Anthelmintic efficacy of an organic fraction from Guazuma ulmifolia leaves and evaluation of reactive oxidative stress on Haemonchus contortus.

This study describes the anthelmintic efficacy of an organic fraction (EtOAc-F) from Guazuma ulmifolia leaves and the evaluation of its reactive oxidative stress on Haemonchus contortus. The first step was to assess the anthelmintic effect of EtOAc-F at 0.0, 3.5, 7.0 and 14 mg kg of body weight (BW) in gerbil's (Meriones unguiculatus) artificially infected with H. contortus infective larvae (L3). The second step was to evaluate the preliminary toxicity after oral administration of the EtOAc-F in gerbils. Finally, the third step was to determine the relative expression of biomarkers such as glutathione (GPx), catalase (CAT), and superoxide dismutase (SOD) against H. contortus L3 post-exposition to EtOAc-F. Additionally, the less-polar compounds of EtOAc-F were identified by gas mass spectrophotometry (GC-MS). The highest anthelmintic efficacy (97.34%) of the organic fraction was found in the gerbils treated with the 14 mg/kg of BW. Histopathological analysis did not reveal changes in tissues. The relative expression reflects overexpression of GPx (p<0.05, fold change: 14.35) and over expression of SOD (p≤0.05, fold change: 0.18) in H. contortus L3 exposed to 97.44 mg/mL of EtOAc-F compared with negative control. The GC-MS analysis revealed the presence of 4-hydroxybenzaldehyde (1), leucoanthocyanidin derivative (2), coniferyl alcohol (3), ferulic acid methyl ester acetate (4), 2,3,4-trimethoxycinnamic acid (5) and epiyangambin (6) as major compounds. According to these results, the EtOAc-F from G. ulmifolia leaves exhibit anthelmintic effect and increased the stress biomarkers on H. contortus.

Effect of an Arthrobotrys musiformis (Fungi: Orbiliales) culture filtrate on the population of gastrointestinal parasitic nematode eggs in faeces of grazing lambs.

This study assessed the anthelmintic activity of the oral administration of a free-spore culture filtrate of the nematophagous fungus (NF) Arthrobotrys musiformis (M-10) on gastrointestinal parasitic nematodes (GIN) in naturally infected lambs. The fungus was grown on potato-dextrose agar plates (PDA) and transferred to a fermented rice medium (FRM). After 40-day incubation the total amount of FRM with the growing fungi was transferred to a flask shaker with distilled water for a 24 h period. The fungus was centrifuged and filtered. Three groups of six naturally-infected lambs (>1000 epg) each were treated once as follows: Group 1) 63.8 mg/kg A. musiformis culture filtrate (CF) (per os); Group 2) Levamisole 7.5 mg/ml (intramuscularly), Group 3) 15 ml of distilled water (per os). Faecal samples were individually collected on days -2, 0, 2, 4, 6, 8 and 10 after treatment. For each experimental group, mean egg shedding was calculated and transformed (log 10 [epg + 1]). Means between the fungal filtrate group and the negative control were analysed using a T-Student Test. The faecal egg count reduction test (FECRT) was performed in groups treated with CF and Levamisole in relation to the control group (water) were 36.8-57.4% and 89-95.4%, respectively., although due to the difference between groups, no statistical significance was found (p > 0.05). The use of A. musiformis CF appears to be a good alternative treatment, although, more studies should be performed to establish the use of these fungal products as potential tools for GIN control.

Haemonchus contortus infection induces a variable immune response in resistant and susceptible Pelibuey sheep.

The immune response and phenotypic characteristics of Pelibuey lambs were analysed after the induction of a Haemonchus contortus trickle infection. Male lambs (n = 29; 20 kg live weight) were infected with 100 H. contortus infective larvae per kg of live weight on day 3, 5 and 7 of the experiment. The number of eggs per gram (epg), seven haematological parameters and the immunoglobulin A (IgA) level were analysed for 56 experimental days. In addition, histopathological samples from the fundic abomasal region and the relative expression of 10 immune-related genes from 15 infected and three non-infected lambs were analysed at day 0 and 49 of the experiment. The epg count and some haematological parameters (leucocytes, red blood cells, haemoglobin and total protein) with statistically significant differences (P < 0.01) were used to identify nine resistant and 20 susceptible lambs (1166 ±â€¯1071 and 3171 ±â€¯1463 epg, respectively). Moreover, acute infiltration of immune cells and parasitic granuloma formation were observed in susceptible lambs; the resistant group had moderate inflammatory cell infiltration. With respect to relative gene expression, resistant lambs showed upregulation (P < 0.001) of 10 genes, from 2.2 to 15.99 fold. Moreover, there was a strong indirect correlation (P < 0.05) between the epg count and interleukin 5 (IL5) gene expression. By contrast, there was an average 0.34 fold downregulation in nine of the immune-related genes (P ≤ 0.05) in susceptible lambs (the only exception was Fc fragment of IgE receptor Ia [FCER1A] upregulation). In addition, there was a direct correlation (P ≤ 0.05) between the epg count and the expression of IL8, which encodes an inflammatory chemokine. In conclusion, this study showed differential IL5 and IL8 gene expression during haemonchosis in resistant and susceptible Pelibuey lambs, respectively, together with a variable immune response based on histopathological and haematological parameters.

Immune and haematological parameters of Blackbelly ewes infected with gastrointestinal nematodes / Parámetros inmunológicos y hematológicos en ovejas Blackbelly infectadas con nematodos gastrointestinales / Parâmetros imunes e hematológicos de ovelhas Blackbelly infectadas por nematoides gastrintestinais

Abstract Background: It is necessary to identify phenotypic traits related to natural resistance against gastrointestinal nematodes (GIN) in order to know the host immunity status in productive ewes. Objetive: To determine haematological and immunological parameters (IgA and IgG) during pregnancy and lactation in Blackbelly ewes naturally infected with GIN Methods: The number of eggs per gram (EPG), packed cell volume (% PCV), plasmatic protein (PP), and peripheral eosinophils were determined during eight months. In addition, sera and saliva samples were collected to establish IgG and IgA kinetics by indirect enzyme-linked immunosorbent assay (ELISA). Results: The results showed 2,592 ± 2,403 EPG and 22.2 ± 4.0% PCV during lactation and 595 ± 901 EPG and 25.1 ± 2.5% PCV during pregnancy. A higher percentage of Trichostrongylus colubriformis larvae were observed in pregnancy (84 to100%) than in lactation (36 to 44%). The IgA activity in serum samples showed a marked reduction (from 80 to 10%) during lambing for both Haemonchus contortus and T. colubriformis antigens. In saliva samples, IgA activity with regard to the standard decreased from 56% at 60 days to 30% at 45 days before lambing and remained low for 45 days during lactation (23 to 32% activity). The eosinophils numbers were 2.0 x 109 cells L-1 in pregnancy and remained low at 0.7 x 109 cells L-1 in lactation. Conclusion: The studied variables reflect the breakdown of immunity against GIN in Blackbelly ewes before and after lambing.

Resumen Antecedentes: La identificación del fenotipo relacionado con la resistencia contra nematodos gastrointestinales (GIN) es necesaria para conocer la inmunidad del huésped en ovejas en producción. Objetivo: Determinar los parámetros hematológicos e inmunológicos (IgA e IgG) en gestación y lactancia en ovejas Blackbelly infectadas naturalmente con GIN. Métodos: Se determinó el número de huevos por gramo de heces (EPG), se identificaron las larvas y se registró el porcentaje del volumen celular aglomerado (% PCV), proteína plasmática (PP) y eosinófilos periféricos durante ocho meses. Además, se colectó suero y saliva para determinar la cinética de IgG e IgA por medio de un ensayo inmuno-enzimático (ELISA) indirecto. Resultados: Los resultados mostraron 2.592 ± 2.403 EPG y 22,2 ± 4,0% PCV durante la lactancia y 595 ± 901 EPG y 25,1 ± 2,5% PCV durante la gestación. Se observó un mayor porcentaje de larvas de Trichostrongylus colubriformis en gestación (84 a 100%) que en lactancia (36 a 44%). La actividad de la IgA en las muestras de suero mostró una marcada reducción después del parto para ambos antígenos de Haemonchus contortus y T. colubriformis (80 a 10%). En saliva, la actividad de la IgA disminuyó de 56 a 30% del día 60 al 45 antes del parto y se mantuvo baja en los primeros 45 días de la lactancia (actividad de 23 a 32%). El número de eosinófilos fue de 2,0 x 109 células L-1 durante la gestación, y se redujo a 0,7 x 109 células L-1 en la lactancia. Conclusión: Las variables estudiadas reflejan la ruptura de la inmunidad contra GIN en ovejas Blackbelly antes y después del parto.

Resumo Antecedentes: A identificação de traços fenotípicos relacionados à resistência natural contra nematóides gastrintestinais (GIN) é necessária para saber a imunidade do hospedeiro em ovelhas produtivas. Objetivo: Determinar parâmetros hematológicos e imunológicos (IgA e IgG) em períodos de gestação e lactação em ovelhas Blackbelly naturalmente infectados com GIN. Métodos: O número de ovos por grama (EPG), volume empacotado de células (% PCV), proteína plasmática (PP) e eosinófilos periféricos foram determinados durante oito meses. Além disso, as amostras de soro e saliva foram recolhidas para determinar a cinética de IgG e IgA por ELISA indireto. Resultados: Os resultados mostraram 2.592 ± 2.403 EPG e 22,2 ± 4,0% PCV durante a lactação e 595 ± 901 EPG e 25,1 ± 2,5% PCV durante a gravidez. A percentagem mais elevada de larvas de Trichostrongylus colubriformis foi observada na gravidez (84 a 100%) do que na lactação (36 a 44%). A atividade de IgA em amostras de soro mostrou uma redução acentuada (80 a 10%) durante o parto nos antígenos de Haemonchus contortus e T. colubriformis. Em amostras de saliva, a atividade de IgA diminuiu de 56 a 30% do dia 60 ao 45 antes do parto e permaneceu baixa por 45 dias durante a lactação (atividade de 23 a 32%). O número de eosinófilos foi de 2,0 x 109 células L-1 na gravidez e manteve-se baixo, com 0,7 x 109 células L-1 na lactação. Conclusaõ: As variáveis estudadas refletem a quebra da imunidade contra GIN em ovelhas Blackbelly antes e depois do parto.

Evaluation of strategic and selective anthelmintic treatments on Pelibuey ewes in Cuba / Evaluación de tratamientos antihelmínticos estratégicos y selectivos en ovejas Pelibuey en Cuba / Avaliação de tratamentos anti-helmínticos estratégicos e seletivos em ovelhas Pelibuey em Cuba

Background: gastrointestinal parasitism is an important limitation for sheep production in Cuba. This situation is worsened by the lack of an effective parasite control strategy. Objective: to assess whether selective and strategic treatments could help achieve a level of control of nematodes similar to systematic drenchings. Methods: an experimental group of 73 Pelibuey ewes was divided into four groups: 1) the Epizoo group (Epizootiological drenching schedule) was dewormed at lambing in the middle of the dry season and at weaning; 2) the FAMACHA© group was treated according to the color of ocular mucosa as indicative of anemia (Categories 4 and 5 of the FAMACHA© color chart); 3) the Chemical group was drenched every three months; and 4) the control group, in which animals remained untreated unless fecal egg count (FEC) was higher than 1500 eggs per gram of feces or the packed cell volume (PCV) was lower than 15%. FECs, body scorings (BCS), hematocrits and color of ocular mucosa (COM) were evaluated monthly. Results: the control group showed the highest FEC and the lowest PCV. We also observed similar levels of FEC and body condition (BCS) in the Epizoo, FAMACHA© and Chemical groups. The PCV in FAMACHA© group was the highest, while no differences were found between the other two groups. Hematocrit had a strong negative correlation (-0.41) with FEC. The FAMACHA© method allowed a dramatic decrease in anthelmintic use. Conclusions: the obtained results suggest that FAMACHA© method could be incorporated as part of an effective parasite control strategy in sheep.

Antecedentes: el parasitismo gastrointestinal constituye una de las principales limitantes para producción de ovejas en Cuba. Esta situación es mucho más grave porque no existe un programa adecuado de control parasitario. Objetivo: evaluar si el uso de un tratamiento antihelmíntico estratégico o selectivo logra reducir los niveles de infestación parasitaria comparado con un sistema de tratamientos químicos sistemáticos. Métodos: se utilizó un rebaño experimental de 73 reproductoras Pelibuey que se dividió aleatoriamente en cuatro grupos: 1) El grupo epizootiológico (Epizoo) se trató al parto, a mediados del periodo seco y al destete, 2) el grupo FAMACHA© se desparasitó en función de la coloración de la mucosa ocular indicativa de anemia (categorías 4 y 5 de la carta de colores FAMACHA©), 3) el grupo Químico: se desparasitó cada tres meses, y 4) grupo control: no recibió tratamiento mientras su hematocrito (PCV) fuera inferior a 15% o la carga parasitaria (FEC) no superara los 1500 huevos por gramo de heces. Con frecuencia mensual se determinó a cada animal el conteo fecal de huevos (FEC), la condición corporal (BCS), el hematocrito (PCV) y la coloración de la mucosa ocular (COM). Resultados: se observaron valores similares para la BCS y el FEC en los grupos Epizoo, FAMACHA© y Químico. El PCV presentó los valores más altos en el grupo FAMACHA© y el grupo Control presentó los más bajos; no se apreciaron diferencias significativas entre los grupos Químico y Epizoo. El grupo control presentó los mayores FEC y los más bajos PCV. El hematocrito tuvo una relación fuerte y negativa (-0,41) con el FEC. El método FAMACHA© propició una disminución en el uso de fármacos antihelmínticos. Conclusiones: los resultados obtenidos sugieren que el método FAMACHA© podría ser incorporado como parte de una estrategia efectiva de control parasitario en ovinos.

Antecedentes: o parasitismo gastrointestinal é uma limitação para a produção de ovinos em Cuba. Esta situação é agravada pela falta de uma estratégia eficaz de controle parasitário. Objetivo: avaliar se os tratamentos seletivos podem ajudar a alcançar um nível de controle de nematóides semelhante a dois tratamentos sistemáticos. Métodos: um rebanho experimental de 73 ovelhas foi dividido em quatro grupos: 1) o grupo Epizoo (Esquema de tratamento epizootiológico) foi tratado no parto, no meio da estação seca e ao desmame, 2) o grupo FAMACHA© foi tratado de acordo com a cor da mucosa ocular (categorias 4 e 5 na cartela de cores FAMACHA©), 3) o grupo químico são vermifugados a cada três meses, e 4) o grupo controle não recebeu tratamento, enquanto seu hematócrito (PCV) fosse inferior a 15% ou a carga parasitária (FEC) não ultrapassasse 1.500 ovos por grama de fezes. A contagem de ovos por grama (FEC) de fezes, o escore da condição corporal (BCS), hematócrito (PCV) e a coloração da mucosa ocular (COM) foram avaliados mensalmente. Resultados: um nível similar de FEC e BCS foram observados nos grupos Epizoo, FAMACHA© y Químico. O PCV apresentou-se maior no grupo FAMACHA© e o grupo Controle teve os valores mais baixos; enquanto nenhuma diferença foi estimada entre os outros dois grupos. O grupo controle apresentou o maior FEC e o menor PVC. O hematócrito teve uma forte correlação negativa (-0,41) com o FEC. O método FAMACHA© permitiu uma diminuição drástica do uso de anti-helmíntico. Conclusões: os resultados sugerem que o método FAMACHA© pode ser incorporado como parte de uma estratégia para o controle de parasitas gastrintestinais em ovelhas.

Variability in the fecal egg count and the parasitic burden of hair sheep after grazing in nematode infected paddocks

This study aimed to evaluate the variability in the fecal egg count (FEC) and the parasitic burden of naive hair sheep after grazing in nematode infected paddocks. The research was carried out in Tabasco, Mexico, during two periods (August and December). In each period 32 lambs were grazed for one month on African star grass (Cynodon plectostachyus) contaminated with gastrointestinal parasitic nematodes. FEC, packed cell volume (PCV) and body weight (BW) were recorded. Gastrointestinal worms were recovered at necropsy. Data were analyzed with the MIXED procedure of SAS using a model of repeated measurements over time. A higher number of Haemonchus contortus worms was found in December (2814±838) than in August (1166±305). The opposite occurred with Cooperia curticei (2167±393 and 3638±441, respectively). The FEC and correlation coefficient in respect to the worm burden were higher in December (6516 ± 1599, r=0.83, respectively) than in August (4364±771, r=0.44, respectively). A high variability in resistance-susceptibility to gastrointestinal nematodes (GIN) occurred in Katahdin × Pelibuey lambs after grazing.

Prevalencia de parásitos gastrointestinales en ovinos sacrificados en un rastro de Tabasco, México / Prevalence of gastrointestinal parasites in slaughtered sheep at a slaughterhouse in Tabasco, Mexico

The objective of this study was to determine the prevalence of gastrointestinal parasitic nematodes in sheep at necropsy at a slaughterhouse in Tabasco, Mexico. Adult parasites from gastrointestinal tract were recovered and preserved in formaldehyde for their later counting and identification. Adult nematodes were separated by species and the final count was transformed to Log + 1 to decrease the variance. GLM procedure using SAS statistic program was performed for data analysis. Provenance, sex, physiological status, and sampling month were considered as the variation sources. From a total sample of 242 slaughtered animals necropsied for monitoring gastrointestinal parasites, 57.4% including Nematoda, Trematoda or Cestoda classes. The main identified species corresponded to Haemonchus contortus in the abomasum. Cooperia curticei, Trichostongylus colubriformis, Strongyloides papillosus and Bunostomum trigonocephalum were found in the small intestine and Oesophagostomum columbianum, and Trichuris ovis in the large intestine. Fasciola hepatica and Moniezia expansa were the Trematoda and Cestoda parasites found in liver and small intestine, respectively with prevalence lower than 7%. The average of the total count of adult nematodes in the gastrointestinal tract of the infested animals was 2175 ± 445. Among the factors studied, the slaughter month and the provenance of the animals affected the prevalence of parasite infestation in sheep at slaughter. The main adult parasites found were H. contortus, C. curticei and T. colubriformis with average counts higher than 1009, 813 and 335, respectively.

El objetivo de esta investigación fue conocer la prevalencia de parásitos en ovinos sacrificados en un rastro en el estado de Tabasco. Se realizó la recuperación y conservación en formol de parásitos adultos presentes en el tracto gastrointestinal, para su posterior conteo e identificación. Los conteos de nematodos adultos por especie se transformaron a Log + 1 para disminuir la varianza, y se realizó el análisis con el procedimiento GLM del SAS en el cual se incluyeron como fuentes de variación: origen, sexo, estado fisiológico y mes de muestreo. De una muestra total de 242 animales sacrificados, se observó que 57.4% se encontraban parasitados con alguna especie de las clases Nematoda, Trematoda o Cestoda. Las principales especies identificadas correspondieron a Haemonchus contortus en abomaso. Cooperia curticei, Trichostongylus colubriformis, Strongyloides papillosus y Bunostomum trigonocephallum en intestino delgado. Oesophagostomum columbianum, y Trichuris ovis en intestino grueso. De los trematodos se encontró Fasciola hepatica en hígado y de los cestodos Moniezia expansa se localizó en intestino delgado con prevalencia menor a 7%. El conteo total de los nematodos adultos en el tracto gastrointestinal de los animales parasitados fue, en promedio, 2175 ± 445. De los factores estudiados, el mes de sacrificio y el origen de los animales afectaron la prevalencia de parasitosis en los ovinos al sacrificio. Los tres principales parásitos fueron: H. contortus, C. curticei y T. colubriformis, con conteos promedio de adultos, superiores a 1009, 813 y 335, respectivamente.