Use of biotherapic in the control of the natural infestation by Boophilus microplus: pilot study

In the control of tick-borne disease (TBD), resistance to chemical products has been reported and a concern with the preservation of the environment requires alternative procedures to control infestation by Boophilus microplus worldwide. The use of biotherapic preparations is one of such alternatives. The aim of this study was to evaluate the effects of a biotherapic mixture including B. microplus in naturally infested cattle. 27 animals were divided in 3 groups (n=9): I – control, received no treatment; II- treated with amitraz dip; III- treated with a standard commercial mixture of biotherapic 12cH, p.o. Group III presented a statistically significant decrease of ticks when compared to the control group (p < 0.05). There was no statistically significant difference between groups II (amitraz) and III (biotherapic) (p < 0.05). These results suggest that biotherapic preparations might be effective in the control of B. microplus in cattle. Keywords: Biotherapic; Tick; Alternative control; Cattle.  Uso do bioterapico no controle da infestação natural por Boophilus microplus: estudo piloto Resumo No controle de enfermidades transmitidas por carrapatos tem sido notada a resisténcia a produtos químicos. A preocupação com a conservação do meio ambiente tem exigido procedimentos alternativos para controlar a infestação por Boophilus microplus, em todo o mundo. O uso de bioterápicos representa uma dessas alternativas. O objetivo deste estudo foi avaliar o efeito de uma mistura de bioterápicos incluindo B. microplus, em gado bovino infestado naturalmente. 27 animais foram divididos em 3 grupos (n=9): I) controle sem tratamento; II) tratado com banho de amitraz; III) tratado com mistura comercial de bioterápicos 12 cH, via oral. O grupo III apresentou uma diminuição estatísticamente significativa de carrapatos, em comparação com grupo controle (p < 0,05). Não houve diferença significativa entre os grupo II e III (p < 0,05). Estes resultados sugerem que os preparados de bioterápicos podem ser eficazes para o controle de B. microplus em bovinos. Palavras-chave: Bioterapicos; Carrapato; Controles alternativos; Bovinos.  Uso de bioterápico en el control de la infestación natural por Boophilus microplus: estudio piloto Resumen En el control de enfermedades trasmitidas por garrapatas, ha sido notada resistencia los productos químicos; la preocupación con la conservación del ambiente exige procedimientos alternativos para controlar la infestación por Boophilus microplus mundialmente. El uso de preparados bioterápicos representa una de esas alternativas. El objetivo de este estudio fue evaluar el efecto de una mezcla bioterápica incluyendo B. microplus en ganado infestado naturalmente. 27 animales fueron divididos en 3 grupos (n=9): I) control, sin tratamiento; II) tratado con banho de amitraz; III) tratado con mezcla comercial de bioterápicos 12cH, vía oral. El grupo III presentó una disminución estadísticamente significativa de garrapatas en comparación al grupo control (p < 0,05). No hubo diferencia significativa entre los grupos II y III (p < 0,05). Estos resultados sugieren que los preparados de bioterápicos pueden ser eficaces para controlar B. microplus en ganado. Palabras-clave: Bioterapicos; Garrapatas; control alternativo; Ganado.  Correspondence author: Alessandro P. Minho. [email protected]; http://www.iapar.br; Instituto Agronômico do Paraná (IAPAR)-PR, Brazil. How to cite this article: Silva NL, Moletta JL, Minho AP, Filippsen LF. Use of biotherapic in the control of natural infestation by Boophilus microplus: pilot study. Int J High Dilution Res [online]. 2008 [cited DD Mmm YYYY]; 7 (22): 36-38. Available from http://journal.giri-society.org/index.php/ijhdr/article/view/251/337. ÂÂÂ

A Simple Micropreparative Gel Electrophoresis Technique For Purification of Nanoscale Materials

Many biochemical, biomedical, and material applications hinge on the ability to effectively separate and purify nanoscale materials. Though this need is largely addressed with biological macromolecules using a variety of chromatographic and electrophoretic purification techniques, such techniques are usually laborious, time-consuming, and often require complex and costly instalments that are inaccessible to most laboratories. Synthetic nanoparticles face similar purification challenges, often relying on techniques that are material-specific. In this work, we introduce a versatile micro-preparative (MP) method based on polyacrylamide gel electrophoresis (PAGE) to purify biological samples containing proteins, nucleic acids, and complex bioconjugates, as well as synthetic nanoparticles based on graphene quantum dots (GQDs). Using a conventional vertical slab PAGE, we demonstrate the extraction of purified DNA, proteins, and DNA-protein bioconjugates from their respective mixtures using MP-PAGE. We apply this system to recover DNA from a ladder mixture with yields of up to 90%, compared to the 58% yield obtained using specialized commercial devices. We also demonstrate the purification of folded enhanced yellow fluorescence protein (EYFP) from crude cell extract with 90% purity, comparable to purities achieved using a two-step size exclusion and immobilized metal-ion affinity chromatography purification procedure. Moreover, we demonstrate the successful isolation of an EYFP-DNA bioconjugate that otherwise could not be processed using the two-step chromatography procedure. Finally, the technique was further extended to demonstrate size-dependent separation of a commercial mixture of GQDs into three different fractions with distinct optical properties. MP-PAGE thus offers a rapid and versatile means of purifying biological and synthetic nanomaterials without the need for specialized equipment.

Behaviour of Titanium Dioxide Particles in Artificial Body Fluids and Human Blood Plasma

The growing application of materials containing TiO2 particles has led to an increased risk of human exposure, while a gap in knowledge about the possible adverse effects of TiO2 still exists. In this work, TiO2 particles of rutile, anatase, and their commercial mixture were exposed to various environments, including simulated gastric fluids and human blood plasma (both representing in vivo conditions), and media used in in vitro experiments. Simulated body fluids of different compositions, ionic strengths, and pH were used, and the impact of the absence or presence of chosen enzymes was investigated. The physicochemical properties and agglomeration of TiO2 in these media were determined. The time dependent agglomeration of TiO2 related to the type of TiO2, and mainly to the type and composition of the environment that was observed. The presence of enzymes either prevented or promoted TiO2 agglomeration. TiO2 was also observed to exhibit concentration-dependent cytotoxicity. This knowledge about TiO2 behavior in all the abovementioned environments is critical when TiO2 safety is considered, especially with respect to the significant impact of the presence of proteins and size-related cytotoxicity.

Assessing the anti-resistance potential of public health vaporizer formulations and insecticide mixtures with pyrethroids using transgenic Drosophila lines

Background Insecticide resistance—and especially pyrethroid resistance—is a major challenge for vector control in public health. The use of insecticide mixtures utilizing alternative modes of action, as well as new formulations facilitating their uptake, is likely to break resistance and slow the development of resistance. Methods We used genetically defined highly resistant lines of Drosophila melanogaster with distinct target-site mutations and detoxification enzymes to test the efficacy and anti-resistance potential of novel mixture formulations (i.e. Fludora® Fusion consisting of deltamethrin and clothianidin), as well as emulsifiable concentrate transfluthrin, compared to alternative, currently used pyrethroid insecticide formulations for vector control. Results The commercial mixture Fludora® Fusion, consisting of both a pyrethroid (deltamethrin) and a neonicotinoid (clothianidin), performed better than either of the single active ingredients against resistant transgenic flies. Transfluthrin, a highly volatile active ingredient with a different molecular structure and primary exposure route (respiration), was also efficient and less affected by the combination of metabolic and target-site resistance. Both formulations substantially reduced insecticide resistance across different pyrethroid-resistant Drosophila transgenic strains. Conclusions The use of mixtures containing two unrelated modes of action as well as a formulation based on transfluthrin showed increased efficacy and resistance-breaking potential against genetically defined highly resistant Drosophila flies. The experimental model remains to be validated with mosquito populations in the field. The possible introduction of new transfluthrin-based products and mixtures for indoor residual spraying, in line with other combination and mixture vector control products recently evaluated for use in public health, will provide solutions for better insecticide resistance management. Graphical abstract

In Vitro Study of the Bioavailability and Bioaccessibility of the Main Compounds Present in Ayahuasca Beverages

Ayahuasca is a psychoactive beverage that contains the psychoactive compound N,N-dimethyltryptamine and β-carboline alkaloids. This study aims at determining in vitro the bioavailability and bioaccessibility of the main compounds present in decoctions of four individual plants, in a commercial mixture and in four mixtures of two individual plants used in the preparation of Ayahuasca. The samples were subjected to an in vitro digestion process, and the Caco-2 cell line was used as an absorption model. The integrity and permeability of the cell monolayer were evaluated, as well as the cytotoxicity of the extracts. After digestion and cell incubation, the compounds absorbed by the cell monolayer were quantified by high-performance liquid chromatography coupled to a diode array detector. The results showed that compounds such as N,N-dimethyltryptamine, Harmine, Harmaline, Harmol, Harmalol and Tetrahydroharmine were released from the matrix during the in vitro digestion process, becoming bioaccessible. Similarly, some of these compounds, after being incubated with the cell monolayer, were absorbed, becoming bioavailable. The extracts did not show cytotoxicity after cell incubation, and the integrity and permeability of the cell monolayer were not compromised.

Stress Effect Induced by Microplastics Coupled with Acid Rain, on Garden Cress, During Short and Long Time: Two Exposures in Comparison

Aim: Plastics, and acid rain, are the results of the anthropogenic activities; although, the injuries caused by acidic precipitation to plants are well known, what happen if acid rain is coupled with microplastics pollution? Methods: In this experiment, we simulated, under controlled condition, a contaminated area from four different kinds of Microplastics (MPs): Polyethylene (PE), Polyvinylchloride (PVC), commercial mixture (PE + PVC) and Polypropylene (PP) subjected to acidic precipitation on Lepidium sativum, both in short then long exposures. At the end of experimentation were analysed: i) plant biometrical parameters (percentage inhibition of seed germination, plant height, leaf number and fresh biomass productions); and ii) oxidative stress (e.g., levels of hydrogen peroxide, glutathione, and ascorbic acid). On plant subjected to long exposure are quantified also: lipidic peroxidation through Malondialdehyde (MDA) concentration, Aminolevulinic Acid (AlA), pigments (Chl-a, Chl-b, carotenoids) and soluble sugars. Results: Our results suggest that root system is more sensitive to MPs coupled with acid rain then above ground biomass. Furthermore, in short exposure toxicity begin with PE+ treatments to shift towards PVC+ toxicity in long exposure at radical level. Conclusion: Our experiment demonstrates that different kinds of microplastics supplied with acid rain are able to interfere in different ways both in short and in long exposure, but also at shoot and root levels, on garden cress.

Stress Effect of Food Matrices on Viability of Probiotic Cells during Model Digestion

The aim of this study was to evaluate the influence of model (alcohol, sugar, salt, protein and acid) and real foods and beverages on the viability of probiotics during incubation and artificial digestion. Viability of monocultures Lactobacillus acidophilus CCM4833 and Bifidobacterium breve CCM7825T, and a commercial mixture of 9 probiotic bacterial strains, was tested by cultivation assay and flow cytometry. In model foods, the best viability was determined in the presence of 0.2 g/L glucose, 10% albumin and 10% ethanol. As the most suitable real food for probiotic survival, complex protein and carbohydrate substrates were found, such as beef broth, potato salad with pork, chicken with rice, chocolate spread, porridge and yoghurt. The best liquid was milk and meat broth, followed by Coca-Cola, beer and coffee. Viability of probiotics was higher when consumed with meals than with beverages only. Addition of prebiotics increased the viability of probiotics, especially in presence of instant and fast foods. Generally, the highest viability of probiotics during artificial digestion was observed in mixed culture in the presence of protein, sugar and fat, or their combination. The increase of cell viability observed in such foods during model digestion may further contribute to the positive effect of probiotics on human health.

Persistent autism-relevant phenotype produced by in utero and lactational exposure of female mice to the commercial PBDE mixture, DE-71

Polybrominated diphenyl ethers (PBDEs) are ubiquitous persistent organic pollutants (POPs) that are known neuroendocrine disrupting chemicals with adverse neurodevelopmental effects. PBDEs may act as risk factors for autism spectrum disorders (ASD), characterized by abnormal psychosocial functioning, although direct evidence is currently lacking. Using a translational exposure model, we tested the hypothesis that maternal transfer of a commercial mixture of PBDEs, DE-71, produces ASD-relevant behavioral and neurochemical deficits in female offspring. C57Bl6/N mouse dams (F0) were exposed to DE-71 via oral administration of 0 (VEH/CON), 0.1 (L-DE-71) or 0.4 (H-DE-71) mg/kg bw/d from 3 wk prior to gestation through lactation. Mass spectrometry analysis indicated in utero and lactational transfer of PBDEs (ppb) to F1 female offspring brain tissue at postnatal day (PND) 15 which was reduced by PND 110. Neurobehavioral testing of social novelty preference (SNP) and social recognition memory (SRM) revealed that adult L-DE-71 F1 offspring display altered short- and long-term SRM, in the absence of reduced sociability, and increased repetitive behavior. These effects were concomitant with reduced olfactory discrimination of social odors. Additionally, L-DE-71 exposure also altered short-term novel object recognition memory but not anxiety or depressive-like behavior. Moreover, F1 L-DE-71 displayed downregulated mRNA transcripts for oxytocin (Oxt) in the bed nucleus of the stria terminalis (BNST) and supraoptic nucleus, vasopressin (Avp) in the BNST and upregulated Avp1ar in BNST, and Oxtr in the paraventricular nucleus. Our work demonstrates that developmental PBDE exposure produces ASD-relevant neurochemical, olfactory processing and behavioral phenotypes that may result from early neurodevelopmental reprogramming within central social and memory networks.

Stress Effect of Food Matrices on Viability of Probiotic Cells During Model Digestion

The aim was to evaluate the influence of model (alcohol, sugar, salt, protein and acid) and real foods and beverages on the viability of probiotics during incubation and artificial digestion. Viability of monocultures Lactobacillus acidophilus CCM4833 and Bifidobacterium breve CCM7825T and commercial mixture of 9 probiotic bacterial strains were tested by cultivation assay and flow cytometry. In model foods, the best viability was determined in the presence of 0.2 g/L glucose, 10% albumin and 10% ethanol. As the most suitable real food for probiotic survival complex protein and carbohydrate substrates were found, such as beef broth, potato salad with pork, chicken with rice, chocolate spread, porridge and yoghurt. The best liquid was milk and meat broth, followed by coca-cola, beer and coffee. Viability of probiotics was higher when consumed with meals than with beverages only. Addition of prebiotics increased the viability of probiotics especially in presence of instant and fast foods. Generally, the highest viability of probiotics during artificial digestion was observed in mixed culture in presence of protein, sugar and fat or their combination. The increase of cell viability observed in such foods during model digestion may further contribute to the positive effect of probiotics on human health.

Establishment of Babesia bovis In Vitro Culture Using Medium Free of Animal Products

Babesia bovis, an etiological agent of bovine babesiosis, causes a significant burden to the cattle industry worldwide. The most efficient method to mitigate bovine babesiosis is a live vaccine produced by serial passage in splenectomized cattle. However, there are several concerns regarding live vaccine production, including variation between batches and the use of many animals. In this study, we report a B. bovis-SF strain continuously cultured in a medium free of components of animal origin enriched with a chemically defined lipid mixture (CD lipid mixture) and the use of a perfusion bioreactor to harvest a large amount of B. bovis. Six culture media were compared, including VP-SFM, CD-CHO, CD-Hydrolyzed, CD-CHO, SFM, and ADMEM/F12. We found that the VP-SFM medium performed the best for B. bovis growth, with a maximum percentage of parasitized erythrocytes (PPE) of 8.6%. The effect of six dilutions of a commercial mixture of CD lipids added to VP-SFM showed that the CD lipid mixture at a dilution of 1:100 had the best B. bovis growth curve, with a maximum PPE of 13.9%. Propagation of the in vitro B. bovis culture was scaled up in a perfusion bioreactor using VP-SFM with a CD lipid mixture, and the PPE reached over 32%. The continuous in vitro B. bovis culture in a medium free of animal origin components could potentially reduce and replace the use of animals to produce a reagent for diagnostics and live vaccines to control bovine babesiosis.