Pharmacopuncture with a low dosage of B complex vitamins in the Da Zhui acupoint (Governing Vessel 14) for controlling gastrointestinal parasitism in periparturient ewes / Farmacopuntura com dose baixa de vitaminas do complexo B no acuponto Da Zhui (Vaso Governador 14) para o controle da parasitose gastrintestinal em ovelhas no periparto

Gastrointestinal parasites are a significant challenge in sheep farming, and periparturient ewes are one of the most susceptible categories in sheep flocks. This study aimed to verify whether the injection of B vitamins at the acupuncture point governing vessel 14 (GV14) can influence the parasitic load or host-parasite interaction in naturally infected ewes during the peripartum period. Four weeks before parturition, 25 ewes were divided into five treatment groups: (1) Dose Control: 0.2 mL of B complex via IM; (2) Drug Control: 2 mL Complex B via IM; (3) Acupoint Control: 0.2 mL of Water at GV14 point; (4) Control; (5) Test group: 0.2 mL of B complex at the GV14 point. Treatments were performed at weeks -3, -1, 2, 4, and 6 about the parturition time. Fecal samples to estimate the parasitic load by fecal egg counts (FEC) and total blood samples for hemogram examination were performed seven days after the treatment, coinciding with weeks -2, 3, 5, and 7 about the parturition time. The number of peripheral eosinophils was higher in group 3 than in group 1 (p <0.05). Although groups 3 and 5 had lower mean values of FEC and higher hematological values at the time of weaning, these differences were not statistically significant (p> 0.05) according to the F test. However, all animals in groups 3 and 5 maintained low parasitism levels until the end of the experiment. Further studies should be performed with larger sample sizes and minor changes to the experimental protocol to elucidate the role of GV14 acupoint stimulation in the host-parasite relationship.(AU)

A parasitose gastrointestinal é o principal desafio sanitário para a criação de ovinos a pasto, e as ovelhas no periparto são uma das categorias mais susceptíveis a esta infeção no rebanho. O objetivo deste trabalho foi verificar se a injeção de vitaminas do complexo B no acuponto Vaso Governador 14 (VG14) poderia influenciar a carga parasitária ou modular a interação hospedeiro-parasito em ovelhas naturalmente infectadas no período de periparto. Quatro semanas antes da data prevista para o parto, 25 ovelhas foram divididas em cinco grupos de tratamento (1) Controle da dose: 0,2 mL de vitaminas do complexo B via IM; (2) Controle do fármaco: 2mL complexo B via IM; (3) Controle do Ponto: 0,2 mL de água destilada no ponto VG14; (4) Controle sem tratamento; (5) Grupo teste: 0,2 mL de complexo B no pontoVG14. Estes tratamentos foram realizados nas semanas -3; -1; 2; 4 e 6 em relação à data do parto. Amostras de fezes para contagem de ovos por grama de fezes (OPG), assim como amostras de sangue para realizar o hemograma, foram obtidas dos animais sete dias após os tratamentos, coincidindo com as semanas -2, 3, 5 e 7 do parto. O número de eosinófilos periféricos foi superior no grupo 3 se comparado ao grupo 1 (p <0,05). Os animais dos grupos 3 e 5 tiveram menores valores de OPG e maior hematócrito, mas estas diferenças não foram consideradas significativas pelo teste F (p> 0,05). No entanto, todas as ovelhas dos grupos 3 e 5 se mantiveram com níveis baixos de parasitismo durante todo o experimento. Sugere-se que novos estudos sejam realizados com algumas modificações deste protocolo a fim de aprofundar o conhecimento do potencial imunomodulador do acuponto VG14.(AU)

Vertical migration of Haemonchus sp. infectant larvae on Stylosanthes spp. and Brachiaria brizantha (Syn. Urochloa brizantha) cv. Marandu forage

The present work aimed to evaluate the influence of Stylosanthes spp. (Campo Grande© styles) and Brachiaria brizantha cv. Marandu in the vertical migration of Haemonchus spp. infective larvae, in the region of Uberlândia, Minas Gerais. Seeds of Stylosanthes spp. and Brachiaria brizantha cv. Marandu were planted in 50 pots of five liters (20 cm diameter) evenly disposed in an area of the Capim Branco Experimental Farm, Federal University of Uberlândia. When both species reached the minimum height of 20 cm, the pots were experimentally contaminated with sheep feces coprocultures. Samplings were performed, with complete removal of the vegetal material contained in the pots, on days one, three, six, 10 and 14 after contamination. Nematoids were recovered from the plant samples by thermo and hydrotropism, counted under an optical microscope and identified. For statistical analysis, the data were analyzed in a completely randomized design (DIC) in a 2X5 factorial scheme (two fodder species X 5 collection dates). No effect of the forage species was observed on the number of L3 of Haemonchus sp. recovered in the feces (p = 0.7790), upper stratum (p = 0.1755) or lower stratum (p = 0.7883) of the forage, however, there was a significant decrease in the number of L3 found with the passing of the days. When comparing the mean L3 recovered in the upper (0.06±0.31 L3) and lower (2.94±0.39) strata, a higher number of larvae was observed in the lower stratum (p < 0.0001). Therefore, in both forage species studied, the upper stratum of the plants, preferably pastured by the animals, remained with less contamination of infective larvae and the contamination tended to decrease over the days.

Economic impact of gastrointestinal nematodes in Morada Nova sheep in Brazil / Impacto econômico de nematódeos gastrintestinais em ovinos Morada Nova no Brasil

Abstract This study evaluated the economic impact of gastrointestinal nematode (GIN) infection in Morada Nova lambs under different parasite chemical control conditions. For this, 246 lambs, in the rainy and dry season, were randomized into groups according to their anthelmintic treatment with levamisole: control (CT: no treatment); routine treatment (RT: treated every 42 days); and targeted selective treatment (TST: treated according to the average daily weight gain, DWG). From 63 days of age (D63) to D210, the lambs were weighed and monitored for GIN infection parameters. Spending on anthelmintics in the production system was 1.3% of the total economic result. The economic result per animal (R$ 5.00 = US$ 1.00) was higher in the RT group, amounting to US$ 6.60 in the rainy and US$ 5.69 in the dry season, due to higher DWG. Thus, RT presented economic results 14.4% and 10.9% higher than CT, and 7.2% and 1.9% higher than TST, in the rainy and dry season, respectively. However, fast development of resistance made RT unfeasible. Here, the economic impact of GIN infection on a national scale is discussed, demonstrating its importance and the impossibility of profitable and sustainable sheep production without adequate control.

Resumo Este estudo avaliou o impacto econômico da infecção por nematoides gastrintestinais (NGI), em cordeiros Morada Nova, sob diferentes condições de controle químico dos parasitas. Para isso, 246 cordeiros, na estação chuvosa e seca, foram randomizados em grupos de acordo com o tratamento com levamisol: controle (TC: sem tratamento); tratamento rotineiro (TR: tratado a cada 42 dias); e tratamento seletivo direcionado (TST: tratado de acordo com o ganho de peso médio diário, GMD). Dos 63 dias de idade (D63) ao D210, os cordeiros foram pesados ​​e monitorados quanto aos parâmetros de infecção por NGI. O gasto com anti-helmínticos no sistema produtivo foi de 1,3% do resultado econômico total. O resultado econômico por animal (R$ 5,00 = US$ 1,00) foi maior no grupo RT, totalizando US$ 6,60 na estação chuvosa e US$ 5,69 na seca, devido ao maior GMD. Assim, o RT apresentou resultados econômicos 14,4% e 10,9% superiores ao TC, e 7,2% e 1,9% superiores ao TST, no período chuvoso e seco, respectivamente. Entretanto o rápido desenvolvimento de resistência inviabiliza o TR. O impacto econômico da infecção por NGI em escala nacional são aqui discutidos, demonstrando sua importância e a impossibilidade de uma ovinocultura lucrativa e sustentável sem o controle adequado.

First Record of Molecular Confirmation, Phylogeny and Haplotype Diversity of Haemonchus contortus from Gaddi (breed) Goats of North India

Abstract: In the present study, molecular identification and genotypic characterization of H. contortus was carried out targeting 28S-18S rRNA intergenic spacer. Faecal samples of Gaddi goats were collected and subjected to qualitative screening. The samples exhibiting the presence of strongyle type eggs were introduced to faecal culturing. The larvae retrieved were molecularly confirmed as of H. contortus species and the phylogenetics was performed. For the estimation of evolutionary divergence in between the present study isolates with the GenBank archived sequences, maximum composite likelihood model was employed. Nucleotide and haplotype diversity indices and Fu's Fs were also estimated. Approximately 260 bp size amplicons retrieved were confirmatory for the presence of H. contortus species. Phylogenetic analysis also accentuated that present parasite isolates were of H. contortus only. The nucleotide diversity (π) obtained was 0.06696, whereas, haplotype diversity was 0.92549 [95% CI: 0.77778-1.0000]. In between the isolates, Fu's Fs statistic value was positive (1.566), evidencing a deficiency of alleles, which would have happened due to recent population bottleneck. The recovered representative sequences were deposited in GenBank under the accession numbers LC600315-LC600317.To the best of our knowledge, the present study is the first report of phylogeny and haplotype diversity of H. contortus isolated from Gaddi goats of North India. The present study would also serve the basis for future detailed molecular epidemiological studies using discriminative markers for the assessment of genetic diversity in different populations of H. contortus in different hosts of the study area.

Eficacia antiparasitaria in vitro del extracto de quebracho (Schinopsis balansae) sobre larvas infectantes de Haemonchus contortus de ovinos / In vitro antiparasitic efficacy of the quebracho extract (Schinopsis balansae) on infecting larvae of Haemonchus contortus of sheep

RESUMEN El objetivo de este trabajo fue evaluar in vitro la eficacia del extracto de quebracho (Schinopsis spp.), rico en taninos condensados, en el control de H. contortus de ovinos, ya que existen evidencias de que estos taninos pueden reducir la excreción de huevos, la fecundidad de las hembras y la carga de parásitos adultos. Para evaluar el efecto antihelmíntico in vitro sobre larvas infectantes de H. contortus susceptibles a todos los grupos químicos, se utilizó el test de inhibición de migración larval (IML) a 3 concentraciones diferentes (5 mg/ml, 15 mg/ml y 30 mg/ml). El efecto de los tratamientos fue analizado mediante un análisis de varianza y la estimación de las diferencias entre grupos se realizó por medio de la prueba LSD Fisher. Los resultados del test in vitro demostraron una reducción de la migración larval que varió entre el 74% y el 80%, a las concentraciones de entre 5 mg/ml y 30 mg/ml. Del análisis de varianza surgen diferencias significativas entre tratamientos (p = 0,0494). Al realizar la prueba de comparación de medias se evidenciaron diferencias significativas (p < 0,05) entre los promedios de migración a las diluciones de 5 mg/ml y 15 mg/ml, y de 5 mg/ml y 30 mg/ml, mientras que no se detectaron diferencias significativas entre la dilución de 15 mg/ml y 30 mg/ml. Estos resultados señalaron que el extracto de quebracho, a las diluciones evaluadas in vitro, presentó actividad antihelmíntica sobre larvas L3 susceptibles de H. contortus. Sin embargo, se requiere ampliar los estudios in vivo para demostrar un efecto antihelmíntico en ovinos.

ABSTRACT The objective of this work was to evaluate in vitro efficacy of the quebracho extract (Schinopsis spp.), rich in condensed tannins, against H. contortus in sheep, since there is evidence that this tanninsthese tannins can reduce egg excretion, fecundity of females and the burden of adult parasites. A larval migration inhibition (IML) test with 3 different concentration (5 mg/ml, 15 mg/ml, and 30 mg/ml) was used to evaluate the in vitro anthelmintic effect upon iInfective H. contortus larvae,from a susceptible strain to all chemical groupswere utilized with 3 diferentconcentration (5mg/ml, 15mg/ml, and 30mg/ml). The effect of the treatments was submitted to a variance analysis and the estimation of the differences between groups was evaluated using LSD Fisher test. Results from the in vitro test, revealed a reduction of the larval migration that varies from 74% to 80%, at the concentrations between 5 mg/ml to 30 mg/ml. From the analysis of variance, significant differences appear between treatments (p = 0,0494). After When performing the mean comparison test were performed, significant differences (p < 0,05) were found between the migration averages at dilutions of 5 mg/ml and 15 mg/ml, and between 5 mg/ml and 30 mg/ml, while were no't detected significant differences between the dilution of 15 mg/ml and 30 mg/ml. These results indicated that quebracho extract at the dilutions evaluated in vitro showed anthelmintic activity on L3 susceptible to H. contortus. However, it is necessary to conduct further studies in vivo to demonstrate an anthelmintic effect in sheep.

Comparative anthelmintic efficacy of Arundo donax, Areca catechu, and Ferula assa-foetida against Haemonchus contortus / Eficácia anthelmíntica comparativa de Arundo donax, Areca catechu e Ferula assa-foetida contra Haemonchus contortus

Abstract In the present study, anthelmintic activities of Arundo (A.) donax L., Areca (Ar.) catechu L., and Ferula (F.) assa-foetida L. were determined. Leaves of A. donax L., latex of F. assa-foetida L. and seeds of Ar. catechu L. in different solvent fractions were subjected to in vitro (egg hatch assay; EHA, and adult motility assay; AMA) and in vivo (faecal egg count reduction test; FECRT) tests of anthelmintic activity using Haemonchus contortus model. In the AMA, crude aqueous methanol extracts (CAME) and ethyl acetate fractions of F. assa-foetida at 10 hr post-treatment showed maximum mortality of H. contortus at 12.5-50 mg mL-1. In the EHA, CAME of F. assa-foetida was identified as a potent ovicide based on its low LC50 (16.9 µg mL-1), followed in order by Ar. catechu and A. donax. Results from the FECRT also showed the extract of F. assa-foetida L. to be more effective than those of Ar. catechu L. and A. donax L., against the gastrointestinal parasitic nematodes. Chloroform and ethyl acetate fractions showed better anthelmintic activities against the adult worms in vitro, while CAME of these plants were better than their crude powders in vivo. It is recommended to document and investigate indigenous knowledge of possible medicinal plants to plan scientific trials that may justify their endorsement.

Resumo No presente estudo, as atividades anti-helmínticas de Arundo (A.) donax L., Areca (Ar.) Catechu L. e Ferula (F.) assa-foetida L. foram determinadas. Folhas de A. donax L., látex de F. assa-foetida L. e sementes de Ar. catechu L. em diferentes frações de solvente foram submetidos a testes in vitro (teste de eclosão de ovos, EHA e ensaio de motilidade em adultos, AMA); e in vivo (teste de redução da contagem de ovos fecais, FECRT) de atividade anti-helmíntica, usando-se Haemonchus contortus. Na AMA, extratos aquosos brutos de metanol (CAME) e frações de acetato de etila de F. assa-foetida. Dez horas pós-tratamento, apresentaram mortalidade máxima de H. contortus em 12,5-50 mg mL-1. No EHA, CAME de F. assa-foetida foi identificado como um ovicida potente baseado em seu baixo LC50 (16,9 µg mL-1), seguido em ordem por Ar. catechu e A. donax. Os resultados do FECRT também mostraram que o extrato de F. assa-foetida L. é mais eficaz do que o de Ar. catechu L. e A. donax L., contra nematoides parasitas gastrointestinais. As frações clorofórmio e acetato de etila mostraram melhores atividades anti-helmínticas contra vermes adultos in vitro, enquanto o CAME dessas plantas foi melhor do que o pó bruto in vivo. Recomenda-se documentar e investigar o conhecimento indígena de possíveis plantas medicinais para planejar ensaios científicos que possam justificar seu endosso.

Cuticular ridge patterns applied for identifying adult female worms of Haemonchus species with various vulvar morphological types in infected native goats in Laos

@#Species of the blood sucking nematode Haemonchus are a main problem in the small ruminant industry worldwide. Haemonchus worms were taken from 68 infected native goats slaughtered in three provinces of Laos in June and July 2019. Cuticular ridge patterns were used for the first time to identify adult female Haemonchus spp. and their vulvar morphs were characterized. The results showed that the variations in vulvar morphology of female Haemonchus spp. presented a knobbed morph as the dominant morphotype and predominant linguiform B subtype was also detected. In total, 270 selected female worms from each vulvar morph type were examined based on their cuticular ridge patterns in cross sections at positions of the esophageal-intestinal junction (EI), the 4 mm region from the anterior end (4 mm), and the mid-body (MB). Only Haemonchus contortus was identified and most worms had constant numbers of ridges at EI, 4 mm, and MB, namely 30, 26, and 22 ridges, respectively, accounting for 99.26%, 97.41%, and 97.04%, respectively, of worms detected, while the lowest variation in the number of ridges was at region EI which is recommended as the single best position. Based on synlophe and ITS2 sequence analysis, it was assumed that H. contortus might dominate in the sample areas with the possible numbers of ridges of H. contortus females in the ranges 29-30, 25-27, and 21-23 for positions EI, 4 mm, and MB, respectively. The cuticular ridge pattern was a useful character for identifying female Haemonchus species in this study and could be utilized as an affordable alternative method for epidemiological studies and as part of parasite control management in native goats of Laos.

Haematological variables of Santa Ines and Ile de France suckling lambs: Influence of Haemonchus contortus infection / Variáveis hematológicas de cordeiros lactentes Santa Inês e Ile de France: influência da infecção por Haemonchus contortus

Haemonchus contortus is the major gastrointestinal parasite of sheep raised in tropical and subtropical areas worldwide. This trial aimed to evaluate the influence of H. contortus infection on the bone marrow response of Santa Ines (SI) and Ile de France (IF) suckling lambs experimentally infected with H. contortus. Fourteen SI lambs and 12 IF lambs were randomized in four groups: infected SI (n=8), non-infected SI (n=6), infected IF (n=8) and non-infected IF (n=4). Lambs of infected groups were submitted to 27 infections, conducted every two days, from 14 to 68 days of age, and each lamb received a total of 5400 H. contortus infective larvae (L3). Ten blood samples were obtained during the experimental period to be used for erythrocyte and leukocyte counts, packed cell volume and total plasma protein estimation. Additionally, it was carried out a differential leukocyte count. Lambs from control groups did not shed eggs in faeces all over the experiment, while infected Santa Ines and Ile de France lambs presented means of 2963 EPG (Eggs Per Gram of faeces) and 8175 EPG in the last sampling (P<0.05), respectively. Infected Santa Ines lambs had an increase in eosinophil release, however differences (P<0.05) on circulation number in comparison with infected Ile de France lambs were identified only in the last sampling (54 days post first infection). The mild H. cortortus infection did not produce significant changes in the blood variables of the Ile de France and Santa Ines suckling lambs.(AU)

Haemonchus contortus é o principal parasita gastrintestinal de ovinos criados em áreas tropicais e subtropicais. Este estudo objetivou avaliar a influência da infecção por H. contortus na resposta medular de cordeiros lactentes das raças Santa Inês (SI) e Ile de France (IF) infectados experimentalmente com H. contortus. Quatorze cordeiros SI e 12 cordeiros IF, foram distribuídos em quatro grupos: SI infectado (n=8), SI não infectado (n=6), IF infectado (n=8) e IF não infectado (n=4). Cordeiros dos grupos infectados foram submetidos a 27 infecções, realizadas a cada dois dias, do 14º até 68º dia de vida, com um total de 5400 larvas infectantes (L3) de H. contortus por animal. Foram realizadas 10 coletas de sangue ao longo do período experimental para determinação do volume globular, proteínas plasmáticas totais, contagem de eritrócitos e leucócitos, além do diferencial de leucócitos. Cordeiros controles não infectados não eliminaram ovos nas fezes durante todo o experimento, enquanto que os cordeiros Santa Ines e Ile de France infectados apresentaram em média 2963 ovos por grama de fezes (OPG) e 8175 OPG na última coleta (P<0,05), respectivamente. Cordeiros Santa Inês infectados tiveram um aumento na produção de eosinófilos, mas diferenças (P<0.05) no número de eosinófilos sanguíneos em comparação a cordeiros Ile de France infectados foram detectadas somente na última coleta (54 dias após a primeira infecção). A infecção leve por H. cortortus não induziu alterações significativas nas variáveis sanguíneas de cordeiros lactentes Ile de France e Santa Inês.(AU)

Hematological, parasitological and biochemical parameters in sheep during the peripartum period / Parámetros hematológicos, parasitológicos y bioquímicos en ovejas durante el periparto / Parâmetros hematológicos, parasitológicos e bioquímicos de ovelhas durante o periparto

Abstract Background: Pregnancy in all animal species requires special care and attention. During this period there is great variation in hematological and biochemical parameters. Many times these parameters are misinterpreted as being obtained from non-pregnant animals with unknown parasite status. Objective: To characterize the hematological, biochemical, and parasitological values within the peripartum period of Santa Ines sheep naturally infected. Methods: For this purpose, 45 ewes were used, with an average body weight of 50 kg. Data were collected during pregnancy, birth and, lactation. Measurements included body weight, body condition score (BCS), evaluation of conjunctive staining, and collection of blood and feces. Results: All variables changed throughout physiological stages. Weight and BCS were different between periods (p<0.05). Fecal egg counts (FEC) were higher in the peripartum period, with a predominance of Haemonchus ssp, 92.85%. Mean hematological and biochemical values varied throughout the study, (p>0.05). Conclusion: The influence of peripartum on the parameters studied should be considered and its use as a reference value. Regarding the physiological variations observed during peripartum, proper animal management could help minimizing the negative effects of parasitic infections on productivity.

Resumen Antecedentes: La preñez en todas las especies animales requiere atención y cuidados especiales. En ese período existe gran variación en los parámetros hematológicos y bioquímicos. Muchas veces estos parámetros son interpretados erróneamente por ser obtenidos de animales no gestantes y con estatus parasitario desconocido. Objetivo: Caracterizar los valores hematológicos, bioquímicos y parasitológicos durante el periparto en ovejas Santa Inés infectadas naturalmente. Métodos: Se utilizaron 45 ovejas, con un peso corporal promedio de 50 kg. la recolección de muestras se realizó durante la gestación, el parto y la lactancia. Se evaluó el peso, la condición corporal, la tinción conjuntiva, y se recolectaron muestras de sangre y heces. Resultados: Todas las variables mostraron variaciones a lo largo de las etapas fisiológicas. La puntuación de peso y condición corporal (BCS) mostró diferencia entre períodos (p<0,05). Los recuentos de huevos fecales (FEC) tuvieron mayores promedios en el periparto, con predominio de Haemonchus ssp, 92,85%. Los valores medios hematológicos y bioquímicos variaron a lo largo del estudio (p<0,05). Conclusiones: Se recomienda considerar la influencia del periparto en los parámetros estudiados y su uso como valor de referencia. Con respecto a las variaciones fisiológicas observadas durante el periparto, el manejo adecuado de los animales podría ayudar a minimizar los efectos negativos de las infecciones parasitarias sobre la productividad.

Resumo Antecedentes: A prenhez em todas as espécies requer atenção e cuidados especiais. Neste período há grande variação dos parâmetros hematológicos e bioquímicos dos animais. Muitas vezes estes parâmetros são mal interpretados na realização de diagnósticos por serem baseados em parâmetros de animais saudáveis não gestantes, desconsiderando também a infecção parasitária. Objetivo: Caracterizar os valores hematológicos, bioquímicos e parasitológicos durante o periparto de ovelhas da raça Santa Inês naturalmente infectadas. Métodos: Foram utilizadas quarenta e cinco ovelhas, com peso corporal médio de 50 kg, os animais foram acompanhados durante a prenhez, parto e lactação. As análises compreendiam: pesagem, condição corporal, Famacha©, sangue e fezes. Resultados: Todas as variáveis apresentaram variações concomitantemente ao avanço da gestação, parto e lactação. O peso (PS) e o escore da condição corporal (ECC) apresentaram diferença estatística entre os períodos (p>0,05). As contagens de ovos por grama de fezes (FEC) apresentaram médias mais elevadas no periparto. Os valores hematológicos e bioquímicos também variaram (p>0,05). Conclusão: Devido ao resultado, sugere-se considerar a influência de periparto sobre os parâmetros estudados e a sua utilização como um valor de referência. Tendo em vista as variações fisiológicas devido o período do periparto e as infecções parasitárias o manejo dos animais deve ter como objetivo minimizar os impactos negativos na produção animal.

In vitro Anthelmintic Activity of Aqueous and Ethanolic Extract of Senna italica (Caesalpiniaceae) on Three-stages of Haemonchus contortus

A phytochemical screening and in vitroanthelminthic activity of aqueous and ethanolic extract of Senna italicaon Haemonchus contortuswere conducted. Polyphenol, tannin and flavonoid contents were determinedby using gallic acid and rutin. egg hatching inhibition test was carried out on fresh eggs; larval mortality test was conducted on infective larvae (L3) and adult worm mortality test was conducted. Eggs, larvae and adults worms were incubated in aqueous and ethanolic extract of S. italicaat different concentrations (0.1; 0.3; 0.5; 0.7 and 1 mg/mL). All extracts of plants showed an effect on all stages of H. contortuswith high efficiency variations depending on the dose used. The inhibition of eggs hatching rate increased from 8.67±1.53% to 65.67±1.15% and from 24.67±1.53% to 80±1.73% respectively for aqueous and ethanolic extract of S. italic.The larval mortality rate increased from 12.22±2.34% to 56.67±4.9% after 24 h and from 45.28±4.11% to 91.25±3.73% after 48 h for aqueous extract and from 30.07±2.84% to 70.83±6.31% after 24 h and from 48.79±3.73% to 96.25±4.79% after 48 h for ethanolic extract. After 24 h of exposure to aqueous and ethanolic extract of S. italica, the adult mortality rate varies from 55.56±9.62% to 83.33±9.62% for aqueous extract and from 61.11±9.62% to 88.89±9.62%.These in vitroresults confirm the use of S. italicain traditional medicine

Anthelmintic activity of nanoencapsulated carvacryl acetate against gastrointestinal nematodes of sheep and its toxicity in rodents / Atividade anti-helmíntica do acetato de carvacrila nanoencapsulado sobre nematoides gastrintestinais de ovinos e toxicidade em roedores

Abstract The objective of this study was to evaluate the efficacy of carvacryl acetate (CVA) and nanoencapsulated CVA (nCVA) on gastrointestinal nematodes of sheep. The CVA was nanoencapsulated with chitosan/gum arabic and the efficacy of nanoencapsulation (EE), yield, zeta potential, nanoparticle morphology and release kinetics at pH 3 and 8 were analyzed. Acute and subchronic toxicity were evaluated in rodents and reduction of egg counts in the faeces (FECRT) of sheep. The sheep were divided into four groups (n = 10): G1, 250 mg/kg CVA; G2, 250 mg/kg nCVA; G3, polymer matrix and G4: 2.5 mg/kg monepantel. EE and nCVA yield were 65% and 57%, respectively. The morphology of the nanoparticles was spherical, size (810.6±286.7 nm), zeta potential in pH 3.2 (+18.3 mV) and the 50% release of CVA at pHs 3 and 8 occurred at 200 and 10 h, respectively. nCVA showed LD50 of 2,609 mg/kg. CVA, nCVA and monepantel reduced the number of eggs per gram of faeces (epg) by 57.7%, 51.1% and 97.7%, respectively. The epg of sheep treated with CVA and nCVA did not differ from the negative control (P>0.05). Nanoencapsulation reduced the toxicity of CVA; however, nCVA and CVA presented similar results in the FECRT.

Resumo O objetivo deste trabalho foi avaliar a eficácia do acetato de carvacrila (ACV) e do ACV nanoencapsulado (nACV) sobre nematóides gastrintestinais de ovinos. O ACV foi nanoencapsulado com quitosana/goma arábica e foi analisada a eficácia de nanoencapsulamento (EE), o rendimento, potencial zeta, morfologia das nanopartículas e cinética de liberação em pH 3 e 8. Foram avaliadas as toxicidades aguda e subcrônica em roedores e a redução da contagem de ovos nas fezes (RCOF) de ovinos. Os ovinos foram divididos em quatro grupos (n = 10): G1, 250 mg/kg ACV; G2, 250 mg/kg de nACV; G3, matriz polimérica e G4: 2,5 mg/kg de monepantel. A EE e o rendimento de nACV foram de 65% e 57%, respectivamente. A morfologia das nanopartículas foi esférica, tamanho (810,6±286,7 nm), potencial zeta no pH 3,2 (+18,3 mV) e a liberação de 50% de CVA nos pHs 3 e 8 ocorreu às 200 e 10 h, respectivamente. nACV apresentou DL50 de 2.609 mg/kg. ACV, nACV e o monepantel reduziram a contagem de ovos por grama de fezes (opg) em 57,7%, 51,1% e 97,7%, respectivamente. A contagem de opg de ovelhas tratadas com ACV e nCVA não diferiu do controle negativo (P>0,05). O nanoencapsulamento reduziu a toxicidade do AVC; no entanto, nACV e ACV apresentaram resultados semelhantes na RCOF.

Castor cake as organic fertilizer to control gastrointestinal nematodes in pasture-raised sheep / Torta de mamona como adubo orgânico para o controle de nematoides gastrintestinais em ovinos criados a pasto

Abstract Gastrointestinal parasitism is one of the factors that discourages farmers from raising small ruminants in cultivated pastures. To validate a soil treatment strategy to control the free-living stages of gastrointestinal nematodes (GIN), castor cake (CC) was used as a fertilizer on a pasture where sheep grazed on guinea grass under continuous stocking. On day zero, the pasture was divided into three paddocks, contaminated by GIN and treated, respectively, with CC divided into two applications (2CC1/2), CC in a single application (CC1) and organic compost in a single application (control). On day 21, eight GIN-free sheep were placed in each paddock. On day 58, significant differences (P<0.05) were observed: reduction of up to 66.10% in larvae.g-1 of dry mass in pastures fertilized with CC, decrease of up to 60.72% in infection rates among the animals in the groups treated with CC, higher average daily weight gain (over 185 g.day-1) and packed cell volume (over 26%) in the groups treated with CC, when compared to the control (128 g.day-1; 20.9%). In view of the results, the use of CC, mainly CC1, as a fertilizer for guinea grass pastures, under continuous stocking, proved to be promising, with 63.41% effectiveness in controlling worm infestations.

Resumo O parasitismo gastrintestinal é um dos fatores que fragiliza a exploração de pequenos ruminantes em pastagens cultivadas. Objetivando validar a estratégia de tratamento do solo para o controle dos estágios de vida livre de nematoides gastrintestinais (NGI), a torta de mamona (TM) foi utilizada como adubo, com ovinos pastejando em capim-tanzânia sob lotação contínua. No dia zero, o pasto foi dividido em três piquetes, contaminados por NGI e tratados, respectivamente, com TM parcelada em duas aplicações (2TM1/2), TM em uma única aplicação (TM1) e composto orgânico em única aplicação (testemunha). No dia 21, cada piquete recebeu oito ovinos livres de NGI. No dia 58, observaram-se diferenças significativas (P<0,05): redução de até 66,10% de larvas.g-1 de massa seca nas pastagens adubadas com TM; redução de até 60,72% da infecção dos animais nos grupos tratados com TM; ganho de peso médio diário (acima de 185 g.dia-1) e volume globular (acima de 26%) superior nos grupos tratados com TM, quando comparados com a testemunha (128 g.dia-1; 20,9%). Diante dos resultados, o uso da TM, principalmente TM1, como adubo em pasto de capim-tanzânia, sob lotação contínua, mostrou-se promissor, com eficácia de 63,41% para controlar a verminose.

Gastrointestinal nematode infections do not hinder the development of Simmental X Nellore crossbred calves raised with a nutritionally enhanced diet / Infecções por nematódeos gastrintestinais não prejudicam o desenvolvimento de bezerros Simental x Nelore criados em plano nutricional elevado

Abstract An evaluation was made of the effect of anthelmintic treatments on the performance of Simmental X Nellore crossbred calves before and after weaning. To this end, the calves were divided into three groups: (1) treated monthly with a low efficacy anthelmintic drug, ivermectin; (2) treated monthly with a highly effective anthelmintic drug, albendazole; and (3) untreated control group. All the groups in this experiment showed an average fecal egg count of less than 400 eggs per gram (EPG), and no clinical signs of parasitic gastroenteritis. The blood variables were within the normal range and no calf presented anemia. In most of the samplings, mean EPGs were significantly lower (P<0.05) in the group treated with albendazole. The calves received dietary supplementation before and after weaning, which enabled them to gain weight in every month of the experiment and reach a body weight of about 250 kg on the last sampling date, before turning one year old. The anthelmintic treatments did not affect body weight gain, leading to the conclusion that, when fed with suitable dietary supplements, Simmental X Nellore crossbred calves are not affected by gastrointestinal nematode parasites acquired by grazing.

Resumo O objetivo do experimento foi avaliar o efeito de tratamentos anti-helmínticos no desempenho de bezerros Simental x Nelore antes e após o desmame. Os bezerros foram alocados em três grupos: (1) tratado mensalmente com anti-helmínticos de baixa eficácia, ivermectina; (2) tratado mensalmente com anti-helmíntico de alta eficácia, albendazol e (3) controle não tratado. A média das contagens de ovos de nematoides durante o experimento foi inferior a 400 ovos por grama (OPG) em todos os grupos sem manifestação clínica de gastroenterite parasitária. As variáveis ​​sanguíneas mantiveram-se dentro dos limites de normalidade e nenhum bezerro apresentou anemia. Na maioria das coletas, as médias de OPG foram significativamente inferiores (P<0,05) no grupo tratado com albendazol. Os bezerros receberam suplementação antes e depois do desmame, o que lhes permitiu ganhar peso em todos os meses do experimento, atingindo peso corporal em torno de 250 kg, ao final do experimento, antes de completarem um ano de idade. Não houve efeito dos tratamentos anti-helmínticos no ganho em peso, o que permitiu concluir que bezerros Nelore x Simental não são afetados pelo parasitismo por nematoides gastrintestinais sob condições de pastejo, quando devidamente suplementados com concentrado.

Anthelmintic activity of Eucalyptus citriodora essential oil and its major component, citronellal, on sheep gastrointestinal nematodes / Atividade anti-helmíntica do óleo essencial de Eucalyptus citriodora e seu componente majoritário, citronelal, sobre nematoides gastrintestinais de ovinos

Abstract This study aimed to evaluate the anthelmintic activity of Eucalyptus citriodora essential oil and citronellal on sheep gastrointestinal nematodes. Essential oil composition was determined by gas chromatography mass spectrometry. The substances were evaluated in vitro using adult worm motility test (AWMT) and transmission electron microscopy (TEM). The acute toxicity test in mice and the fecal egg count reduction test (FECRT) in sheep were performed. Citronellal was confirmed as the essential oil major constituent (63.9%). According to the AWMT, 2 mg/mL of essential oil and citronellal completely inhibited Haemonchus contortus motility at 6 h post exposure. H. contortus exposed to essential oil and citronellal exhibited internal ultrastructural modifications. The lethal dose 50 values in mice were 5,000 and 2,609 mg/kg for essential oil and citronellal, respectively. E. citriodora essential oil reduced sheep epg at 14 days post treatment by 69.5% (P<0.05). No significant differences were observed in epg between the citronellal and negative control groups (P>0.05). The interaction between citronellal and other constituents in the essential oil may be relevant for its in vivo anthelmintic activity. Thus, E. citriodora essential oil and citronellal pharmacokinetic studies may help elucidate the anthelmintic activity of these compounds.

Resumo Este trabalho objetivou avaliar a atividade anti-helmíntica do óleo essencial de Eucalyptus citriodora e citronelal sobre nematoides gastrintestinais de ovinos. A composição do óleo essencial foi determinada por cromatografia gasosa acoplada à espectrometria de massas. As substâncias foram avaliadas in vitro utilizando-se teste de motilidade de vermes adultos (AWMT) e microscopia eletrônica de transmissão (TEM). Teste de toxicidade aguda em camundongos e teste de redução da contagem de ovos fecais (FECRT) em ovinos foram realizados. Citronelal foi confirmado como componente majoritário do óleo essencial (63,9%). No AWMT, 2 mg/mL de óleo essencial e citronelal inibiram completamente a motilidade de H. contortus 6 h pós-exposição. H. contortus expostos ao óleo essencial e citronelal exibiram modificações ultraestruturais internas. Os valores da dose letal 50 em camundongos foram 5.000 e 2.609 mg/kg para óleo essencial e citronelal, respectivamente. Óleo essencial de E. citriodora reduziu opg de ovinos 14 dias pós-tratamento em 69,5% (P<0,05). Não houve diferença significativa de opg entre grupo controle negativo e citronelal (P>0,05). A interação entre citronelal e outros constituintes do óleo essencial pode ser relevante na atividade anti-helmíntica in vivo. Portanto, avaliação farmacocinética do óleo essencial de E. citriodora e citronelal pode auxiliar a elucidar a atividade anti-helmíntica desses compostos.

Losses caused by gastrointestinal nematode infections in Dorper lambs under two nutritional status / Prejuízos causados pelas infecções por nematoides gastrintestinais em cordeiros Dorper submetidos a dois manejos nutricionais

Abstract The aim of this study was to evaluate the effect of two nutritional statuses on the productive performance of Dorper lambs naturally infected with gastrointestinal nematodes. Thirty-two lambs, grazing together on the same pasture, were allocated into four experimental groups: (G1) infected-supplemented diet, (G2) control-supplemented diet, (G3) infected-basal diet, and (G4) control-basal diet. Control animals received suppressive treatment with monepantel every two weeks, while precautionary anthelmintic treatments were given to all lambs of the infected groups with packed cell volume (PCV) <23%. There was reduction in the PCV means of all groups, which was more pronounced in the infected lambs that also presented reduction in total plasma protein values in comparison with the controls. Weight gain was affected by diet and infection status (P < 0.05). Daily body weight gain was 0.170 kg in the G1, 0.205 kg in the G2, 0.085 kg in the G3, and 0.116 kg in the G4. The cold carcass weight was 4.1% and 13.7% higher in controls in comparison with infected lambs, respectively, in the supplemented and basal diets. The infected groups, despite receiving precautionary anthelmintic treatments to prevent deaths due to haemonchosis, presented reduction in the production parameters in comparison with the controls.

Resumo O experimento teve por objetivo determinar o efeito de dois níveis de nutrição no desempenho produtivo de cordeiros Dorper naturalmente infectados por nematoides gastrintestinais. Trinta e dois cordeiros, mantidos juntos na mesma pastagem, foram alocados em quatro grupos experimentais: (G1) infectado-suplementado, (G2) controle-suplementado, (G3) infectado-dieta basal e (G4) controle-dieta basal. Os cordeiros suplementados receberam diariamente concentrado em quantidade equivalente a 2% do peso corporal (PC), enquanto na dieta basal receberam apenas uma pequena quantidade de concentrado (0,35% do PC). Os animais controles receberam tratamento supressivo com anti-helmíntico a cada duas semanas e os infectados foram tratados individualmente quando apresentaram volume globular (VG) <23%. Houve redução nas médias de VG em todos os grupos, as quais foram mais pronunciadas nos animais dos grupos infectados, que também apresentaram redução nos valores de proteína plasmática total em comparação com os controles. Houve efeito significativo da dieta e da infecção no ganho de peso (P <0,05). O ganho em peso diário foi de 0,170 kg no G1, 0,205 kg no G2, 0,085 kg no G3 e 0,116 kg no G4. Os grupos infectados, apesar de receberem tratamentos anti-helmínticos preventivos que evitaram mortes por haemonchose, apresentaram redução nos parâmetros produtivos em comparação com os controles.

Cross Anthelmintic Resistance in Goats of Unorganized Sector in Haryana

The present study was conducted to detect the status of anthelmintic resistance of fenbendazole and morantel against gastrointestinal nematodes in goats of village Badhra, district Charkhi dadri, Haryana. Forty five goats with eggs per gram of more than or equal to 150 were divided into three groups i.e. G1, G2 and G3 of 15 animals each. Group G1 and G2 were treated with fenbendazole @ 10 mg/kg b.wt. orally and morantel @ 20 mg/ kg b.wt. orally, respectively. Group G3 served as untreated control. Faecal egg count of goats was ascertained on day of treatment (0 day) and 12th day post treatment (PT) of all groups by the modified McMaster technique. Pooled faecal cultures were made to recover infective larvae on day 0 and 12 PT. Results revealed that fenbendazole (G1) and morantel (G2) reduced the faecal egg counts by 76.47% and 70.80% on 12th day PT with upper and lower confidence levels as 89.30% and 48.23% and 82.43% and 51.48%, respectively indicating moderate anthelmintic resistance against both drugs. The post-treatment coproculture showed larvae of Haemonchus contortus and Strongyloides sp. Thus, the present study revealed presence of cross anthelmintic resistance against fenbendazole and morantel in goats of unorganized sector in Charkhi Dadri district of Haryana

Cross Anthelmintic Resistance in Haemonchus contortus in Sheep of Unorganized Sector in Haryana

The present study was conducted to detect the status of anthelmintic resistance of commonly used anthelmintic drugs viz. fenbendazole and morantel against gastrointestinal nematodes in sheep of village Badhra, district Charkhi dadri, Haryana. Forty five sheep with egg per gram of more than or equal to 150 were divided into three groups i.e. S1, S2 and S3 of 15 animals each. Group S1 and S2 were treated with fenbendazole @ 5 mg/kg b.wt. orally and morantel @ 10 mg/ kg b.wt. orally, respectively. Group S3 served as untreated control. Faecal samples were collected on the day of treatment (zero day) and 12th day post treatment (PT), from sheep of all groups and individual FEC were determined by the modified McMaster technique. Pooled faecal cultures were made to recover infective larvae on day 0 and 12 PT. Results revealed that fenbendazole (S1) reduced the FEC by 74.77% on 12th day PT with upper and lower confidence levels as 86.41% and 53.15%, respectively indicating moderate anthelmintic resistance. Morantel (S2) caused 49.84% reduction in FEC with upper and lower confidence levels as 65.04% and 28.04%, respectively indicating severe anthelmintic resistance. Thus, the present study revealed presence of cross anthelmintic resistance against fenbendazole and morantel in Haemonchus population in sheep of unorganized sector in Haryana as the post-treatment coproculture showed only Haemonchus contortus larvae

Analysis of genome-wide SNPs based on 2b-RAD sequencing of pooled samples reveals signature of selection in different populations of Haemonchus contortus

The antitumor effect of calycosin has been widely studied, but the targets of calycosin against glioblastomas are stillunclear. In this study we focused on revealing c-Met as a potential target of calycosin suppressing glioblastomas. In thisstudy, suppressed-cell proliferation and cell invasion together with induced-cell apoptosis appeared in calycosin-treatedU251 and U87 cells. Under treatment of calycosin, the mRNA expression levels of Dtk, c-Met, Lyn and PYK2 wereobserved in U87 cells. Meanwhile a western blot assay showed that c-Met together with matrix metalloproteinases-9(MMP9) and phosphorylation of the serine/threonine kinase AKT (p-AKT) was significantly down-regulated by calycosin.Furthermore, overexpressed c-Met in U87 enhanced the expression level of MMP9 and p-AKT and also improved cellinvasion. Additionally, the expression levels of c-Met, MMP9 and p-AKT were inhibited by calycosin in c-Met overexpressed cells. However, an AKT inhibitor (LY294002) only effected on MMP9 and p-AKT, not on c-Met. These datacollectively indicated that calycosin possibility targeting on c-Met and exert an anti-tumor role via MMP9 and AKT.

Anthelmintic effect of Cymbopogon citratus essential oil and its nanoemulsion on sheep gastrointestinal nematodes / Efeito anti-helmíntico do óleo essencial de Cymbopogon citratus e sua nanoemulsão sobre nematoides gastrintestinais de ovinos

Abstract The anthelmintic resistance stimulated the search for strategies for controlling gastrointestinal nematodes, including the use of free essential oils or its nanoemulsion. This study evaluated the anthelmintic efficacy of Cymbopogon citratus essential oil (CcEO) and C. citratus essential oil nanoemulsion (CcEOn). Pysicochemical analyses were performed. The in vitro effect was determined using the egg hatch test (EHT) on Haemonchus contortus and in vivo effect was evaluated in sheep infected with gastrointestinal nematodes. The animals were treated with CcEO (500 mg/kg) or CcEOn (450 mg/kg) for the fecal egg count (FEC) and the determination of worm burden. The main component of CcEO was citral. The CcEO content in the nanoemulsion was 20% (v/v), and the mean particle size was 248 nm. In EHT, CcEO and CcEOn (1.25 mg/mL) inhibited larval hatching by 98.4 and 97.1%, respectively. Three animals treated with CcEO died whereas in the group treated with CcEOn one animal died. The FEC and total worm burden of the treated groups did not differ from the negative control (p>0.05). The CcEOn showed efficacy only on H. contortus (p<0.05). In conclusion, nanoencapsulation reduced toxicity and increased efficacy on H. contortus.

Resumo A resistência anti-helmíntica estimulou a busca por estratégias de controle de nematoides gastrintestinais, incluindo óleos essenciais livres ou em nanoemulsão. Este estudo avaliou a eficácia anti-helmíntica do óleo essencial de Cymbopogon citratus (OECc) e da nanoemulsão do óleo essencial de C. citratus (nOECc). Análises físico-químicas foram realizadas. O efeito in vitro foi determinado no teste de eclosão de ovos (TEO) sobre Haemonchus contortus e o efeito in vivo foi avaliado em ovinos infectados com nematoides gastrintestinais. Os animais foram tratados com OECc (500 mg/kg) ou nOECc (450 mg/kg) para determinação do número de ovos por grama de fezes (OPG) e carga parasitária. O principal constituinte do OECc foi citral. O conteúdo de OECc na nanoemulsão foi 20% e o tamanho médio de partícula foi 248 nm. No TEO, OECc e nOECc (1,25 mg/mL) inibiram 98,4 e 97,1% da eclosão larvar, respectivamente. Três animais tratados com o OECc morreram, enquanto um animal do grupo tratado com a nOECc morreu. O OPG e a carga parasitária total dos grupos tratados não diferiram do controle negativo (p>0.05). A nOECc apresentou eficácia somente sobre H. contortus (p<0.05). Como conclusão, o nanoencapsulamento reduziu a toxicidade e aumentou a eficácia sobre H. contortus.

Crop residues activity against the free-living stages of small ruminant nematodes / Atividade de resíduos de culturas contra os estágios de vida livre de nematoides de pequenos ruminantes

Abstract The nematicidal effect of different organic materials was evaluated in order to develop a non-chemical alternative soil treatment for control of the free-living stages of small ruminant gastrointestinal nematodes. The selected organic materials were residues from the juice industry of acerola, cashew, grape, guava, papaya and pineapple, as well as castor residue from the biodiesel industry. LC90 results showed that pineapple residue was the most efficient inhibitor of larval development, followed by castor, grape, cashew, acerola, guava and papaya. Castor residue was also a good source of nitrogen and was used in a greenhouse experiment to prevent larval development in contaminated goat faeces that was deposited in pots containing the grasses Brachiaria brizantha (var. Paiaguás) or Megathyrsus maximus x M. infestum (var. Massai). Castor residue caused a significant (P < 0.05) reduction (85.04%) in Paiaguás grass contamination (L3.dry mass-1) and a reduction of 17.35% in Massai grass contamination (P > 0.05), with an increase in the biomass production of Massai (251.43%, P < 0.05) and Paiaguás (109.19%, P > 0.05) grasses. This strategy, called Econemat®, with good results in vitro shows to be promising on pasture increasing phytomass production.

Resumo O efeito nematicida de diferentes materiais orgânicos foi avaliado, com o objetivo de desenvolver um tratamento alternativo não químico do solo para o controle dos estágios de vida livre de nematoides gastrintestinais de pequenos ruminantes. Os materiais orgânicos selecionados foram resíduos da indústria de suco de acerola, caju, uva, goiaba, mamão e abacaxi, além do resíduo de mamona da indústria de biodiesel. Os resultados da LC90 mostraram que o resíduo de abacaxi foi o mais eficiente inibidor do desenvolvimento larval, seguido pela mamona, uva, caju, acerola, goiaba e mamão. O resíduo de mamona também se mostrou uma boa fonte de nitrogênio, sendo usado em experimento em casa de vegetação para prevenir o desenvolvimento larval em fezes contaminadas, depositadas em vasos, contendo as gramíneas Brachiaria brizantha (var. Paiaguás) ou Megathyrsus maximus x M. infestum (var. Massai). O resíduo de mamona reduziu (85,04%) significativamente (P < 0,05) a contaminação no capim-Paiaguás (L3.dry massa-1), 17,35% no capim Massai (P> 0,05), e aumentou a produção de biomassa das gramíneas Massai (251,43%, P <0,05) e Paiaguás (109,19%, P> 0,05). Essa estratégia, chamada Econemat®, com bons resultados in vitro, mostra-se promissora nas pastagens aumentando a produção de fitomassa.