Survey on plants used by the population of Fez City (central Morocco) as bioinsecticides in the control of insects responsible for vector-borne diseases

1 Laboratory of Biotechnology and Preservation of Natural Resources, Faculty of Sciences Dhar El Mahraz, Sidi Mohamed Ben Abdellah University, Fez, Morocco. 2 Biotechnology, Environmental Technology and Valorization of Bio-Resources Team, Department of Biology, Faculty of Science and Technology Al-Hoceima, Abdelmalek Essaadi University, Tetouan, Morocco. 3 Physico-chemical laboratory of inorganic and organic materials, Materials Science Center (MSC), Ecole Normale Supérieure , Mohammed V University in rabat, Morocco. 4 Regional Diagnostic Laboratory of Epidemiological and Environmental Health, Regional Directorate of Health, EL Ghassani Hospital, Morocco. 5Institute of Nursing Professions and Health Techniques Fez, Regional Health Directorate, EL Ghassani Hospital, Fez, Morocco.


INTRODUCTION
Medicinal and aromatic plants have been considered and always remain as a source of care, medical remedy, and control against various diseases, particularly in developing countries in the absence of a modern and appropriate medicinal system (Tabuti et al., 2003), whose effectiveness is very often discussed (Salhi et al., 2010).
Morocco's geographical position, which is one of the Mediterranean countries, offers a very great floristic and ecological biodiversity. It is fortunate to have such a diversity of biotopes wherein almost any plant can grow and be cultivated economically: out of the 7,000 species and subspecies existing varieties, 537 are endemic in the country and 1,625 are rare or endangered (Benabid, 2000).
Currently, medicinal and aromatic plants have been considered and still remain as a source of care and are used in the fight against various pathologies, especially in developing countries in the absence of a modern and appropriate medicinal system. In Fez, traditional herbal medicines have always held a strong position. According to the bibliography and field researches, we can assume that the population in Fez city has long used very important plant species (Zeggwagh et al., 2013). It must be noted that the Quarawiyine University in Fez is the mean academic center of Africa and includes the medical section based on plants' use (Bellakhdar, 1985(Bellakhdar, , 1997. Also, the region of Fez includes a National Institute of Medicinal and Aromatic Plants in the city of Taounate, located 70 km of Fez city. Several ethnobotanical studies, aiming at the therapeutics, the cosmetics, the treatment of diseases of the digestive system, the treatment of diabetes, and cardiac and renal diseases, have been conducted in various regions of Morocco by various authors (Bellakhdar, 1987(Bellakhdar, , 1997Ben akka et al., 2015;Benkhnigue et al., 2011, El hilah andZidane, 2014;Hassan et al., 2001;Hseini et al., 2007;Kahouadji, 1995;Mehdioui and Kahouadji, 2007;Salhi et al., 2010;Slimani et al., 2016). However, the surveys on medicinal plants used as complementary medicine and prevention of vector-borne disease are virtually nonexistent.
It is within this framework that the objective of this work was registered. Indeed, the purpose of this research was to investigate and inventory the medicinal and aromatic plants used traditionally by herbalists in Fez (Central Morocco) as complementary medicine and prevention of vector-borne diseases.
In the absence of ethnobotanical studies on the national or international level on the use of the population of aromatic and medicinal plants to fight against mosquitoes, we assume that this study would be an essential means of research carried out on bioinsecticides to fight against vector-borne diseases. Bioinsecticides will be very useful as alternatives to synthetic insecticides in the control of mosquitoes and disease vectors. It would also be crucial for other future studies concerning different Moroccan regions.

The study region
The study area (Fez) is located in the northern half of Morocco between 5°07′00″ and 4°55′00″ W and 34°00′00″ and 34°05′00″ to the north latitude (Fig. 1). The urban perimeter of the city covers an area of approximately 450 km 2 . The climate of Fez is Mediterranean; winter is mild and humid and becomes extremely dry from June to September.

Studied population
According to the sampling plan, an ethnobotanical study was carried out in the city of Fez on medicinal and aromatic plants, using a questionnaire having exhaustive information on local traditional uses of medicinal and aromatic plants, particularly used as bioinsecticides to control the vector-borne disease. For this, our study sample comprised 250 herbalists (n = 250).

Studied variables in ethnobotanical survey
The study, which is the subject of the questionnaire, is part of an ethnobotanical survey. An interview grid was developed, including questions about the plant used as a bioinsecticide to control vector-borne diseases, other uses of the plant, the part used, the form of function, the dose, and the duration of use.

Statistical analysis
Pearson's chi-squared test was used at a 95% confidence level to test the significance between the percentages. We used version 20.0 of the Statistical Package for the Social Sciences software for statistical analyses. Excel analysis software was used for data entry and processing.

RESULTS
This study consisted of a sample of 250 herbalists aged between 18 and 70 years. Our sample was taken at random and consisted of people of different and varied intellectual levels, which has allowed realizing our questionnaires on the application and uses of local aromatic plants as complementary medicine and prevention of vector-borne diseases.
A total of 250 herbalists, containing 150 women with a percentage of 60% and 100 men with a percentage of 40%, were interviewed. The difference between men and women was statistically significant (χ 2 = 10; p-value = 0.002). Results have also shown that 83% of interviewed herbalists have a university level of study against 12% and 5% of herbalists with secondary and primary levels, respectively (χ 2 = 281.69; p-value = 0.002).

Plant type
The result of the analysis grids of our ethnobotanical survey unveiled that most of the plants used for medicinal plants to control the vector-borne disease are cultivated plants (74%), against 26% of wild plants.

Mode of preparation
Infusion was the most used mode of preparing plants with 68%, followed by decoction (20%). Furthermore, the rest of the plants are used in cooked and raw forms with 7% and 5%, respectively. This difference between percentages of preparing mode is statistically significant (χ 2 = 274.18; p-value < 0.001)

Used dose
The interviewees indicated that the used dose was in the form of handfuls. It has been represented with a majority percentage of 81% and pinched by 11%, while 8% was attributed to other dosage forms (Fig. 5). Percentages are statistically different (χ 2 = 236.5; p-value < 0.001).

Duration of use of plants
As shown in Figure 6, we observed that the duration of the plants used as medicinal plants to control vector-borne diseases varies from 1 day to another, which is represented by 76%, for 1 week (19%), and up to 1 month (5%). Significant difference between percentages was observed (χ 2 = 210.86; p-value < 0.001).

DISCUSSION
Traditional phytotherapy has always been solicited by a population which generally does not have the means of supporting the methods and costs of care in modern medicine.
Currently, there is a resurgence of interest in alternative medicine and traditional pharmacopeia. The fight against disease vectors is an integral part of it, as vector-borne diseases represent an important part of the infectious diseases.
Many of the technical solutions have been developed by research to improve the efficiency of the management of mosquito vectors of diseases (chemical control, biological control, and varietal resistance), but their appropriation has remained limited.
According to the World Health Organization, these repellents have been recognized as a disease prevention tool to supplement insecticide-based means of vector control. For public use, PMD is the only herbal repellant advocated by the Centers for Disease Control, which is safe for human health (El Ouali Lalami et al., 2016a;Zielinski-Gutierrez et al., 2010).
The species we have listed, namely, Lamiaceae and Asteraceae, have also been the most represented among families identified by several realized studies in Morocco (Benkhnigue et al., 2011;Hseini et al., 2007;Salhi et al., 2010), relating to the flora and ethnobotanical studies on medicinal plants in the western region (City of Kenitra and Rabat).
According to the analysis of our data, the most cited plants in the fight against insects belong to the family of Lamiaceae, Asteraceae, and Rutaceae. El-Akhal et al. (2014 reported that the larvicidal action of the essential oils of T. vulgaris and O. majorana (Lamiaceae) had a significant effect against mosquito larvae in particular (Diptera: Culicidae). Other plants, such as Mentha pulegium, Salvia officinalis, and O. basilicum, belonging to the family Lamiaceae, and Ammi visnaga, belonging to the family Apiaceae, have also been studied as complementary medicine and in the prevention of vector-borne disease El-Akhal et al., 2016b, 2016cEz Zoubi et al., 2016). Among these plants, there are some who were described and mentioned by the herbalists interviewed during our ethnobotanical survey.
The other plant species cited in this study will serve us later for the achievement of mosquito larvicide tests by using plant extracts (either essential oils or aqueous extract).
The results of this type of study will also enable us to identify effective herbal medicine against mosquito vectors of disease and/or liable in nuisance and can be used instead of chemical insecticides widely incriminated in the phenomenon resistance observed in mosquitoes, as well as environmental pollution problems.
Note that we have not carried out the survey on either on a national or international level or on medicinal and aromatic plants used as complementary medicine and in the prevention of vector-borne disease. This has limited the comparison of the results found in our investigation.
This study will contribute to the knowledge of medicinal plants in the north-central region of Morocco and precisely to explore Moroccan medicinal flora, which is poorly known as medicinal plants to control the vector-borne diseases. We also consider that this survey would have a great interest for future work concerning other Moroccan regions.

CONCLUSION
This work was conducted for the purpose of identifying plants used traditionally by the herbalists in the city of Fez as complementary medicine and in the prevention of vector-borne diseases.
The analysis of data obtained from a survey carried out allowed us to report that 22 plant species belonging to 13 botanical families were mostly used, with a clear dominance of the Lamiaceae family. The leaves, stems, and flowers were the most utilized parts.
By observing the results obtained in this study and the richness of our traditional pharmacopeia, it seems interesting from one hand to extend this kind of investigation to other parts of the country in order to identify other plants to use them as medicinal plants and to control the vector-borne diseases. On the other hand, it is necessary to validate experimentally the plant extracts and preparations identified by adequate scientific testing.

FUNDING
This research did not receive specific funding, but it was carried out within the framework of the activities of the authors and our laboratories.

CONFLICT OF INTERESTS
The authors declare that they have no conflict of interests.

ACKNOWLEDGMENT
The authors thank everyone who contributed to this work.

AUTHORS' CONTRIBUTIONS
Fouad El Akhal conducted the ethnobotanical survey and drafted the manuscript. Yassine Ez Zoubi identified the scientific name of plants and read the manuscript. Mouhcine Fadil conducted the statistical data analysis. Raja Guemmouh participated in the configuration and reading of the manuscript. Abdelhakim El Ouali Lalami contributed to the conception and design of the study and helped in the English writing of the manuscript. All authors read and approved the final manuscript.