Haematological and biochemical blood study in baby alpaca with diarrhoea

The aim of this study was to determine the haematological value and biochemical blood in baby alpacas with enteric disorder. A total of 30 blood and serum samples were collected from alpacas of 1 month old with diarrhoea and 5 blood samples of clinically healthy baby alpacas (controls). The animals were from communities in the central Andes from Peru. About haematology were determined haematocrit, haemoglobin concentration, red blood count and white blood count that were not significantly different between control animals and animals with diarrhoea. Moreover, biochemical blood parameters as total protein, albumin and calcium decrease significantly (p<0.05). We conclude that our results could be considered as factors in the mortality of baby alpaca by infectious diarrhoea.

Actividad anti-giardia in vitro de los compuestos de foeniculum vulgare y citrus aurantifolia

Giardia lamblia es un protozoario parásito que habita el intestino delgado de humanos y otros vertebrados además de ser el agente responsable de la giardiasis. El fármaco de primera línea para tratar esta parasitosis es el metronidazol, el cual posee efectos adversos considerables, presenta potencial teratogénico y embriotóxico y está considerado como posible carcinógeno en humanos. Los productos naturales son una alternativa eficaz y con menos efectos secundarios para el tratamiento de la giardiasis. En el presente trabajo se determinó la Concentración Inhibitoria media (CI50) de los extractos hexánicos de Foeniculum vulgare y Citrus aurantifolia y algunos de sus constituyentes principales en contra de los trofozoítos de G. lamblia utilizando la técnica del microensayo. También se evaluó la citotoxicidad de los compuestos más activos sobre células Vero empleando el método de exclusión con azul de tripano. Contribuciones y Conclusiones: Los extractos hexánicos de F. vulgare (CI50 89.33 μg/ml) y C. aurantifolia (CI50 185.78 μg/ml) presentaron actividad anti-Giardia in vitro. Los compuestos puros más activos presentes en el extracto hexánico de F. vulgare son Trans,trans-2,4- undecadienal (CI50 72.11 μg/ml) , (+)-Canfeno (CI50 181.13 μg/ml), p-Anisaldehído (CI50 196.78 μg/ml) y (-)-Carvona (CI50 207.01 μg/ml). Mientras que los compuestos puros más activos presentes en el extracto hexánico de C. aurantifolia son Citral (CI50 58.44 μg/ml), Geraniol (CI50 229.01 μg/ml), 3-Metil-1,2-ciclopentanediona (CI50 207.01 μg/ml), 4-Hexen-3-ona (CI50 34.35 μg/ml) y (-)-Carvona (CI50 207.01 μg/ml). De todos ellos el 4-hexen-3ona es el compuesto puro más activo y con el mejor índice de selectividad (IS 19.6820). Ninguno de los compuestos fue tan activo como el metronidazol, sin embargo, ninguno fue tan citotóxico como este.

Deconjugated Bile Salts Produced by Extracellular Bile-Salt Hydrolase-Like Activities from the Probiotic Lactobacillus johnsonii La1 Inhibit Giardia duodenalis In vitro Growth

Giardiasis, currently considered a neglected disease, is caused by the intestinal protozoan parasite Giardia duodenalis and is widely spread in human as well as domestic and wild animals. The lack of appropriate medications and the spread of resistant parasite strains urgently call for the development of novel therapeutic strategies. Host microbiota or certain probiotic strains have the capacity to provide some protection against giardiasis. By combining biological and biochemical approaches, we have been able to decipher a molecular mechanism used by the probiotic strain Lactobacillus johnsonii La1 to prevent Giardia growth in vitro. We provide evidence that the supernatant of this strain contains active principle(s) not directly toxic to Giardia but able to convert non-toxic components of bile into components highly toxic to Giardia. By using bile acid profiling, these components were identified as deconjugated bile-salts. A bacterial bile-salt-hydrolase of commercial origin was able to mimic the properties of the supernatant. Mass spectrometric analysis of the bacterial supernatant identified two of the three bile-salt-hydrolases encoded in the genome of this probiotic strain. These observations document a possible mechanism by which L. johnsonii La1, by secreting, or releasing BSH-like activity(ies) in the vicinity of replicating Giardia in an environment where bile is present and abundant, can fight this parasite. This discovery has both fundamental and applied outcomes to fight giardiasis, based on local delivery of deconjugated bile salts, enzyme deconjugation of bile components, or natural or recombinant probiotic strains that secrete or release such deconjugating activities in a compartment where both bile salts and Giardia are present.

Evaluation of five commercial methods for the extraction and purification of DNA from human faecal samples for downstream molecular detection of the enteric protozoan parasites Cryptosporidium spp., Giardia duodenalis, and Entamoeba spp

High quality, pure DNA is required for ensuring reliable and reproducible results in molecular diagnosis applications. A number of in-house and commercial methods are available for the extraction and purification of genomic DNA from faecal material, each one offering a specific combination of performance, cost-effectiveness, and easiness of use that should be conveniently evaluated in function of the pathogen of interest. In this comparative study the marketed kits QIAamp DNA stool mini (Qiagen), SpeedTools DNA extraction (Biotools), DNAExtract-VK (Vacunek), PowerFecal DNA isolation (MoBio), and Wizard magnetic DNA purification system (Promega Corporation) were assessed for their efficacy in obtaining DNA of the most relevant enteric protozoan parasites associated to gastrointestinal disease globally. A panel of 113 stool specimens of clinically confirmed patients with cryptosporidiosis (n = 29), giardiasis (n = 47) and amoebiasis by Entamoeba histolytica (n = 3) or E. dispar (n = 10) and apparently healthy subjects (n = 24) were used for this purpose. Stool samples were aliquoted in five sub-samples and individually processed by each extraction method evaluated. Purified DNA samples were subsequently tested in PCR-based assays routinely used in our laboratory. The five compared methods yielded amplifiable amounts of DNA of the pathogens tested, although performance differences were observed among them depending on the parasite and the infection burden. Methods combining chemical, enzymatic and/or mechanical lysis procedures at temperatures of at least 56 °C were proven more efficient for the release of DNA from Cryptosporidium oocysts.