A Review of the Phytochemistry, Traditional Uses, and Biological Activities of the Genus Ballota and Otostegia.

The 2 genera Ballota and Otostegia, belonging to the Lamiaceae family, are closely related taxonomically and found mainly in the Mediterranean area, Middle East, and North Africa. Since ancient times, they have been largely employed in traditional medicine for their biological properties such as antimicrobial, anti-inflammatory, antispasmodic, insecticidal, anti-malaria, etc. Phytochemical investigations of Ballota and Otostegia species have revealed that diterpenoids are the main constituents of the genera. A large number of flavonoids and other metabolites were also identified. This review, covering literature from 1911 up to 2018, includes traditional uses, chemical profiles (both of volatile and nonvolatile metabolites), and biological properties of all the taxa of these 2 genera studied to date.

ANTIOXIDANT ACTIVITY OF TISSUE CULTURE-RAISED BALLOTA NIGRA L. PLANTS GROWN EX VITRO.

Antioxidant properties and total phenolic and flavonoid contents were evaluated in methanolic extracts of shoots from Ballota nigra plants initiated in vitro (from nodal explants) and in vivo (from seeds). The plants were grown in greenhouse and in the field, and were analyzed at the vegetative and flowering stages. The shoot extract of wild-grown plants of B. nigra was also investigated. The results indicate that antioxidant potential of the B. nigra extracts seems to be due to their scavenging of free radicals (DPPH assay) and metal reducing (FRAP test), while they were less effective at the prevention of linoleic acid peroxidation (LPO test). The extracts from shoots of in vitro derived plants were found to exhibit the greatest antioxidant properties. The extracts were also characterized by the highest content of phenolic compounds and their level was affected by plant developmental stage. The extracts of shoots collected at the flowering period exhibited higher amounts of phenolics and flavonoids than in the extracts of immature plants. A close correlation between the total phenolic content and flavonoid content and antioxidant activity using the DPPH and FRAP assays was obtained. The results of the present study suggest the use in vitro-derived plants of B. nigra instead of using wild plants for pharmaceutical purposes.

Essential oil composition and antifungal activity of aerial parts of Ballota nigra ssp foetida collected at flowering and fruiting times.

The present study reports the results of gas chromatographic-mass spectrometric (GC/MS) analyses of the essential oils from the aerial parts of Ballota nigra L. ssp foetida (Lamiaceae) collected at flowering and fruiting times, as well as their in vitro antifungal activity against nine plant pathogenic fungi. Moreover, the essential oils were evaluated for their antifungal activity using the agar dilution method, and also MICs (minimum inhibitory concentrations) and MFCs (minimum fungicidal concentrations) were determined. The major compounds identified in the flowering and fruiting aerial parts oils respectively were beta-caryophyllene (22.6% and 21.8%), caryophyllene oxide (18.0% and 20.5%) and germacrene-D (16.5 and 13.1%). The oils showed in vitro antifungal activity against some species of Fusarium, Botrytis cinerea, and Alternaria solani. Our study indicates that the oil of B. nigra ssp foetida could be used as a control agent for plant pathogenic fungi in natural formulations.

In vitro antimicrobial and antiprotozoal activities, phytochemical screening and heavy metals toxicity of different parts of Ballota nigra.

The study was done to assess the phytochemicals (flavonoids, terpenoids, saponins, tannin, alkaloids, and phenol) in different parts (root, stem, and leaves) of Ballota nigra and correlated it to inhibition of microbes (bacteria and fungi), protozoan (Leishmania), and heavy metals toxicity evaluation. In root and stem flavonoids, terpenes and phenols were present in ethanol, chloroform, and ethyl acetate soluble fraction; these were found to be the most active inhibiting fractions against all the tested strains of bacteria, fungi, and leishmania. While in leaves flavonoids, terpenes, and phenols were present in ethanol, chloroform, and n-butanol fractions which were the most active fractions against both types of microbes and protozoan (leishmania) in in vitro study. Ethanol and chloroform fractions show maximum inhibition against Escherichia coli (17 mm). The phytochemical and biological screenings were correlated with the presence of heavy metals in selected plant Ballota nigra. Cr was found above permissible value (above 1.5 mg/kg) in all parts of the plant. Ni was above WHO limit in B. nigra root and leaves (3.35 ± 1.20 mg/kg and 5.09 ± 0.47 mg/kg, respectively). Fe was above permissible value in all parts of B. nigra (above 20 mg/kg). Cd was above permissible value in all parts of the plant (above 0.3 mg/kg). Pb was above WHO limit (above 2 mg/kg) in all parts of Ballota nigra.

Two new ballonigrin-type diterpenoids from the roots of Ballota limbata.

Two new ballonigrin type lactone diterpenoids, named ballonigrin lactone A and B, have been isolated from the roots of Ballota limbata. Structure elucidation of the isolated compounds was based on spectroscopic {IR, 1H- and 3C-NMR, and 2D-NMR (HMQC, HMBC, COSY and NOESY} and EI-MS data.

Investigation of antitussive and toxicological activity of Ballota limbata in mice.

CONTEXT: Ballota limbata Benth. (Lamiaceae) (syn, Otostegia limbata Hook.f.) is a species grown in the North West Frontier Province and the lower hills of West Punjab, Pakistan. Ballota species are renowned for their antispasmodic, antiulcer, diuretic, vermifuge, and especially sedative effects. However, little is known about the biological activity of B. limbata. OBJECTIVE: Evaluation of antitussive activity and safety profile of dried B. limbata extract. MATERIALS AND METHODS: Whole air-dried plants were partitioned with various solvents and the butanol fraction was subjected to antitussive evaluation using a sulfur dioxide (SO(2))-induced cough model in mice. Codeine and dextromethorphan were used as positive control. Safety profile of the testing material was established using standard toxicity tests. RESULTS: B. limbata extract inhibited cough provoked by SO(2) gas in mice in a dose-dependent manner. The extract exhibited maximum protection against SO(2)-induced cough after 60 min of administration. B. limbata offered maximum cough suppressive effects, that is, number of coughs during 60 min was 11.66 ± 1.2 (mean ± SEM), after s.c. administration of 800 mg/kg, as compared with codeine 10 mg/kg, s.c., dextromethorphan 10 mg/kg, s.c., and saline showing a frequency of cough of 11.75 ± 1.18, 12.25 ± 0.83, and 46.25 ± 1.52, respectively. LD(50) value of B. limbata was greater than 5000 mg/kg. No sign of neural impairment was observed at antitussive doses and the extract has been well-tolerated at higher doses. DISCUSSION AND CONCLUSION: This study demonstrates that the extract of B. limbata has shown strong cough suppressive effect in mice without yielding any notable toxicity.

Radical quenching activity, ferric-reducing antioxidant power, and ferrous ion-chelating capacity of 16 Ballota species and their total phenol and flavonoid contents.

The ethyl acetate, methanol, and water extracts of 16 Ballota species (Family Lamiaceae)-Ballota acetabulosa, Ballota antalyanse, Ballota cristata, Ballota glandulosissima, Ballota inaequidens, Ballota larendana, Ballota latibracteolata, Ballota macrodonta, Ballota nigra ssp. anatolica, B. nigra ssp. foetida, B. nigra ssp. nigra, B. nigra ssp. uncinata, Ballota pseudodictamnus ssp. lycia, Ballota rotundifolia, Ballota saxatilis ssp. brachyodonta, and B. saxatilis subsp. saxatilis-were screened for their 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical quenching, ferric-reducing antioxidant power, and ferrous ion-chelating capacity at 1mg/mL. Hispanolone, a major diterpene found in the Ballota genus, was also tested in the same manner. Total phenol and flavonoid contents of the extracts were determined by Folin-Ciocalteau and AlCl(3) reagents, respectively. The extracts showed insignificant quenching activity against DPPH radical, but they had moderate antioxidant activity (0.597 ± 0.03 to 1.342 ± 0.01) in the ferric-reducing test compared to chlorogenic acid (the reference compound) (3.618 ± 0.01). All of the extracts (ranging from 65.1 ± 0.64% to 96.3 ± 0.09%) and hispanolone (97.31 ± 0.30%) exerted a remarkable ferrous ion-chelating effect. The highest total phenol (gallic acid equivalent) and flavonoid (quercetin equivalent) contents were found in the ethyl acetate extract of B. glandulosissima (393.7 ± 3.03 and 140.6 ± 1.97 mg/g of extract, respectively). Therefore, Ballota species could be a good source of natural preservatives in foodstuffs.

Essential oil composition and antimicrobial activity of Ballota nigra L. ssp foetida.

The chemical composition of the essential oil of Ballota nigra L. ssp foetida obtained from the flowering aerial parts was analyzed by GC/MS. From the 37 identified constituents of the oil, beta-caryophyllene (20.0%), germacrene D (18.0%) and caryophyllene oxide (15.0%) were the major components. The oil was active against both Gram-negative and Gram-positive bacteria as well as against three Candida species.

Determination of the antimicrobial and antioxidative properties and total phenolics of two "endemic" Lamiaceae species from Turkey: Ballota rotundifolia L. and Teucrium chamaedrys C. Koch.

This study was designed to examine the in vitro antimicrobial and antioxidant activities and the amount of total phenolics of the methanol extracts of Ballota rotundifolia L. and Teucrium chamaedrys C. Koch. In the case of antimicrobial activity tests, polar sub-fractions of the methanol extracts of plant species exhibited weakest antimicrobial activity when compared with the non-polar ones. While, non-polar sub-fraction of B. rotundifolia showed moderate activity against A. lwoffii, C. perfringens and the yeasts, T. chamaedrys performed excellent activity pattern against all of the tested microorganisms. The sub-fractions were also screened for their possible antioxidant activities by two complementary tests, namely DPPH free radical-scavenging and beta-carotene/linoleic acid assays. Non-polar extracts of the plant species remained inactive in both test systems. On the other hand polar extracts showed remarkable antioxidant activities. In DPPH system, free radical scavenging effect of T. chamaedrys was measured as 18.00 +/- 1.42 microg.mg(-1). It is extremely important to point out that, polar sub-fraction of T. chamaedrys is found as effective as the positive control BHT. Non-polar sub-fraction of T. chamaedrys found to have the highest total phenolic amount (97.12 +/- 1.28 microg/mg). Results obtained from this experiment confirm the relationship between the amount of phenolics and biological activities.

Antimicrobial activity of the essential oil obtained from roots and chemical composition of the volatile constituents from the roots, stems, and leaves of Ballota nigra from Serbia.

The chemical composition of essential oils obtained from the roots, stems, and leaves of Ballota nigra, growing in Serbia, was investigated by gas chromatography/mass spectrometry analyses. Kovats indices, mass spectra, and standard compounds were used to identify a total of 115 individual compounds. The plant produces two types of essential oils. Oils derived from stems and leaves were sesquiterpene rich (78.17% and 88.40%, respectively), containing principally beta-caryophyllene, germacrene D, and alpha-humulene, present in appreciable amounts. In contrast, oil derived from the root was dominated by p-vinylguiacol (9.24%), borneol (7.51%), myrtenol (7.13%), trans-pinocarveol (5.22%), pinocarvone (4.37%), 2-methyl-3-phenylpropanal (4.32%), and p-cymen-8-ol (4.30%). Essential oil obtained from the roots was evaluated for the antimicrobial activity against seven bacterial species and one fungi.

Antioxidant properties of Ballota species growing in Turkey.

In the present study, the antioxidant properties of ethanol extracts of 16 Ballota species belonging to the Lamiaceae family and growing in Turkey on superoxide anion formation and lipid peroxidation were investigated. All extracts inhibited superoxide anion formation but to different extents. The extracts of Ballota antalyense, Ballota macrodonta, Ballota glandulosissima, Ballota larendana, Ballota pseudodictamnus, Ballota nigra subsp. anatolica, Ballota saxatilis subsp. saxatilis, Ballota rotundifolia, and Ballota saxatilis subsp. brachyodonta exhibited remarkable anti-superoxide anion formation with IC(50) values from 0.50 to 0.87 mg/ml. The extracts of Ballota antalyense, Ballota macrodonta and Ballota glandulosissima have the lowest IC(50) values (0.50, 0.51 and 0.51 mg/ml, respectively) which were rather close to the well-known superoxide anion scavenger alpha-tocopherol (IC(50): 0.22 mg/ml). The extracts of Ballota inaequidens, Ballota glandulosissima, Ballota saxatilis subsp. saxatilis, Ballota macrodonta and Ballota antalyense inhibited lipid peroxidation with IC(50) values from 12 to 20 mg/ml. The extracts of Ballota inaequidens (IC(50): 12 mg/ml) and Ballota glandulosissima (IC(50): 15 mg/ml) exhibited the strongest inhibitory effects on lipid peroxidation compared with alpha-tocopherol (IC(50): 3 mg/ml). The results show that Ballota glandulosissima is the best antioxidant source among these 16 Ballota species.