Study ofthe comparative efficacy of the phytotherapeutic complex against nematodes of mice

The purpose of the research is to studythe efficacy of phytotherapeutic, and homeopathic drugs. Phytotherapeutic drugs have been widely used in parasitology for many years. Also, homeopathic drugs such as Cina have anti-helminthicproperties. The possibility of using phytotherapeutic and high dilutiondrugs during the treatment of nematodedisease has been studied. Two experimental parasitological disease models (aspiculuriosis and trichinellosis) were used.Trichinellosisis a common model of nematodesinmice [1,2].Aspiculuris tetrapteraa pinworm of mice is an important parasite in institutions with mice colonies for both research and teaching purposes. The infection is generally asymptomatic. This study aimed to assess the protective efficiency of a homeopathic drug such as Cina C6cH, sphagnum,and its complex against experimental trichinosis and aspiculuriosis. Methods:An assay was carried out on 80 white outbred mice. These were divided into 6 groups of ten mice each. Groups1 and 4 had sphagnum q.s. per os; groups2 and 5 -desoldering dissolved in water Cina C6cH one time a day per os, the groups3 and 6 received complex sphagnum+Cina C6cHone time a day per os. Groups1-3 were inoculated with a dose of 80 ± 5 units of T. spiralislarva per mouse, groups 4-6 had spontaneous aspiculuriosis, and group 7 had T. spiralis, group 8 had Aspiculuris tetraptera. After 60 days of trichinellosis incubation, process the mice were euthanized and dissected for evaluation.Results:Maximum protection was obtained in the group of mice 3: T. spiralisdetected larvae in animals was 650.5±25.1 larva/animal. Group N2 (Cina C6cH) presented 2840.5±183.3 larva/animal. This was less than the control group (4485±430.6 larva/mouse). Also,groups 4-6 of mice with aspiculuriosis showed 75%, 80%,and 86% efficacy against nematodes.

Captación de ácido siálico por larvas musculares de Trichinella spiralis durante incubación in vitro / Sialic acid capture by Trichinella spiralis muscle larvae during incubation in vitro / Captação de ácido siálico por larvas musculares de Trichinella spiralis durante a incubação in vitro

Se ha comunicado que los eritrocitos (GR) incubados in vitro con larvas infectantes (LM) de T. spiralis presentan mayor agregación que los GR Controles incubados con solución salina, lo que indica que el parásito capta el ácido siálico globular. El objetivo del trabajo fue estudiar la captación de ácido siálico por LM durante incubación in vitro. Se incubaron 30, 60, 90 y 180 larvas con 30 mL del sedimento de GR en 1 mL de solución salina durante 24 y 48 h (37 °C). Se aplicó el Método de Agregación por Polibrene y se calculó Tïtulo, Score Total y CexpCASP. Los resultados mostraron una disminución significativa de estos 3 valores en relación al Control, excepto en el cultivo con 30 LM donde solo se observó un descenso moderado del Score Total y CexpCASP. La captación fue máxima a las 24 h, sin diferencias con los valores obtenidos a las 48 h. Posteriormente se incubaron 60 LM durante 1, 2 y 3 h. Se observó que no hubo captación de ácido siálico en la primera hora de incubación y que fue moderada a las 3 h. La experiencia demostró la captación de ácido siálico globular por las LM in vitro, lo que sugiere que podrían secuestrarlo del miocito con el objeto de evadir y/o interferir la respuesta inmune a los fines de asegurar su permanencia en el hospedador.

It has been reported that erythrocytes (RBC) incubated with infective larvae of T. spiralis (ML) exhibit higher aggregation than Control RBC incubated with saline solution, indicating that the parasite captures erythrocyte sialic acid. The objective of this work was to study sialic acid capture by ML during in vitro incubation. A total of 30, 60, 90 and 180 larvae were incubated with 30 mL of GR sediment in 1 mL of saline solution for 24 to 48 hours (37 °C). Aggregation by Polybrene Method was used, and Titre, Total Score and CexpCASP were calculated. The results showed a significant decrease of these three values compared to Control except in the culture with 30 ML where only a moderate decrease of Total Score and CexpCASP were observed. The capture was maximal at 24 hours, with no difference with the values obtained at 48 hours. Then, 60 ML were incubated for 1, 2 and 3 hours. It was noted that ML did no capture sialic acid in the first hour of incubation and the capture was moderate at 3 hours. The experience showed globular sialic acid capture by ML in vitro, suggesting that they could sequester it from the myocyte in order to evade and/or interfere with the immune response for the purposes of assuring their permanence in the host.

Foi informado que os eritrócitos (GV) incubados in vitro com larvas infetantes (LM) da T. spiralis apresentam maior agregação que os GV Controles incubados com solução salina, indicando que o parasita capta o ácido siálico globular. O objetivo do trabalho foi estudar a captação de ácido siálico por LM durante a incubação in vitro. Foram incubadas 30 , 60 , 90 e 180 larvas com 30 μL do sedimento de GV em 1 mL de solução salina durante 24 a 48 horas (37 °C). Foi aplicado o Método de Agregação por Polibrene e se calculou o Título, Pontuação Total e CexpCASP. Os resultados mostraram uma diminuição significativa desses três valores em relação ao Controle, exceto na cultura com 30 LM, onde foi observada apenas uma redução moderada da Pontuação Total e CexpCASP. A captação foi máxima às 24 horas, sem diferença com os valores obtidos às 48 horas. Posteriormente 60 LM foram incubadas durante 1, 2 e 3 horas. Observou-se que não houve captação alguma de ácido siálico na primeira hora de incubação e que foi moderada às 3 horas. A experiência mostrou captação de ácido siálico globular pelas LM in vitro, sugerindo que poderiam sequestrá-lo do miócito visando a evadir e/ou interferir a resposta imune, para garantir sua permanência no hospedeiro.

Alteración de la agregación eritrocitaria por larvas recién nacidas de Trichinella spiralis / Alteration of the erythrocyte aggregation by newborn larvae of Trichinella spiralis / Alteração do agregação eritrocitária por larvas recém-nascidas da Trichinella spiralis

El objetivo del trabajo fue estudiar las alteraciones en la agregación eritrocitaria producidas por recién nacidas (LRN) de T. spiralis. Se usaron concentrados de larvas LRN incubados en partes iguales con glóbulos rojos (GR) Grupo O (GR Tratados) durante 2 horas, con y sin agitación controlada, tomando muestras al tiempo inicial, 60 y 120 minutos. Los Controles fueron incubados con igual volumen de solución salina. Se aplicó Análisis Digital de Imágenes para estudiar la distribución de los agregados eritrocitarios y calcular el valor de coeficiente de células aisladas (CCA) y la Técnica de Titulación de la Agregación para determinar el Título y el CexpST. Se utilizó ANOVA bifactorial para analizar el efecto de la agitación y del tiempo de incubación en los valores de CCA. Los resultados mostraron que el aumento del tiempo de tratamiento produjo la disminución de las células aisladas y los pequeños rouleaux, y el aumento de los agregados formados por 5 o más glóbulos, lo cual incrementó significativamente el valor de CCA. El análisis estadístico determinó que la agregación de los GR Tratados a los 60 minutos fue mayor que al tiempo 0, y a los 120 minutos mayor que a los otros dos tiempos. La Titulación de la agregación mostró la disminución del CexpST y del Título de los GR Tratados. Las metodologías empleadas no mostraron diferencias significativas en tratamientos con y sin agitación. Se concluye que la disminución de carga eritrocitaria producida por LRN podría provocar alteraciones hemorreológicas en el hospedador.

The aim of this paper was to study the alterations in the erythrocyte aggregation produced by newborn larvae (NL) of T. spiralis. Work was performed with NL concentrates, which were incubated with an equal volume of O Group RBC (Treated RBC) for 2 hours, with and without controlled agitation, taking samples at the initial time, at 60 and 120 minutes. RBC Controls were incubated with equal volume of saline solution. Digital Image Analysis was applied to study the distribution of erythrocyte aggregates and to calculate ICC values, and the Aggregation Titration Technique was used to determine the Title and TSexpC. Two-factor ANOVA was used to analyze the effect of agitation and incubation time on the ICC values. The results showed that at higher treatment time, there was a decrease of isolated cells and small rouleaux and an increase of the aggregates formed by 5 or more cells, with a significant increase of ICC values. Statistical analysis determined that Treated RBC aggregation at 60 minutes was higher than at initial time and that at 120 minutes it was higher than the other two times. Aggregation Titration showed a decrease in the Treated RBC Title and TSexpC. The methodologies employed showed no significant differences in treatments with and without agitation. It is concluded that the decrease in erythrocyte charge produced by NL could cause hemorrheologic alterations in the host.

O objetivo foi estudar as alterações na agregação de eritrócitos produzidas por larvas recém-nascidas (LRN) da T. spiralis. O trabalho foi feito com concentrados de LRN incubados em partes iguais com glóbulos vermelhos (GV) Grupo O (GV Tratados), durante 2 horas, com e sem agitação controlada, levando as amostras ao tempo inicial, 60 e 120 minutos. Os controles foram incubados com igual volume de solução salina. Análise Digital de Imagens foi aplicada para estudar a distribuição dos agregados de eritrócitos e calcular o valor do Quoficiente de Células Isoladas (QCI) e a Técnica de Titulação da Agregação para determinar o Título e CexpST. Para analisar o efeito da agitação e do tempo de incubação sobre os valores do QCI foi utilizada ANOVA bifatorial. Os resultados mostraram que o aumento do tempo de tratamento produziu a diminuição das células isoladas e os pequenos rouleaux, e o aumento dos agregados formados por 5 ou mais glóbulos, aumentando o valor de QCI significativamente. A análise estatística determinou que a agregação de GV Tratados aos 60 minutos foi maior que no tempo inicial e, aos 120 minutos era maior que durante os outros dois tempos. A Titulação da Agregação mostrou a diminuição do CexpST e do Título de GV Tratados. As metodologias usadas não mostraram diferenças significantes em tratamentos com e sem agitação. Conclui-se que a diminuição de carga de eritrócitos produzida por LRN poderia resultar em alterações hemorreológicas no hospedeiro.

Desenmascaramiento del antígeno T por larvas de Trichinella spiralis y Ascaris lumbricoides / T antigen unmasking by Trichinella spiralis and Ascaris lumbricoides larvae / Desmascaramento do antigeno T por larvas de Trichinella spiralis e Ascaris lumbricoid

La activación T es causada por cambios en la estructura de la membrana de los glóbulos rojos (GR), que producen la aglutinación de esas células transformadas con la mayoría de los sueros de adultos ABO compatible. La unión del antígeno T con su anticuerpo específico desencadena poliaglutinación, hemólisis, trombocitopenia y trombosis. El objetivo del trabajo fue estudiar el efecto de larvas de A. lumbricoides y T. spiralis sobre el desenmascaramiento del antígeno T eritrocitario. Se trabajó con 3 concentrados de larvas L1/ L2 de A. lumbricoides (CLAL), y 6 de Larvas Musculares de T. spiralis (LM): CLAL1 y LM1: 450-500 larvas/mL; CLAL2 y LM2: 900-1000 larvas/mL; CLAL3 y LM3: 1800-2000 larvas/mL; LM4: 3000-3500 larvas/mL; LM5: 7500-8000 larvas/mL; LM6: 20.000 larvas/mL. Se utilizaron GR Grupo O en medio enzimático. Los GR se incubaron con igual volumen de CLAL/ LM y los GR Controles con solución fisiológica durante 120 minutos a 37 ºC. Se realizaron Pruebas de Aglutinación en Placa y en Tubo, enfrentando GR Tratados y Controles a sueros de adulto y de cordón. Los resultados mostraron que 2 de las 5 suspensiones de GR Tratados con CLAL3 y 1 de las 5 Tratadas con LM6, aglutinaron con suero de adulto, pero no con suero de cordón. Los GR incubados con los concentrados restantes y los Controles no aglutinaron con ninguno de los sueros. Se concluye que es importante continuar estos estudios para relacionar la activación T con las dosis infectantes en ascariosis y triquinosis, particularmente en patologías que cursen con déficit de ácido siálico.

T activation is caused by changes in the structure of red blood cells (RBC) membrane producing the agglutination of these transformed cells with the majority of adult ABO compatible sera. The union of T antigen with its specific antibody unleashes polyagglutination, haemolysis, thrombocytopenia, and thrombosis. The aim of the present work was to study the effect of A. lumbricoides and T. spiralis larvae on erythrocyte T antigen unmasking. Work was performed on 3 L1/ L2 larvae concentrates of A. lumbricoides (ALLC) and 6 muscle larvae concentrates of T. spiralis (ML): ALLC1 and ML1: 450-500 larvae/ mL; ALLC2 and ML2: 900-1,000 larvae/ mL; ALLC3 and ML3: 1,800-2,000 larvae/ mL; ML4: 3,000-3,500 larvae/ mL; ML5: 7,500-8,000 larvae/ mL; ML6: 20,000 larvae/ mL. Group O RBC in enzymatic medium were used. RBC were incubated with an equal volume of ALLC/ ML and the Controls with physiological solution for 120 minutes at 37 ºC. Plate and Tube Agglutination Tests were made, facing Treated RBC and Controls against adult and cord human sera. The results showed that 2 of the 5 RBC suspensions treated with ALLC3 and 1 of the 5 RBC suspensions treated with LM6 agglutinated with serum from adult, but not cord serum. RBC incubated with the remaining concentrates and Control suspensions were not agglutinated with any of the sera. It can be concluded that it is important to continue these studies to correlate T activation with infective dose in ascariosis and trichinosis, particularly in pathologies that course with sialic acid deficiency.

Ativação T é causada por alterações da estrutura do membrana dos glóbulos vermelhos (GV), que produz a agregação dessas células transformadas com a maioria dos soros de adultos ABO compatível. A União de antígeno T com o anticorpo específico desencadeia poliaglutinação, hemólise, trombocitopenia e trombose. O objetivo do trabalho foi estudar o efeito de larvas da A. lumbricoides e T. spiralis sobre o desmascaramento do antígeno T eritrocitário. Trabalhou-se com 3 concentrados de larvas L1 / L2 da A. lumbricoides (CLAL) e 6 de larvas musculares da T. spiralis (LM): CLAL1 y LM1: 450-500 larvas/mL; CLAL2 LM2: 900-1000 larvas/mL; CLAL3 y LM3: 1800-2000 larvas/mL; LM4: 3000-3500 larvas/mL; LM5: 7500-8000 larvas/mL; LM6: 20.000 larvas/mL. Foram utilizados eritrócitos Grupo O em meio enzimático de bromelina O sedimento globular foi incubado com igual volume de CLAL / LM e o de GR Controles com solução fisiológica durante 120 minutos a 37 ºC. Foram realizados Testes de Aglutinação em Placa e em Tubo, enfrentando os GV Tratados e Controles a soros humanos de adulto e de cordão. Se utilizaron GV Grupo O en medio enzimático. Os GV foram incubados com igual volume de CLAL/ LM e os GV Controles com solução fisiológica durante 120 minutos a 37 ºC. Realizaram-se Provas de Aglutinação em Placa e em Tubo, enfrentando GV Tratados e Controles a soros de adulto e de cordão. Os resultados mostraram que 2 das 5 suspensões de GV Tratadas com CLAL3 e 1 das 5 Tratadas com LM6, aglutinaram com soro de adulto, mas não com soro de cordão. Os GV incubados com as concentrações restantes e os Controles não aglutinaram com nenhum dos soros. Conclui-se que é importante continuar estes estudos para relacionar a ativação T com as doses infectantes em ascaríase e triquinose, particularmente em patologias que cursem com déficit de ácido siálico. controles, não aglutinaram com nenhum dos soros.

Efecto del albendazol en ratas long evans gestantes e infectadas con T. spiralis / Albendazol effect in Long Evans Rats pregnant and infected with T. spiralis

El Albendazol es el antihelmíntico de primera elección para varias parasitosis, entre ellas la Trichinellosis, sin embargo, el tiempo de tratamiento debe ser por periodos prolongados. El objetivo del trabajo fue determinar el efecto de este medicamento sobre ratas Long Evans gestantes e infectadas con T. spiralis. Se sincronizaron las ratas para gestarlas e infectarlas, el tratamiento fue de uno a catorce días, se utilizó la técnica de Hematoxilina-Eosina para evaluar el efecto sobre el hígado y el aparato reproductor, se determino el número de crías por tratamiento y las características de las crías obtenidas, los resultados indican una degeneración de los tejidos conforme aumenta el tiempo de tratamiento además se observa que el numero de crías es afectado directamente al aumentar el tiempo de tratamiento

The Albendazol is the antihelmintic of first election for several parasitism, among them trichinellosis, nevertheless, the time of treatment must be per prolonged periods. The aim of this work was to determine the effect of this medicine on Long rats Evans pregnant and infected with T. spiralis. We synchronize the rats to develop them and to infect them, the treatment went of one to fourteen days, used the Hematoxilina-Eosin technique to verify the effect on the liver and the reproductive apparatus and verified the number of young for each treatment and if correspondence between the obtained characteristics of the reproductive apparatus and young existed, the results indicate a degeneration of weaves as increases the time of treatment in addition is observed that I number of young is directly affected when increasing the treatment

Detección de los cambios fenotípicos en productos de ratas long evans infectadas con trichinella spiralis y tratadas con albendazol / Phenotypic changes detection in new born Long Evans Rat infected with T. spiralis and Albendazol treated

Trichinella spiralis es uno de los agentes causantes de la Trichinellosis, es una zoonosis de distribución mundial, afecta a gran número de mamíferos así como al humano. En México la causa principal de infección se asocia con el consumo de carne de cerdo infectada con T. spiralis. En este trabajo se detectaron los cambios fenotípicos en productos de ratas Long Evans infectas con T. spiralis y tratadas con Albendazol. Utilizamos ratas cepa Long Evans en edad reproductiva. Divididas en grupos, incluidos controles y con tratamiento de uno a catorce días de tratamiento, para sincronizar la gestación en las ratas se realizaron frotis vaginales. Para la detección del parásito se utilizaron técnicas directas de compresión en placa y digestión artificial y la técnica indirecta de MIDD, para los cambios fenotípicos de crías obtenidas se realizo análisis detallado con microscopio estereoscópico

Trichinella spiralis is one of the causative agents of the Trichinellosis, is a zoonosis of world-wide distribution, affects to big number of mammals like this as to the human. In Mexico the main cause of infection associate with the consumption of pig infected meat with T. spiralis. In this work detected the phenotypical changes in long Evans rats products infected with Trichinells spiralis and threated with albendazol medication. Use rats cepa long evans ina reproductive age. Divided in groups, included controls and with treatment varying of one to fourteen days of treatment, to synchronize the management in the rats realized vaginal.smears For the detection of the parasites used direct techniques of understanding in plate and artificial digestion and the indirect technician of MIID, for the phenotypical changes of baby rats obtained realized a detailed analysis with stereoscopic microscope

OBSERVATION OF THE KINETIC CHANGE ON IgG,T-CELL SUBSETS,IL-2 LEVEL OF MICE INFECTED TRICHINELLA SPIRALIS / 中国人兽共患病学报

AimTo explore the kinetic change of the IgG,T-cell subsets and IL-2 level from the mice infected with T. Spiralis. Methods The level of specific IgG A band IL-2 was determined by ELISA,the percentage of CD4+and CD8+ T-cells were examined by flow cytometry on 7, 14,21,28,35 days after mice infected with T. spiralis respectively. ResultAfter infected with T. spiralis,the level of IgG in mice rised gradually,and reached its peak on the 35th day. The change of T-cell subsets showed that CD4+T cells decreased,which CD8+Tcells increased. The ration of CD4+/CD8+cells decreased,and which was the obvious on the 14thday. It did not recover to normal level even on the 35th day. The IL-2 of level reached the peak on the 7th day after infected,then IL-2 level decreased quickly and lower than that of normal mice on 35th day after infected. Conclusion When the acute phase of T. spiralis infection,the immune function of host was inhibited. The protective immunity of against T. spiralis infection was cellular immunity mainly ,in cooperation with humoral immunity.