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

Epidemiología de tumor venéreo transmisible canino en Reynosa Tamaulipas México

The CTVT is transmitted mainly through sexual contact, with females being the most exposed to the pathology; vincristine sulfate is currently the most effective therapy. In the present work, 28 canines were analyzed, mostly of indefinite breed. A diagnosis was made by complete physical examination and sur­veys were carried out on their owners to determine risk factors. The proportion of the disease was similar in females and males under 5 years of age, but considerably higher in adult males than in females; only 25% of those older than 5 years were female. The incidence of this tumor in non-sterilized animals was 28% higher than in sterilized ones. The presence of the tumor was considerably higher in non-rescued males compared to females in the same condition (73 and 27%). Finally, 89% of the reported cases presented lesions in the genital region. The objective of this work was to identify factors associated with the persistence of CTVT in the city of Reynosa Tamaulipas.El CTVT por sus siglas en inglés (Canine Transmissible Venereal Tumor) se transmite principalmente a través de contacto sexual siendo las hembras las más expuesta a la patología; el sulfato de vincristina es actualmente la terapia más eficaz. En el presente trabajo se analizaron 28 caninos, en su mayoría de raza indefinida; se realizó diagnóstico por reconocimiento físico completo y se realizaron encuestas a sus propietarios para determinar factores de riesgo. La proporción de la enfermedad fue similar en hembras y machos menores de 5 años pero considerablemente mayor en machos adultos con respecto a hembras; solo el 25% de los mayores de 5 años fueron hembras. La incidencia de este tumor en animales no esterilizados fue un 28% mayor con respecto a los esterilizados. La presencia del tumor fue considerablemente más alta en machos no rescatados con respecto a hembras en la misma condición (73 y 27%). Finalmente, el 89% de los casos reportados presentaron las lesiones en la región genital. El objetivo del presente trabajo fue identificar factores asociados a persistencia de CTVT en la ciudad de Reynosa Tamaulipas

Health Seeking Behaviour and Treatment Intentions of Dengue and Fever:A Household Survey of Children and Adults in Venezuela

BACKGROUND:Dengue in Venezuela is a major public health problem with an increasing incidence of severe cases. Early diagnosis and timely treatment influences the outcome of dengue illness, as delay in care-seeking is significantly associated with complications leading to severe dengue. We aimed to understand patterns of health seeking behaviour (HSB) in individuals exposed to high dengue incidence in order to improve early attendance to health centres. METHODS:Between September 2013 and February 2014 a cross-sectional household survey was performed in Maracay, Venezuela. Intended HSB of adults and children's parents/guardians was assessed with respect to fever or suspected dengue. Data was collected through structured questionnaires from 105 individuals. RESULTS:Most individuals felt at risk of dengue and believed it could be a deadly disease. In the case of suspected dengue, the majority (60%) would choose to first seek medical help versus first treating at home, in contrast to 11% in the case of fever. Amongst those who decided to visit a doctor, a suspected dengue infection would prompt them to search medical help earlier than if having only fever (p<0.001). Multivariate analysis modelling showed that the independent factors associated with the intention to firstly visit a doctor versus treating at home in the case of dengue were feeling at risk (OR = 3.29; p = 0.042) and being an adult (as opposed to caring for a child as a parent/guardian; OR = 3.33, p = 0.021), while having had a previous dengue infection (OR = 0.29; p = 0.031) and living in the neighbourhood Caña de Azúcar (OR = 0.28, p = 0.038) were negatively associated with seeking medical care as their first action. CONCLUSION:Knowledge of HSB related to dengue is scarce in the Americas, our study attempts to contribute to a better understanding of HSB in this region. Improving early dengue disease recognition and awareness may enhance prompt attendance to medical care in affected populations and thereby reduce mortality and severity of dengue. Especially for those with a previous dengue infection, efforts have to be made to promote prompt health centre attendance

Formulación y ajuste de estructuras reguladoras para el manejo de aguas en las sabanas de Arauca y su efecto sobre la producción en el sistema de explotación de bovinos de cría: informe anual

Las actividades de este proyecto se desarrollaron en la vereda Bogotá, a los 6° 51' de latitud norte y 70° 24 de longitud oeste, a una altura de 128 msnm, municipio de Arauca, en la llanura aluvial de desborde, durante el penado comprendido entre enero de 1998 y diciembre de 2001.El estudio agroeconómico realizado en el área de influencia del proyecto, indica que el 70% de los propletanos son onundos del muniCipio de Arauca, predomina la unión libre Las diez familias encuestadas suman un total de 56 personas El 80% de los usuarios tienen educación primaria y 20% secundaria El 100% de los productores encuestados son propietarios y administran personalmente sus fincas.Ganadería bovin

Isospin dependence of electromagnetic transition strengths among an isobaric triplet

Electric quadrupole matrix elements, M, for the J=2→0, ΔT=0, T=1 transitions across the A=46 isobaric multiplet Cr-V-Ti have been measured at GSI with the FRS-LYCCA-AGATA setup. This allows direct insight into the isospin purity of the states of interest by testing the linearity of M with respect to T. Pairs of nuclei in the T=1 triplet were studied using identical reaction mechanisms in order to control systematic errors. The M values were obtained with two different methodologies: (i) a relativistic Coulomb excitation experiment was performed for Cr and Ti; (ii) a “stretched target” technique was adopted here, for the first time, for lifetime measurements in V and Ti. A constant value of M across the triplet has been observed. Shell-model calculations performed within the fp shell fail to reproduce this unexpected trend, pointing towards the need of a wider valence space. This result is confirmed by the good agreement with experimental data achieved with an interaction which allows excitations from the underlying sd shell. A test of the linearity rule for all published data on complete T=1 isospin triplets is presented.Peer Reviewe

Study of Isomeric States in ^{198,200,202,206}Pb and ²⁰⁶Hg Populated in Fragmentation Reactions

Isomeric states in isotopes in the vicinity of doubly-magic 208Pb were populated following reactions of a relativistic 208Pb primary beam impinging on a 9Be fragmentation target. Secondary beams of 198;200;202;206Pb and 206Hg were isotopically separated and implanted in a passive stopper positioned in the focal plane of the GSI Fragment Separator. Delayed γ rays were detected with the Advanced GAmma Tracking Array (AGATA). Decay schemes were re-evaluated and interpreted with shell-model calculations. The momentum-dependent population of isomeric states in the two-nucleon hole nuclei 206Pb/206Hg was found to differ from the population of multi neutron-hole isomeric states in 198;200;202Pb