The Phlebotomine sand fly fauna of Switzerland revisited.

Sand flies (Diptera: Psychodidae, Phlebotominae; Newstead, 1911) are widespread in Europe, being particularly common in the Mediterranean region but rare north of the Alps. Thus, Switzerland is an opportune place to investigate the sand fly fauna on both sides of the Alpine crest, in southern sub-Mediterranean climate and northern oceanic temperate climate. We reinvestigated the Swiss sand fly fauna with the aim to assess changes in composition, altitudinal distribution, abundance and seasonality. Thirty-eight sites were investigated with light traps and/or interception sticky traps in 4 years. Ninety and 380 specimens were caught by light traps and sticky traps, respectively, at 15 collecting sites. Four species were identified. Phlebotomus mascittii (Grassi, 1908), Phlebotomus perniciosus (Newstead, 1911) and Sergentomyia minuta (Rondani, 1843) were confirmed in Ticino, and P. mascittii for the first time in neighbouring Grisons. Also, Phlebotomus neglectus (Tonnoir, 1921) is for the first time reported, though at a very low density compared to P. perniciosus at the same site. Its presence in Ticino supports the northward spread observed in Italy. Sand flies were detected north of the Alps at one site only, endorsing a historical report. Overall, the low density of P. perniciosus and very low density of P. neglectus suggest that canine leishmaniosis may not be an important disease risk in Switzerland.

Recent progress on heterologous protein production in methylotrophic yeast systems.

Recombinant protein production technology is widely applied to the manufacture of biologics used as drug substances and industrial proteins such as recombinant enzymes and bioactive proteins. Various heterologous protein production systems have been developed using prokaryotic and eukaryotic hosts. Especially methylotrophic yeast in eukaryotic hosts is suggested to be particularly valuable because such systems have the following advantages: protein secretion into culture broth, eukaryotic quality control systems, a post-translational modification system, rapid growth, and established recombinant DNA tools and technologies such as strong promoters, effective selection markers, and gene knock-in and -out systems. Many methylotrophic yeasts such as the genera Candida, Ogataea, and Komagataella have been studied since methylotrophic yeast was first isolated in 1969. The methanol-consumption-related genes in methylotrophic yeast are strongly and strictly regulated under methanol-containing conditions. The well-regulated gene expression systems under the methanol-inducible gene promoter lead to the potential application of heterologous protein production in methylotrophic yeast. In this review, we describe the recent progress of heterologous protein production technology in methylotrophic yeast and introduce Ogataea minuta as an alternative production host as a substitute for K. phaffii and O. polymorpha.

Characterization of the physico-chemical properties of the natural habitat and in vitro culture effects on the biochemistry, proliferation and morphology of Lemna minuta.

In this study, the ecological conditions of the natural habitat of Lemna minuta Kunth in Morocco were investigated, and the impact of five synthetic growth media (Murashige-Skoog (MS), Schenk-Hildebrand (SH), Hoagland medium (HM), 10X Algal Assay Procedure (AAP), and Swedish Standard Institute medium (SIS)) on the morphophysiological and biochemical parameters was analysed. The morphophysiological parameters included root length, frond surface area, and fresh weight, while the biochemical parameters included photosynthetic pigments, carbohydrates, and protein content. The study was conducted in vitro in two phases: an uncontrolled aeration system (Phase I) and a controlled aeration system (Phase II).The results showed that the pH, conductivity, salinity, and ammonium levels in the natural habitat were within the optimal range for duckweed growth. The measured orthophosphate concentrations were higher compared to previous observations, while the recorded chemical oxygen demand values were low. The study also revealed a significant effect of the culture medium composition on the morphophysiological and biochemical parameters of the duckweed. The fresh weight biomass, relative growth rate in fronds, relative growth rate in surface area, root length, protein content, carbohydrates, chlorophyll (a), chlorophyll (b), total chlorophyll, carotenoids, and the chlorophyll (a/b) ratio were all affected by the culture medium.The most accurate regression models described the growth index GI(F) based on time and in vitro culture conditions in both phases. In Phase I, the best models for MS, SIS, AAP, and SH media were linear, weighted quadratic, cubic, and weighted cubic, respectively. In Phase II, the best models for all growth media were linear. The time coefficients (in days) for Phase II were 0.321, 0.547, 1.232, 1.470, and 0.306 for AAP, HM, MS, SH, and SIS, respectively.Comparing the morphophysiological and biochemical parameters of fronds from different media and analysing the regression model results showed that the SH and MS media were the best among the tested media for the in vitro culture of L. minuta in controlled aeration conditions. However, further research is needed to develop new synthetic media that best promote the growth and maintenance of this duckweed in long-term culture.

Assessments of Alpha-Amylase Inhibitory Potential of Tagetes Flavonoids through In Vitro, Molecular Docking, and Molecular Dynamics Simulation Studies.

Diabetes is a chronic fast-growing metabolic disorder that is characterized by high blood glucose levels. Tagetes minuta L. has been used as a traditional remedy for various illnesses for many years, and, furthermore, its oil is used in the perfume and flavor industries. T. minuta contains various metabolites, such as flavonoids, thiophenes, terpenes, sterols, and phenolics, with varied bioactivities. Flavonoids can inhibit carbohydrate-digesting enzymes, such as alpha-amylase, which is a convenient dietary strategy for controlling hyperglycemia. In the current investigation, the isolated flavonoids quercetagetin-6-O-(6-O-caffeoyl-ß-D-glucopyranoside), quercetagetin-7-O-ß-D-glucopyranoside, quercetagetin-6-O-ß-D-glucopyranoside, minutaside A, patuletin-7-O-ß-D-glucopyranoside, quercetagetin-7-methoxy-6-O-ß-D-glucopyranoside, tagenols A and B, quercetagetin-3,7-dimethoxy-6-O-ß-D-glucopyranoside, patuletin, quercetin-3,6-dimethyl ether, and quercetin-3-methyl ether from T. minuta were assessed for their alpha-amylase inhibition (AAI) efficacy using an in vitro assay, as well as molecular docking, dynamics simulation, and ADMET analyses. Our findings show that quercetagetin-6-O-(6-O-caffeoyl-ß-D-glucopyranoside) (1), quercetagetin-7-O-ß-D-glucopyranoside (2), quercetagetin-6-O-ß-D-glucopyranoside (3), minutaside A (4), patuletin-7-O-ß-D-glucopyranoside (5), and quercetagetin-7-methoxy-6-O-ß-D-glucopyranoside (6) had a notable AAI capacity (IC50s ranged from 7.8 to 10.1 µM) compared to acarbose (IC50 7.1 µM). Furthermore, these compounds with the highest binding affinity among the tested flavonoids revealed high docking scores for AA (ranging from -12.171 to 13.882 kcal/mol) compared to that of acarbose (-14.668 kcal/mol). In MDS, these compounds were observed to show maximum stability and the greatest binding free energy, suggesting that they may contend with native ligands. In addition, the ADMET analysis showed that these active compounds had a broad span of drug-like, pharmacokinetic, and physicochemical features and did not possess any considerable undesired effects. The current results suggest the potential of these metabolites as AAI candidates. However, further in vivo and mechanistic studies are warranted to specify the efficacy of these metabolites.

Kovacikia minuta sp. nov. (Leptolyngbyaceae, Cyanobacteria), a new freshwater chlorophyll f-producing cyanobacterium.

A few groups of cyanobacteria have been characterized as having far-red light photoacclimation (FaRLiP) that results from chlorophyll f (Chl f) production. In this study, using a polyphasic approach, we taxonomically transferred the Cf. Leptolyngbya sp. CCNUW1 isolated from a shaded freshwater pond, which produces Chl f under far-red light, to the genus Kovacikia and named this taxon Kovacikia minuta sp. nov. This strain was morphologically similar to Leptolyngbya-like strains. The thin filaments were purplish-brown under white light but became grass green under far-red light. The 31-gene phylogeny grouped K. minuta CCNU0001 into order Synechococcales and family Leptolyngbyaceae. Phylogenetic analysis based on 16S rRNA gene sequences further showed that K. minuta CCNU0001 was clustered into Kovacikia with similarities of 97.2-97.4% to the recently reported type species of Kovacikia muscicola HA7619-LM3. Additionally, the internal transcribed spacer region between 16S-23S rRNA genes had a unique sequence and secondary structure compared with other Kovacikia strains and phylogenetically related taxa. Draft genome sequences of K. minuta CCNU0001 (8,564,336 bp) were assembled into one circular chromosome and two circular plasmids. A FaRLiP 20-gene cluster comprised two operons with the unique organization. In sum, K. minuta was established as a new species, and it is the first species reported to produce Chl f and for which a draft genome was produced in genus Kovacikia. This study expanded our knowledge regarding the diversity of Chl f-producing cyanobacteria in far-red light-enriched environments and provides important foundational information for future investigations of FaRLiP evolution in cyanobacteria.

Stomatal density and mechanics are critical for high productivity: insights from amphibious ferns.

Angiosperm dominance in terrestrial landscapes is partially attributable to high photosynthetic capacities. Angiosperms benefit from diverse anatomical and physiological adaptations, making it difficult to determine which factors may have been prerequisites for the evolution of enhanced photosynthetic rates in this group. We employed a novel approach to this problem: comparisons between angiosperms and Marsileaceae, a family of semi-aquatic ferns that are among the only land plants to match angiosperm photosynthetic rates. We found that Marsileaceae have very high stomatal densities and, like angiosperms but unlike all other ferns previously studied, exhibit wrong-way stomatal responses to excision. These results suggest that stomatal density and a little-studied angiosperm trait, the capacity for lateral displacement of guard cells into neighboring epidermal cells, are crucial for facilitating high rates of gas exchange. Our analysis also associates these adaptations in Marsileaceae with an increased risk of excessive water loss during drought. Our findings indicate that evolution in stomatal physiology was a prerequisite for high photosynthetic capacities in vascular plants and a key driver of the abrupt Cretaceous rise of the angiosperms.

Photodynamic activity of Tagetes minuta extracts against superficial fungal infections.

Candida and dermatophyte species are the most common causes of superficial mycoses because their treatment can be difficult due to limitations of current antifungal drugs in terms of toxicity, bioavailability, interactions, narrow-spectrum activity, and development of resistance. Photodynamic therapy (PDT) involves the topical administration of a photosensitizer in combination with light of an appropriate wavelength and molecular oxygen that produces reactive oxygen species (ROS), which promote damage to several vital components of the microorganism. Tagetes species are known as a source of thiophenes, biologically active compounds whose antifungal activity is enhanced by irradiation with UVA. The present investigation evaluated Tagetes minuta extracts as a photosensitizer on growth of Candida and dermatophytes and their effect on Candida virulence factors. T. minuta root hexane and dichloromethane extracts demonstrated high photodynamic antifungal activity. Bioautographic assays and chromatographic analysis revealed the presence of five thiophenes with reported photodynamic antifungal activities under UVA. Analysis of ROS production indicated that both type I and II reactions were involved in the activity of the extracts. In addition, the extracts inhibited virulence factors of Candida, such as adherence to epithelial surfaces and germ tube formation and showed efficacy against different Candida morphologies: budding cells, cells with germ tube and biofilms. Results suggested that PDT with T. minuta extracts might become a valuable alternative to the already established antifungal drugs for the treatment of superficial fungal infections.

Bloodstream Infection of a Cancer Patient by Cystobasidium minutum: A Case Report and Literature Review.

Emergent fungal infections are uncommon conditions which frequently lead to death. To our knowledge, only a few cases of invasive infection by Cystobasidium minutum (previously known as Rhodotorula minuta) have been reported. Moreover, several factors are responsible for deep site infections, such as catheter-related fungemia. This report describes the first case report of Cystobasidium minutum causing fungemia in Brazil. The pathogens fungemia was demonstrated by catheter and blood culture-proven, and both yeasts were identified by sequences of D1/D2 rDNA region. After the end of antifungal therapy and catheter removal, a second blood culture was found to be negative and the clinical signs and symptoms of the patient improved.

PER1, GUP1 and CWH43 of methylotrophic yeast Ogataea minuta are involved in cell wall integrity.

In eukaryotes, the glycosylphosphatidylinositol (GPI) modification of many glycoproteins on the cell surface is highly conserved. The lipid moieties of GPI-anchored proteins undergo remodelling processes during their maturation. To date, the products of the PER1, GUP1 and CWH43 genes of the yeast Saccharomyces cerevisiae have been shown to be involved in the lipid remodelling. Here, we focus on the putative GPI remodelling pathway in the methylotrophic yeast Ogataea minuta. We found that the O. minuta homologues of PER1, GUP1 and CWH43 are functionally compatible with those of S. cerevisiae. Disruption of GUP1 or CWH43 in O. minuta caused a growth defect under non-permissive conditions. The O. minuta per1Δ mutant exhibited a more fragile phenotype than the gup1Δ or cwh43Δ mutants. To address the role of GPI modification in O. minuta, we assessed the effect of these mutations on the processing and localization of the O. minuta homologues of the Gas1 protein; in S. cerevisiae, Gas1p is an abundant and well-characterized GPI-anchored protein. We found that O. minuta possesses two copies of the GAS1 gene, which we designate GAS1A and GAS1B. Microscopy and western blotting analysis showed mislocalization and/or lower retention of Gas1Ap and Gas1Bp within the membrane fraction in per1Δ or gup1Δ mutant cells, suggesting the significance of lipid remodelling for GPI-anchored proteins in O. minuta. Localization behaviour of Gas1Bp differed from that of Gas1Ap. Our data reveals, for the first time (to our knowledge), the existence of genes related to GPI anchor remodelling in O. minuta cells.

Effects of Christensenella minuta lipopolysaccharide on RAW 264.7 macrophages activation.

Christensenella minuta (C. minuta) is a gram-negative gastrointestinal bacterium associated with weight loss. However, recent studies have shown that C. minuta might be a potential pathogen and thus limited its application in the control of obesity. Research into the genetic characteristics and pathogenicity of C. minuta remain elusive. As a major virulence factor of gram-negative bacteria, lipopolysaccharide (LPS) can induce various diseases. In this study, we report the complete genome sequence of C. minuta and have also identified some genes related to LPS biosynthesis. The structure of C. minuta LPS, detected by SDS-PAGE, was different from that of Escherichia coli (E. coli) LPS. The incubation of RAW 264.7 macrophages with C. minuta LPS resulted in lower levels of cellular proliferation, phagocytosis and nuclear factor-kappa B (NF-κB) activation as compared to incubation with E. coli LPS. Furthermore, the expression of pro-inflammatory cytokines, as well as nitric oxide and reactive oxygen species production, was induced in C. minuta LPS-treated cells but to a much lower extent than that by E. coli LPS. These findings show that C. minuta LPS acts as a weak agonist for RAW 264.7 macrophages and can only trigger a weak inflammatory response through the NF-κB signalling pathway. In conclusion, these results suggest that the toxicity of C. minuta LPS is significantly attenuated due to its atypical structure and weak agonist activity for RAW 264.7 macrophages.

Tagetes spp. Essential Oils and Other Extracts: Chemical Characterization and Biological Activity.

Tagetes (marigold) is native to America, and its cultivation currently extends to other countries in Africa, Asia, and Europe. Many species of this genus, such as T. minuta, T. erecta, T. patula, and T. tenuifolia, are cultivated as ornamental plants and studied for their medicinal properties on the basis of their use in folk medicine. Different parts of the Tagetes species are used as remedies to treat various health problems, including dental, stomach, intestinal, emotional, and nervous disorders, as well as muscular pain, across the world. Furthermore, these plants are studied in the field of agriculture for their fungicidal, bactericidal, and insecticidal activities. The phytochemical composition of the extracts of different Tagetes species parts are reported in this work. These compounds exhibit antioxidant, antiinflammatory, and enzyme inhibitory properties. Cultivation and the factors affecting the chemical composition of Tagetes species are also covered. In the current work, available literature on Tagetes species in traditional medicine, their application as a food preservative, and their antimicrobial activities are reviewed.

Chemical constituents, antibacterial and antioxidant properties of the essential oil flower of Tagetes minuta grown in Cala community Eastern Cape, South Africa.

BACKGROUND: Tagetes minuta has a long record of human use for the treatment of stomach and intestinal diseases. Most drugs used for diseases treatment are less efficacious with side effects and this brought the search for new treatment regimens mainly from medicinal plants. METHOD: The essential oil (EO) was extracted by Clevenger's-type apparatus and its chemical composition, antioxidant and antibacterial properties were determined by GC-MS, spectrophotometric and broth dilution methods respectively. S. uberis, E. cloacae, S. aureus, M. smegmatis, L. ivanovii, Vibrio spp. and E. coli bacteria strains were used as test bacteria. RESULTS: GC-MS analysis revealed 98 compounds in the EO flower of T. minuta and ß-Ocimene (14. 40%) was the major chemical constituents. The EO exhibited highest inhibitory effect against DPPH radical, followed by its effect on ABTS, while LP radical showed the least sensitivity with IC50 values of 2.45 mg/mL, 2.76 mg/mL and 3.23 mg/mL respectively. The EO showed antibacterial activities against all test organisms with MIC value for S. aureus, M. smegatis and S. uberis at 0.125 mg/mL and for L. ivanovii, Vibrio spp., E. cloacae and E. coli at 0.06 mg/mL. The EO showed MBC against E. cloacae and E. coli at 0.06 mg/mL at 0.5 mg/mL for S. uberis and 0.125 mg/mL for Vibrio spp. CONCLUSION: Findings from this study suggest that the EO of T. minuta flower may be a useful candidate in the search for lead constituents for the synthesis of new potent antibacterial and antioxidant agent.

First report of human infection by Christensenella minuta, a gram-negative, strickly anaerobic rod that inhabits the human intestine.

Christensenella minuta is a Gram-negative strictly anaerobic short rod that inhabits the human gut. This bacterium was isolated in a mixed infection with Desulfovibrio desulfuricansfrom the blood of a patient with a diagnosis of acute appendicitis. The strain was identified by 16S rRNA sequence analysis. As far as we know, this is the first time C.minuta has been isolated from a human clinical specimen.

Phytoremediation applications in natural condition and in mesocosm: The uptake of cadmium by Lemna minuta Kunth, a non-native species in Italian watercourses.

Metal pollution in water and soil is an environmental and public health issue. Cadmium (Cd) is included in the list of priority hazardous substances in the European Water Framework Directive. Phytoremediation system is a cost-effective, plant-based approach that takes advantage of the ability of plants to concentrate elements and compounds from the environment and to metabolize various molecules in their tissues. We studied the presence and the importance of an invasive species, such as Lemna minuta, in the environment and the effects of Cd pollution on this species. Growth, removal, and tolerance were evaluated for different Cd concentrations and different times of plant exposure. Overall, the results show that L. minuta has a good capacity of growth, metal bioconcentration, and tolerance up to 3 days of exposure at 0.5 and 1.5 mg L-1 of Cd. In particular, L. minuta was able to accumulate Cd up to 3771 mg kg-1 on dry mass basis. We can conclude that L. minuta possesses a great capability of Cd absorption and accumulation, thus supporting a potential use of this species in designing a metal bioremediation system in phytoremediation field.

Mitigating arsenic contamination in rice plants with an aquatic fern, Marsilea minuta.

Dangers of arsenic contamination are well known in human civilization. The threat increases when arsenic is accumulated in food and livestock through irrigated crops or animal food. Hence, it is important to mitigate the effects of arsenic as much as possible. This paper discusses a process for reducing the level of arsenic in different parts of rice plants with an aquatic fern, Marsilea minuta L. A pot experiment was done to study the possibility of using Marsilea minuta as a phytoremediator of arsenic. Rice and Marsilea minuta were allowed to grow together in soils. As a control, Marsilea minuta was also cultured alone in the presence and absence of arsenic (applied at 1 mg/L as irrigation water). We did not find any significant change in the growth of rice due to the association of Marsilea minuta, though it showed a reduction of approximately 58.64% arsenic accumulation in the roots of rice grown with the association of fern compared to that grown without fern. We measured a bioaccumulation factor (BF) of > 5.34, indicating that Marsilea minuta could be a good phytoremediator of arsenic in rice fields.

Aliidiomarina minuta sp. nov., a haloalkaliphilic bacterium that forms ultra-small cells under non-optimal conditions.

An aerobic haloalkaliphilic bacterium, designated strain MLST1(T), was isolated from filtered (0.22 µm) Mono Lake (USA) waters. The isolate was observed to grow primarily on yeast extract, peptone and tryptone. Optimal growth occurred in media at pH 9.5 containing 5-11 g/l yeast extract, and 70-100 g/l NaCl. When in log phase of growth, cells were found to be mostly curved motile rods (1-3 µm length by 0.4-1 µm diameter). Phylogenetic analysis of the 16S rRNA gene and chemotaxonomic data revealed that the isolate belonged to the family Idiomarinaceae, and is closely related to Aliidiomarina maris (96.67 % sequence similarity). The major fatty acids were identified to be iso-C17:1 ω9c (27.1 %), iso-C17:0 (21.3 %) and iso-C15:0 (12.2 %). Predominant polar lipids included phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, and the major respiratory quinone was identified as Q8. The DNA base composition was 46.3 mol% G+C. Survival studies indicated that strain MLST1(T) remains viable after exposure to adverse conditions, particularly in the prolonged absence of a carbon source, at low temperatures and with no NaCl. Under these conditions, the cells shrunk to around 0.2 µm in length by 0.1 µm in diameter and passed through 0.22 µm filters. The ultra-small cells could only be resuscitated in media with low levels of yeast extract, up to 0.6 g/l. Once resuscitated, cells were able to grow to full size. Strain MLST1(T) is clearly a unique bacterium in the waters of Mono Lake and the name Aliidiomarina minuta sp. nov. is proposed. The type strain is MLST1(T) (=JCM 17425(T) = KCTC 23357(T)).

Opinion of the Scientific Committee on Consumer Safety (SCCS) - Opinion on the fragrance ingredients Tagetes minuta and Tagetes patula extracts and essential oils (phototoxicity only) in cosmetic products.

CONCLUSION OF THE OPINION: The SCCS considers a maximum level of 0.01% Tagetes minuta and Tagetes patula extracts and essential oils in leave-on products (except sunscreen cosmetic products) as safe, provided that the alpha terthienyl (terthiophene) content of the Tagetes extracts and oils does not exceed 0.35%. The Tagetes extracts and oils should not be used as ingredients of sunscreen products.

First molecular detection of Leishmania tarentolae-like DNA in Sergentomyia minuta in Spain.

Phlebotomine sand flies (Diptera, Psychodidae) are vectors of multiple Leishmania species, among which Leishmania infantum stands out as a being frequently pathogenic to humans and dogs in Mediterranean countries. In this study, Sergentomyia minuta sand flies were collected using CDC miniature light traps in different 431 biotopes from Southwest Spain. A total of 114 females were tested for the presence of Leishmania DNA by targeting ITS-1 and cyt-B sequences by PCR. Leishmania DNA was detected in one S. minuta. Characterization of the obtained DNA sequences by phylogenetic analyses revealed close relatedness with Leishmania tarentolae Wenyon, 1921 as well as with both human and canine pathogenic strains of Asian origin (China), previously described as Leishmania sp. To our knowledge, this is the first report of phlebotomine sand flies naturally infected with L. tarentolae-like in Spain. The possible infection of sand flies with novel Leishmania species should be taken into consideration in epidemiological studies of vector species in areas where leishmaniosis is endemic.

Effects of co-cropping Bidens pilosa (L.) and Tagetes minuta (L.) on bioaccumulation of Pb in Lactuca sativa (L.) growing in polluted agricultural soils.

Polluted agricultural soils are a serious problem for food safety, with phytoremediation being the most favorable alternative from the environmental perspective. However, this methodology is generally time-consuming and requires the cessation of agriculture. Therefore, the purpose of this study was to evaluate two potential phytoextractor plants (the native species Bidens pilosa and Tagetes minuta) co-cropped with lettuce growing on agricultural lead-polluted soils. The concentrations of Pb, as well as of other metals, were investigated in the phytoextractors, crop species, and in soils, with the potential risk to the health of consumers being estimated. The soil parameters pH, EC, organic matter percentage and bioavailable lead showed a direct relationship with the accumulation of Pb in roots. In addition, the concentration of Pb in roots of native species was closely related to Fe (B. pilosa, r = 0.81; T. minuta r = 0.75), Cu (T. minuta, r = 0.93), Mn (B. pilosa, r = 0.89) and Zn (B. pilosa, r = 0.91; T. minuta, r = 0.91). Our results indicate that the interaction between rhizospheres increased the phytoextraction of lead, which was accompanied by an increase in the biomass of the phytoextractor species. However, the consumption of lettuce still revealed a toxicological risk from Pb in all treatments.

Efficacy of Tagetes minuta (Asteraceae) essential oil against Rhipicephalus sanguineus (Acari: Ixodidae) on infested dogs and in vitro.

Ticks from Rhipicephalus sanguineus complex are widely distributed in the world and one species from this complex is the most common tick on dogs in Brazil, notably in urban areas. This tick is a vector of several diseases. Among others it transmits the agent of canine Ehrlichiosis, a major dog infectious disease and the agent of Rocky Mountain spotted fever. This tick can spread rapidly and develop intolerable infestations within no time. Currently tick control is done with acaricides and demand for such drugs has grown fast. However, R. sanguineus has already developed resistance to the main active compounds and the development of new acaricides is necessary. Many essential oils of plants have acaricidal effect and may be an important source of molecules for the synthesis of new acaricide products. In this study, we evaluated the effectiveness of a new herbal phytotherapic, consisting of the essential oil of Tagetes minuta L., against R. sanguineus in vitro and on dogs undergoing experimental infestations. The product displayed 100% efficacy against larvae, nymphs and adults of the tick on all tested conditions.