Acaricidal activity of synthetic spilanthol derivative against ticks of medical and veterinary importance.

The ANESPSAT, a synthetic spilanthol derivative, and its nanoformulation were evaluated against Rhipicephalus microplus and Amblyomma sculptum ticks. ANESPSAT activity was compared with spilanthol and derivatives (ANESPE and others). The compound was synthesized in a gram-scale by a 2-step process, comprising a direct ester amidation and a Horner-Wadsworth- Emmons reaction. The nanoemulsions were produced by coarse homogenization followed by high-energy ultrasonication, in which hydrodynamic diameter, polydispersity index, and zeta potential remained stable. The spilanthol-eugenol hybrid derivatives did not show significant acaricidal activity. ANESPE killed 83% of the R. microplus larvae at 30 mg.mL-1, while ANESPSAT killed 97% at 0.5 mg.mL-1, showing to be the most active compound. Spilanthol and ANESPSAT had similar high mortality rates for tick larvae, with LC50 values of 0.10 and 0.14 mg.mL-1 for R. microplus larvae, and 0.04 and 0.48 mg.mL-1 for A. sculptum larvae, respectively. The efficacy of spilanthol was lower against R. microplus engorged females when compared with ANESPSAT, which was highly effective (>98%) against R. microplus engorged females. The nanoemulsion with ANESPSAT was effective against tick females, preventing egg laying and achieving 100% efficacy at 2.5 mg.mL-1. Spilanthol had only 59% efficacy at 10 mg.mL-1. The results suggest that ANESPSAT, a natural product derivative, could be used in novel formulations for tick management that might be safer and environmentally friendly.

Efficacy of essential oils of Egletes viscosa and Lippia schaueriana on the reproductive biology of Rhipicephalus sanguineus sensu lato engorged females.

The study analyzed the chemical composition and the acaricide effect of Egletes viscosa Less (macela-da-terra) and Lippia schaueriana Mart. (lipia-da-serra) essential oils (EOs) on Rhipicephalus sanguineus s. l. (Acari: Ixodidae) engorged females. The chemical analysis (GC-MS and GC-FID) identified 27 components in E. viscosa EO and 18 in L. schaueriana EO, which comprise more than 98% of its constituents. The effects of the oils on the reproductive biology of R. sanguineus ticks were assessed by adult immersion test. Both EOs significantly reduced (p < 0.05) the egg production index when the females were exposed to 25 and 50 mg/mL, also affecting the egg viability. During the laying process, the eggs produced by the females exposed to the EO showed several morphological alterations such as dehydrated, darkened, and disaggregated, and these alterations were more severe as the concentrations increased. The mortality percentages were 58.9%, 70.8% and 92.7% when the ticks were exposed to 12.5, 25 and 50 mg/mL of E. viscosa oil, respectively. In the same concentrations, the efficacy of L. schaueriana was 39.3%, 53.4%, and 84.6%. Therefore, it can be concluded that the essential oils of E. viscosa and L. schaueriana have acaricidal effect in females of R. sanguineus s.l ticks.

Acaricidal potential of volatile oils from Croton species on Rhipicephalus microplus

ABSTRACT The objective of this study was to evaluate the acaricidal activity of the volatile oils of three species of Croton, Euphorbiaceae, against the cattle tick Rhipicephalus microplus. The volatile oils were obtained by hydrodistillation, analyzed by GC-MS and GC-FID and their acaricidal activity was evaluated by the larval packet test and adult immersion test. The volatile oils from Croton conduplicatus Kunth, Croton pulegiodorus Baill., and two different collections of Croton grewioides Baill. (CG1 and CG2) showed eucalyptol (24.09%), p-cymene (23.13%) and methyl chavicol (83.59% and 95.38%) as the major compounds, respectively. All the volatile oils tested in this study showed efficacy against larvae and engorged females of Rhipicephalus microplus. Therefore, Croton pulegiodorus volatile oil is promising for a potential acaricidal formulation because of the best activity against both stages of the cattle tick.

Effect of Alpinia zerumbet essential oil on the shelf life of tambaqui fillets during short-term refrigerator storage

We investigated the effect of Alpinia zerumbet essential oil on the quality and shelf life of tambaqui (Colossoma macropomum) fillets stored under refrigeration (10.0 ± 0.5 °C) for 14 days. The treatments were A. zerumbet essential oil at 0.75% v v-1 (AEO 0.75%), A. zerumbet essential oil at 1.5% v v-1 (AEO 1.5%) and a control (no essential oil). The sample quality and shelf life were determined by the total psychrotrophic count (TPC) and chemical parameters (pH, total volatile basic nitrogen, centesimal composition and thiobarbituric acid reactive substances - TBARS) at zero, seven and 14 days of storage time. The TPC decreased significantly (p < 0.05) with an A. zerumbetessential oil level of 1.5% until seven days of storage. The concentration of A. zerumbet essential oil at 0.75% resulted in lower pH, TBARS, and TVBN values in comparison with the other treatment and the control. Thus, A. zerumbet essential oil was efficient in extending the shelf life of refrigerated tambaqui fillets up to approximately seven days. (AU)

Morphological alterations in the ovaries of Amblyomma cajennense semi-engorged ticks exposed to ethanolic extract of Acmella oleracea.

The present study analyzed the effects of different concentrations of Acmella oleracea crude ethanolic extract (EEAO) on the development of germ cells from semi-engorged Amblyomma cajennense females in order to evaluate the potential of this natural chemical as a strategy to control these important ectoparasites. A hundred semi-engorged females were divided into five groups (duplicates) (10 animals/group): Control 1 (distilled water); Control 2 (solvent ethanol 50% and DMSO 1%); and Treatment I to III (3.1, 6.2, and 12.5 mg/mL of EEAO, respectively). For the exposure of the ticks to the extract was used the Adult Immersion Test. After the exposition, the ovaries were removed and submitted to histological analysis using Harris hematoxylin and aqueous eosin. The histochemical tests were performed using PAS and Bromophenol blue staining techniques, for the detection of total polysaccharides and total protein, respectively. The extract caused significant alterations in the oocytes, including changes in the shape of the cells, disorganization, and cytoplasmic vacuolation, decrease in the number of yolk granules and germ vesicle fragmentation. These alterations were more intense in the oocytes in initial developmental stages (I and II). The results obtained in this study confirm the cytotoxic potential of the ethanolic extract of A. oleracea on the germ cells of A. cajennense females, opening up the possibility to use this extract as an alternative to control these ectoparasites.

In vitro efficacy of essential oils with different concentrations of 1, 8-cineole against Rhipicephalus (Boophilus) microplus / Eficácia in vitro de óleos essenciais com diferentes concentrações de 1, 8-cineol contra Rhipicephalus (Boophilus) microplus

Abstract The aim of this study was to evaluate the acaricidal activity of essential oils from three species of plants with intermediary concentrations of 1,8-cineole against the tick species Rhipicephalus (Boophilus) microplus. For this purpose, five serial concentrations (100.0, 50.0, 25.0, 12.5, 6.2 mg/mL) of essential oils from Mesosphaerum suaveolens (L.) Kuntze, Ocimum gratissimum L. and Alpinia zerumbet (Pers.) B. L. Burtt & R. M. Sm. were used on larval packet and adult immersion tests. The essential oils were analysed by gas chromatography-mass spectrometry (GC/MS) and gas chromatography-flame ionization detection (GC-FID), being detected 35.8, 24.7 and 24.0% of 1.8-cineol in the oils of M. suaveolens, O. gratissimum and A. zerumbet, respectively. The lethal concentration (LC 50) of each oil for larvae and engorged females was calculated through Probit analysis. All essential oils showed high efficacy (≥ 95.0%) on engorged females at the 100.0 mg/mL concentration. In regards to larvae, O. gratissimum (LC 50 = 11.9 mg/mL) was the most potent, followed by the A. zerumbet (LC50 = 19.7 mg/mL) and the M. suaveolens (LC50 = 51.6 mg/mL) essential oils. These results show that other compounds interfere with 1,8-cineole action.

Resumo O objetivo deste estudo foi avaliar a atividade acaricida de óleos essenciais de três espécies de plantas com concentrações intermediárias de 1,8-cineol contra o carrapato Rhipicephalus (Boophilus) microplus . Dessa forma, cinco concentrações diferentes (100,0; 50,0; 25,0; 12,5; 6,2 mg/mL) de óleos essenciais de Mesosphaerum suaveolens (L.) Kuntze , Ocimum gratissimum L. e Alpinia zerumbet (Pers.) B. L. Burtt & R. M. Sm. foram avaliadas pelos testes de pacote de larvas e de imersão de adultos. Os óleos essenciais foram analisados pela cromatografia gasosa acoplada a espectometria de massa (GC/MS) e cromatografia gasosa acoplada a detector de ionização de chama (GC-FID), sendo detectados 35,8, 24,7 e 24,0% de 1,8-cineol nos óleos de M. suaveolens, O. gratissimum e A. zerumbet , respectivamente. A concentração letal (CL50) de cada óleo essencial para larvas e fêmeas ingurgitadas foi calculada por meio da análise de Probit. Todos os óleos essenciais na concentração de 100,0 mg/mL apresentaram elevada eficácia (≥ 95,0%) sobre fêmeas ingurgitadas. Com relação as larvas, o óleo essencial de O. gratissimum (CL50 = 11,9 mg/mL) foi o mais potente, seguido pelos óleos de A. zerumbet (LC50 = 19,7 mg/mL) e M. suaveolens (LC50 = 51,6 mg/mL). Estes resultados demonstram que outros compostos interferem na eficácia de 1,8-cineol.

In vitro efficacy of essential oils with different concentrations of 1,8-cineole against Rhipicephalus (Boophilus) microplus.

The aim of this study was to evaluate the acaricidal activity of essential oils from three species of plants with intermediary concentrations of 1,8-cineole against the tick species Rhipicephalus (Boophilus) microplus. For this purpose, five serial concentrations (100.0, 50.0, 25.0, 12.5, 6.2 mg/mL) of essential oils from Mesosphaerum suaveolens (L.) Kuntze, Ocimum gratissimum L. and Alpinia zerumbet (Pers.) B. L. Burtt & R. M. Sm. were used on larval packet and adult immersion tests. The essential oils were analysed by gas chromatography-mass spectrometry (GC/MS) and gas chromatography-flame ionization detection (GC-FID), being detected 35.8, 24.7 and 24.0% of 1.8-cineol in the oils of M. suaveolens, O. gratissimum and A. zerumbet, respectively. The lethal concentration (LC 50) of each oil for larvae and engorged females was calculated through Probit analysis. All essential oils showed high efficacy (≥ 95.0%) on engorged females at the 100.0 mg/mL concentration. In regards to larvae, O. gratissimum (LC 50 = 11.9 mg/mL) was the most potent, followed by the A. zerumbet (LC50 = 19.7 mg/mL) and the M. suaveolens (LC50 = 51.6 mg/mL) essential oils. These results show that other compounds interfere with 1,8-cineole action.

Dynamics of natural infection by Babesia bovis and Babesia bigemina in dairy cattle from an enzootic instability area in Northeastern Brazil.

This study aimed to determine the dynamics of natural infection in the transmission of Babesia spp. to cattle in an enzootic instability area in Northeastern Brazil. Blood samples were collected from 30 calves located on two dairy farms to determine the packed cell volume (PCV) and the timing of the primo-infection using polymerase chain reaction (PCR) and their association with climatic factors and management practices. On Farm A, the determination of primo-infection was observed on average at 249.4 (±24.42) days of age for B. bigemina and at 252.6 (±17.07) days of age for B. bovis; there was no significant difference between the times of infection (P> 0.05). The infection coincided with a period of high rainfall in the region. On Farm B, primo-infection infection was not observed. There was no infection by Babesia spp. on Farm B due to the intensive use of acaricides that led to an absence of ticks. There was no significant difference between the average PCV of animals from Farms A and B (P> 0.05). The management practices on the properties, in addition to the weather conditions influenced the exposure of the animals to disease vectors and may have contributed to the maintenance of this enzootic area in Northeastern Brazil.

Dynamics of natural infection by Babesia bovis and Babesia bigemina in dairy cattle from an enzootic instability area in Northeastern Brazil / Dinâmica da infecção natural por Babesia bovis e Babesia bigemina em bovinos leiteiros de uma área de instabilidade enzoótica no Nordeste do Brasil

Abstract This study aimed to determine the dynamics of natural infection in the transmission of Babesia spp. to cattle in an enzootic instability area in Northeastern Brazil. Blood samples were collected from 30 calves located on two dairy farms to determine the packed cell volume (PCV) and the timing of the primo-infection using polymerase chain reaction (PCR) and their association with climatic factors and management practices. On Farm A, the determination of primo-infection was observed on average at 249.4 (±24.42) days of age for B. bigemina and at 252.6 (±17.07) days of age for B. bovis; there was no significant difference between the times of infection (P> 0.05). The infection coincided with a period of high rainfall in the region. On Farm B, primo-infection infection was not observed. There was no infection by Babesia spp. on Farm B due to the intensive use of acaricides that led to an absence of ticks. There was no significant difference between the average PCV of animals from Farms A and B (P> 0.05). The management practices on the properties, in addition to the weather conditions influenced the exposure of the animals to disease vectors and may have contributed to the maintenance of this enzootic area in Northeastern Brazil.

Resumo Este estudo teve como objetivo determinar a dinâmica da infecção natural na transmissão de Babesia spp. em bovinos de uma área de instabilidade enzoótica no Nordeste do Brasil. Foram coletadas amostras de sangue de 30 bezerras, proveniente de duas propriedades leiteiras para determinação do volume globular e da primo-infecção por meio da reação em cadeia da polimerase associando aos fatores climáticos e medidas de manejo. Na fazenda A, o período médio da primo-infecção para B. bigemina, determinado por meio da PCR, foi de 249,4 (±24,42) dias de idade, enquanto que para B. bovis foi aos 252,6 (±17,07) dias de idade, não existindo diferença estatística. A infecção coincidiu com o período de alta precipitação pluviométrica na região. Não houve infecção por Babesia spp. na fazenda B, na qual o uso intensivo de acaricidas determinou ausência de carrapatos. Não houve diferença significativa entre médias de VG dos animais das fazendas A e B. O manejo adotado nas fazendas estudadas, associado às condições climáticas, interferem na exposição dos animais aos vetores, podendo favorecer a manutenção de uma área de instabilidade enzoótica no Nordeste do Brasil.

Toxic action of Acmella oleracea extract on the male reproductive system of Amblyomma cajennense ticks.

The present study evaluated through morphohistological and histochemical techniques the effects of different concentrations of crude ethanolic extract of A. oleracea (EEAO) (Jambu) on the male reproductive system of Amblyomma cajennense sensu stricto (s.s.) ticks. The toxicity of this natural chemical was stablished, signalizing the promising potential of the compound as a strategy to control ectoparasites in the near future. For the experiment, 100 males fed on host rabbits with homogeneous weight (p>0.05) were used. The ticks were divided into five groups (10 animals each): Control 1-exposed to distilled water; Control 2-exposed to ethanol 50% and DMSO 1%; Treatment 1-3-exposed to the concentrations of 6.2, 12.5 and 25mg/mL of the EEAO, respectively, diluted in ethanol 50% and DMSO 1%, with exposure by immersion. After exposure, the males were dissected for the removal of the reproductive system and subjected to routine histological analysis with HE staining and histochemical techniques (PAS for the detection of neutral polysaccharides and Bromophenol blue to detect total proteins). The exposed individuals showed alterations in the glandular complex cells; however, the testes remained intact. The secretory cells of the multilobulated accessory glands presented intense cytoplasmic vacuolation. Additionally, the synthesis and secretion were reduced in the secretion granules, mainly concerning the polysaccharides, glyco- and lipoprotein elements, substances that will constitute the seminal fluid and enable the capacitation of spermatozoa in the female genital tract and also necessary for the formation of the spermatophore, which will encapsulate the mature spermatids. The alterations were dose-dependent, i.e., more intense and severe as the concentration of the product increased. .This experiment confirmed the cytotoxic potential of A. oleracea ethanolic extract in the concentrations of 6.2, 12.5 and 25mg/mL on the reproductive system of A. cajennense s.s. male ticks.

In vitro effects of Pilocarpus microphyllus extracts and pilocarpine hydrochloride on Rhipicephalus (Boophilus) microplus.

The aim of this study was to assess the activity of aqueous (AE) and ethanolic extracts (EE) and pilocarpine hydrochloride, which were extracted and isolated from Pilocarpus microphyllus (Jaborandi), respectively, on Rhipicephalus (Boophilus) microplus. High performance liquid chromatography (HPLC) was performed to quantify these compounds. Larval packet and adult immersion tests were conducted with different concentrations. Five AE and EE concentrations, ranging from 6.2 to 100.0 mg mL-1, and six concentrations of pilocarpine hydrochloride, ranging from 0.7 to 24.0 mg mL-1, were tested. The lethal concentration (LC50) of each extract for larvae and engorged females was calculated through Probit analysis. The concentration of pilocarpine hydrochloride obtained from the EE and the AE was 1.3 and 0.3% (m/m), respectively. Pilocarpine hydrochloride presented the highest acaricidal activity on larvae (LC50 2.6 mg mL-1) and engorged females (LC50 11.8 mg mL-1) of R.(B.) microplus, followed by the EE which presented LC50 of 56.4 and 15.9 mg mL-1, for larvae and engorged females, respectively. Such results indicate that pilocarpine hydrochloride has acaricidal activity, and may be the primary compound responsible for this activity by P. microphyllus EE.

In vitro effects of Pilocarpus microphyllus extracts and pilocarpine hydrochloride on Rhipicephalus (Boophilus) microplus / Efeitos in vitro do extrato de Pilocarpus microphyllus e do cloridrato de pilocarpina sobre Rhipicephalus (Boophilus) microplus

Abstract The aim of this study was to assess the activity of aqueous (AE) and ethanolic extracts (EE) and pilocarpine hydrochloride, which were extracted and isolated from Pilocarpus microphyllus (Jaborandi), respectively, on Rhipicephalus (Boophilus) microplus. High performance liquid chromatography (HPLC) was performed to quantify these compounds. Larval packet and adult immersion tests were conducted with different concentrations. Five AE and EE concentrations, ranging from 6.2 to 100.0 mg mL–1, and six concentrations of pilocarpine hydrochloride, ranging from 0.7 to 24.0 mg mL–1, were tested. The lethal concentration (LC50) of each extract for larvae and engorged females was calculated through Probit analysis. The concentration of pilocarpine hydrochloride obtained from the EE and the AE was 1.3 and 0.3% (m/m), respectively. Pilocarpine hydrochloride presented the highest acaricidal activity on larvae (LC50 2.6 mg mL–1) and engorged females (LC50 11.8 mg mL–1) of R.(B.) microplus, followed by the EE which presented LC50 of 56.4 and 15.9 mg mL–1, for larvae and engorged females, respectively. Such results indicate that pilocarpine hydrochloride has acaricidal activity, and may be the primary compound responsible for this activity by P. microphyllus EE.

Resumo O objetivo desse estudo foi avaliar a atividade dos extratos aquoso (AE) e etanólico (EE) e do cloridrato de pilocarpina, que foram, respectivamente, extraídos e isolado de Pilocarpus microphyllus (Jaborandi), sobre Rhipicephalus (Boophilus) microplus. Cromatografia líquida de alta eficiência foi realizada para quantificação dos compostos. Testes de pacote de larvas e de imersão de adultos foram realizados com diferentes concentrações. Cinco concentrações do AE e EE variando de 6,2 a 100,0 mg mL–1 e seis concentrações do cloridrato de pilocarpina variando de 0,7 a 24,0 mg mL–1 foram testadas. A concentração letal (CL50) de cada extrato para larvas e fêmeas ingurgitadas foi estimada por meio da análise Probit. A concentração de cloridrato de pilocarpina obtida do EE e AE foi de 1,3 e 0,3% (m/m), respectivamente. O cloridrato de pilocarpina apresentou a maior atividade carrapaticida sobre larvas (CL50 2,6 mg mL–1) e fêmeas ingurgitadas (CL50 11,8 mg mL–1) de R. (B.) microplus, seguido do EE que apresentou CL50 de 56,4 e 15,9 mg mL–1, para larvas e fêmeas ingurgitadas, respectivamente. Tais resultados indicam que o cloridrato de pilocarpina apresenta atividade carrapaticida e pode ser o principal responsável pela atividade acaricida do EE de P. microphyllus.

Composição química e eficácia do óleo essencial e do extrato etanólico de Alpinia zerumbet sobre Staphylococcus aureus / Chemical composition and efficacy of essential oil and ethanol extract of Alpinia zerumbet on Staphylococcus aureus

O presente trabalho descreve a atividade antibacteriana do óleo essencial e do extrato etanólico das folhas de Alpinia zerumbet (colônia) sobre cepas de Staphylococcus aureus isoladas de vacas com mastite subclínica e cepas padrão ATCC 29213 e ATCC 25923, por meio do método de difusão em ágar. Foram utilizados 10 tratamentos contendo diferentes concentrações do óleo essencial e do extrato etanólico (100,0; 50,0; 25,0; 12,5 e 6,3 mg.mL-1) e o grupo controle (álcool etílico a 50% e Tween a 1%). Os constituintes majoritários do óleo essencial foram p -cimeno (32,72%), 1,8-cineol (24,05%) e 4-terpineol (20,23%), sendo esses determinados por cromatografia a gás acoplada a espectrometria de massas e cromatografia a gás com detector de ionização de chama (CG-EM/DIC). No extrato etanólico foi detectado o ácido elágico e três flavonoides: rutina, quercetina e campferol, por meio de cromatografia a líquido de alta eficiência acoplada a detector de arranjo de diodo (CLAE-DAD). Todas as cepas apresentaram sensibilidade aos tratamentos com óleo essencial e extrato etanólico. A melhor resposta foi obtida com o óleo essencial de A. zerumbet que, na concentração de 100 mg.mL-1 proporcionou inibição total do crescimento bacteriano. Esses resultados sugerem o potencial antibacteriano do óleo essencial e do extrato etanólico de A. zerumbet no controle da mastite bovina.(AU)

The present work describes the antibacterial activity of the essential oil and the ethanol extract from leaves of Alpinia zerumbet (colônia) on Staphylococcus aureus strains isolated from cows with subclinical mastitis and standard strains ATCC 29213 and ATCC 25923, using the agar diffusion method. Ten treatments containing different concentrations of essential oil and ethanol extract (100.0; 50.0; 25.0; 12.5 and 6.3 mg.mL-1) and the control group (50% ethyl alcohol and 1% Tween solutions) were used for antimicrobial testing. The major constituents of the essential oil were p -cimeno (32.72%), 1.8-cineol (24.05%) and 4-terpineol (20.23%), which were determined by gas chromatographymass spectrometry and gas chromatography - flame ionization detector (CG-MS/FID). Ellagic acid and three flavonoids (rutin, quercetin and campferol) were detected in the ethanol extract by means of high performance liquid chromatography-photodiode array detector (HPLC-PDA). All strains showed sensitivity to the treatments with essential oil and the ethanol extract. The best response was obtained with A. zerumbet essential oil at a 100 mg.mL-1, showing complete inhibition of bacterial growth. These results demonstrate the antibacterial potential of essential oil and ethanol extract of A. zerumbet in the control of bovine mastitis.(AU)