Pathogenicity of an Indigenous Strain of the Entomopathogenic Fungus Metarhizium anisopliae (Hypocreales: Clavicipitaceae) (MET-GRA4 Strain) as a Potential Biological Control Agent Against the Red Palm Weevil (Coleoptera: Dryophthoridae)

Author(s):  
I Ishak ◽  
L C Ng ◽  
M Haris-Hussain ◽  
J Jalinas ◽  
A B Idris ◽  
...  

Abstract Metarhizium anisopliae Metchnikoff (Hypocreales: Clavicipitaceae) is a fungal pathogen that causes disease in various insect pests, and it can be exploited and developed as a biological control agent to combat the red palm weevil, Rhynchophorus ferrugineus Olivier (Coleoptera: Dryophthoridae). The study on indigenous isolates is crucial especially for development of bioinsecticides in the future. The M. anisopliae strain called MET-GRA4 was tested for pathogenicity against adult red palm weevil and treated in vitro with different spore viabilities. The isolates exhibited pathogenicity with 100% mortality 21 d postinfection. The median lethal time (LT50) for 85% viable spores was 8.6 d, while 39% viable spores had an LT50 value of 21.37 d, with 92 and 16.6% mycosis, respectively. The species MET-GRA4 strain was molecularly characterized using ITS1 and ITS4 from pure culture (Isolate A), mass-produced spores (Isolate B), and infected red palm weevil cadavers (Isolate C). The DNA sequences obtained matched M. anisopliae sequences, with 99% similarity. This new isolate of M. anisopliae has potential as a targeted bioinsecticide for management of red palm weevil.

2011 ◽  
Vol 43 (2) ◽  
pp. 269
Author(s):  
Behzad Habibpour ◽  
Amir Cheraghi ◽  
Mohammad Saeed Mossadegh

This article is the first report on the promising effect of an entomopathogenic fungus, <em>Metarhizium anisopliae</em> (Metschnikoff) Sorokin to control populations of <em>Microcerotermes diversus </em>Silvestri. Biological control is an alternative to the long-term usage of chemical pesticides.<em> M. anisopliae</em>, the causal agent of green muscardine disease of insects, is an important fungus in biological control of insect pests. Bait systems can eliminate entire colonies of subterranean termites. Baiting reduces adverse environmental impacts caused by organochlorine and organophosphate pesticides in the control of termites and creates sustainable protection of buildings against their invasion. Treated-sawdust bait was applied by two methods: a) combination of treated sawdust and untreated filter paper, and b) combination of treated sawdust and untreated sawdust. When combinations of treated sawdust and untreated sawdust were used, LC50 and LC90 were 8.4&times;106 and 3.9&times;107 (spore/ml), respectively. With the use of improved bait formula and more virulent strains, we hope to achieve better control of termite colonies and enable pathogens to become a useful element in the Integrated Pest Management system.


2005 ◽  
Vol 40 (1) ◽  
pp. 67-73 ◽  
Author(s):  
L. A. Rodríguez-del-Bosque ◽  
F. Silvestre ◽  
V. M. Hernández ◽  
H. Quiroz ◽  
J. E. Throne

Five isolates of Beauveria bassiana (Balsamo) Vuillemin and 3 isolates of Metarhizium anisopliae (Metschnikoff) Sorokin were tested against third-instar larvae of Phyllophaga crinita (Burmeister) and Anomala flavipennis Burmeister under laboratory conditions using the “maximum challenge test” protocol. The M. anisopliae strains were more virulent than the B. bassiana strains, with the isolates MAGL3N and MAGL4N of M. anisopliae causing the highest mortality in both white grub species. Regardless of scarab species, mortality caused by MAGL3N was &gt;63% after 4 days and &gt;96% after 10 days. The other strain of M. anisopliae, MAGC2N, also caused high mortality in A. flavipennis, but at a slower rate than MAGL3N. Median lethal time (LT50) for MAGL3N was 2.9 days for P. crinita and 3.0 d for A. flavipennis. The LT50 for MAGL4N was 5.3 d for P. crinita and 7.6 d for A. flavipennis while the LT50 for MAGC2N was 4.4 d for A. flavipennis. Metarhizium anisopliae is a potential biological control agent for P. crinita and A. flavipennis and should be further investigated for possible development.


2004 ◽  
Vol 94 (12) ◽  
pp. 1305-1314 ◽  
Author(s):  
O. Carisse ◽  
D. Rolland

Field and in vitro trials were conducted to establish the influence of the biological control agent Microsphaeropsis ochracea on the ejection pattern of ascospores by Venturia inaequalis and on apple scab development, and to establish the best timing of application. The ejection pattern of ascospores was similar on leaves sprayed with M. ochracea and on untreated leaves. Fall application of M. ochracea combined with a delayed-fungicide program was evaluated in orchards with intermediate and high scab risk. For both orchards, it was possible to delay the first three and two infection periods in 1998 and 1999, respectively, without causing significant increase or unacceptable leaf and fruit scab incidence. To evaluate the best timing of application, sterile leaf disks were inoculated with V. inaequalis and then with M. ochracea 0, 2, 4, 6, 8, 10, 12, 14, and 16 weeks later. After incubation under optimal conditions for pseudothecia development, the number of ascospores was counted. Similarly, M. ochracea was sprayed on scabbed leaves on seven occasions from August to November 1999 and 2000. Leaves were overwintered on the orchard floor and ascospore production was evaluated the following spring. Ascospore production was reduced by 97 to 100% on leaf disks inoculated with M. ochracea less than 6 weeks after inoculation with V. inaequalis, but ascospore production increased with increasing period of time when M. ochracea was applied 8 to 16 weeks after the inoculation with V. inaequalis. In the orchard, the greatest reduction in production of ascospores (94 to 96% in 2000 and 99% in 2001) occurred on leaves sprayed with M. ochracea in August. The production of ascospores was reduced by 61 to 84% in 2000 and 93% in 2001 on leaves sprayed with M. ochracea in September, reduced by 64 to 86% in 2000 and 74 to 89% in 2001 on leaves sprayed in October, and reduced by 54 and 67% in 2000 and 2001, respectively, on leaves sprayed in November. It was concluded that M. ochracea should be applied in August or September and that ascospore maturation models and delayed-fungicide program could be used in orchards treated with this biological control agent.


2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
M. M. Rahman ◽  
M. E. Ali ◽  
A. A. Khan ◽  
A. M. Akanda ◽  
Md. Kamal Uddin ◽  
...  

A total of 91 isolates of probable antagonistic bacteria of potato soft rot bacteriumErwinia carotovorasubsp.carotovora(Ecc) were extracted from rhizospheres and endophytes of various crop plants, different soil varieties, and atmospheres in the potato farming areas of Bangladesh. Antibacterial activity of the isolated probable antagonistic bacteria was testedin vitroagainst the previously identified most common and most virulent soft rot causing bacterial strain Ecc P-138. Only two isolates E-45 and E-65 significantly inhibited thein vitrogrowth of Ecc P-138. Physiological, biochemical, and carbon source utilization tests identified isolate E-65 as a member of the genusBacillusand the isolate E-45 asLactobacillussp. The stronger antagonistic activity against Ecc P-138 was found in E-65in vitroscreening and storage potatoes. E-65 reduced the soft rot infection to 22-week storage potatoes of different varieties by 32.5–62.5% in model experiment, demonstrating its strong potential to be used as an effective biological control agent for the major pectolytic bacteria Ecc. The highest (62.5%) antagonistic effect of E-65 was observed in the Granola and the lowest (32.7%) of that was found in the Cardinal varieties of the Bangladeshi potatoes. The findings suggest that isolate E-65 could be exploited as a biocontrol agent for potato tubers.


2002 ◽  
Vol 68 (9) ◽  
pp. 4383-4389 ◽  
Author(s):  
Pingsheng Ji ◽  
Mark Wilson

ABSTRACT Bacterial speck of tomato, caused by Pseudomonas syringae pv. tomato, was used to determine whether similarity in carbon source utilization between a preemptive biological control agent and the pathogen was significant in determining the ability of the bacterium to suppress disease. Similarity in carbon source utilization was quantified as the ratio of the number of tomato carbon sources utilized in vitro by the biological control agent to the number of tomato carbon sources utilized in vitro by the target pathogen (the niche overlap index [NOI]). Suppression of the disease was quantified as the percent reduction in disease severity compared to the pathogen-only control when nonpathogenic bacteria were applied to foliage 48 h prior to the pathogen. In the collection of 36 nonpathogenic bacterial strains, there was a significant (P < 0.01), but weak (r2 = 0.25), correlation between reduction in disease severity and similarity in carbon source utilization, suggesting that similarity in carbon source use was significant in determining ability to suppress disease. The relationship was investigated further using catabolic mutants of P. syringae strain TLP2, an effective biological control agent of speck. Catabolic mutants exhibited lower levels of similarity (NOI = 0.07 to 0.90) than did wild-type TLP2 (NOI = 0.93). With these catabolic mutants there was a significant (P < 0.01), and stronger (r2 = 0.42), correlation between reduction in disease severity and similarity in carbon source utilization. This suggests that similarity in carbon source utilization was a more important component of biological control ability for the catabolic mutants than for the nonpathogenic bacteria. Together, these studies indicate that suppression of bacterial speck of tomato was correlated with nutritional similarity between the pathogenic and nonpathogenic bacteria and suggest that preemptive utilization of carbon sources was probably involved in the biological control of the disease by both the naturally occurring nonpathogenic bacteria and the catabolic mutants.


2012 ◽  
Vol 61 (3) ◽  
pp. 185-193 ◽  
Author(s):  
K.P. Singh ◽  
S.S. Vaish ◽  
Niranjan Kumar ◽  
K.D. Singh ◽  
Minakshi Kumari

2021 ◽  
Vol 13 (2) ◽  
pp. 237-242
Author(s):  
Dyah Rini Indriyanti ◽  
Siti Harnina Bintari ◽  
Ning Setiati ◽  
Jamil Maulana Zahriyan Alfiyan

Metarhizium anisopliae is a parasitic fungus on insects, and thus called entomopathogenic fungus. This fungus is used as a biological control agent for insect pests. Fungal propagation can be done using a variety of media. The purpose of this study was to analyze the growth of M. anisopliae on four types of media, with conidial density and viability as the growth parameters. This research was conducted at the Laboratory of Microbiology, Universitas Negeri Semarang. This study was an experimental research used a Completely Randomized Design (CRD) with one factor and four treatments: Control (PDA/Potato Dextrose Agar medium), Treatment I (ELSA/Extract Larvae Sucrose Agar medium), Treatment II (CWSA/Coconut Water Sucrose Agar medium), Treatment III (CWELSA/Coconut Water and Extract Larvae Sucrose Agar medium). The and results showed that there was an effect of growth media on the density and viability of M. anisopliae conidia. CWELSA media had the highest conidial density (2.91 x 108 cfu/mL) and viability (97.17%). CWSA media had  2.82 x 108 cfu/mL and 95.33%. PDA media had 2.25 x 108 cfu/mL and 92.83%. ELSA media had 1.64 x 108 cfu/mL and 90.83%. The high conidial density and viability of M. anisopliae is CWELSA  medium. This study is as an alternative growth medium to improve the quality of M. anisopliae propagation.


2020 ◽  
Vol 87 (1) ◽  
Author(s):  
Thao D. Tran ◽  
Celia Del Cid ◽  
Robert Hnasko ◽  
Lisa Gorski ◽  
Jeffery A. McGarvey

ABSTRACT Listeria monocytogenes is a foodborne pathogen that causes high rates of hospitalization and mortality in people infected. Contamination of fresh, ready to eat produce by this pathogen is especially troubling because of the ability of this bacterium to grow on produce under refrigeration temperatures. In this study, we created a library of over 8,000 plant phyllosphere-associated bacteria and screened them for the ability to inhibit the growth of L. monocytogenes in an in vitro fluorescence-based assay. One isolate, later identified as Bacillus amyloliquefaciens ALB65, was able to inhibit the fluorescence of L. monocytogenes by >30-fold in vitro. B. amyloliquefaciens ALB65 was also able to grow, persist, and reduce the growth of L. monocytogenes by >1.5 log CFU on cantaloupe melon rinds inoculated with 5 × 103 CFU at 30°C and was able to completely inhibit its growth at temperatures below 8°C. DNA sequence analysis of the B. amyloliquefaciens ALB65 genome revealed six gene clusters that are predicted to encode genes for antibiotic production; however, no plant or human virulence factors were identified. These data suggest that B. amyloliquefaciens ALB65 is an effective and safe biological control agent for the reduction of L. monocytogenes growth on intact cantaloupe melons and possibly other types of produce. IMPORTANCE Listeria monocytogenes is estimated by the Centers for Disease Control and Prevention and the U.S. Food and Drug Administration to cause disease in approximately 1,600 to 2,500 people in the United States every year. The largest known outbreak of listeriosis in the United States was associated with intact cantaloupe melons in 2011, resulting in 147 hospitalizations and 33 deaths. In this study, we demonstrated that Bacillus amyloliquefaciens ALB65 is an effective biological control agent for the reduction of L. monocytogenes growth on intact cantaloupe melons under both pre- and postharvest conditions. Furthermore, we demonstrated that B. amyloliquefaciens ALB65 can completely inhibit the growth of L. monocytogenes during cold storage (<8°C).


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