatory infiltrate that forms well-defined lymphoid follicles. Toluidine blue is a thiazide, acidophilic, and metachromatic dye used in vivo to selectively stain the acidic components of tissues such as sulfates, carboxylates, and phosphate radicals that are incorporated into DNA and RNA. It is necessary to develop a method that allows detecting, on clinical grounds the area of the lesion in which it is more feasible to find such structures. Thus to increase the sensitivity of the biopsy, in AP cheilitis to accurately identify where the lymphoid follicles reside, based on the higher concentration of DNA in such structures and thus confirm the diagnosis. In this study, staining was positive in 85% of patients with AP cheilitis, in 14 of whom 82% lymphoid follicles were observed by histopathology. One of the pathologist’s problems in establishing the diagnosis of AP is that the main histopathological characteristics are not always identified in the submitted samples because it is not easy to clinically identify the most representative site of the lesion selected for performing a biopsy. Based on our results, we propose using toluidine blue as an auxiliary method to choose a tissue sample to facilitate the diagnosis and allow clinicians to make clinical correlations between the histopathological and therapeutic findings

Autonomus Motivation as a Mediator Between an Empowering Climate and Enjoyment in MAle Volleyball Players

The objective of this work was to analyze a mediation model concerning the perception ofan empowering climate generated by a coach and enjoyment through the autonomous motivation ofathletes. The sample consisted of 71 elite male volleyball players from six countries. The age rangewas 14 to 18 years (M=16.5, SD=0.96). The relationships between the perception of an empoweringclimate, autonomous motivation, and enjoyment were positive and significant. The mediation modelshowed that autonomous motivation acts as a mediator in the relationship between the perception ofan empowering climate generated by the coach and the enjoyment reported by the athletes

Ulcera lingual como signo único de infección recurrente por micobacteria en un paciente con VIH/SIDA

Se describe un paciente con VIH/SIDA en el que se identificó una infección por micobacteria en la mucosa bucal, probablemente tuberculosis, en un centro de referencia para VIH/SIDA de la Ciudad de México. El propósito del presente informe es describir los hallazgos clínicos e histológicos en un paciente con VIH/SIDA, quien después de haber sido tratado exitosamente para tuberculosis ganglionar 4 años antes, presentó una úlcera lingual como único signo que sugirió recurrencia de infección por micobacteria, probablemente tuberculosis. Hombre de 39 años de edad, atendido desde 1991 en el Instituto Nacional de Ciencias Médicas y Nutrición 'Salvador Zubirán', por el diagnóstico de infección con VIH. En 1999, el paciente presentó tuberculosis ganglionar, recibiendo tratamiento antifímico con involución de las adenopatías y desaparición de los síntomas sistémicos. En mayo del 2003 acudió a consulta por presentar una úlcera superficial en lengua, dolorosa, de 4 meses de evolución, de 0.7 cm. de diámetro, bien circunscrita, crateriforme, con bordes ligeramente elevados, irregulares e indurados. El estudio histopatológico mostró inflamación granulomatosa crónica con células gigantes multinucleadas sugestivas de infección por mi-cobacteria, lo cual hizo pensar en recurrencia de tuberculosis, por lo que se indicó rifampicina, pirazinamida, etambutol y estreptomicina. En junio del 2003 el paciente inició TARAA, que incluyó dos ITRAN y un ITRNN. La lesión lingual evolucionó favorablemente, con cicatrización parcial a la primera semana y remisión total a los 45 días del inicio del tratamiento antifímico; a los 7 meses de seguimiento permanece sin lesión. El presente caso tiene la particularidad de que la úlcera lingual fue la única manifestación de infección por micobacteria, sugestiva de tuberculosis, en un paciente con VIH/SIDA, que pudo ocurrir como resultado de la recurrencia del episodio previo de TB ganglionar.The report describes an HIV/AIDS patient seen at a referral center in Mexico City, in whom a mycobacterial infection in the oral mucosa, probably tuberculosis (TB) was identified. The purpose is to describe the clinical and histological findings in an HIV-infected patient, who after being treated successfully for tuberculous lymphangitis 4 years ago, presented with a lingual ulcer as the only suggestive sign of recurrence of mycobacterial infection, probably M. tuberculosis. A 39-year-old man seen inthe HIV clinic of the Instituto Nacional de Ciencias Médicas y Nutrición 'Salvador Zubirán' in Mexico City since 1991 for HIV infection. In 1999 the patient developed tuberculous lymphangitis; he was managed with a 4-drug regimen for 12 months, with improvement of local and systemic symptoms. In May of 2003, the patient presented a painful superficial lingual ulcer, 0.7 cm in diameter, well circumscribed, crateriform with slightly elevated, irregular and indurated borders, of 4 months duration. The histopathological examination showed chronic granulomatous inflammation with giant multinucleated cells, suggestive of mycobacterial infection, and recurrence of TB was considered. Rifampin, isoniazide, pyrazinamide, ethambutol and streptomycin were administered. The lingual lesion improved with partial healing at the first week and total remission at 45 days after the beginning of the antituberculous treatment. In June, 2003, the patient began highly active antiretroviral therapy (HAART) that included two NRTIs and one NNRTI. At 7 months of follow-up, the patient remains free of lingual lesions. The particularity of the present case is that the lingual ulcer was the only sign of infection by mycobacteria, suggestive of TB, in an HIV/AIDS patient that probably represented a recurrence of a previous episode

Bovipain-2, the falcipain-2 ortholog, is expressed in intraerythrocytic stages of the tick-transmitted hemoparasite Babesia bovis

<p>Abstract</p> <p>Background</p> <p>Cysteine proteases have been shown to be highly relevant for Apicomplexan parasites. In the case of <it>Babesia bovis</it>, a tick-transmitted hemoparasite of cattle, inhibitors of these enzymes were shown to hamper intraerythrocytic replication of the parasite, underscoring their importance for survival.</p> <p>Results</p> <p>Four papain-like cysteine proteases were found to be encoded by the <it>B. bovis </it>genome using the MEROPS database. One of them, the ortholog of <it>Plasmodium falciparum </it>falcipain-2, here named bovipain-2, was further characterized. Bovipain-2 is encoded in <it>B. bovis </it>chromosome 4 by an ORF of 1.3 kb, has a predicted molecular weight of 42 kDa, and is hydrophilic with the exception of a transmembrane region. It has orthologs in several other apicomplexans, and its predicted amino acid sequence shows a high degree of conservation among several <it>B. bovis </it>isolates from North and South America. Synteny studies demonstrated that the <it>bovipain-2 </it>gene has expanded in the genomes of two related piroplasmids, <it>Theileria parva </it>and <it>T. annulata</it>, into families of 6 and 7 clustered genes respectively. The <it>bovipain-2 g</it>ene is transcribed in <it>in vitro </it>cultured intra-erythrocyte forms of a virulent and an attenuated <it>B. bovis </it>strain from Argentina, and has no introns, as shown by RT-PCR followed by sequencing. Antibodies against a recombinant form of bovipain-2 recognized two parasite protein bands of 34 and 26 kDa, which coincide with the predicted sizes of the pro-peptidase and mature peptidase, respectively. Immunofluorescence studies showed an intracellular localization of bovipain-2 in the middle-rear region of <it>in vitro </it>cultured merozoites, as well as diffused in the cytoplasm of infected erythrocytes. Anti-bovipain-2 antibodies also reacted with <it>B. bigemina</it>-infected erythrocytes giving a similar pattern, which suggests cross-reactivity among these species. Antibodies in sera of two out of six <it>B. bovis</it>-experimentally infected bovines tested, reacted specifically with recombinant bovipain-2 in immunoblots, thus demonstrating expression and immunogenicity during bovine-infecting stages.</p> <p>Conclusions</p> <p>Overall, we present the characterization of bovipain-2 and demonstrate its <it>in vitro </it>and <it>in vivo </it>expression in virulent and attenuated strains. Given the involvement of apicomplexan cysteine proteases in essential parasite functions, bovipain-2 constitutes a new vaccine candidate and potential drug target for bovine babesiosis.</p

Time evolution of activity concentration of natural emitters in a scenario affected by previous phosphogypsum contamination

The estuary formed by the confluence of Tinto and Odiel river-mouths is located in the South of Spain, close to Huelva town. This estuary has been deeply studied through the years because it has a double particularity. On one hand, since the beginning of the 1960s, the estuary has been affected by direct and indirect phosphogypsum (pg.) releases from two phosphoric acid and fertilizers factories that are working in the area. On the other hand, the pyrite mining operations upstream the Odiel and Tinto rivers has caused historically the formation of H2SO4, through oxidation of the natural sulphur deposits, the acidification of the waters and the consequent mobilisation of heavy metals from the mining area to the Huelva estuary. As a consequence, enhancement contamination levels in natural emitters from the ^'*U series were found in the surroundings of the factories in the previous years to 1998. However, in 1998 the management policy of waste releases drastically changed in the area, and direct discharges to Tinto and Odiel River had to be ceased. A thorough study of the affected zone is being carried out. Riverbed sediments and water samples have been analyzed from four different sampling campaigns in the estuary during the years 1999, 2001, 2002 and 2005. Different radioanalytical techniques have been employed to obtain the activity concentrations of U-isotopes, Th-isotopes, ^^^Ra, ^'"Pb and ^'"Po. Furthermore, the results for the rates of de-contamination of the area are presented. This data will be discussed in order to establish the present status of the contamination in the area, and moreover, to predict the time-evolution of the self-cleanin

Development and standardization of a Loop-mediated isothermal amplification (LAMP) test for the detection of Babesia bigemina

Bovine babesiosis is a tick-borne disease caused by protozoan parasites of the genus Babesia. Babesia bigemina is one of the most prevalent and economically important parasite species that infects cattle because of its impact on the meat and milk production industry. Effective disease control strategies should include detection of reservoir animals and early and specific pathogen detection using rapid, economical, sensitive, and specific detection techniques. The loop-mediated isothermal amplification technique (LAMP) is a one-step molecular reaction that amplifies DNA sequences with high sensitivity and specificity under isothermal conditions and requires no special equipment. The results can be observed by the naked eye as color changes. The aim of this work was to develop and standardize the LAMP technique for B. bigemina detection and its visualization using hydroxynaphtol blue. For this situation, primers were designed from the conserved sequences of the B. bigemina ama-1 gene. The results showed that at 63 °C in 1 h and under standardized conditions, this technique could amplify B. bigemina DNA as indicated by the characteristic colorimetric change. Sensitivity evaluation indicated that DNA was amplified at a 0.00000001% parasitemia, and it was demonstrated that this technique specifically amplified the DNA of B. bigemina. Additionally, this technique could amplify DNA from 10 strains of B. bigemina from three different countries. It is concluded that the LAMP technique as modified in our case could specifically amplify B. bigemina DNA and shows high sensitivity, does not cross-react with related organisms, and the product is observed by 60 min of reaction time based on color changes. This report is the first LAMP report that uses sequences that are conserved between strains of the ama-1 gene, demonstrates the results by color changes using hydroxynaphtol blue. We propose LAMP as a rapid and economical alternative method for the molecular detection of B. bigemina.Fil: Lizarazo Zuluaga, Andrea P.. Universidad Autonoma de Queretaro.; MéxicoFil: Carvajal Gamez, Bertha I.. Universidad Autonoma de Queretaro.; MéxicoFil: Wilkowsky, Silvina Elizabeth. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Cravero, Silvio Lorenzo Pedro. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Trangoni, Marcos David. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Mosqueda, Juan. Universidad Autonoma de Queretaro.; Méxic

Salicylic acid and nutrient immersion to maintain apple quality and bioactive compounds in postharvest

The world production of apples in the 2019 cycle reached 7´620,288 tonnes. For marketing purposes and to supply the demand, apple fruits need to be stored for different periods under refrigerated conditions. However, in the market, the shelf life of the fruit is short, the quality decreases in postharvest due to the dynamic changes of its physicochemical properties, which cannot be stopped, but can be slowed down to improve its shelf life. Postharvest treatments by immersing apple fruit in salicylic acid (SA) and nutrients are an innovative technological alternative to maintain their quality. In this study, 5 concentrations were tested for the immersion of apple fruits cv ‘Golden Delicious’, using a 56 factorial arrangement delimited to 25 treatments, using the Taguchi L25 structure: SA 0 - 1.440 mM, potassium (K) 0 - 2.250, calcium (Ca) 0 - 31.500 mM, cobalt (Co) 0 - 0.180 mM, molybdenum (Mo) 0 - 0.0900 mM and magnesium (Mg) 0 - 0.0900 mM. The study was conducted in the municipality of Cuauhtémoc, Chihuahua, Mexico. After 7 months of storage and 13 days of shelf life, the combination of K, Ca, SA and Co with the appropriate concentration values can maintain the quality variables and bioactive compounds at the desired optimum. It is concluded that the quality variables; firmness, juice percentage, juice density, titratable acidity and total soluble solids and the bioactive compounds; total phenols and antioxidant capacity can be maintained at the desired optimum