In vitro evaluation of Sporothrix brasiliensis biotherapic

Sporothrix brasiliensisis one of themost virulent zoonosis which affects animals and humans. This fungus is responsible for subcutaneous infection and its contamination is possible through trauma to the skin. Sporotrichosis is highly prevalent in feline. And Rio de Janeiro appears to have the highest occurrence of cases. Objectives: This study aims to evaluate the in vitroefficacy of Sporothrix brasiliensisbiotherapic, with and without an association to allopathic medicine commonly used in the treatment.Methodology: Conidiumcells of Sporothrix brasiliensiswill be cultured in Potato dextrose agar (PDA) for 5 to 7 days and yeast cells in Brain heart infusion (BHI) for 3 to 5 days. After incubation, the cells will be scraped with a drigalski handle and filtered using cells strainer into a tube and centrifuge for 5 minutes at 3000 RPM. The cells will be resuspended with Phosphate buffered saline (PBS), centrifuge again,and finally resuspended in PBS. After preparing the inocule, the microplates will be prepared. There will be 5groups in vitro. The first one will be the control group, only fungi. The second will be the treatment of fungi with homeopathic medicine (Sporothrix brasiliensis30DH). The third group will be the homeopathic medicine in association with itraconazole. The fourth will be the treatment with itraconazole only. And the last group will be the fungi with dynamized distilled water 30DH. Sporothrix brasiliensis30DHwill be prepared according to Brazilian Homeopathic Pharmacopeia. Results and discussions: The experiments are still in progress and the results will be analyzed through Analysis of variance (ANOVA) to determine statistically significant differences. Previous articles based on biotherapic treatments demonstrated successful results, so our research group is conducting these experiments to evaluate the effect in this model. Conclusion: Experiments will be made to verify the efficacy of biotherapic on sporotrichosis treatments.

Biotherapic 200dH is harmful to acute murine infection with Trypanosoma cruzi

Biotherapics employed to treat mice infected by Trypanosoma cruzi were carried out with encouraging results. The aim of this study was evaluated the effect of biotherapic of Trypanosoma cruzi 200dH, using two different schedules of treatment. Swiss male mice, aged 56 days-old were infected intraperitonealy with 1,400 blood trypomastigotes of Trypanosoma cruzi Y strain and were divided into groups: C.I.- infected animals, E.D. â€" Infected animals treated from the day 1 until the end of the experiment; 200dH S.D. â€" Infected animals treated on the day 1. Parasitological, clinical and immunological parameters were evaluated. The group of animals that received the medicine in a single dose presented higher value to total parasitaemia and lower value of pre patent period compared to control untreated group (p<0.05), as well as the number of amastigotes which was also higher for this group (S.D.) (p<0.05). Clinically, the S.D.group presented more stable temperature (p<0.05) but not presented another clinical difference among treatments. IL-6 and TNF-α presented similar dosage among treated groups as well as IL-4 and IL-10. IL-17A and INF-γ, presented highest values to S.D. group (p<0.05). All animals died until the 20th day of infection. The lack of improvement in clinical and parasitological parameters, the untimely death and the immunological imbalance display the harmful evolution of the experimental infection by T. cruzi using biotherapic 200dH. The results could be useful for homeopathic physicians. In human clinical use, the choice of dynamizations and treatment schedule should consider acute and chronic diseases to achieve the expected results. (AU)

Efeito de bioterápico na eosinofilia durante a SLMV experimental / Biotherapic effect in eosinophilia during experimental VLMS

O Toxocara canis (Tc) é um parasito pertencente ao filo Nematódeo que possui como hospedeiro definitivo os cães, O homem é hospedeiro paratênico e contamina-se acidentalmente ao ingerir ovos contendo larvas infectantes (L3) do parasito, as quais são liberadas e atravessam a mucosa intestinal, atingem a circulação, Durante este processo migratório, antígenos de excreção e secreção (TES) são liberados provocando intensa reação inflamatória, do tipo Th2, caracterizando a síndrome, denominada Larva Migrans Visceral (SLMV), As principais características desta doença crônica são as eosinofilias sanguínea e tecidual persistentes, Desse modo, torna-se importante a busca por terapias que contribuam com a redução dos quadros inflamatórios com intensa eosinofilia, Assim, o uso deste bioterápico, produzido a partir do extrato antigênico de ovos e larvas de (Tc), e seu efeito no recrutamento de leucócitos totais, células mononucleares e eosinófilos no sangue, para o espaço broncoalveolar e para a cavidade peritoneal de camundongos infectados pelo (Tc) foi investigado, Foram utilizados camundongos fêmeas (Swiss), divididos nos grupos: Controle (C), Infectado (Tc), Imunizado (Im+Tc) e Tratado (Tc+Bio), Os animais Tc, Im+Tc e Tc+Bio receberam 500 ovos/animal por gavagem, Posteriormente, os animais foram eutanasiados no 18º dia da infecção e o número das células nos compartimentos foi determinado, Os resultados obtidos demonstraram que, Im+Tc, assim como nos Tc+Bio tiveram redução significativa dessas células nos compartimentos analisados quando comparados grupo Tc, Assim, sugeriu-se que a bioterapia modulou negativamente o recrutamento de células inflamatórias, principalmente eosinófilos no sangue, pulmão e intestino demonstrando um potencial anti-inflamatório desse bioterápico na SLMV experimental...

The Toxocara canis (Tc) is a parasite that belongs to the nematode phylum and has dogs as definitive host. The men can be accidentally contaminated by ingesting eggs containing infective larvae of the parasite. These larvae, when ingested, pass through the intestinal mucosa, reach the portal circulation and migrate through different tissues of the host. During this process, excretory-secretory antigens (TES) are released causing an intense inflammatory reaction, the Th2 type, characterizing the syndrome, called Visceral Larva migrans (VLMS). The main features of this chronic disease are blood and tissue eosinophilias. Thus, it is important to search for therapies that may contribute to the reduction of inflammatory conditions with intense eosinophilia. In this study, we investigated the use of a biotherapic produced from the antigenic extract from eggs and larvae (Tc) and its effect on the recruitment of total leukocyte, mononuclear cells and eosinophils in blood, bronchoalveolar space and peritoneal cavity of mice infected with (Tc). Female mice (Swiss) were used divided in three groups: control (C), Infected (Tc) Immunized (Im + Tc) and Treaty (Tc + Bio). The animals Tc, Im + Tc and Tc + Bio received 500 eggs / animal by gavage. Subsequently, the animals were euthanized on day 18 after infection and the number of cells in the compartments was determined. Our results showed that, Im + Tc, as in Tc + Bio had reduced these cells in compartments analyzed compared to Tc group. Thus, it was suggested that the biotherapy negatively modulated the recruitment of inflammatory cells, particularly eosinophils in blood, lung and intestine demonstrating an anti-inflammatory potential of the biotherapic in the experimental VLMS...

Cell alterations induced by a biotherapic for influenza / Cell alterations induced by a biotherapic for influenza

Introduction: Influenza viruses have been responsible for highly contagious acute respiratory illnesses with high mortality, mainly in the elderly, which encourages the development of new drugs for the treatment of human flu. The biotherapics are medicines prepared from biological products, which are not chemically defined. They are compounded following the homeopathic procedures indicated for infectious diseases with known etiology [1]. Aim: The purpose of the present study is to verify cellular alterations induced by a biotherapic prepared from the infectious influenza A virus. Methodology: This biotherapic was prepared for this study in the homeopathic potency of 30X according to the Brazilian Homeopathic Pharmacopeia [2]. The concentration of 10% was not cytotoxic to cells, as verified by neutral red assay. The cellular alterations observed in MDCK cells were analyzed by optical microscopy for the quantification of mitosis, nucleoli and lipid bodies. The mitochondrial activity was assessed by MTT assay and the phosphosfructokinase-1 (PFK-1) enzyme activity was analyzed on the MDCK cells treated for 5, 10 and 30 days. Macrophages J778.G8 were treated with this biotherapic to evaluate the immunostimulatory cytokine release. Results: The cellular alterations observed in MDCK cells were verified by optical microscopy. The number of lipid bodies present in MDCK cells stimulated for 10 days was significantly lower (p <0.05) when compared to controls. The biotherapic significantly increased (p <0.05) the number of mitosis and the mitochondrial activity of MDCK cells stimulated for 10 and 30 days. These changes were confirmed by a significant reduction (p <0.05) on the PFK-1 activity. These results suggest that the biotherapic was able to activate the Krebs cycle and pentosephosphate metabolism to the generation of amino acids and nucleotides, situations common to cells whose rate of mitosis is increased. The quantification of immunostimulatory cytokines by macrophages J774.G8 indicated that the tumor necrosis factor (TNF-?) production was higher (p <0.05) in the supernatant of the macrophages pre-treated with this biotherapic and infected with influenza virus, suggesting an activation of the macrophages by this biotherapic. Conclusion: This biotherapic is able to induce some cellular alterations, which show strong evidence that it might be a promising option for the human flu. New experiments are being developed to understand the mechanisms of action of this biotherapic.

Efficiency of tick biotherapic on the control of infestation by rhipicephalus (boophilus) microplus in dutch dairy cows / Eficiência de bioterápico de carrapato no controle da infestação por rhipicephalus (boophilus) microplus em vacas leiteiras de raça holandesa / Eficiencia de bioterápico de garrapata para control de la infestación por Rhipicephalus (Boophilus) microplus en vacas lecheras de raza holandesa

Background: cattle tick Rhipicephalus (Boophilus) microplus poses serious problems for farmers in Brazil, especially because the parasite easily develops resistance to pesticide agents. For this reason, together with other factors including environmental, human and animal contamination and costs, alternative approaches have been sought for. Aims: this study sough to evaluate the efficiency of a tick biotherapic on tick-infested cows. Methods: 34 dairy Dutch cows were divided in 2 groups: one group received 100g/day of mineral salt supplement impregnated with tick biotherapic 12cH for 6 months, and then in alternate days with tick biotherapic 30cH to complete 28 months of treatment; the other group (control) received only the mineral salt supplement. After 28 months of treatment, engorged Rhipicephalus (boophilus) microplus females were collected in both groups, counted and weighed; in vitro tests were carried out to assess mass of ticks; egg mass; egg-hatching rate; and reproductive efficiency. Results: There was significant difference between both groups for all parameters evaluated; tick-mass (p = 0.0008); egg mass (p=0.0044); egg-hatching rate (p= 0.0017); and reproductive efficiency (p = 0.0044). Conclusion: treatment with tick biotherapic significantly decreased the mass of engorged females, deposition and hatching rate of eggs, resulting consequently in the decrease of the reproductive efficiency of ticks.

Introdução: os carrapatos do gado Rhipicephalus (Boophilus) microplus representam um sério problema para os criadores brasileiros, especialmente porque desenvolvem rapidamente resistência aos pesticidas. Por isso, junto a outros fatores incluindo contaminação ambiental, humana e animal, assim como os custos, têm sido procuradas abordagens alternativas. Objetivos: avaliar a eficiência de um bioterápico desenvolvido a partir de carrapatos em vacas infestadas. Métodos: 34 vacas leiteiras de raça holandesa foram divididas em 2 grupos; um recebeu 100 mg/dia de suplemento salino mineral impregnado com o bioterápico 12cH durante 6 meses e após 30cH em dias alternos até completar 28 meses de tratamento; o outro grupo (controle) recebeu só o suplemento mineral. Após 28 meses de tratamento foram coletadas fêmeas ingurgitadas do carrapato em ambos os grupos, contadas e pesadas, e foram realizados testes in vitro para determinar a massa de parasitas; massa de ovos; taxa de eclosão dos ovos; e eficiência reprodutiva. Resultados: houve diferença significativa entre ambos os grupos em todos os parâmetros avaliados ? massa de parasitas (p=0,0008); massa de ovos (p=0,0044); taxa de eclosão de ovos (p=0,0017) e eficácia reprodutiva (0,0044). Conclusão: o tratamento com bioterápico de carrapato diminuiu significativamente a massa de fêmeas ingurgitadas e a deposição e taxa de eclosão, resultando, consequentemente, na diminuição da eficiência reprodutiva dos carrapatos.

Introducción: la garrapata del ganado Rhipicephalus (Boophilus) microplus produce serios problemas a los hacenderos brasileños especialmente porque desarrolla rápidamente resistencia a los agentes pesticidas. Por este motivo, asociado a otros factores como contaminación ambiental, humana y animal y los costos, se buscan abordajes alternativos. Objetivos: este estudio buscó evaluar la eficiencia de un bioterápico preparado de garrapatas en vacas infestadas. Métodos: 34 vacas lecheras de raza holandesa fueron divididas en 2 grupos; uno recibió 100 mg/día de suplemento mineral impregnado con bioterápico 12cH durante 6 meses y después, en días alternados hasta completar 28 meses de tratamiento; el otro recibió apenas el suplemento mineral (control). Después de 28 meses de tratamiento, garrapatas hembra ingurgitadas fueron recogidas en ambos grupos, contadas y pesadas; fueron realizados pruebas in vitro para medir la masa de garrapatas; la masa de huevos, la tasa de eclosión de huevos y la eficiencia reproductiva. Resultados: hubo diferencias significativas entre los 2 grupos en todos los parámetros evaluados ? masa de garrapatas (p=0,0008), masa de huevos (p=0,0044), tasa de eclosión de huevos (p=0,0017) y eficiencia reproductiva (p=0,0044). Conclusión: el tratamiento con bioterápico de garrapata redujo significativamente la masa de hembras ingurgitadas y la deposición y eclosión de huevos, resultando en disminución de la eficacia reproductiva de las garrapatas.

Influência do grupo genético, condição sexual e tratamento antiparasitário nas medidas de área de olho do lombo e espessura de gordura in vivo e na carcaça de bovinos de corte / Influence of breed, gender condition, and anti-parasitic treatment on the measurements of in vivo rib eye area and fat thickness and on the carcass of beef cattle

Foram estudados 48 bovinos machos oriundos de inseminação artificial, criados em pasto, sendo 24 (12 Nelore e 12 F1 ½ Red Angus-Nelore ) tratados com antiparasitários alopáticos e 24 (mesmo número de puros e cruzados) tratados com o antiparasitário bioterápico Fator C&MC. Os animais foram desmamados aos oito meses, metade de cada subgrupo genético (6) foi castrado aos 13 meses e todos abatidos aos 32 meses, com o objetivo de verificar a influência do tratamento antiparasitário, do grupo genético e da condição sexual sobre as medidas de área de olho de lombo (AOL) e espessura de gordura de lombo (EGL). Usaram-se medidas de ultrassonografia no animal vivo (AOLU e EGLU) e na carcaça, plástico quadriculado e paquímetro (AOLC e EGLC). Os animais F1, os inteiros e os tratados com alopatia apresentaram peso vivo maior quando comparados aos Nelores, castrados e tratados com bioterápicos. Não houve diferença da AOLU e AOLC entre os grupos genéticos. EGLC foi mais alta nos cruzados. Os animais inteiros apresentaram AOLU e AOLC maiores que os castrados, e EGLU e EGLC menores. Foram altamente significativos os coeficientes de correlação entre as medidas por ultrassom e na carcaça para área de olho de lombo (0,87) e espessura de gordura do lombo (0,95).

Forty-eight male bovines, products of artificial insemination and raised on pasture, were studied, being 24 (12 Nelore, 12 F1 ½ Nelore ½ Red Angus) treated with allopathic antiparasitic drugs and 24 (same number of pure and crossbred) treated with a biotherapic antiparasitic drug Factor C&MC. Animals were weaned at eight months of age and half of each genetic subgroup (six) was castrated at 13 months of age. All animals were slaughtered at 32 months of age, in an attempt to evaluate the influence of antiparasitic treatment, genetic group, and gender condition in the measurements of rib eye area (AOL) and fat thickness (EGL) of loin. Measurements of ultrasonography were used for live animals (AOLU and EGLU), whereas a direct plastic grid reading of the eye muscle and a pachymeter (AOLC and EGLC) were used for carcasses. F1 animals, non-castrated, and those treated with allopathic drugs showed higher live weight when compared with Nelore, castrated, and biotherapic treated animals. There were no differences between genetic groups of AOLU and AOLC. EGLC was higher in crossbred animals. Non-castrated animals showed higher AOLU and AOLC when compared with castrated animals, and lower EGLU and EGLC. Correlation coefficients for ultrasound and carcass measurements were highly significant for rib eye area (0.87) and fat thickness (0.95).