Application of Herbal Essential Oil Extract Mixture for Honey Bees (Apis mellifera L.) Against Nosema ceranae and Nosema apis

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Plant properties and bioactivities considered in preparing HEOEM

Species	Major components	Bioactivities	References
Rumex acetosella	Oxalate (binoxalate) of potassium, tannic acid, and nitrogenous matter	Refrigerant, asidic and diuretic	https://www.henriettes-herb.com/eclectic/kings/rumex-acet.html
Achillea millefolium	Artemisia ketone, camphor, linalyl acetate and 1,8-cineole	Anti-inflammatory and disinfectant	Ghanbari et al., 2017
Plantago lanceolata	Flavonoids, alkaloids, terpenoids, phenolic compounds (caffeic acid derivatives), iridoid glycosides, fatty acids, polysaccharides and vitamins	Anti-inflammatory antimicrobial activity, antioxidant and cytotoxic activity, anti-tumoural activity and antispasmodic	Bajer et al., 2016
Salvia officinalis	Cis-thujone, camphor, cineole, humulene, trans-thujone, camphene, pinene, limonene, bornyl acetate and linalool	Antibacterial, antioxidant, antimicrobial, insecticidal and allelopathic activities antitumoral antidiabetic and antioxidant	Khedher et al., 2017
Thymus vulgaris	Geraniol, linalool, gamma-terpineol, carvacrol, thymol and trans-thujan-4-ol/terpinen-4-ol, p-cymene, γ-terpinene and thymol	Antimicrobial, antitussive, antibroncholitic, antispasmodic, anthelmintic, carminative, antiseptic, antioxidant, diuretic	Borugă et al., 2014
Rosmarinus officinalis	Rosmarinic acid, caffeic acid, ursolic acid, betulinic acid, carnosic acid and carnosol, camphor, 1,8-cineole, α-pinene, borneol, camphene, β-pinene and limonene	Antibacterial, antiviral, antioxidant and anti-inflammator, antidiabetic anti-inflammatory, antitumor and antioxidant	Andrade et al., 2018
Laurus nobilis	1,8-cineole, sabinene, and linalool	Antibacterial, antioxidant, antifungal, cytotoxic	Caputo et al., 2017

Differences between groups regarding application doses classified by Duncan test

Subgroups	Subset for alpha = .05	The number of Nosema spp. spores/bee±SE
Cages	N (Sampling days)	a*	b*	c*	d*	*e
ExpNa(2)	8	428703±21.3
ExpNc(2)	8	471887±24.2
ExpNa(1)	8		518812±28.1
ExpNc(1)	8			639750±24.6
PCNa	8				3337500±126.3
PCNc	8				3750000±131.7
NCNa	8					41519226±336.3
NCNc	8					40621143±321.8

Comparison of numbers of Nosema spp_ spores following varying HEOEM application methods analysed by Duncan test

Subgroups	Subset for alpha = .05	The number of Nosema spp. spores/bee (×106)±SE
Appl. methods	N (apiaries)	a*	b*	c*	d*
SW	9	1.23±0.1
SS	9		2.27±0.4
FS	9		2.60±0.2
PC	9			4.48±0.2
MVSN	9				10.26±1.8

Number of dead bees and final mortality rates (%) during the cage experiments

		Sampling Days							Total number of dead bees	Final mortality rates(%)
Cages	Cage number	3	6	9	12	15	18	21		(190 newly emerged bees at the beginning point x10 cages)
ExpNa(1)	1–10	5	15	33	48	63	85	93	342	18.0
ExpNa(2)	1–10	5	15	17	29	34	51	64	215	11.3
PCNa	1–2	0	6	15	11	20	27	53	132	34.7
NCNa	1–2	1	11	20	31	59	95	117	334	87.8
ExpNc(1)	1–10	8	14	32	39	85	108	165	451	23.7
ExpNc(2)	1–10	8	11	29	33	51	81	102	315	16.5
PCNc	1–2	0	13	15	22	30	48	70	198	52.1
NCNc	1–2	15	25	46	59	70	105	59	380	100
Native	1–2	1	5	8	10	12	10	15	61	16.0