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Article

In Vitro Activity of Ethanolic Extract and Essential Oil of Achyrocline satureioides Against Larvae of the Tick Rhipicephalus sanguineus

by
Rafaela Regina Fantatto
,
Flávio Augusto Sanches Politi
,
Rodrigo Sorrechia
and
Rosemeire Cristina Linhari Rodrigues Pietro
*
Departament of Drugs and Medicines, São Paulo State University UNESP, Rodovia Araraquara-Jaú Km 1, Araraquara 14800-903, SP, Brazil
*
Author to whom correspondence should be addressed.
Parasitologia 2025, 5(4), 60; https://doi.org/10.3390/parasitologia5040060
Submission received: 1 July 2025 / Revised: 25 September 2025 / Accepted: 20 October 2025 / Published: 7 November 2025
(This article belongs to the Special Issue Exploring Natural Products as Antiparasitic Agents: Efficacy Against Parasites of Veterinary and Public Health Significance)
Versions Notes

Abstract

The tick Rhipicephalus sanguineus is the most prevalent ectoparasite in dogs, causing discomfort to the animals and acting as a vector for several pathogens, including the bacterium Ehrlichia canis and the protozoa Babesia canis, Babesia gibsoni, and Hepatozoon canis. Control of this parasite is traditionally carried out with synthetic chemical acaricides. However, due to the increasing number of cases of resistance, phytotherapy has been increasingly investigated as a promising alternative. In this study, the larvicidal activity of the crude ethanolic extract and essential oil obtained from the inflorescences of Achyrocline satureioides was evaluated, whose constituents were identified through phytochemical analyses and gas chromatography. The analyses revealed that the extract is rich in flavonoids, tannins, and saponins, while the essential oil is composed mainly of terpenes. In contact tests with impregnated paper, the extract at 100 mg/mL showed a mortality rate of 32.2% in R. sanguineus larvae with LC50 calculated at 249.62 mg/m., while the essential oil, at the same concentration, resulted in 56.55% mortality, and the LC50 and LC90 were 119.73 mg/mL and 185.53 mg/mL, respectively. These results indicate that the essential oil of A. satureioides has significant larvicidal activity and has potential for use as an alternative, alone or in combination with other extracts or synthetic acaricides.

1. Introduction

The tick Rhipicephalus sanguineus is considered the most important and incident ectoparasite in dogs worldwide [1], and, in light of this, it has attracted great attention from the veterinary pharmaceutical industry, which is dedicated to finding new molecules and ways to control this parasite. Several products and formulations designed for its control can be found commercially, representing a considerable portion of the revenue in the veterinary line of pharmaceutical industries. In addition to the direct damage caused by parasitism, such as anemia, discomfort, and lesions, it is also a vector of pathogens such as the bacterium Ehrlichia canis and the protozoa Babesia canis, Babesia gibsoni, and Hepatozoon canis [2,3,4]. Furthermore, it may be involved in the transmission of Rickettsiae to humans in Europe, especially Rickettsia conorii [5], and also in Mexico and the United States, it has been cited as a vector of Rickettsia rickettsii, the etiological agent of Rocky Mountain spotted fever [6,7]. Some studies also suggest the possibility of its participation in the transmission of the causative agent of canine visceral leishmaniasis, although this association has not been scientifically proven [8]. Effective management of Rhipicephalus sanguineus s.l. involves multiple control methods but primarily relies on the use of chemical acaricides for both on-host and off-host treatments [9]. Permethrin and fipronil are two common active ingredients found in various commercial products targeting ticks [10,11]. However, studies by Miller et al. (2001) [12] and Estrada-Peña (2005) [13] have reported resistance to all tested products, with the exception of fipronil and amitraz. Due to the high incidence of resistance, phytotherapy has been investigated and considered a potential alternative for tick control. The use of natural compounds containing bioactives (active chemical substances) from plant extracts, with the potential to control different pests, presents itself as a possible alternative to chemical acaricides [14]. Essential oils (EOs) and ethanolic extracts extracted from different parts of plants present themselves as a source of environmentally friendly and effective insecticide/acaricide alternatives to synthetic chemical products [15]. The species Achyrocline satureioides, popularly known as macela and considered an invasive plant due to its ability to grow in vacant lots and abandoned pastures, has several uses for its healing properties and is widely used in folk medicine in countries where it is native, such as Brazil, Argentina, Uruguay, and Paraguay [16]. Studies carried out by Dal Magro et al., 1998 [17], demonstrated that this same species has several activities reported as a repellent to Simulids (rubber), where the flower extract showed 98.5% efficiency. Rojas de Arias et al. (1995) [18] show trypanocidal (Trypanossoma cruzi) and insecticidal (Triatoma infestans) as effective, and previous studies carried out by Fantatto et al., 2022 [19], demonstrated the potential of the essential oil and ethanolic extract of this species against the cattle tick Rhipicephalus microplus. Thus, the species A. satureioides in this study was the target of acaricidal activity tests against larvae of the tick R. sanguineus with the objective of controlling this parasite.

2. Materials and Methods

2.1. Obtaining Plant Material

Inflorescences of A. satureioides were obtained through an agreement signed with the Pluridisciplinary Center for Chemical, Biological and Agricultural Research (CPQBA) of the University of Campinas, Brazil under the geographical coordinates 22°48′ S; 47°07′ W, climate (Koeppen) Cwa: tropical with humid summers and dry winters, typical red soil and deposited under voucher number 308. The species A. satureioides was registered and has authorization for use by SISGEN (National System for the Management of Genetic Heritage and Associated Traditional Knowledge) under registration number AD73F75. Before use, the plant was preserved in a dry and ventilated place with no signs of pests.

2.2. Preparation of the Ethanolic Extract

Ethanol extracts were prepared from the inflorescences using the maceration method. A total of 100 g of inflorescences obtained from the aerial parts were crushed in a knife mill and the powder was placed in an amber bottle, where 2 L of pure ethanol was added. The pharmacogen remained in contact with the solvent for seven days; afterwards, the solution was filtered and concentrated on a rotary evaporator at reduced pressure and totally dried in a chemical hood. The extracts were also prepared in the same way but using 50% ethanol. Then, the extracts were adequately solubilized to achieve the desired concentrations in the tests.

2.3. Essential Oil Extraction

The essential oil was obtained through the hydrodistillation technique using a modified Clevenger-type device (Unividros®, Ribeirão Preto, Brazil) with heating blanket with temperature regulator Q321A28 (Quimis®, Diadema, Brazil) [15]). At each extraction, 100 g of inflorescences was placed in a round-bottomed flask with 500 mL of water, which remained in contact with the heating blanket and coupled to the Clevenger apparatus for about 4 h. The extracted oil was collected using ethyl ether to completely remove the oil from the device. The recovered content was dried with anhydrous sodium sulfate to separate the oil from the residual water. The extracted oil, whose yield was 1% (% v/w, mL of oil per 100 g of plant material), was kept in a sealed glass bottle under refrigeration.

2.4. Preliminary Phytochemical Screening

Preliminary phytochemical screening of crude drug powder was performed as per the standard procedure described by Harbone (1998) [20] for various phyto-constituents such as steroids, terpenoids, alkaloids, tannins, phenolic compounds, flavonoids, carbohydrates, and amino acids.

2.5. Gas Chromatography–Mass Spectrometry (GC-MS) Analysis

GC-MS analyses were performed on the Shimadzu QP-2010 gas chromatograph (Shimadzu Corporation, Kyoto, Japan) equipped with an automatic AOC-5000 Shimadzu injector (Shimadzu Corporation, Kyoto, Japan) and interface with a mass spectrometer. The column used was a Phenomenex ZB-5MS (30 m × 0.25 × 0.25 mm; Phenomenex Inc., Torrance, CA, EUA), GCMS solutions Ver. 2.5. The GC-MS analysis was carried out with the following oven program: 140 °C held for 3 min, ramped at 3 °C/min to 32 °C, and held for 10 min. The injector temperature was 260 °C. Injection mode: splitless/split. Splitless start, 0.75 min split (1/50), 2 min split 1/20, helium carrier gas (99.999%) at a constant flow of 1.3 mL/min, and the sample volume injected was 1 μL. Pressure: 114.9 kPa, linear speed: 43.1 cm/sec, and total flow: 17.3. MS conditions: ion source temperature and interface of 250 °C, electron impact mode at 70 eV, and range of acquisition masses of m/z 40–650 Daltons.

Data Processing and Analysis

The chromatogram of the GC-MS analysis was integrated and the Rt (retention time) and peaked areas tabulated. The calculation of the linear retention index was calculated using a homologous series of n-alkanes C8–C40 (Sigma-Aldrich, St. Louis, MO, USA). The retention index of an analyte is a number, obtained by interpolation, relating the retention time of the analyte of interest with the retention time of two standards (homologous series of hydrocarbons) eluted before and after the peak of the compound of interest, according to Equation (1):
RI = 100 × n + 100 Rtx − Rtn
Rtn + 1 − Rtn
where IR is the retention index, n is the number of carbons of the n-alkane prior to the analyte, tRx is the retention time of the analyte of interest, tRn is the retention time of the n-alkane eluted before the analyte, and tRn + 1 is the retention time of the n-alkane eluted after the analyte. The CG-MS retention indexes were compared with the National Institute of Standards and Technology (NIST) [21] database and Golm Metabolome Database (Max Planck Institute). The retention index is the number obtained by interpolating the retention time of the component of interest with the retention times of two patterns of the homologous series of alkanes eluted immediately before and after the component of interest. Retention rates were compared with the NIST database.

2.6. Mortality of Larvae in Patch Test on Impregnated Paper

The sensitivity tests of R. sanguineus larvae were performed according to the technique developed by FAO [22]. Rhipicephalus sanguineus larvae were used for the tests at 15 days post-hatching. At this stage, the larvae exhibit full motility and a consistent response to stimuli, ensuring the reliability of the acaricidal tests. Larvae used in this study were provided through a research partnership with Embrapa Pecuária Sudeste, São Carlos, São Paulo, Brazil. In this methodology, approximately 100 larvae of the species were placed on filter paper measuring approximately 10 × 8 cm and impregnated with 0.3 mL of the different concentrations of extract, ranging from 100 mg/mL to 0.78 mg/mL, always falling in half. The test also included controls on the solvents used (ethanol in the case of the extract and Tween® 80 in the case of oil). Each of these papers impregnated with the extracts was folded to form a “sandwich” and sealed with plastic pegs. The envelopes were placed in an oven at 27 °C and 80% relative humidity, and the triplicates were read after 24 h of incubation with the help of a vacuum compressor adapted with a pipette differentiating live and dead larvae.

2.7. Statistical Analysis

The results were expressed as mean ± standard deviation, calculated in Microsoft Excel (Office 2013), and presented in tables. Statistical analyses were performed using analysis of variance (ANOVA), with comparison of means using the Tukey–Kramer test (p < 0.05) using Assistat software version 7.6 beta. To estimate the 50% and 90% lethal concentrations (LC50 and LC90) of samples as well as their respective confidence intervals, the Probit procedure of the Statistical Analysis System (SAS) software (version 2002/2003) was used.

3. Results

3.1. Preliminary Phytochemical Screening Results

Preliminary phytochemical screening of inflorescences of the species A. satureioides revealed the presence of flavonoids, tannins, and saponins, as can be seen in Table 1.

3.2. CG-EM Analysis

A. satureioides essential oil showed 122 peaks, among which 58 substances have been identified based on the largest peaks and retention times in minutes, which are basically terpenes such as α pinene, β mycrene, d-limonene, eucalyptol, β ocimene, verbenol, borneol, terpineol, limonene oxide, α muurelene, α gurgugene, aromadendrene, humulene, patchoulene, azulene, α-campholene aldehyde, trans carveol, caryono, nonyne, copaeno, α copalleno, α guraryene, β germene, humulene, azulene, cyclodecadiene, delta cadinene, patchouli, cis limonene oxide, nerodiol, and cedrol. These results are summarized in Table 2.

3.3. Mortality of Larvae in Patch Test on Impregnated PaperResults

The contact test on paper impregnated with the essential oil and ethanolic extract of A. satureioides was performed with the species R. sanguineus, popularly known as dog tick (Table 3).

4. Discussion

Due to the high pathogenicity and prevalence of tick Rhipicephalus sanguineus, the search for safer and more sustainable control methods has intensified. The phenomenon of resistance in ticks represents a global problem that threatens both the livestock economy and health of humans and animals, as it hinders the effective control and prevention of ectoparasites and the diseases they transmit. Due to their curious and exploratory behavior, dogs are particularly susceptible to tick infestation, becoming hosts for specific or all developmental stages of these parasites present in the environment [23,24]. Therefore, as dog ownership increases globally, so does the need to protect animals and their owners from these troublesome parasites that pose health risks [25].
Botanical repellents have gained prominence due to their low toxicity and reduced environmental impact. Plants, commonly found in nature, produce a variety of primary and secondary metabolites. Primary metabolites, such as carbohydrates, amino acids, and lipids, are essential for basic metabolism, while secondary metabolites such as essential oils, flavonoids, terpenoids, alkaloids, and tannins are often associated with plant defense and can exhibit medicinal or toxic effects [26]. Consequently, essential oils and plant extracts, rich in these compounds, are considered ecological alternatives to synthetic acaricides [27,28].
In light of this, the present study aimed to produce, characterize, and evaluate the ethanolic extract and essential oil of A. satureioides (macela) against R. sanguineus larvae through in vitro tests. For characterization of the ethanolic extract, phytochemical prospecting was performed to identify the classes of secondary metabolites present in the species. As an alternative to chromatography, which can be unfeasible for small producers or remote laboratories, preliminary phytochemical screening was conducted using colorimetric reactions. The inflorescences of A. satureioides revealed the presence of flavonoids, tannins, and saponins, corroborating results reported by Marques-Junior et al. (2009) [29] in commercial and household samples from the city of Uruguaiana, RS, Brazil. Despite the ease and low cost of colorimetric methods, they are not recommended for essential oil analysis, which requires more robust techniques such as gas chromatography.
Liquid chromatography analysis of the essential oil of A. satureioides revealed 122 peaks, with 58 compounds identified based on the retention time and main peaks. The predominant compounds were terpenes, including α-pinene, β-myrcene, d-limonene, eucalyptol, verbenol, borneol, terpineol, humulene, patchoulene, cedrol, and others. Similar findings have been reported by Sierra et al. (2015) [30] in Colombia and Lorenzi et al. (2008) [31], with variations attributed to factors such as analysis timing, storage, or cultivation conditions. Terpenes, widely used in the cosmetic and food industries, have also attracted pharmaceutical interest due to their biological properties, including antimicrobial, antifungal, antiviral, anti-inflammatory, and antiparasitic activities (Paduch et al., 2007) [32].
In the present study, in vitro tests with R. sanguineus larvae showed that the ethanolic extract, initially tested with 2% Tween, did not exhibit significant activity. However, when solubilized in 50% ethanol, the extract demonstrated larvicidal effects, with the highest efficacy at 100 mg/mL (32.21%) and an LC50 of 249.62 mg/mL. The essential oil solubilized in 2% Tween induced significant mortality at the three highest concentrations tested (100, 50, and 25 mg/mL), with the highest mortality (56.55%) observed at 100 mg/mL. The LC50 and LC90 were 119.73 mg/mL and 185.53 mg/mL, respectively.
Previous studies on the same tick species using another Asteraceae plant, Tagetes patula, demonstrated that the 70% ethanolic extract of the aerial parts exhibited larval LC50 and LC95 values of 7.43 mg/mL and 49.4 mg/mL, respectively (Politi et al., 2012) [33]. For essential oils, T. patula oil induced 100% mortality at a 20% concentration in R. sanguineus larvae (da Silva et al., 2016) [34]. A study by Fantatto et al. (2024) [35], using the ethanolic extract of A. satureioides in formulation form, demonstrated that acaricidal effects against R. microplus are enhanced when the extract is incorporated into a hydrogel. At the highest concentration evaluated (20 mg/mL), larval mortality reached 91.48%, while even at half that concentration (10 mg/mL), mortality remained high at 78.13%. The lowest concentrations tested exhibited mortality rates of 12.94% and 20.81%, respectively, relative to the highest concentration. These findings highlight the significant acaricidal potential of A. satureioides, its extracts, and essential oil. To date, no study has investigated the use of A. satureioides extracts or essential oil against R. sanguineus, which gives this work an innovative character and positions it as a basis for future research aimed, for example, at developing different formulations.

Author Contributions

Conceptualization, R.R.F. and R.C.L.R.P.; Methodology, R.R.F., F.A.S.P., and R.S.; Formal analysis, R.R.F. and R.C.L.R.P.; Investigation, R.R.F., F.A.S.P., and R.S.; Resources, R.C.L.R.P.; Data curation, R.R.F. and R.C.L.R.P.; Writing—original draft, R.R.F. and R.C.L.R.P.; Writing—review and editing, R.R.F. and R.C.L.R.P. All authors have read and agreed to the published version of the manuscript.

Funding

Conselho de Desenvolvimento Científico e Tecnológico (CNPq) for scholarship to R.R.F. (Process 141451/2017-3); CAPES PrInt (R.S. scholarship 88887.716768/2022-00); Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)—Finance Code 001, and Fundação de Amparo à Pesquisa do Estado de São Paulo, FAPESP (Process FAPESP N° 2018/14116-4).

Institutional Review Board Statement

In this study, only in vitro assays were performed, specifically with the objective of reducing the use of vertebrate animals in research. For these assays, engorged females were collected directly from the environment and left to oviposit in the laboratory; therefore, ethics committee approval does not apply to this work.

Informed Consent Statement

Not applicable.

Data Availability Statement

The raw data supporting the conclusions of this article will be made available by the authors on request.

Acknowledgments

The authors express their gratitude to the School of Pharmaceutical Sciences of Araraquara (UNESP); Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq); Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)—Finance Code 001, and Fundação de Amparo à Pesquisa do Estado de São Paulo, FAPESP (Process FAPESP N° 2018/14116-4).

Conflicts of Interest

The authors declare no conflicts of interest.

Abbreviations

The following abbreviations are used in this manuscript:
A. satureioidesAchyrocline satureioides
ASEtOH50% ethanolic extract of Achyrocline satureioides
CPQBAPluridisciplinary Center for Chemical, Biological and Agricultural Research
EmbrapaBrazilian Agricultural Research Corporation
EosEssential oils
FAOFood and Agriculture Organization of the United Nations
GC-MS Gas chromatography–mass spectrometry
OEASEssential oil of Achyrocline satureioides
R. sanguineusRhipicephalus sanguineus
RSRio Grande do Sul
RTRetention time
RIRetention index
SISGENNational System for the Management of Genetic Heritage and Associated Traditional Knowledge
T. patulaTagetes patula
UnespSão Paulo State University

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Table 1. The phytochemical constituents in the pulverized inflorescences of Achyrocline satureioides.
Table 1. The phytochemical constituents in the pulverized inflorescences of Achyrocline satureioides.
Phyto-ConstituentsReagents/ChemicalsObservationsIf Positive, It Presents
Shinoda+Pinkish or red coloration
Taubock+Yellowish-green fluorescence under UV light
FlavonoidsPew+Red coloration
Ferric chloride+Green, yellow, or purplish coloration
Aluminum chloride+Yellowish-green coloration under UV light
Gelatin+Precipitate formation
TanninsIron salts
Lead acetate		+
+		Green or blue coloration
SaponinsPermanent foaming+Persistence of foam
Legal−Intense red coloration
Cardiotonic glycosidesKedde+Reddish-violet to brown coloration
Perez−Red coloration
Keller–Kiliani+Reddish-brown ring and greenish-blue acetic layer
AnthraquinoneLieber−Pink, blue, or green coloration
Borntrager−Pink aqueous layer
Bertrand−Reddish coloration
AlkaloidsBouchadart−Brown coloration
Dradendorff−Reddish coloration
Mayer−Whitish-yellow precipitate
CoumarinsNaOH−Fluorescence under UV light
Table 2. Substances identified in Achyrocline satureioides essential oil listed in order of column elution.
Table 2. Substances identified in Achyrocline satureioides essential oil listed in order of column elution.
Proposed IdentificationtR (min)Area (%)IS
(%)		RI (Calculated)RI (Literature)
dimethyl butane3.853121,13791-474.2
octane4.54445,64191-786
alpha pinene5.37251,60781-948.55
alpha pinene5.66863,748,56897-948.55
bicyclo[2.2.1]heptane, 2,2-dimethyl-3-methylene- (1R)6.005112,83281-935.2
beta myrcene6.062806,07197-1056
benzene6.848816,876971.3231119
d-limonene7.125515,844961.3341173
eucalyptol8.2762,188,093971.3791266
beta ocimene8.4913,003,501931.3871204
3oxatricyclo[4.1.1.0(2,4)]ocane, 2,7,7-trimethyl8.5413,341,050871.3891215
nonal8.663321,303941.3941240
cis limonene oxide11.007260,031931.471-
-11.12093,261861.475-
α-campholene aldehyde11.412869,110851.4841384
bicyclo[3.1.1]heptan-3-ol, 6,6-dimethyl-2-methylene-11.790105,599-1.497-
α-campholeno aldehyde12.044121,69784
1.5051486
bicyclo[3.1.1]heptan-3-ol, 6,6-dimetil-2-methylene-12.702316,037921.5251615
verbenol12.9161,227,261911.5311665
bicyclo[2.2.1]heptan-2-ol, 2,3,3-trimethyl-13.20165,429811.5401540
borneol13.948249,212921.5621652
terpineol14.327176,388851.5741585
benzene methanol14.560126,305-1.581-
bicyclo[3.1.1]hept-2-ene-2-methanol, 6,6-dimethyl-15.06480,579841.5961202
trans carveol16.01278,153781.6241225
carvone16.94223,028801.6511705
nonyne17.05850,490941.6551134
limonene oxide19.525567,157941.7281468
verbenol20.120355,314841.7451680
alpha muurolene22.441375,249861.815-
copaene22.5591,283,759901.8181487
alpha copaene22.83222,021,156961.8271533
methanol23.437116,884-1.845-
caryophyllene23.996467,100901.8621632
alpha gurgunene24.154486,222921.8671549
beta caryophyllene24.67688,485,233971.8831612
germacrene D25.016493,124831.8931739
aromadendrene25.4114,903,702951.9051643
humulene26.09967,048,398971.9271671
alpha morulene26.8788,775,615-1.952-
naphthalene27.0291,189,497961.9571815
cycloprop27.7113,411,429941.9781719
naphthalene27.8463,656,600911.9821746
naphthalene27.988362,880 1.987
azulene28.3936,999,597932.0001664
cyclodecadiene28.6497,396,568862.0081854
cadinene delta28.776318,952882.0121752
patchouli29.15583,682832.0251793
beta selinen29.176114,700822.0251727
caryophyllene30.275550,433902.0622032
cis limonene oxide31.8521,376,176872.114-
nerodiol32.226414,492552.127-
cedrol32.470860,640822.135-
butanone33.2881,221,586712.1631854
2-naphhtalenemetanol33.375508,516832.1662226
alpha calacoreno33.7971,808,840772.180-
naphtalene tms34.135561,124672.1922203
hexadecanal34.7121,338,546862.2122119
Table 3. Percentage of mortality (%) and standard deviation of R. sanguineus larvae in contact with paper impregnated with ethanolic extract (ASEtOH) and essential oil (OEAS) from A. satureioides.
Table 3. Percentage of mortality (%) and standard deviation of R. sanguineus larvae in contact with paper impregnated with ethanolic extract (ASEtOH) and essential oil (OEAS) from A. satureioides.
Concentration (mg/mL)ASEtOH (%)OEAS (%)
100.032.2 a ± 4.8756.55 a ± 0.06
50.01.28 b ± 0 36.91 ab ± 2.14
25.00 b ± 020.04 ab ± 1.39
12.50 b ± 00 b ± 0
6.250 b ± 00 b ± 0
3.120 b ± 00 b ± 0
1.560 b ± 00 b ± 0
0.780 b ± 00 b ± 0
Control *1.30 b ± 00 b ± 0
(ASEtOH) 50% ethanolic extract of A. satureioides, (OEAS) essential oil of A. satureioides. Control *: ethanol for the extract and Tween® 80 for the oil. Tukey test at 5% probability level was approved. The means followed by the same letter do not differ statistically from each other.
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Fantatto, R.R.; Politi, F.A.S.; Sorrechia, R.; Pietro, R.C.L.R. In Vitro Activity of Ethanolic Extract and Essential Oil of Achyrocline satureioides Against Larvae of the Tick Rhipicephalus sanguineus. Parasitologia 2025, 5, 60. https://doi.org/10.3390/parasitologia5040060

AMA Style

Fantatto RR, Politi FAS, Sorrechia R, Pietro RCLR. In Vitro Activity of Ethanolic Extract and Essential Oil of Achyrocline satureioides Against Larvae of the Tick Rhipicephalus sanguineus. Parasitologia. 2025; 5(4):60. https://doi.org/10.3390/parasitologia5040060

Chicago/Turabian Style

Fantatto, Rafaela Regina, Flávio Augusto Sanches Politi, Rodrigo Sorrechia, and Rosemeire Cristina Linhari Rodrigues Pietro. 2025. "In Vitro Activity of Ethanolic Extract and Essential Oil of Achyrocline satureioides Against Larvae of the Tick Rhipicephalus sanguineus" Parasitologia 5, no. 4: 60. https://doi.org/10.3390/parasitologia5040060

APA Style

Fantatto, R. R., Politi, F. A. S., Sorrechia, R., & Pietro, R. C. L. R. (2025). In Vitro Activity of Ethanolic Extract and Essential Oil of Achyrocline satureioides Against Larvae of the Tick Rhipicephalus sanguineus. Parasitologia, 5(4), 60. https://doi.org/10.3390/parasitologia5040060

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