Synergistic Interaction Effect of Artemisia cina n-hexane Extract and Tagetes lucida Ethyl Acetate Extract on Haemonchus Contortus

Acta Parasitologica (2024) 69:1132–1140 https://doi.org/10.1007/s11686-024-00839-6 ORIGINAL PAPER Synergistic Interaction Effect of Artemisia cina n-hexane Extract and Tagetes lucida Ethyl Acetate Extract on Haemonchus Contortus Itzel Santiago-Figueroa1 · Manases González-Cortazar3 · Julieta Gertrudis Estrada-Flores4 · Jorge Alfredo Cuéllar-Ordaz1 · María Eugenia López-Arellano2 · Francisco Javier González-Reyes5 Agustín Olmedo-Juárez2 · Rosa Isabel Higuera-Piedrahita1 · Received: 16 October 2023 / Accepted: 12 March 2024 / Published online: 3 April 2024 © The Author(s) 2024 Abstract Purpose We analysed the possible synergistic activity among active extracts from Artemisia cina and Tagetes lucida combinations on Haemonchus contortus, a nematode parasitising sheep. Methods The work was carried out in vitro on eggs and infective larvae (L3) of H. contortus. The results were analysed with SAS 9.1, applying the ANOVA and Tukey test, and the lethal concentration (LC) values LC50 and LC90 were determined with regression analysis, employing Proc Probit of SAS 9.1. Additionally, the lethal concentration (LC) was calculated with LC50 and LC90 to determine the synergistic effect. Results The results demonstrated a high efficacy of the two plants studied on both nematode eggs and L3 larvae as well as of their combinations. The highest egg hatching inhibition was obtained with a 50/50 combination, and the best larvae mortality was obtained with 25% A. cina and 75% T. lucida at 10 mg/mL. Additionally, this combination showed a synergistic effect. Conclusion The two plant species studied here can be applied as natural anthelmintic alternatives due to their high bioactive effect and synergistic response. Keywords Gastroenteric Nematodes · Small Ruminants · Pharmacology Interaction · Bioactive Plants · Sustainable Anthelmintic Agustín Olmedo-Juárez Rosa Isabel Higuera-Piedrahita Itzel Santiago-Figueroa Manases González-Cortazar 1 Facultad de Estudios Superiores Cuautitlán, Universidad Nacional Autónoma de México, Carr. Cuautitlán- Teoloyucan Km 2.5, Col. San Sebastián Xhala, CP 54714 Cuautitlán, México 2 Centro de Investigación Disciplinaria en Salud Animal e Inocuidad, Agrícolas y Pecuarias (INIFAP), Instituto Nacional de Investigaciones Forestales, Carr. Fed. Cuernavaca-Cuautla No. 8534, CP 62550 Jiutepec, México 3 Centro de Investigación Biomédica Del Sur, Instituto Mexicano del Seguro Social (IMSS), Argentina No. 1, 62790 Xochitepec, CP, México 4 Instituto de Ciencias Agropecuarias y Rurales (ICAR), Universidad Autónoma del Estado de México (UAEM), Estado de México, Campus UAEM El Cerrillo, El Cerrillo Piedras Blancas, 50090 Toluca, México 5 Universidad Autónoma Chapingo, Carr. México-Texcoco Km 38.5, C.P. 56230 Chapingo, Estado de México, México Jorge Alfredo Cuéllar-Ordaz María Eugenia López-Arellano Francisco Javier González-Reyes 13 Acta Parasitologica (2024) 69:1132–1140 Introduction Gastroenteric helminths are the cause of major parasitic diseases in grazing ruminants. These disorders cause damage to animal health and welfare and reduce productive performance such as milk production, weight gain, carcass quality and fertility, among others [1]. Haemonchosis is a parasitic disease with high importance in small ruminants. Haemonchus contortus feeds on the blood of its host, resulting in anaemia, weight loss, hyperacute infections and sudden death [2]. Due to the rapid development and increase of anthelmintic resistance (AR) in nematode populations worldwide and to diminish the damage caused by these parasites in production systems, the search for pharmacological and non-pharmacological tools is necessary [3, 4]. Within these management strategies, the use of selective deworming with the aid of indicators such as the body condition score [5] and the FAMACHA colour chart [5]. Other practices are focused on population resistance, such as using secondary metabolites from biological organisms, including nematophagous fungi [6] or ruminal bacteria [7]. Plants with secondary metabolites display important anthelmintic activity against gastrointestinal nematodes, including H. contortus [8]. Also, various plants have been used in the treatment of anxiety and depression [9], inflammation [10], cancer [9, 11–13], fibromyalgia disorders [14], immunomodulatory control [15], as antioxidants [16] and as pain modulators [17] among others. Flavonoids, as the major compounds of some plants, play a role in the treatment of Parkinson’s disease [18]. The family Asteraceae contains numerous species with benefits for human and animal health. These plants contain several secondary compounds such as terpenoids, flavonoids and other molecules with anticancer, antimalarial and anthelmintic properties [19–21]. Artemisinin is a sesquiterpene lactone obtained from Artemisia annua with demonstrated efficacy against malaria, which has been resistant to chloroquine since 1950, along with other properties [19, 21, 22]. Artemisia cina, commonly known as Levant wormseed, is a herbaceous plant that has been traditionally used as an anthelmintic agent in many parts of the world, especially in the Mediterranean and Middle Eastern regions. Several studies have shown the efficacy of A. cina in controlling various parasitic infections. For instance, a study found that the aqueous and ethanolic extracts of A. cina significantly reduced the number of eggs of gastrointestinal nematodes in sheep [23, 24]. Similarly, another study reported that the ethanolic extract of A. cina was highly effective in controlling gastrointestinal nematodes in goats [25]. Moreover, A. cina has also been found to be effective against other parasitic infections such as malaria, 1133 leishmaniasis and schistosomiasis [26, 27]. The active compounds present in A. cina, such as santonin, artemisinin and their derivatives, are the main contributors to its anthelmintic activity [28]. A study with an A. cina aqueous extract, which was administered to sheep naturally infected with Moniezia spp., recorded a 100% efficacy faecal egg count reduction) on at 9th day post-administration [29]. Additionally, an A. cina n-hexane extract was assessed against H. contortus infective larvae and caused a mortality of 80% at 2 mg/mL. In this same research, the authors administered the A. cina n-hexane at 2 mg/kg to gerbils artificially infected with H. contortus, resulting in a 100% efficacy [23]. Tagetes lucida is another genus (Asteraceae) extensively studied as an anthelmintic since it is rich in some phenolic acids and other metabolites such as coumarins and terpenes [30]. This plant, commonly known as Mexican “Pericón,” is a flowering plant traditionally used for various medicinal purposes, including its cytotoxic activity, which can result in anthelmintic potential [31]. Studies have reported the efficacy of T. lucida in controlling parasitic infections. For example, the authors of one study found that the ethanolic extract of T. (...truncated)