The use of Eucalyptus staigeriana nanoemulsion for control of sheep haemonchosis / Uso da nanoemulsão de Eucalyptus staigeriana no controle da hemoncose em ovinos

Sustainable control of gastrointestinal nematodes (GIN) in small ruminants has been based on the use of alternative methods, including targeted selective treatment, such as FAMACHA. Another GIN control alternative is the use of herbal medicines, although in many cases their use is based on empirical knowledge. Biopolymer nanoformulations has been investigated to maximize the essential oil effects against sheep gastrointestinal nematodes. The aim of the present study was to combine a Eucalyptus staigeriana essential oil nanoemulsion (EsNano) with FAMACHA as an alternative control for sheep haemonchosis. The study was performed over six months at a commercial sheep farm located in a semiarid region of Northeast Brazil. Initially, a fecal egg count reduction test (FECRT) in sheep with levamisole, ivermectin and oxfendazole in sheep was performed used to determine the most effective anthelmintic to use as the positive control. Levamisole has been selected because it showed efficacy superior to 95%. EsNano was obtained and then its physicochemical properties were characterized. The average (±SE) size of the particles in the nanoemulsion was 276.8 (±12.3) nm with bimodal distribution and polydispersity. Nine visits were performed, from April to September 2013, with an interval of 17 days. One hundred sixty-two male and female sheep were divided into three groups (n=54 each) and were treated when FAMACHA score was 3, 4, or 5: G-EsNano 250mg kg-1 EsNano; G-Lev 7.5mg kg-1 levamisole (positive control), and G-Neg was not treated (negative control). Feces from sheep were collected to quantify the number of eggs per gram of feces (epg) and to identify nematode genera. Sheep weight gain was monitored. The epg data for each group and the average sheep weight gains were analyzed by variance analysis and compared with the Tukey's test (P<0.05). Significant difference between the number of animals treated with EsNano and levamisole was not observed in any visit (P>0.05). The epg variation was similar in the G-EsNano and G-Lev groups on visits (P>0.05), except the second and fifth evaluation in the epg groups were significantly different (P<0.05). Haemonchus spp. was the most prevalent nematode. There was no significant weight gain in any of the treated groups (P<0.05). The combination of phytotherapy and FAMACHA can be an alternative to minimize the use of synthetic anthelmintics to control resistant GIN populations of small ruminants.(AU)

O controle sustentável de nematoides gastrintestinais (NGI) em pequenos ruminantes tem sido baseado na utilização de métodos alternativos, incluindo o tratamento alvo-seletivo, tal como o FAMACHA. Outra alternativa de controle de NGI é o uso de plantas medicinais, embora, em muitos casos, a sua utilização seja baseada no conhecimento empírico. Nanoformulações biopoliméricas tem sido investigadas para maximizar os efeitos de óleos essenciais sobre nematoides gastrointestinais em ovinos. O objetivo do presente estudo foi combinar a nanoemulsão do óleo essencial de Eucalyptus staigeriana (EsNano) com o método FAMACHA como uma alternativa para o controle da hemoncose em ovinos. Este estudo foi realizado ao longo de seis meses em uma fazenda comercial de ovinos localizada em uma região semiárida do Nordeste do Brasil. Inicialmente, um teste de redução da contagem de ovos nas fezes (FECRT) em ovinos com levamisol, ivermectina e oxfendazole foi realizado para determinar o anti-helmíntico mais eficaz, para posterior uso como controle positivo. Levamisol foi selecionado porque mostrou eficácia superior a 95%. EsNano foi obtido e, em seguida, as suas propriedades físico-químicas foram caracterizadas. O tamanho médio (±SE) das partículas na nanoemulsão foi 276,8 (±12,3) nm, com distribuição bimodal e polidispersividade. Foram realizadas nove visitas, de abril a setembro de 2013, com um intervalo de 17 dias. Cento e sessenta e dois ovinos machos e fêmeas foram divididos em três grupos (n=54 cada) e foram tratados quando o FAMACHA foi 3, 4 ou 5: G-EsNano 250 mg kg-1 EsNano; G-Lev 7,5mg kg-1 de levamisol (controlo positivo), e G-Neg não foi tratado (controle negativo). As fezes dos ovinos foram coletadas para quantificar o número de ovos por grama de fezes (opg) e identificar os gêneros de nematóides. O ganho de peso de ovinos foi monitorado. Os dados do opg de cada grupo e os ganhos de peso médio dos ovinos foram analisados por análise de variância e comparadas com o teste de Tukey (P<0,05). Diferença significativa entre o número de animais tratados com EsNano e levamisol não foi observada em nenhuma visita (P>0,05). A variação opg foi semelhante para os grupos G-EsNano e G-Lev nas vistas (P>0,05), excetuando a segunda e quinta avaliação em que os opg dos grupos foram significativamente diferentes (P>0,05). O nematoide Haemonchus spp. foi o mais prevalente. Não houve aumento significativo de peso em qualquer um dos grupos tratados (P>0,05). Assim, a combinação de fitoterapia e FAMACHA pode ser uma alternativa para minimizar o uso anti-helmínticos sintéticos para controlar populações resistentes NGI em pequenos ruminantes.(AU)

In vitro activity of Lantana camara, Alpinia zerumbet, Mentha villosa and Tagetes minuta decoctions on Haemonchus contortus eggs and larvae.

The resistance of gastrointestinal nematodes to anthelmintics has increased the need to evaluate natural products that can replace or assist current strategies to control gastrointestinal nematodes. The objective of this study was to evaluate the effect of decoctions of Lantana camara (DLc), Alpinia zerumbet (DAz), Mentha villosa (DMv) and Tagetes minuta (DTm) on Haemonchus contortus by two in vitro tests. The effects of increasing concentrations of lyophilized decoctions (0.31 to 10mg/ml) were assessed using the egg hatch test (EHT). The decoctions were then tested in the larval artificial exsheathment assay. H. contortus third stage larvae (L3) were exposed to 0.31 mg/ml A. zerumbet and M. villosa decoctions and 0.62 mg/ml T. minuta and L. camara decoctions for 3h and then exsheathment procedure at 10 min intervals. An inhibitor of tannins, polyvinyl polypyrrolidone (PVPP), was used to study if tannins were responsible for the inhibitory effect on hatching and exsheathment of larvae. A. zerumbet, M. villosa and T. minuta showed a dose-dependent effect in the EHT, which did not disappear after the addition of PVPP. No effect was observed for L. camara in the EHT. However, the decoctions inhibited the process of larval exsheathment, which may be related to tannin action because the addition of PVPP reversed the inhibitory effect. A. zerumbet, M. villosa and T. minuta decoctions showed inhibitory activity on H. contortus larvae hatching and exsheathing. The decoctions of these plants could be used to control gastrointestinal nematodes following confirmation of their anthelmintic activity in vivo.

Anthelmintic effect of Eucalyptus staigeriana essential oil against goat gastrointestinal nematodes.

The aim of this study was to evaluate the anthelmintic efficacy of Eucalyptus staigeriana essential oil (EsEO). The in vitro effects of EsEO were determined through the egg hatching test and the inhibition of larval development of Haemonchus contortus. The oil was subjected to acute and subacute toxicity rodents. The in vivo effects were determined through fecal egg count reduction test in goats. 1.35 and 5.4 mgml(-1) EsEO inhibited 99.27 and 99.20% H. contortus egg hatching and larval development. In subacute toxicity of EsEO, all parameters were found to be in the normal range, and histopathological analysis of organs did not present alterations. EsEO efficacy against goat gastrointestinal nematodes was 76.57% at 15th day after treatment. E. staigeriana essential oil showed in vitro and in vivo anthelmintic activity.

[Ovicidal and larvicidal activity in vitro of Eucalyptus globulus essential oils on Haemonchus contortus]. / Atividade ovicida e larvicida in vitro do óleo essencial de Eucalyptus globulus sobre Haemonchus contortus.

The objective of this work was to evaluate ovicidal and larvicidal effects of Eucalyptus globulus essential oil (EGEO) on Haemonchus contortus. The chemical composition determination of EGEO was through gas chromatography and mass spectrometry. Egg hatch test (EHT) was performed in concentrations 21.75; 17.4; 8.7; 5.43 e 2.71 mg x mL(-1). In larval development test (LDT) were used the concentrations 43.5; 21.75; 10.87; 5.43 e 2.71 mg x mL(-1). Each trial was conducted by negative control with Tween 80 (3%) and positive control, 0.02 mg x mL(-1) of thiabendazole in EHT and 0.008 mg x mL(-1) of ivermectin in LDT. The maximum effectiveness of EGEO on eggs was 99.3% in concentration of 21.75 mg x mL(-1) and on larvae was 98.7% in concentration 43.5 mg x mL(-1). The concentration of EGEO that inhibits 50% of the eggs and larvae was 8.3 and 6.92 mg x mL(-1), respectively. The oil chemical analysis identified as main component the monoterpen 1,8-cineol. EGEO presented ovicidal and larvicidal activities in vitro, revealing a good potential for use in the control of sheep and goat gastrointestinal nematodes.

Atividade ovicida e larvicida in vitro do óleo essencial de Eucalyptus globulus sobre Haemonchus contortus / Ovicidal and larvicidal activity in vitro of Eucalyptus globulus essential oils on Haemonchus contortus

O objetivo deste trabalho foi avaliar o efeito do óleo essencial de Eucalyptus globulus (OEEG) sobre a eclosão e desenvolvimento de larvas de Haemonchus contortus. A determinação da composição química do OEEG foi feita por meio de cromatografia gasosa acoplada à espectrometria de massa. No teste de eclosão de ovos (TEO), utilizaram-se as concentrações 21,75; 17,4; 8,7; 5,43 e 2,71 mg.mL-1; e no de desenvolvimento larvar (TDL) as concentrações foram 43,5; 21,75; 10,87; 5,43 e 2,71 mg.mL-1. Cada ensaio foi acompanhado por um controle positivo, 0,02 mg.mL-1 tiabendazol, no TEO e 0,008 mg.mL-1 ivermectina no TDL, e por um controle negativo com o diluente Tween 80 (3 por cento). A eficácia máxima obtida pelo OEEG sobre ovos foi de 99,3 por cento na concentração de 21,75 mg.mL-1 e, sobre larvas, 98,7 por cento na concentração de 43,5 mg.mL-1. A concentração do OEEG, que inviabilizou 50 por cento dos ovos e das larvas do parasito, foi de 8,3 e 6,92 mg.mL-1, respectivamente. A análise química do óleo identificou como principal componente o monoterpeno 1,8-cineol. O OEEG apresentou atividade ovicida e larvicida in vitro sobre H. contortus, determinando um bom potencial para utilização no controle de nematóides gastrintestinais de ovinos e caprinos.

The objective of this work was to evaluate ovicidal and larvicidal effects of Eucalyptus globulus essential oil (EGEO) on Haemonchus contortus. The chemical composition determination of EGEO was through gas chromatography and mass spectrometry. Egg hatch test (EHT) was performed in concentrations 21.75; 17.4; 8.7; 5.43 e 2.71 mg.mL-1. In larval development test (LDT) were used the concentrations 43.5; 21.75; 10.87; 5.43 e 2.71 mg.mL-1. Each trial was conducted by negative control with Tween 80 (3 percent) and positive control, 0.02 mg.mL -1 of thiabendazole in EHT and 0.008 mg.mL-1 of ivermectin in LDT. The maximum effectiveness of EGEO on eggs was 99.3 percent in concentration of 21.75 mg.mL-1 and on larvae was 98.7 percent in concentration 43.5 mg.mL-1. The concentration of EGEO that inhibits 50 percent of the eggs and larvae was 8.3 and 6.92 mg.mL-1, respectively. The oil chemical analysis identified as main component the monoterpen 1,8-cineol. EGEO presented ovicidal and larvicidal activities in vitro, revealing a good potential for use in the control of sheep and goat gastrointestinal nematodes.