Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati

Toxocariasis is a zoonotic disease caused mainly by Toxocara canis and Toxocara cati and diagnosis in dogs and cats is an important tool for its control. For this reason, a new coprological loop-mediated isothermal amplification (LAMP) assay was developed for the simultaneous detection of these species. The primer set was designed on a region of the mitochondrial cox-1 gene. Amplification conditions were evaluated using a temperature gradient (52°C to 68°C), different incubation times (15-120 min), and different concentrations of malachite green dye (0.004-0.4% w/v). The analytical sensitivity was evaluated with serial dilutions of genomic DNA from T. canis and T. cati adult worms, and with serial dilutions of DNA extracted from feces using a low-cost in-house method. The specificity was evaluated using genomic DNA from Canis lupus familiaris, Felis catus, Escherichia coli, Toxascaris leonina, Ancylostoma caninum, Echinococcus granulosus sensu stricto and Taenia hydatigena. The LAMP assay applied to environmental fecal samples from an endemic area showed an analytical sensitivity of 10-100 fg of genomic DNA and 10-5 serial dilutions of DNA extracted from feces using the low-cost in-house method; with a specificity of 100%. Additionally, the total development of the assay was carried out in a basic laboratory and per-reaction reagent cost decreased by ~80%. This new, low-cost tool can help identify the most common agents of toxocariasis in endemic areas in order to manage prevention strategies without having to rely on a laboratory with sophisticated equipment.Fil: Avila, Héctor Gabriel. Universidad Católica de Cuyo. Facultad de Ciencias Químicas y Tecnológicas. Laboratorio Provincial de Zoonosis Provincial; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis; ArgentinaFil: Risso, Marikena Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones en Microbiología y Parasitología Médica. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones en Microbiología y Parasitología Médica; ArgentinaFil: Ruybal, Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones en Microbiología y Parasitología Médica. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones en Microbiología y Parasitología Médica; ArgentinaFil: Repetto, Silvia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones en Microbiología y Parasitología Médica. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones en Microbiología y Parasitología Médica; ArgentinaFil: Butti, Marcos Javier. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Laboratorio de Parasitosis Humanas y Zoonosis Parasitarias; ArgentinaFil: Trangoni, Marcos David. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; ArgentinaFil: Grune Loffler, Sylvia. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Patobiología Veterinaria - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Patobiología Veterinaria; ArgentinaFil: Perez, Veronica Mirtha. Gobierno de la Provincia de San Juan. Ministerio de Salud Publica. Direccion de Epidemiologia. Seccion Rabia y Zoonosis.; ArgentinaFil: Periago, Maria Victoria. Fundación Mundo Sano; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Efecto de la sombra de un rodal de Eucapyptus en verano sobre el comportamiento y el peso de terneros al destete en un sistema de cría bovina en el norte de Buenos Aires.

Shade helps to reduce the negative effects of heat stress, modifies animal behavior, and contributes to increase production rates in beef and dairy cattle. More information is needed on these effects in the north of the province of Buenos Aires, which has a temperate climate. Animal behavior, and weaning weight were evaluated in a herd of 50 pregnant Aberdeen angus cows divided in two groups. Each group was placed in a 35 ha plot. One of them was shaded (S) with 4.5 ha of Eucalyptus sp. and the other one with no access to shade (NS). The states of ingestive and thermoregulatory behavior of cows and calves were recorded using 20-min instantaneous scan sampling on each group, at three times of the day, on 10 days with THI>80 in the summer (2021 & 2022). Calves from each group were weighed at the time of weaning. A greater ingestive behavior with shade was demonstrated (S=826; NS=733; p=0.0005), in cows (S=442; NS=375; p=0.0270) and calves (S=384; NS =358; p=0.0046). The thermoregulation was higher in the NS treatment (S=624; NS=717; p=0.0065), verified by resting in the drinking area at noon and afternoon. A higher herd weight (kg b.w.) of S calves was recorded (S=11,544; NS=11,157; p>0.05). The S females had a higher average of kg b.w. (S=226.25; NS=214.64; p0.05)Instituto de Recursos BiológicosFil: Mujica, Gerardo Oscar. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Recursos Biológicos; ArgentinaFil: Padín, Martín. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Recursos Biológicos; ArgentinaFil: Mc Dermott, Augusto. Productor, Establecimiento "La Negra"; ArgentinaFil: Trangoni, Marcos David. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentin

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

Changes in hematological, biochemical, and blood gases prameters in response to progressive inclusion of nitrate in the diet of Holstein calves

Background and Aim: Nitrate (NO3-) reduces enteric methane emissions and could be a source of non-protein nitrogen in ruminant feeds. Nonetheless, it has a potential toxic effect that could compromise animal health and production. The purpose of this study was to determine the effects of progressive inclusion of NO3- in the diet on the hematological, biochemical, and blood gases parameters, in turn, the effects on feed intake and live weight gain (LWG) in Holstein calves. Materials and Methods: Eighteen Holstein heifers and steers (nine animals/treatment) were maintained in individual pens for 45 days. Animals were randomly allocated to either a control or nitrate diet (ND) (containing 15 g of NO3-/kg of dry matter [DM]). The biochemical parameters and blood gases were analyzed only in the NO3- group on days: -1, 1, 7, 13, 19, and 25 corresponding to 0, 20, 40, 60, 80, and 100% of the total inclusion of NO3- in the diet, respectively. In addition, DM intake (DMI) and LWG were evaluated among dietary treatments. Results: Feeding the ND did not influence DMI or LWG (p>0.05). Methemoglobin (MetHb) and deoxyhemoglobin increased according to the NO3- concentrations in the diet (p0.05). However, glucose, urea, aspartate aminotransferase (AST), and retinol concentrations increased (p<0.05) according to the NO3- concentrations in the diet.Instituto de PatobiologíaFil: Ortiz Chura, Abimael. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; ArgentinaFil: Ortiz Chura, Abimael. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Marcoppido, Gisela Ariana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; ArgentinaFil: Marcoppido, Gisela Ariana. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Gere, José. Universidad Tecnológica Nacional. División Investigación y Desarrollo de Ingenierías; ArgentinaFil: Gere, José. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Depetris, Gustavo. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; ArgentinaFil: Stefañuk Bahamonte, Francisco José. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; ArgentinaFil: Trangoni, Marcos David. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular. ArgentinaFil: Trangoni, Marcos David. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Cravero, Silvio Lorenzo Pedro. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Cravero, Silvio Lorenzo Pedro. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Faverin, Claudia. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; ArgentinaFil: Cataldi, Angel Adrian. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Cataldi, Angel Adrian. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Ceron Cucchi, Maria Esperanza. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; ArgentinaFil: Ceron Cucchi, Maria Esperanza. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

A closed-tube loop-mediated isothermal amplification assay for the visual endpoint detection of Brucella spp. and Mycobacterium avium subsp. paratuberculosis

LAMP (loop-mediated isothermal amplification) is an isothermal nucleic acid amplification technique that is characterized by its efficiency, rapidity, high yield of final product, robustness, sensitivity, and specificity, with the blueprint that it can be implemented in laboratories of low technological complexity. Despite the conceptual complexity underlying the mechanistic basis for the nucleic acid amplification, the technique is simple to use and the amplification and detection can be carried out in just one step. In this chapter, we present a protocol based on LAMP for the rapid identification of isolates of Brucella spp. and Mycobacterium avium subsp. paratuberculosis, two major bacterial pathogens in veterinary medicine.Instituto de BiotecnologíaFil: Trangoni, Marcos David. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; ArgentinaFil: Gioffre, Andrea Karina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Cravero, Silvio Lorenzo Pedro. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentin

Cytokine expression profile of B. melitensis-infected goat monocyte-derived macrophages

Brucella parasitize the macrophage where is able to replicate and modulate the immune response in order to establish a chronic infection. The most adequate response to control and eliminate Brucella infection is a type 1 (Th1) cell-mediated effector immunity. Research in immune response of B. melitensis-infected goats is relatively scarce. In this study, we first evaluated changes in the gene expression of cytokines, a chemokine (CCL2) and the inducible nitric oxide synthase (iNOS) of goat macrophage cultures derived from monocytes (MDMs) infected for 4 and 24 h with Brucella melitensis strain 16 M. TNFα, IL-1β and iNOS, and IL-12p40, IFNγ and also iNOS were significantly expressed (p < 0.05) at 4 and 24 h respectively, in infected compared to non-infected MDMs. Therefore, the in vitro challenge of goat MDMs with B. melitensis promoted a transcriptional profile consistent with a type 1 response. However, when the immune response to B. melitensis infection was contrasted between MDM cultures phenotypically restrictive or permissive to intracellular multiplication of B. melitensis 16 M, it was observed that the relative IL-4 mRNA expression was significantly higher in permissive macrophage cultures with respect to restrictive cultures (p < 0.05), independently of the time p.i. A similar trend, although non-statistical, was recorded for IL-10, but not for pro-inflammatory cytokines. Thus, the up-expression profile of inhibitory instead of pro-inflammatory cytokines could explain, in part, the difference observed in the ability to restrict intracellular replication of Brucella. In this sense, the present results make a significant contribution to the knowledge of the immune response induced by B. melitensis in macrophages of its preferential host species.Instituto de PatobiologíaFil: Maurizio, Estefania. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria; Argentina.Fil: Maurizio, Estefania. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Rossi, Ursula. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria; ArgentinaFil: Rossi, Ursula. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Trangoni, Marcos David. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular. ArgentinaFil: Trangoni, Marcos David. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Rossetti, Carlos Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria; Argentina.Fil: Rossetti, Carlos Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Characterization of innate immune response to Brucella melitensis infection in goats with permissive or restrictive phenotype for Brucella intramacrophagic growth

Caprine brucellosis is a chronic, world-wide distributed disease which causes reproductive failure in goats and Brucella melitensis, its causative agent, bears a great zoonotic potential. There is evidence suggesting that some cattle and pigs have an innate ability to resist Brucella infection, but this has not yet been investigated in goats. In this study, we compared caprine macrophages that exhibit extreme restriction and permissiveness to B. melitensis’ intracellular growth in vitro. Monocyte derived macrophages (MDMs) from 110 female goats were cultured and challenged in vitro with B. melitensis 16 M. After initial screening, 18 donor goats were selected based on their macrophages ability to restrict or allow bacterial intracellular growth and some elements of humoral and cellular immunity were studied in depth. MDMs that were able to restrict the pathogen’s intracellular growth showed enhanced bacterial internalization, although there were no differences between groups in the production of reactive oxygen and nitrogen intermediates following 48 h treatment with heat-killed B. melitensis. Moreover, there were no differences between groups in the level of antibodies reacting with keyhole limpet hemocyanin (natural antibodies, NAbs) or with Brucella LPS antigens (cross-reacting antibodies, CrAbs), although a strong positive correlation between individual levels of IgM NAbs and IgM CrAbs was detected. Altogether, these results represent an initial step in understanding innate primary host response to B. melitensis, and deciphering which mechanisms may determine a successful outcome of the infection in goats.Instituto de PatobiologíaFil: Maurizio, Estefania. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; ArgentinaFil: Maurizio, Estefania. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Trangoni, Marcos David. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular. ArgentinaFil: Trangoni, Marcos David. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Rossi, Ursula. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; ArgentinaFil: Rossi, Ursula. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Dunleavy, Mariana Victoria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; ArgentinaFil: Dunleavy, Mariana Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Colato, Claudio. Cabaña Nuevo Milenium; ArgentinaFil: Rossetti, Carlos Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; ArgentinaFil: Rossetti, Carlos Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Brucella canis Group 2 isolated in Argentina = Brucella canis Grupo 2 aislado en Argentina

The aim of this study was to estimate the diversity and prevalence of both groups of Brucella canis 1 and 2 with and without deletion respectively in different areas of Argentina. A total of 104 bacterial cultures were typed as B. canis strains using the classical biotyping method. Two PCR assays were performed to confirm that all isolates were B. canis and not Brucella suis. The differentiation between groups 1 and 2 was achieved using another PCR assay and the diversity of B. canis isolates was assessed with four MLVA_16 markers. All strains belonged to Group 2. Bruce 09 marker (MLVA_16 assay) showed the greatest diversity. Only Group 2 of B. canis was identified among the strains evaluated. The markers chosen from the MLVA_16 allowed us to detect genetic diversity among the strains of B. canis studied.El objetivo de este trabajo fue estimar la diversidad y la prevalencia de ambos grupos de Brucella canis 1 y 2 (con y sin deleción, respectivamente) en diferentes áreas de Argentina. Un total de 104 cultivos bacterianos se tipificaron como cepas de B. canis usando biotipado clásico. Se realizaron dos ensayos de PCR para confirmar que todos los aislamientos eran B. canis y no Brucella suis. La diferenciación entre los grupos 1 y 2 se logró con otro ensayo de PCR, y la diversidad entre las cepas de B. canis se obtuvo mediante el empleo de cuatro marcadores del ensayo de MLVA_16. Todas las cepas pertenecieron al grupo 2. El marcador Bruce 09 (ensayo MVLA-16) mostró la mayor diversidad. Sólo se halló el Grupo 2 de B. canis entre las cepas estudiadas. Los marcadores del MLVA_16 permitieron detectar la presencia de diversidad genética entre las cepas de B. canis analizadas.Instituto de BiotecnologíaFil: Boeri, Eduardo Jorge. Instituto de Zoonosis Luis Pasteur; ArgentinaFil: Madariaga, María Julia. Instituto de Zoonosis Luis Pasteur; ArgentinaFil: Dominguez, María Luz. Instituto de Zoonosis Luis Pasteur; ArgentinaFil: Teijeiro, María Luisa. Instituto de Zoonosis Luis Pasteur; ArgentinaFil: Fernandez, Natalia Mercedes. Instituto de Zoonosis Luis Pasteur; ArgentinaFil: Elena, Sebastián. Servicio Nacional de Sanidad y Calidad Agroalimentaria. Dirección General de Laboratorio y Control Técnico. Laboratorio de Referencia de la OIE para Brucelosis; ArgentinaFil: Trangoni, Marcos David. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular. ArgentinaFil: Trangoni, Marcos David. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Development of a New LAMP Assay for the Detection of Ancylostoma caninum DNA (Copro-LAMPAc) in Dog Fecal Samples

Ancylostoma caninum is a zoonotic nematode which is able to affect animals and humans. Diagnosis in the definitive host and environmental detection are key to prevent its dissemination and achieve control. Herein, a new coprological LAMP method for the detection of A. caninum (Copro-LAMPAc) DNA was developed. DNA extraction was performed using a low-cost method and a fragment of the cox-1 gene was used for primer design. The analytical sensitivity, evaluated with serial dilutions of genomic DNA from A. caninum adult worms, was 100 fg. A specificity of 100% was obtained using genomic DNA from the host and other pathogens. The Copro-LAMPAc was evaluated using environmental canine fecal samples. When compared with gold standard optical microscopy in epidemiological studies, it proved to be more sensitive. This new LAMP assay can provide an alternative protocol for screening and identification of A. caninum for epidemiological studies in endemic areas.Fil: Avila, Héctor Gabriel. Gobierno de la Provincia de San Juan. Ministerio de Salud Pública. Dirección de Epidemiología. Laboratorio Provincial de Zoonosis; Argentina. Universidad Católica de Cuyo. Facultad de Ciencias Químicas y Tecnológicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Juan; ArgentinaFil: Risso, Marikena Guadalupe. Universidad de Buenos Aires. Facultad de Medicina. Departamento de Microbiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones en Microbiología y Parasitología Médica. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones en Microbiología y Parasitología Médica; ArgentinaFil: Cabrera, Marta. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud "Dr. C. G. Malbran". Departamento de Parasitología; ArgentinaFil: Ruybal, Paula. Universidad de Buenos Aires. Facultad de Medicina. Departamento de Microbiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones en Microbiología y Parasitología Médica. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones en Microbiología y Parasitología Médica; ArgentinaFil: Repetto, Silvia Analía. Universidad de Buenos Aires. Facultad de Medicina. Departamento de Microbiología; Argentina. Universidad de Buenos Aires. Facultad de Medicina. Hospital de Clínicas General San Martín; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones en Microbiología y Parasitología Médica. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones en Microbiología y Parasitología Médica; ArgentinaFil: Butti, Marcos Javier. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Laboratorio de Parasitosis Humanas y Zoonosis Parasitarias; ArgentinaFil: Trangoni, Marcos David. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Nacional de Investigaciones Agropecuarias; ArgentinaFil: Santillan, Graciela Ines. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud "Dr. C. G. Malbran". Departamento de Parasitología; ArgentinaFil: Perez, Veronica Mirtha. Gobierno de la Provincia de San Juan. Ministerio de Salud Pública. Dirección de Epidemiología. Laboratorio Provincial de Zoonosis; Argentina. Universidad Católica de Cuyo. Facultad de Ciencias Químicas y Tecnológicas; ArgentinaFil: Periago, Maria Victoria. Fundación Mundo Sano; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Development of a copro-LAMP assay for detection of several species of Echinococcus granulosus sensu lato complex

Cystic echinococcosis represents a significant problem in human and animal health and constitutes one of the most severe Neglected Tropical Diseases prioritized by the World Health Organization. The etiological agent is the complex Echinococcus granulosus sensu lato (s. l.), composed of several species/genotypes. Diagnosis in the definitive host and molecular epidemiology studies are important points for cystic echinococcosis control. Here we developed a new copro-LAMP assay, LAMP EGSL, for diagnosis in the definitive host for simultaneous detection of Echinococcus granulosus sensu stricto (s. s.), Echinococcus ortleppi, and Echinococcus canadensis species. Also, the analytical sensitivity, specificity and plausibility of performance in a rural context of a previously reported species-specific LAMP reaction, was evaluated. Both reactions showed high analytical sensitivity values (10 fg-100 fg DNA) and did not show cross reaction with DNA from host or other helminthic parasites. LAMP EGSL was performed with samples from an endemic area. In addition, the alkaline hydrolysis of one E. granulosus s. s. adult parasite followed by specific LAMP to E. granulosus s. s. was performed in a laboratory with low resources from another cystic echinococcosis endemic area. The results obtained suggest that LAMP EGSL represents a potential tool for canine diagnosis that could be useful for cystic echinococcosis control programs. In addition, we showed that LAMP reaction for E. granulous s. s., E. ortleppi and E. canadensis specific detection, could be useful for molecular epidemiology studies applicable to the definitive host. Both reactions were performed in endemic, rural areas without sophisticated equipment.Instituto de BiotecnologíaFil: Avila, Héctor Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones en Microbiología y Parasitología Médica. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones en Microbiología y Parasitología Médica; Argentina. Centro de Investigación en Zoonosis (Chubut); ArgentinaFil: Mozzoni, Cecilia. Santa Cruz (Argentina : provincia). Ministerio de Salud y Ambiente. Hospital Zonal Caleta Olivia; ArgentinaFil: Trangoni, Marcos David. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular. Laboratorio de Brucella, Campylobacter & Microbiota; Argentina.Fil: Cravero, Silvio Lorenzo Pedro. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular. Laboratorio de Brucella, Campylobacter & Microbiota; ArgentinaFil: Pérez, Verónica Mirtha. San Juan (Argentina : provincia). División Zoonosis; ArgentinaFil: Valenzuela, Federico. San Juan (Argentina : provincia). División Zoonosis; ArgentinaFil: Gertiser, María Laura. Centro de Investigación en Zoonosis (Chubut); ArgentinaFil: Butti, Marcos Javier. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Cátedra de Parasitología Comparada. Laboratorio de Parasitosis Humanas y Zoonosis Parasitarias; ArgentinaFil: Kamenetzky, Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones en Microbiología y Parasitología Médica. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones en Microbiología y Parasitología Médica; Argentina.Fil: Jensen, Oscar. Centro de Investigación en Zoonosis (Chubut); ArgentinaFil: Rosenzvit, Mara Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones en Microbiología y Parasitología Médica. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones en Microbiología y Parasitología Médica; Argentina