Annals of Agrarian Science: N.L. Ghazanchyan, M.H. Kinosyan, P.E. Tadevosyan, N.S. Khachaturyan, E.G. Afrikian T
Annals of Agrarian Science: N.L. Ghazanchyan, M.H. Kinosyan, P.E. Tadevosyan, N.S. Khachaturyan, E.G. Afrikian T
Annals of Agrarian Science: N.L. Ghazanchyan, M.H. Kinosyan, P.E. Tadevosyan, N.S. Khachaturyan, E.G. Afrikian T
A R T I C LE I N FO A B S T R A C T
Keywords: The paper discusses the main features of Brevibacillus laterosporus bacilli as bioinsecticides for pest control, as
Entomopathogenes well as perspective sources for searching the novel cancerolytic substances of microbial origin. It has been
Brevibacillus laterosporus established that the culture of B. laterosporus is widespread in Armenia as a highly active producer of protein
Bacillus thuringiensis inclusions against leaf beetles. Isolated strains of B. laterosporus can be used as an alternative to biological
Bioinsecticides
larvicides, which are produced on the basis of B. thuringiensis and B. sphaericus.
Parasporins
Antimicrobial activity
https://doi.org/10.1016/j.aasci.2018.03.011
Received 18 February 2018; Accepted 29 March 2018
Available online 11 July 2018
1512-1887/ © 2018 Agricultural University of Georgia. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/BY-NC-ND/4.0/).
N.L. Ghazanchyan et al. Annals of Agrarian Science 16 (2018) 413–415
Antimicrobial activity microorganisms has been carried out. According to the results of this
study, B. laterosporus strains have demonstrated antagonism in relation
Antagonistic features of B. laterosporus strains in relation to gram to other species of Bacillus, as well as gram positive and gram negative
positive and gram negative bacteria as well as to fungi were studied by bacteria (see Table 2).
using the following test strains: Micrococcus luteus 5270, Bacillus subtilis The tested strains of B. laterosporus did not have influence on the
1820, B. cereus 2111, Staphylococcus aureus 5233, Mycobacterium sp. growth of E. coli 5002 and C. albicans 8013, fungi A. flavus, P. expansum
5253, Serratia marcescens 5251, E. coli 5002, Candida albicans 10231, and A. alternata. The antagonistic action of B. laterosporus strains can be
Streptococcus faecalis 5254, Pseudomonas fluorescens 5248, Aspergillus explained by produced metabolites and enzymes such as endotoxins,
unilateralis 10648, A. flavus 10676, Penicillium funiculosum 8189, P. bacteriocins, antibiotics, phospholipases, proteases, etc. [12,13] (see
expansum 8134, Alternaria alternata 8126 (MDC, Table 3).
SPC″Armbiotechnology” NAS RA). Insecticidal features. The insecticidal activity of B. laterosporus strains
Test culture suspension was added to nutrient agar and placed in was tested on leaf beetle of Coleoptera detachment, of Chrysomelidae
Petri dishes. B. laterosporus strains were sown on the agar surface by family. The results of testing demonstrated that influence of the strain
loop. 199-3 caused 100% death of the insect. The insecticidal activity of B.
laterosporus towards Coleoptera, but not towards Lepidoptera was shown
earlier [14,15].
Results and analysis
Today the great interest has been given to the parasporal bodies
possessing anticancer activity. Such parasporal bodies are produced by
During long-term work the vast collection of entomopathogenic
species and subspecies of Bacillus thuringiensis, L. sphaericus and related
bacilli (more than 5000 strains) isolated from various sources has been
species [16,17]. Our investigations of entomopathogenic Bacilli have
created in MDC. The physiological, biochemical and genetic char-
revealed that formation of crystalline protein bodies (parasporins) is
acteristics of these strains were thoroughly studied. The number of
indicative characteristic for these bacteria. Searching for new para-
strains were used in industrial production of insecticides. The collection
sporins derived from bacteria of genus Bacillus, having specific effect on
includes entomopathogenic strains active against a wide range of pest
certain tumor cell lines was provided [18].
and infection spreading mosquitoes, mites and other insects.
Study of parasporins produced by entomopathogenic bacteria paves
The new species of enthomopathogenic bacilli producing inclusions
the way to reveal novel anticancer drugs.
of protein nature were revealed by serological and modern methods of
molecular genotyping. 30 strains were isolated from various samples.
Isolated bacteria were identified as Brevibacillus laterosporus according
Conclusion
to morphological and physiological features and molecular genotyping.
All strains did not produce crystalline inclusions.
The species specificity of entomopathogenic Bacilli producing crys-
Morphological features. B. laterosporus is a rod-shaped, endospore-
talline toxins reflects their genetic patterns and is of basic importance
forming bacterium morphologically characterized by the production of
for directed screening of the strains needed. So, the strategy for ob-
a typical canoe-shaped parasporal body (CSPB) firmly attached to one
taining of new insecticides from Bacilli has to be based on isolation and
side of the spore, which determines its lateral position in the spor-
use of their new species and subspecies.
angium [10,11]. On agar surface B. laterosporus forms flat, smooth
Nevertheless among bacilli there are many other species to be stu-
(plain), white or beige colonies with rhizoid margins.
died and used as perspective source of new insecticides: many sub-
Utilization of carbohydrates. The utilization of 24 carbohydrates by
species or serovars of L. sphaericus, B. thuringiensis, B. lentimorbus, B.
strains of B. laterosporus was studied (Table 1).
laterosporus and related Bacilli producing specific entomocidal crystal-
According to auxanographic data, all tested strains of B. laterosporus
line toxins of proteinous nature or having infectious action on the in-
could not utilize 17 carbohydrates from 24: L-, D-arabinose, L-, D-lyxose,
sect.
D-fucose, L-rhamnose, L-sorbose, sucrose, D-melibiose, D-melezitose, D-
Today entomopathogenic aerobic spore-forming B. laterosporus
galactose, L-lactose, D-raffinose, inulin, D-sorbitol, D-mannitol, D-xy-
strains having high level of parasporal body production are very at-
litol. All tested strains were able to utilize fructose, glucose, trehalose,
tractive from the scientific and industrial point of view. The wide range
maltose. Four strains could not utilize ribose and mannose. The ob-
of entomopathogenic features of these strains should be used for pest
tained results have indicated the diversity of B. laterosporus species.
control. Biocide effect (antibacterial, fungicidal) allows recommending
Antagonistic features. Since the data on antagonistic features of B.
B. laterosporus strains as producers of biological means of plant pro-
laterosporus are limited, the study B. laterosporus strains antimicrobial
tection.
activity and spectrum of their influence on the other groups of
Table 1
Utilization of carbohydrates by B. laterosporus.
Strain number D- ribose D- glucose D- fructose D- mannose D- cellobiose D - trehalose D- maltose
59–1 – 2+ + + 3+ + +
69–1 2+ 2+ 3+ 3+ 3+ 3+ 3+
69-3-5 – + + + + + +
101–5 + 2+ 2+ + + + +
233–2 – 2+ 2+ – – + +
670–1 + + 2+ 2+ – + +
688–2 2+ 2+ 2+ + + 2+ 2+
194–1 2+ 2+ 3+ 2+ – 2+ 2+
199–3 + + + + + 2+ +
105–1 2+ + 2+ 2+ – 2+ 3+
209–3 2+ 2+ 3+ 2+ + 2+ 2+
186–1 – 2+ 2+ – 2+ + 2+
186–2 + 2+ 2+ – 2+ + –
Notes: ”+” - growth 1–10 mm; “2+” - 11–20 mm; “3+” - more than 21 mm; ” - ” no growth.
414
N.L. Ghazanchyan et al. Annals of Agrarian Science 16 (2018) 413–415
Table 2
Antimicrobial activity of B. laterosporus species.
No. of Species and No. of test-cultures
B. laterosporus
strains E.coli 5002 St.aureus S.marcescens Myc.sp. St.faecalis Ps. fluorescens B.subtilis B.cereus Mic.luteus C.albicans
5233 5251 5253 5254 5248 1820 2111 5270 8013
186–1 – 2+ + 3+ 3+ + + 3+ 3+ –
688–2 – 2+ + 3+ 3+ 2+ + 3+ 3+ –
69–1 – 2+ + 3+ 3+ + + 3+ 3+ –
184–2 – 2+ + 3+ 3+ 3+ 2+ 3+ 3+ –
194–1 – 2+ + 3+ 3+ 3+ + 3+ 3+ –
105–1 – 2+ + 3+ 3+ 2+ 2+ 2+ 2+ –
200–4 – + + 2+ 2+ + + – + –
199–3 – + – 2+ 3+ + + 3+ 2+ –
59–1 – + + 2+ 2+ 3+ + – + –
233–2 – + + 2+ 2+ 3+ + – + –
561–1 – + + 2+ 2+ 3+ + – + –
55–2 – + + 2+ 2+ 3+ + – – –
69-3-5 – 2+ + 2+ 2+ + 3+ 2+ 2+ –
209–3 – + + 2+ 2+ 3+ 3+ 2+ 2+ –
Notes: growth inhibition: “+” - 1–3 mm, “2+” - 4–7 mm, “3+” - 8–10 mm; no inhibition: “-”
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