There Is Not Only Cupressus sempervirens L.: A Review on the Phytochemistry and Bioactivities of the Other Cupressus L. Species

This review article reports for the first time phytochemistry, ethnobotanical uses and pharmacological activities of all Cupressus L. species other than Cupressus sempervirens L. Indeed, the literature survey showed how many other Cupressus species are rich of important phytochemical compounds, widely used in the ethnobotanical field for several purposes and endowed with interesting biological activities, even if they are somehow neglected by the scientific community. This review aims to continue the study of these other Cupressus species and promote more research on them.


Introduction
Cupressus L. (common name Cypress) is a genus of evergreen trees belonging to the Cupressaceae within the Gymnosperms. From a morphological standpoint, they are characterized by short scale-like leaves when the trees are adult, whereas they are bigger and needle-like when the trees are young (up to two years). They are arranged in opposite decussate pairs in both cases. The cones are quite long, globose or ovoid, and are mature in 18-24 months from pollination. The seeds are quite small with one narrow wing along each side ( Figure 1) [1]. Cupressus species are native to some temperate regions of the Northern Hemisphere, including western North America, Central America, northwest Africa, the Middle East, the Himalayas, southern China and northern Vietnam, even if they also are widely cultivated [2]. The most important species of the genus is surely Cupressus sempervirens L., whose phytochemistry, ethnobotanical uses, and biological activities have already been widely studied and discussed in detail [3][4][5].
The aim of this paper is to present a detailed review on the phytochemistry, the ethnopharmacological uses and the biological activities of all the other Cupressus species as reported in literature. To the best of our knowledge, this represents the first study ever performed on this subject. In addition, a deep chemotaxonomic analysis of the genus is given. The data were collected from several bibliographic sources, i.e., Pub-Chem, Scopus, Google Scholar using the following keywords alone or in combination: Cupressus L., phytochemistry, ethnobotany, ethnopharmacological uses, biological activities and the names of all the existing species of the genus except C. sempervirens, as reported in www.theworldfloraonline.org (accessed on 14 May 2022). The papers reporting studies

Chemotaxonomy
The chemotaxonomy of Cupressus genus must be first considered in the general context concerning Gymnospermae [135][136][137]. In particular, on the basis of the phytochemical compounds evidenced, three main compositionally distinct groups can be distinguished within that. The first one regards the non-conifers, i.e., the divisions Ginkgophyta and Cycadophyta that mainly contain gums but biosynthesise other compounds, too. The second one regards the division Gnetophyta which, considering their intermediate position to angiosperms [138,139], must be separately considered as also evidenced by the anomalous presence of alkaloids and the recent isolation of stilbenoids [140]. The third one regards the division Pinophyta which comprises species that mainly biosynthesize volatile-rich exudates and phenolics contained in gums and resins. Cupressus genus, being a member of the Cupressaceae family, belongs to this latter group together with Araucariaceae and Podocarpaceae families. In fact, Cupressus species have evidenced volatile components and phenolic compounds as their major phytochemical classes. Going in major detail, monoterpenes, sesquiterpens and diterpenes mainly characterize the volatile profiles of Cupressus species. Yet, these compounds are very common in the plant kingdom having been identified in many other families like Apiaceae, Araucariaceae, Boraginaceae, Lamiaceae, Podocarpaceae, Rutaceae, Scrophulariaceae and Verbenaceae [141][142][143][144][145][146][147][148][149] and for this, individuating a specific chemotaxonomic marker is not easy. Nevertheless, one chemotaxonomic marker could be found, i.e., bakerol, the occurrence of which seems to be limited to only C. bakeri [43,44]. Indeed, for what concerns the non-volatile constituents, simple flavonoids and bioflavonoids are predominant in Cupressus species, but other classes of nonvolatile compounds were also found like fatty acids, diterpenoids, triterpenoids, alkaloids, and simple organic acids. In this respect, Cupressus species present several phytochemical similarities with the genera Wollemia W.G.Jones, K.D.Hill & J.M.Allen, Araucaria Juss. and Agathis Salisb., all belonging to the Auracariaceae family, which is considered to be taxonomically close to Cupressaceae [148]. In particular, these phytochemical similarities mainly regard the contents in biflavonoids and diterpenoids with compounds that had been previously considered as chemotaxonomic markers of the Cupressus genus like cupressuflavone and its derivatives and cupressic acid and its derivatives and that have been later found in Wollemia, Araucaria and Agathis genera, thus losing this characteristic. All these facts are extremely interesting and urge new phytochemical studies on Cupressus species in this sense but confirm the previous subdivision.

Ethnobotanical Uses
Several Cupressus species are employed in the traditional medicine of several countries all around the world. The most common utilizations regard C. lusitanica and C. macrocarpa. The following sections explore the ethnobotanical uses of Cupressus species other than C. sempervirens as reported in literature. It is important to underline that not all the Cupressus species possess ethnobotanical uses and that not all the reported ethnobotanical uses are accompanied by a strong phytochemical analysis capable to explain the relative utilizations.

C. cashmeriana
In Kashmir, the species is mainly an ornamental plant [150].

C. chengiana
This species has been traditionally used in Chinese folk medicine for the treatment of rheumatoid arthritis, pruritus, rheumatism, and pertussis [53].

C. dupreziana
In the Tassili N'ajjer desert in Sahara, the infusion of the leaves is drunk to treat fever [151].

C. funebris
In Vietnam, the cones are used to treat fever and physical injuries [152].

C. goveniana
In Myanmar, this species is said to be used for medicinal purposes, but the exact uses are not provided [153].

C. x leylandii
The leaves are used in Cameroon for screening or hedging [154].

C. lusitanica
The leaves and cones of this species are used in Kenya to treat liver, spleen, kidney, bladder, bone and joint diseases as well as antitumoral [155]. In Cameroon, this species is used in postpartum and against hair loss [156]. The decoction of the leaves is used to treat toothache in Ethiopia [157]. In the Bambotous department of Cameroon, the decoction of the leaves, whole plant and stems is used against postpartum pains and hair loss [158]. In Guatemala and Mexico, the crude extract of this species is used against cancer [159]. The decoction of the leaves is used in Ethiopia for the treatment of toothache [160].

C. macrocarpa
The leaves are employed by Baduga sub-tribe in India for the treatment of rheumatic muscular pains both externally and internally [161]. In Ecuador, the leaves and cones are used as infusions, vapors and plasters to treat wounds, cardiovascular and gastric problems, respiratory problems, inflammations, diarrhea, varix, phlebitis and as tonic [162].

C. nootkaensis
In the state of Washington, the branches are sued in baths to treat arthritis and rheumatism while their infusion is used to wash sores and swellings [124].

C. sargentii
The decoction of the branch apexes is used in Bolivia to treat cough [163]. In the same country, the species is also used to treat asthma [164].

C. torulosa
The seeds are used in Nepal against sinusitis and gingivitis [165]. In Kashmir, the species wood is used for general constructions and to make pencils and incense. In addition, its essential oil is used to cure inflammatory wounds and as an antiseptic [150].

Biological Activities
Several biological studies have been performed on Cupressus species other than C. sempervirens, evidencing interesting biological activities of their essential oils, extracts and derived compounds. These biological activities are mainly insecticidal, antibacterial, and antifungal, even if other properties have also been reported. The most extensive reports are on C. arizonica, C. lusitanica and C. marocarpa. The following sections explore the biological activities associated to Cupressus species other than C. sempervirens as reported in literature. Additionally in this case, it is important to underline that not all the Cupressus species have been studies in this sense and that not all the reported biological uses are accompanied by a strong phytochemical analysis capable to explain them.

Essential Oil
The essential oil from the leaves, branches and cones collected in Tunisia shows modest activity against Escherichia coli ATCC 25922, Salmonella typhimurium ATCC 19430, Staphylococcus aureus ATCC 25923, Enterococcus faecalis ATCC 29212, Klebsiella pneumoniae and Streptococcus pneumoniae, with MIC values ranging from 0.37 to 11.8 µg/mL. All these values are generally much lower than the standard compound levofloxacin in the relative assays (MIC values from 0.3 to 4.88 µg/mL) [21]. The essential oil from the cones collected from a cultivated population in Iran exerts strong growth inhibiting effects against Aspergillus flavus Link in a concentration-dependent manner according to the disc diffusion medium [22]. The essential oil from the leaves and cones collected from a cultivated population in Iran shows a modest antioxidant activity, according to the DPPH + assay, at the concentration of 4 µL/mL, with inhibition percentage values of 21% and 25.7%, respectively. The same essential oil shows very poor antioxidant activity according to the deoxyribose degradation assay. Lastly, the same essential oils show a moderate antioxidant activity according to the non-enzymatic lipid peroxidation test with inhibition percentages values of 27.1% and 31.6%, respectively, at the concentration of 0.05 µL/mL [23]. The essential oil from its leaves exerts strong larvicidal effects on Anopheles stephensi with a LC 50 value of 79.30 µg/mL [25]. The essential oil from the needles collected in Mississippi exhibits no significant antifungal, antibacterial, antimalarial and antileishmanial activities [26]. The essential oil from the leaves collected in Greece exerts medium larvicidal effects against third to fourth instar larvae of Aedes albopictus with LC 50 value of 64.8 mg/L. The same essential oil exerts weak repellent activity at the concentration of 0.2 mg/cm 2 [27]. The essential oil from the leaves collected in a Tunisian botanical garden exerts strong herbicidal activity. At the doses of 0.8 and 1.0 mg/mL, it totally inhibits the shoot and the root growth of Sonchus arvensis L., whereas only at the dose of 1.0 mg/mL does it totally inhibit the shoot and root growth of Trifolium campestre Schreb. At the same doses, it partially reduces the germination and the shoot and root growth of Phoenix canariensis Chabaud and Lolium rigidum Gaudin, too. The cone and stem essential oils were much less active [28]. The essential oil from the leaves of this species collected in Tunisia exhibits good antifungal properties against Candida albicans ATCC 18804, Candida glabrata 8D, Candida dubliniensis CIPO 82, Candida parapsilosis 28B, Candida tropicalis IGC3097, Candida bracarensis NCYC 3133 and Saccharomyces cerevisiae BY4741 with MIC values from 0.1 to 0.05 µg/mL. The mechanism of action is via an oxidative process leading to increased intracellular oxidation and DNA [29]. The essential oil from the cones collected in Lebanon exerts moderate antibacterial activities against Escherichia coli K12 strain carrying plasmid DH5-alpha/pTY250 (PTA-4079) and Staphylococcus epidermidis ATCC 14990 with MIC values of 200 µg/mL in both cases [166]. The essential oil from the leaves collected in Iran exhibit effects against two spotted spider mites on Alii fig leaf with 100% mortality starting from the concentration of 2.5 mg/L. This same essential oil also shows minimal toxic effects on humans and environment [167]. The essential oil of the leaves of this species obtained from a botanical garden in Iran shows moderate feeding deterring effects on Callosobruchus maculatus Fabricus, Sitophilus granarius L. and Oryzaphilus surinamensis L. with various percentages (10.66, 10.66 and 11.33%, respectively), as well as good inhibitory effects on their F1-progeny with various percentages (50.81, 59.64 and 49.99%, respectively) [31]. The essential oil of this species collected in Morocco exerts a high anticorrosion effect on carbon steel in 1.0 M HCl solution which is dependent on the temperature. The maximum efficiency of 97% is reached at the concentration of the essential oil equal to 500 ppm and at 25 • C. The mechanism of action is through the formation of a protective film on the carbon steel surface from the adsorbed components of the essential oil [34]. The essential oil of this species exhibits contact insecticidal activity on the larvae of the 3rd, 4th and 5th instars of Tortrix viridana. The concentration of 0.5% showed the best results with MMT of 1h 31 min 12 s [168]. The essential oil from the aerial parts collected in Iran exerts high toxic fumigant effects against adult rice weevil (Sitophilus oryzae L.) with a LC 50 value of 172.36 µL/L air [36]. The essential oil from the cones collected in Iran exerts strong antinociceptive and anti-inflammatory activities in mice. In particular, according to the formalin test, it is able to reduce the nociceptive responses especially at the concentration of 0.5 g/Kg in the late phase with values similar to those of diclofenac acting at the level of opioid and benzodiazepine receptors. With respect to the anti-inflammatory activity, according to the carrageenan induced paw edema assay, at the doses of 1, 0.5 and 0.25 g/Kg, the essential oil is able to significantly decrease the paw edema especially after 4 h from carrageenan injection. The values of these reduction are very similar among them and also with respect to diclofenac [37].

Solvent Extracts
The methanolic extracts of the leaves and cones of this species obtained from a botanical garden in Iran show moderate feeding deterring effects on Callosobruchus maculatus Fabricus, Sitophilus granarius L. and Oryzaphilus surinamensis L. with various percentages (13.66, 13.66 and 14.66%, respectively) as well as good inhibitory effects on their F1-progeny with various percentages (49.74, 47.24 and 50.65%, respectively) [31]. The ethanolic extract from the wood knots collected from a cultivated population in Iran exhibits good DPPH .+ effects with a percentage of inhibition equal to 47.99 at the volume of 0.1 mL and DPPH .+ concentration of 0.5 mg/mL. This value is comparable to that of BHA (57.96%). It also exerts very strong Fe 2+ chelating capacities with a percentage of 100% at the volume of 150 µL and Fe 2+ concentration of 2 mg/mL [33].

C. arizonica var. glabra
The essential oils from the female cones, male cones, needle twigs and wood barks of this species collected in South Carolina show good to moderate toxicity activity against first-instar larvae of Aedes aegypti at 24 h post treatment, with LD 50 values ranging from 33.7 to 55.5 ppm. Indeed, only the needle twig and wood bark essential oils show good deterrent activities on Aedes aegypti females, at 100 µg/cm 2 , with BDI values of 1.04 and 1.01, respectively. Lastly, only the wood bark essential oil exerts modest activity against Colletotrichum acutatum, Colletotrichum fragariae and Colletotrichum gloeosporioides with inhibition zone diameter values equal to 4.0, 4.5 and 4.0 mm, respectively, at the concentration of 320 µg/spot [39].

Essential Oil
The essential oil from the leaves of this species collected in Morocco has shown significant bacteriostatic action against Klebsiella pneumoneae, Proteus mirabilis, Escherichia coli and Staphylococcus aureus bacterial strains with diameters of inhibition zones between 17.6 and 28.4 mm and presented MIQ values between 80 and 140 µL [63]. The essential oils derived from the cones, branches and sawdust of this species collected in Morocco show some concentration-dependent antibacterial, antifungal and antimould effects on Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Micrococcus luteus, Aspergillus niger, Penicillium digitatum, Penicillium expansum, Coniophora puteana, Coriolus versicolor, Gloeophyllum trabeum and Poria placenta [64]. The essential oils from the leaves, twigs, cones and heartwood of this species collected in Morocco show moderate to weak antifungal activities against Gloeophyllum trabeum, Oligoporus placenta, and Coniophora puteana with MIC values from 900 to 1900 ppm obtained by micro atmosphere technique on Malt-Agar medium [65].

Solvent Extracts
The polyphenol rich extract (10% w/v) from the plant pruning wastes recovered from leaves, branches twigs, needles and cones collected in Morocco exerts good antifungal activity. At the concentration of 4%, it inhibits the Fusarium oxysporum f.sp. albedinis mycelial growth by about 61% as well as its spore germination by about 68% and its sporulation by about 45%. The same extract shows high antioxidant effects according to the TEAC and FRAP methods with values equal to about 25,000 and 35,000 µTE/100g DW, respectively [169].

Essential Oil
The essential oil of the wood purchased in China shows high repellency effects on Amblyomma americanum and Ixodes scapularis nymphs with an EC 95 value of 0.43 mg oil/cm 2 filter paper. DEET has an EC 95 value of 0.68 mg oil/cm 2 filter paper. At the concentration of 0.103 mg oil/cm 2 filter paper, the essential oil repels all I. scapularis nymphs. At the concentration 0.827 mg oil/cm 2 filter paper, the essential oil repels more than 80% A. americanum 4 h after application. The same essential oil causes a mortality of 80% to Aedes aegypti larvae at 250 ppm. This concentration is much higher than that obtained for permethrin (0.25 ppb with mortality of 67%) [70] (Carrol et al., 2001).

Solvent Extracts
The methanolic, hydro-methanolic and aqueous extracts of this species collected in Vietnam exert very poor antiproliferative effects on human HT-1080 fibrosarcoma cells with EC 50 values much above 100 µg/mL [152].

C. × leylandii
Several aqueous extracts from the leaves of this species, macerated for different periods, are able to delay the germination of Trifolium repens L. by at least 1.5 days as well as the germination of Lepidium sativum L. by at least 2 days. The same macerates can also slow down the appearance of their seedlings [79]. 5.6. C. lusitanica 5.6.1. Essential Oil The essential oil from the leaves and the cones of this species collected in Cameroon shows a dose-dependent anti-dermatophytic activity on Microsporum audouinii (cp22), Microsporum langeronii (cp46), Trichophyton rubrum (cp21) and Trichophyton mentagrophytes (cp38). In addition, the leaf essential oil highly prevents the growth of Microsporum audouinii (cp22), Microsporum langeronii (cp46), Trichophyton rubrum (cp21) at the doses of 500 and 1000 µg/mL. At the dose of 500 µg/mL, it also shows fungicidal effect while at the dose of 275 µg/mL, it is fungistatic [81]. The essential oil from the leaves collected in Costa Rica exerts good antibacterial and antifungal effects against Bacillus cereus ATCC14579 and Aspergillus niger ATCC1688 with MIC values equal to 78 µg/mL both [84]. The essential oil from the cones collected in India shows moderate antibacterial effects on Staphylococcus aureus, Bacillus subtilis, Escherchia coli and Salmonella typhi with MIC values above 125 µg/mL.
Amoxycillin trihydrate show much lower MIC values (3.125, 6.25, 6.25 and 50, respectively). The same essential oil also has moderate antifungal effects on Candida albicans and Candida neoformans with MIC values of 250 µg/mL, compared to Amphotericin B which has much lower MIC values (12.5 and 6.25, respectively). This essential oil also has a big effect on excision wound with infection with a percentage of wound contraction after 8 days from incision of 51.0% and after 12 days of 84.4%. The healing is characterized by deposition of collagen fibers and maturation of fibrous connective tissues with no hair follicles in the healed area and with a big keratinization of grown epidermis [170]. The essential oil from its leaves collected in Brazil showed moderate activity against the growth of its endophytic fungi associated Xylaria sp1 (MIC = 500 ppm), Guignardia sp. and Xylaria sp2 (MIC = 250 ppm) [87]. The essential oil from the leaves of this species collected in Cameroon exhibits good concentration dependent effects against  [87]. The essential oil from whole plant, leaf and cone extracts of this species collected in Cameroon exert very poor effect against Salmonella typhi ATCC 6539 with MIC values above 2048 mg/mL [156]. The leaf essential oil of this species collected in Kenya is a strong insect pest contact toxicant depending on the insect species, duration of exposure and concentration according to the instant toxicity assay. In particular, it is highly toxic on Sitotroga cerealella and Acanthoscelides obtectus with LC 50 values of 0.05 and 0.11% v/w after 24 h from contact, respectively, and less toxic on Tribolium castaneum and Sitophilus zeamais with LC 50 values of 0.11 and 0.13% v/w after 168 h from contact, respectively. The same essential oil is also a very good space fumigant with LC 50 values of 4.08 and 4.71 µL/L air against A. obtectus and S. cerealella, respectively, 24 h post fumigation and LC 50 values of 13.54 and 15.28 µL/L air against S. zeamais and T. castaneum, respectively, 168 h post fumigation. This essential oil is also a strong repellent to T. castaneum at 0.20% v/w with a value of 92.5% and decreases the PR values also for A. obtectus, S. cerealella and S. zeamais, 24 h after exposure [88].

Solvent Extracts
Several fractions derived from n-hexane leaf extract of this species collected inCameroon and differently enriched in essential oil compounds, show antifungal activity against Microsporum audouinii, Microsporum langeronii, Microsporum canis, Trichophyton rubrum and Trichophyton tonsurans with MIC values ranging from 125 to 500 µg/mL [82]. The chloroform, methanolic and ethyl acetate extracts from the leaves collected in Tanzania show weak or very good antibacterial and antifungal activities against Escherichia coli ATCC11775, Klebsiella pneumoniae ATCC13883, Pseudomonas aeruginosa ATCC27853, Salmonella typhi ATCC6539, Klebsiella oxytoca, Salmonella kisarawe, Candida albicans and Cryptococcus neoformans with MIC values ranging from 25.0 to 1.56 mg/mL. The chloroform, methanolic and ethyl acetate extracts from the covers of the seeds collected in Tanzania show weak or good antibacterial and antifungal activities against the same strains with MIC values ranging from 25.0 to 6.25 mg/mL. The chloroform and methanolic extracts from the seeds collected in Tanzania show weak or moderate antibacterial and antifungal activities against the same strains with MIC values ranging from above 25.0 to 3.12 mg/mL [171]. The ethanol extract from the leaves of this species collected in Cameroon shows high antifungal activity on Colletotrichum lindemuthianum and different Fusarium spp., especially at the concentration of 8 mg/mL, which resulted in 100% growth inhibition. The aqueous extract was not so active against the same fungi giving 86.08% of growth inhibition at the concentration of 30 mg/mL [172].

C. lusitanica var. benthamii
The essential oil from the leaves of this species collected in Greece exerts moderate larvicidal effects against third to fourth instar larvae of Aedes albopictus with LC 50 value of 37.5 mg/L. The same essential oil exerts strong repellent activity at the concentration of 0.2 mg/cm 2 [27].

Essential Oil
The essential oil from the leaves collected in Egypt shows good DPPH .+ antioxidant effects with an inhibition percentage of 68%. This value is comparable to that of BHA (87%). In addition, the same essential oil exerts good antibacterial and antifungal effects on Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Aspergillus niger, Aspergillus parasiticus and Candida albicans with MIC values from 0.65 to 0.75 mL/100 mL. The ethanol and dichloromethane extracts of the same leaves show good DPPH .+ antioxidant effects with inhibition percentages of 70 and 65%, respectively. The same extracts also show good antibacterial and antifungal effects on the same strains with MIC values from 0.55 to 0.8 mL/100 mL [96]. The essential oil derived from the cones of this species collected in India exerts potent antimicrobial and antifungal properties against Bacillus coagulans, Bacillus megaterium, Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Kleibseilla pneumoniae, Pseudomonas aeruginosa, Salmonella typhii. Aspergillus flavus, Candida albicans, Trichoderma lignorium and Cryptococcus neoformans with inhibition zone values between 11 and 17 mm [97]. The essential oil from the leaves of this species collected in Egypt exerts toxicity effect against the third instar larvae of Synthesiomyia nudiseta exposed for 24 h with an LC 50 value of 1.11%. In addition, it is able to decrease the percentage of pupation by 91.57%. The treatment with this volatile oil decreases the survival of the larvae and leads to noticeable larval, pupal, and adult abnormalities, especially at the level of the integument, muscles, fat body cells, midgut and salivary gland [98]. The essential oil from the leaves of this species collected in Greece exerts medium larvicidal effects against third to fourth instar larvae of Aedes albopictus with LC 50 value of 54.6 mg/L. The same essential oil exerts strong repellent activity at the concentration of 0.08 mg/cm 2 [27]. The essential oil from the leaves of this species shows very good antifungal activity against Alternaria alternata, Botrytis cinerea and Fusarium oxysporum with EC 50 values of 182, 181 and 109 mg/L respectively. The same essential oil shows moderate antibacterial activity against Agrobacterium tumefaciens and Erwinia carotovora var. carotovora. with MIC values of 600 and 525 mg/L, respectively. This essential oil also shows good and moderate inhibitory effects on the spore germinations of Alternaria alternata, Fusarium oxysporum and Fusarium solani with EC 50 values of 318, 199 and 485 mg/L, respectively [100]. The essential oil from the leaves collected in Egypt exerts mild fumigant toxicity effects against the fourth larval stage of Spodoptera littoralis with a LC 50 value equal to 9.91 µL/L air. The same essential oil exerts modest fumigant toxicity effects against the adults of Theba pisana with a LC 50 value equal to 11.67 µL/L air [103]. The essential oil from the leaves of this species collected in Mauritius shows good antibacterial effects against Escherichia faecalis, Escherichia coli ATCC25922, Staphylococcus aureus ATCC25923, Pseudomonas aeruginosa ATCC27853, Staphylococcus epidermidis ATCC12228 with inhibition zone diameters ranging from 16.1 for the first one to 24.0 for the second one. The same essential oil shows also fungicidal activity against Candida albicans ATCC10231, Candida tropicalis ATCC750, Aspergillus niger ATCC16404 and Trichophyton mentagrophytes ATCC9533. The same essential oil shows weak anti-tyrosinase activity with an IC 50 value of 70.98 µg/mL [104]. The essential oil from its leaves collected in Lebanon exerts good antimicrobial properties against Trichophyton rubrum SNB-TR1, Trichophyton mentagrophytes SNB-TM1, Trichophyton soudanense SNB-TS1, Trichophyton violaceum SNB-TV1 and Trichophyton tonsurans SNB-TT1 with MIC values of 64, 64, 32, 32 and 64 µg/mL, respectively [104]. The essential oil from the leaves of this species collected in Egypt exhibit promising concentration dependent effects against Fusarium solani, Fusarium oxysporum NRRL 28184, Aspergillus niger NRRL 599, Candida albicans NRRLY-477, Pseudomonas aeruginosa NRRLB-23, Staphylococcus aureus NRRLB-313, Escherichia coli NRRLB-210 and Bacillus subtilis NRRL-543 with inhibition zone diameters ranging from 10.5 to 21 mm [105]. The essential oil from the leaves collected in Egypt exerts good toxic effects on Culex pipiens adults after 24 and 48 h of exposure with LC 50 values of 0.12 and 0.11 mg/L, respectively. Indeed, the same essential oil exerts very poor toxic effects against fourth instar larvae of Culex pipiens after 24 and 48 h of treatment with LC 50 values of 67.37 and 46.07 mg/L, respectively [173]. The essential oil from the leaves collected in Egypt possesses good antioxidant activities according to the DPPH .+ radical scavenging and β-carotene-linoleic acid methods, with IC 50 values of 6.1 and 4.2 mg/L, respectively. BHT shows IC 50 values of 2.9 and 2.6 mg/L, respectively. The same essential oil also shows good antibacterial activities against Bacillus cereus ATCC14579, Micrococcus flavus ATCC10240, Listeria monocytogenes (clinical isolate), Staphylococcus aureus ATCC6538, Pseudomonas aeruginosa ATCC 27853, Dickeya solani DS0432-1 and Escherichia coli ATCC35210 with MIC values from 0.63 to 0.38 mg/mL. The same essential oil also shows good antifungal activities against Aspergillus flavus ATCC9643, Aspergillus ochraceus ATCC12066, Aspergillus niger ATCC 6275, Penicillium ochrochloron ATCC 48663, Penicillium funiculosum ATCC56755 and Candida albicans ATCC 12066 with MIC values from 1.53 to 0.37 mg/mL. Lastly, the same essential oil exerts moderate antitumoral activity against MCF-7, HeLa and Jurkat with IC 50 values of 25.4, 24.16 and 30.54 µg/mL, respectively, as well as weak antitumoral activities against HT-29 with an IC 50 value of 124.8 µg/mL [107]. The essential oil from the branchlets of this species collected in Egypt shows medium antibacterial and antifungal effects on Agrobacterium tumefaciens, Bacillus cereus, Dickeya solani, Escherichia coli, Pectobacterium atrosepticum, Pectobacterium carotovorum and Staphylococcus aureus with MIC values between 0.07 and 0.31 mg/mL. The same essential oil exerts only weaker antifungal effects on Aspergillus flavus, Aspergillus ochraceus, Aspergillus niger, Candida albicans, Fusarium oxysporum, Penicillium funiculosum and Penicillium ochrochloron with MIC values between 0.29 and 3.21 mg/mL. In addition, it exhibits good DPPH antioxidant activities, with an IC 50 value equal to 6.1 µg/mL, which is, anyway, lower than BHT (IC 50 = 2.9 µg/mL) [108]. The essential oil of the leaves collected in Mauritius shows moderate antioxidant effects according to several assay such as DPPH .+ , ABTS, xanthine oxidase, nitric oxide, hydroxyl radical, FRAP and ORAC with IC 50 values of 3.667 µg/mL, 1.068 µg/mL, 1.546 µg/mL, 1.258 µg/mL, 106.87 µM Fe 2+ /mg EO and 0.925 g TE/µg EO, respectively. In addition, its TPC value is 1223.17 µg GAE/µg EO, and it shows modest antiglycation properties with an IC 50 value of 451.53 µg/mL [109]. The essential oil from the leaves collected from Egypt exerts decent acaricidal effects against the adults of Tetranychus urticae with a LC 50 value equal to 5.69 mg/L air after 24 h of treatment using a fumigant assay. This activity is also confirmed by the slide dip technique assay with a LC 50 value equal to 170.07 mg/L after 24 h of treatment and 22.76 after 48 h of treatment. The same essential oil also exerts moderate inhibitory effects against AChE from the adults of Tetranychus urticae with a IC 50 value equal to 10.0 mg/L [110]. The essential oil from the leaves of this species obtained from an Egyptian botanical garden shows moderate insecticidal activity against Sitophilus oryzae and Tribolium castaneum with LC 50 values of 55.2 and 255.4 µg/cm 3 , in the 3 days post exposure test. The same essential oil exerts quite weak acetylcholinesterase inhibitory activity with an IC 50 value of 350 ppm [111]. The essential oil from the leaves of this species collected in Turkey as a dietary supplement exerts stress-relieving effects in common carp at low concentrations (0.5%) by increasing the total protein, albumin concentrations and growth performance parameters as well as by reducing the serum triglycerides, cholesterol levels and the activities of the serum liver enzymes (GOT, GPT, lactate dehydrogenase) [114]. The essential oil from the leaves of this species collected in Cameroon exerts medium larvicidal and antiplasmodial effects with LD 50 and IC 50 values of 60.45 and 147.29 µg/mL, respectively [115].

Solvent Extracts
The methanolic extracts of the leaves, barks and roots collected in Cameroon exert from poor to moderate cytotoxic effects against THP-1, DU145, HeLa, MCF-7 and HepG2 cancer cell lines with IC 50 values from above 400 to 60.8 µg/mL. These values are much higher than those reported for doxorubicin (from 5.5 to below 3.3 µg/mL). The same extracts also show poor anti-gonorrheal effects against different Neisseria gonorrhoeae strains with MIC values from above 512 to 128 µg/mL. These values are much higher than those reported for gentamicin (from 32 to 0.5 µg/mL). Lastly, the same extracts show poor reverse transcriptase activity with inhibition percentages equal to 16.75, 6.07 and 8.16%, respectively, at the concentration of 200 µg/mL. These values are much lower than those reported for doxorubicin (91.22%) [85]. In rats, the combined methanol and petroleum ether extracts of the leaves and the methanol extract of the roots of this species collected in Egypt exhibit strong protective effects on kidney after gentamicin induced nephrotoxicity. Indeed, they are able to significantly reduce the serum blood urea nitrogen and creatinine levels, to enhance the gene expression of AMPK-α1, to downregulate iNOS and to decrease NF-κB [112]. The methanolic extract of the leaves of this species collected in Egypt shows weak DPPH .+ free radical scavenging activity, with a value of 0.20 mg AAE/g [117]. The petroleum ether fraction of its leaves collected in Algeria has good acetylcholinesterase inhibitory properties with an IC 50 value of 88.79 µg/mL. The same petroleum ether fraction exerts modest antibacterial activity against Methicillin-resistant Staphylococcus aureus (MRSA) ATCC33591 with an IC 50 value of 123.0 µg/mL [117]. The methanolic and diethyl ether extracts of the leaves collected in Egypt exert from strong to moderate antibacterial effects against 41 MRSA isolates with MIC values from 2 to 8 µg/mL for the latter and 256 to 1024 µg/mL for the former, to be compared to vancomycin that shows MIC values from 0.5 to 4 µg/mL. The same diethyl ether extract is able to reduce the growth of all the isolates by 48.78% and the efflux pump of 12 of them as well as to produce a certain degradation of the cell walls. The mechanism of action is in vitro via a significant down expression of norA and norB genes and in vivo by regenerating the epidermis, maturing the granulation tissue and reducing the inflammatory cell infiltration in rats. In rats, this extract is also able to contract wounds by 83.9% and to reduce the size of the injured area by regenerating thin epidermis after 7 days of treatment. The same extract has modest cytotoxic effects on HSF cell line with an IC 50 value of 21.3 µg/mL. Doxorubicin shows a value of 4.36 µg/mL [119]. The methanolic extract of the roots collected in Egypt exerts from good to moderate antimicrobial effects on several Salmonella enterica isolates with MIC values from 64 to 1024 µg/mL. This extract significantly decreases the bacterial membrane of all the isolates as well as the membrane depolarization (40%) and efflux activity. It also significantly increases the inner and outer membrane permeability of all the isolates. The reduction of the biofilm formation at concentration from 32 to 512 µg/mL occurs only in eight isolates. According to the castor-oil induced diarrheal model, the same extract also shows good antidiarrheal effects by reducing the rate of defecation (85.9% at 100 mg/Kg body weight, 88% at 200 mg/Kg body weight, 93.5% at 100 mg/Kg body weight). The value of this rate for loperamide is 91.5% at 3 mg/Kg body weight. This extract has an inhibitory effect on the intestinal transit of charcoal in a dose-dependent manner (64.95% at 100 mg/Kg and 80.24% at 200 mg/Kg) and increases the in vivo antidiarrheal index in the same way [120].

C. nootkaensis
The methanol extract from the outer bark shows high antibacterial effect against Mycobacterium tuberculosis H37Rv with a percentage of inhibition equal to 98% [124]. The essential oil from the leaves of this species collected in Greece exerts medium larvicidal effects against third to fourth instar larvae of Aedes albopictus with LC 50 value of 57.1 mg/L. The same essential oil exerts weak repellent activity at the concentration of 0.2 mg/cm 2 [27]. The essential oil from the leaves showed only moderate antibacterial activities against different bacterial strains such as Aspergillus niger, Aspergillus terreus, Candida spp., Trichophyton rubrum, Trichophyton mentagrophytes, Microsporum canis, Pencillium crysogensum, Pencillium expansum and Pencillium griseofulvum with inhibition zone values equal or above 10 mm. Indeed, it showed good antibacterial effects against Microsporum audouinii with an inhibition zone value of 7 mm [129]. Various solvent extracts and the essential oil from the leaves of this species collected in India possess medium to high antibacterial effects against Micrococcus luteus, Staphylococcus aureus, Campylobactor coli, Bacillus subtilis, Bacillus cereus, Alcaligenes denitrificans, Pseudomonal aeruginosa and Pseudomonas alcaligenes with mean diameters of zone inhibition from 5 to 21 mm at the concentration of 50 mg/mL, being the essential oil the most powerful. All these values are comparable to those of chloramphenicol. The same extracts and essential oil also have antifungal activity against Alternaria alternata, Curvularia lunata and Bipolaris specifera with IC 50 values ranging from 0.19 to 4.99 mg/mL. These values are in any case lower than amphotericin B, which is used as a standard [131].

Solvent Extracts
Different extracts of the needles of this species collected in India exert moderate to good antioxidant activities according to the DPPH, ABTS and SSA assay with IC 50 values from 34.93 to 83.83 µg/mL [130]. The methanolic extract of the leaves collected in Himalaya exerts modest antibacterial activities against Agrobacterium tumefaciens MTCC609, Escherichia coli MTCC40 and Xanthomonas phaseoli with MIC values of 250, 500 and 500 µg/mL, respectively [174]. The leaf ethanolic extract of this species collected in India shows effects against Pectobacterium carotovorum subsp. carotovorum with a diameter of inhibition zone equal to 18.66 mm according to the agar well diffusion method and a diameter of inhibition zone equal to 13 mm according to the disc diffusion method [175]. The methanol extract from the leaves collected in Himalaya exerts modest antifungal activities against Alternaria alternata, Colletotrichum falcatum, Fusarium oxysporum, Pyricularia oryzae and Sclerotinia rolfsii with inhibition percentages of 49, 34, 33, 33 and 49%, respectively. The ethanol extract from the same leaves exerts moderate antifungal activities against Alternaria alternata, Colletotrichum falcatum, Pyricularia oryzae and Sclerotinia rolfsii with inhibition percentages of 46, 40, 27 and 51%, respectively. The chloroform extract from the same leaves exerts good antifungal activities against Alternaria alternata, Colletotrichum falcatum, Fusarium oxysporum and Sclerotinia rolfsii with inhibition percentages of 46, 47, 32 and 63%, respectively. Lastly, the n-hexane extract from the same leaves exerts modest antifungal activities against Alternaria alternata, Colletotrichum falcatum and Sclerotinia rolfsii with inhibition percentages of 39, 51 and 42%, respectively [176]. The methanolic extract of the leaves collected in Nepal show good antibacterial effects against Staphylococcus aureus, Streptococcus pyogenes, Acinetobacter calcoaceticus and Escherechia coli with inhibition zone diameter values of 14, 15, 12 and 14.5 mm, respectively, at the concentration of 200 mg/mL. The ethyl acetate extract of the same leaves shows moderate antibacterial effects against the same strains with inhibition zone diameter values of 12, 7.5, 7 and 10.5 mm, respectively, at the same concentration. Lastly, the n-hexane extract of the same leaves shows moderate antibacterial effects against only Staphylococcus aureus, Streptococcus pyogenes and Acinetobacter calcoaceticus with inhibition zone diameter values of 7, 9 and 10 mm, respectively, at the same concentration [132].

Conclusions
In this review article, the phytochemical analyses, ethnobotanical uses and pharmacological activities relative to Cupressus species other than Cupressus sempervirens were described. Several phytochemical compounds have been evidenced in these other Cupressus species mainly essential oil components, diterpenoids and flavonoids. For what concerns the traditional medicine uses, only ten species are reported to be employed in this sense for a certain variety of ailments including rheumatisms and fevers. As for the pharmacological activities associated to these species, only ten species have shown to have several activities including antibacterial, antiviral, anti-inflammatory and antitumoral. Additionally, some compounds isolated from these species have been shown to possess important biological effects (mainly anti-inflammatory and antitumoral), which may justify the ethnobotanical uses of the species where they have been evidenced. However, there is still a large absence of knowledge of these species in several fields and this review article may represent a further reason to study all the Cupressus species in major detail.