National Center for Animal and Plant Health

San José de las Lajas, Cuba

National Center for Animal and Plant Health

San José de las Lajas, Cuba
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Sanchez Calero J.,National Center for Animal and Plant Health | Young L.C.,University of Strathclyde | Marrero Faz E.,National Center for Animal and Plant Health | L. Harvey A.,University of Strathclyde
Revista Cubana de Farmacia | Year: 2014

Introduction: Allophylus cominia (L.) Sw is a Cuban medicinal plant used by traditional medicine for the treatment of diabetes with unknown mechanisms of action. Objective: to evaluate the effect of Allophylus cominia (L.) Sw leaves aqueous extract and its fractions on protein tyrosine phosphatase 1B (PTP1B) and dipeptidyl peptidase IV (DPPIV) enzymatic activity, as therapeutic targets of type 2 diabetes. Methods: the aqueous extract of A. cominia leaves was successively partitioned with organic solvents mixtures, thus increasing polarity in order to obtain ten fractions. The extract and its fractions were tested for their possible antidiabetic activity on therapeutic targets of type 2 diabetes: PTP1B and DPPIV. The enzymatic inhibition assays were performed and the inhibitory activity was calculated with the fluorescence values using an excitation wavelength of 360 nm and an emission wavelength of 460 nm. Results: the aqueous extract from A. cominia inhibited the enzymatic activity of PTP1B and DPPIV according to the concentration, being IC50 values equal to 0.69 μg/mL and 344.3 μg/mL, respectively. Several fractions were detected as potent PTP1B inhibitors. The most polar fractions AcF9 and AcF10 were more active, showing IC50 values of 4.4 µg/mL and 3.8 µg/mL respectively. The fractions showed a slight DPPIV inhibition, being fractions AcF6, AcF9 and AcF10 the most active, exhibiting inhibition percentages of 52.0 %, 39.0 % and 40.0 % respectively. Conclusions: A. cominia aqueous extract and its polar fractions (AcF9 and AcF10) have antidiabetic properties in vitro and are promissory candidates for development of new drugs with inhibitory activity of PTP1B and DPPIV for type 2 diabetes treatment. © 2015 1999, Editorial Ciencias Médicas.

PubMed | University of Buenos Aires, Instituto Nacional de Tecnologia Agropecuaria, CIRAD UMR 15 UMR CIRAD INRA 1309 controle des maladies animales exotiques et emergentes, The Conservation Land Trust and 2 more.
Type: Journal Article | Journal: Parasites & vectors | Year: 2016

Anaplasma marginale is a well-known cattle pathogen of tropical and subtropical world regions. Even though, this obligate intracellular bacterium has been reported in other host species different than bovine, it has never been documented in Myrmecophaga tridactyla (giant anteater) or Hippocamelus antisense (taruca), which are two native endangered species.Samples from two sick wild animals: a Myrmecophaga tridactyla (blood) and a Hippocamelus antisense (blood and serum) were studied for the presence of A. marginale DNA through msp5 gene fragment amplification. Further characterization was done through MSP1a tandem repeats analysis and MLST scheme and the genetic relationship among previously characterized A. marginale sequences were studied by applying, eBURST algorithm and AMOVA analysis.Anaplasma marginale DNA was identified in the Myrmecophaga tridactyla and Hippocamelus antisense samples. Through molecular markers, we identified an identical genotype in both animals that was not previously reported in bovine host. The analysis through eBURST and AMOVA revealed no differentiation between the taruca/anteater isolate and the bovine group.In the present publication we report the identification of A. marginale DNA in a novel ruminant (Hippocamelus antisense) and non-ruminant (Myrmecophaga tridactyla) host species. Genotyping analysis of isolates demonstrated the close relatedness of the new isolate with the circulation population of A. marginale in livestock. Further analysis is needed to understand whether these two hosts contribute to the anaplasmosis epidemiology.

PubMed | Center for Genetic Engineering and Biotechnology, National Center for Animal and Plant Health, Laboratory for Avian Investigations and Diagnostic LIDA and Ibero-American University of the Dominican Republic
Type: Journal Article | Journal: Open veterinary journal | Year: 2015

In this study we demonstrated that the vaccine candidate against avian influenza virus H5N1 based on the hemagglutinin H5 (HA) fused to the chicken CD154 (HACD) can also be used for differentiating infected from vaccinated animals (DIVA). As the strategy of DIVA requires at least two proteins, we obtained a variant of the nucleoprotein (NP49-375) in E. coli. After its purification by IMAC, the competence of the proteins NP49-375 and HACD as coating antigens in indirect ELISA assays were tested by using the sera of chickens immunized with the proteins HA and HACD and the reference sera from several avian influenza subtypes. Together with these sera, the sera from different species of birds and the sera of chickens infected with other avian viral diseases were analyzed by competition ELISA assays coated with the proteins NP49-375 and HACD. The results showed that the segment CD154 in the chimeric protein HACD did not interfere with the recognition of the molecule HA by its specific antibodies. Also, we observed variable detection levels when the reference sera were analyzed in the ELISA plates coated with the protein NP49-375. Moreover, only the antibodies of the reference serum subtype H5 were detected in the ELISA plates coated with the protein HACD. The competition ELISA assays showed percentages of inhibition of 88-91% for the positives sera and less than 20% for the negative sera. We fixed the cut-off value of these assays at 25%. No antibody detection was observed in the sera from different species of birds or the sera of chickens infected with other avian viral diseases. This study supported the fact that the ELISA assays using the proteins NP49-375 and HACD could be valuable tools for avian influenza surveillance and as a strategy of DIVA for counteracting the highly pathogenic avian influenza virus H5N1 outbreaks.

Soler D.M.,National Center for Animal and Plant Health | Rodriguez Y.,National Center for Animal and Plant Health | Correa H.,National Center for Animal and Plant Health | Moreno A.,Venezuelan Institute for Scientific Research | Carrizales L.,Venezuelan Institute for Scientific Research
Radiation Physics and Chemistry | Year: 2012

This study is aimed of producing pilot batches of hydrogel wound dressings by gamma radiation and evaluating their shelf stability. Six batches of 3L capacity were prepared based on poly(vinyl pyrrolidone), agar and polyethylene glycol and they were dispensed in polyester trays, covered with polyester films and sealed in two types of materials: polyethylene bags and vacuum polyethylene bags. Dressings were formed in a single step process for the hydrogel formation and sterilization at 25-30. kGy gamma radiation dose in a JS-9500 Gamma Irradiator (Nordion, Canada). The six batches were initially physicochemical characterized in terms of dimensions and appearance, gel fraction, morphology analysis, hydrogel strength, moisture retention capability and swelling capacity. They were kept under two storage conditions: room temperature (T: 30±2 °C/RH: 70± 5%) and refrigerated temperature (T: 5±3 °C) during 24 months and sterility test was performed. The appearance of membranes was transparent, clear, uncut and flexible; the gel fraction of batches was higher than 75% and the hydrogel surface showed a porous structure. There was a slow decrease of the compression rate 20% until 7. h and about 70% at 24 h. Moisture retention capability in 5. h was similar for all the batches, about 40% and 60% at 37 °C and at room temperature respectively. The swelling of hydrogels in acidic media was strong and in alkaline media the weight variation remains almost stable until 24. h and then there is a loss of weight. The six batches remained sterile during the stability study in the conditions tested. The pilot batches were consistent from batch to batch and remained stable during 24 months. © 2012 Elsevier Ltd.

Perez K.A.,Las Tunas University | Pinol B.,National Center for Animal and Plant Health | Rosete Y.A.,Rothamsted Research | Rosete Y.A.,CABI Inc | And 3 more authors.
Journal of Phytopathology | Year: 2010

Transmission tests were conducted with field-collected Bunchy Top Symptoms (BTS) phytoplasma-infected specimens of Empoasca papayae. BTS developed in all eight inoculated papayas 3 months later. The BTS phytoplasma was identified in six of eight inoculated papayas, whose partial 16S rRNA sequence (GenBank Accession no. FJ6492000) was 99.9% identical with those from the collected papayas (GenBank Accession no FJ649198) and E. papayae (GenBank Accession no. FJ649199), all of which are members of group 16SrII, '. Candidatus Phytoplasma aurantifolia'. Results confirmed the ability of E. papayae to transmit the BTS phytoplasma. © 2009 Blackwell Verlag GmbH.

PubMed | Federal Rural University of Rio de Janeiro, University of Habana and National Center for Animal and Plant Health
Type: Journal Article | Journal: Ticks and tick-borne diseases | Year: 2016

Canine cyclic thrombocytopenia, an infectious disease caused by Anaplasma platys is a worldwide dog health problem. This study aimed to detect and characterize A. platys deoxyribonucleic acid (DNA) in dogs and ticks from Cuba using molecular methods. The study was conducted in four cities of Cuba (Habana del Este, Boyeros, Cotorro and San Jos de las Lajas). Blood samples were collected from 100 dogs in these cities. The animals were inspected for the detection of tick infestation and specimens were collected. Genomic DNA was extracted from dog blood and ticks using a commercial kit. Genomic DNA samples from blood and ticks were tested by a nested polymerase chain reaction (nPCR) to amplify 678 base pairs (bp) from the 16S ribosomal DNA (rDNA) of A. platys. Positive samples in nPCR were also subjected to PCR to amplify a fragment of 580bp from the citrate synthase (gltA) gene and the products were sequenced. Only Rhipicephalus sanguineus sensu lato (s.l.) was found on dogs, and 10.20% (n=5/49) of these ticks plus sixteen percent (16.0%, n=16/100) of dogs were considered positive for A. platys by nPCR targeting the 16S rDNA gene. All analyzed gltA and 16S rDNA sequences showed a 99-100% identity with sequences of A. platys reported in around the world. Phylogenetic analysis showed two defined clusters for the 16S rDNA gene and three defined clusters for the gltA gene. Based on the gltA gene, the deduced amino acid sequence showed two mutations at positions 88 and 168 compared with the sequence DQ525687 (GenBank ID from Italian sample), used as a reference in the alignment. A preliminary study on the epidemiological aspects associated with infection by A. platys showed no statistical association with the variables studied (p>0.05). This is the first evidence of the presence of A. platys in dogs and ticks in Cuba. Further studies are needed to evaluate the epidemiological aspects of A. platys infection in Cuban dogs.

Rodriguez-Roche R.,Institute of Tropical Medicine | Sanchez L.,Institute of Tropical Medicine | Burgher Y.,National Center for Animal and Plant Health | Rosario D.,Institute of Tropical Medicine | And 5 more authors.
Vector-Borne and Zoonotic Diseases | Year: 2011

Dengue epidemics in Cuba have repeatedly demonstrated a month-to-month increase in clinical severity during secondary infections. The dengue 2 outbreak that occurred in Santiago de Cuba in 1997 was accompanied by the most severe intraepidemic increase in disease severity reported to date. It was initially proposed that the appearance of neutralization escape mutants during the course of the epidemic might explain this phenomenon. Recent studies have revealed that during the course of this epidemic, nucleotide substitutions appeared only in nonstructural (NS) genes, most of which were silent, except for one change in the NS1 gene. To study whether or not variation in the NS1 gene might be associated with increased disease severity during the epidemic, this gene was partially sequenced from 15 isolates obtained at different times during the 1997 epidemic. Early epidemic isolates differed from those obtained later by replacement only of threonine with serine at position 164 in the NS1 protein, an amino acid rarely found in any genotype of dengue 2 virus. All viruses isolated from patients located in Health Districts, where dengue 2 transmissions occurred late in the epidemic, contained Serine at position 164, indicating that this change was fixed within a few months. Here we argue that this single mutation contributes to viral survival or replication efficiency, resulting in enhanced infection in the presence of enhancing antibodies, a phenomenon that we term increased virus "fitness" in contrast to "virulence," an intrinsic property of the virus. © Copyright 2011, Mary Ann Liebert, Inc.

Toledo J.R.,University of Concepción | Toledo J.R.,Galactous Biotech Ltda | Barrera M.,National Center for Animal and Plant Health | Farnos O.,Center for Genetic Engineering and Biotechnology | And 8 more authors.
Vaccine | Year: 2010

Subunit vaccines are a suitable alternative for the control of classical swine fever. However, such vaccines have as the main drawback the relatively long period of time required to induce a protective response, which hampers their use under outbreak conditions. In this work, type I interferon is used as an immunostimulating molecule in order to increase the immunogenicity of a vaccine candidate based on the E2-CSFV antigen produced in goat milk. Pigs vaccinated with E2-CSFV antigen co-formulated with recombinant human alpha interferon were protected against clinical signs and viremia as early as 7 days post-vaccination. It was also demonstrated that interferon stimulates a response of specific anti-CSFV neutralizing antibodies. The present work constitutes the first report of a subunit vaccine able to confer complete protection by the end of the first week after vaccination. These results suggest that the E2-CSFV antigen combined with type I interferons could be potentially used under outbreak conditions to stop CSFV spread and for eradication programs in CSF enzootic areas. © 2010 Elsevier Ltd.

Sanchez J.,National Center for Animal and Plant Health | Oliva Y.,National Center for Animal and Plant Health | Marrero E.,National Center for Animal and Plant Health
Boletin Latinoamericano y del Caribe de Plantas Medicinales y Aromaticas | Year: 2014

Allopfiylus cominia (L) Sw (Sapindaceae), is one of the most popular medicinal plant in Cuba. It is traditionally used in the treatment of diabetes. The aim of this study was to investigate the effect of A cominia leaves aqueous extract in a type 2 diabetes model induced by streptozotocin treatment in neonatal rats. Two experiments was executed: at 6 weeks old rats, before starting the plant evaluation, and at 11 weeks old rats, after the oral administration daily of three doses 0.25, 0.5 and 1.0 g/kg of A cominia aqueous extract during 3 weeks and plasma blood glucose level was evaluated. At 6 weeks old rats, the group of diabetic animals showed significant decrease in blood glucose and body weight values compared to the control group (p 0.05). At 11 weeks old rats, the groups of diabetic animals treated with A cominia aqueous extract significantly decreased blood glucose values compared to the untreated diabetic group (p 0.05). Only the groups treated with 0.5 and 1.0 g/kg of A cominia extract showed a recovery of normal glycaemic values. Hence, it can be concluded that aqueous extract from A cominia leaves has antidiabetic properties and may be effective in the type 2 diabetes treatment. © 2014.

Ramos O.S.,University of Concepción | Pose A.G.,Center for Genetic Engineering and Biotechnology | Gomez-Puerta S.,University of Concepción | Gomez J.N.,National Center for Animal and Plant Health | And 5 more authors.
Comparative Immunology, Microbiology and Infectious Diseases | Year: 2011

Recombinant adenoviral vectors have emerged as an attractive system for veterinary vaccines development. However, for poultry vaccination a very important criterion for an ideal vaccine is its low cost. The objective of this study was to test the ability of chicken CD154 to enhance the immunogenicity of an adenoviral vector-based vaccine against avian influenza virus in order to reduce the amount of antigen required to induce an effective immune response in avian. Chickens were vaccinated with three different doses of adenoviral vectors encoding either HA (AdHA), or HA fused to extracellular domain chicken's CD154 (AdHACD). Hemagglutination inhibition (HI) assay and relative quantification of IFN-γ showed that the adenoviral vector encoding for the chimeric antigen is able to elicit an improved humoral and cellular immune response, which demonstrated that CD154 can be used as a molecular adjuvant allowing to reduce in about 50-fold the amount of adenoviral vector vaccine required to induce an effective immune response. © 2010 Elsevier Ltd.

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