Ex vivo development of Phasmarhabditis spp. associated with terrestrial molluscs

2022 ◽  
Vol 96 ◽  
Author(s):  
A. Pieterse ◽  
S. Haukeland ◽  
V. Půža ◽  
J.L. Ross ◽  
A.P. Malan

Abstract The success of Phasmarhabditis hermaphrodita (Schneider) Andrássy (Rhabditida: Rhabditidae) as a biological control agent of molluscs has led to a worldwide interest in phasmarhabditids. However, scant information is available on the lifecycle development of species within the genus. In the current study, the development of P. hermaphrodita, Phasmarhabditis papillosa, Phasmarhabditis bohemica and Phasmarhabditis kenyaensis were studied using ex vivo cultures, in order to improve our understanding of their biology. Infective juveniles (IJs) of each species were added to 1 g of defrosted homogenized slug cadavers of Deroceras invadens and the development monitored after inoculated IJ recovery, over a period of eight–ten days. The results demonstrated that P. bohemica had the shortest development cycle and that it was able to produce first-generation IJs after eight days, while P. hermaphrodita, P. papillosa and P. kenyaensis took ten days to form a new cohort of IJs. However, from the perspective of mass rearing, P. hermaphrodita has an advantage over the other species in that it is capable of forming self-fertilizing hermaphrodites, whereas both males and females are required for the reproduction of P. papillosa, P. bohemica and P. kenyaensis. The results of the study contribute to the knowledge of the biology of the genus and will help to establish the in vitro liquid cultures of different species of the genus.

Author(s):  
Júlia Gabriela Aleixo Vieira ◽  
Alexandra Peter Krüger ◽  
Tiago Scheuneumann ◽  
Maíra Chagas Morais ◽  
Hugo Julio Speriogin ◽  
...  

Abstract Spotted-wing Drosophila, Drosophila suzukii (Matsumura, 1931), is an economic pest of thin-skinned fruit crops. Its control has commonly been carried out through chemical methods. However, given the need to develop safer and environmentally friendly management alternatives, the pupal endoparasitoid Trichopria anastrephae Lima stands out as a potential biological control agent. However, the lack of information on the mass rearing of this parasitoid limits its use. Thus, the objective of our study was to provide information that is useful for rearing T. anastrephae using D. suzukii as a host. The effects of pupal age, exposure time, and pupal density on the parasitism rate were examined, as was the effect of honey provision for extending adult parasitoid longevity. Exposing 15 12-h-old pupae per female for 24 h resulted in higher parasitism rates and a greater number of emerged parasitoids. Males and females of T. anastrephae fed with pure honey (100%) or honey diluted to 50% in water lived longer compared to those fed 10% honey, no food, or only water.


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.


2021 ◽  
Vol 285 ◽  
pp. 03010
Author(s):  
Irina Agasyeva

Ectoparasite Habrobracon hebetor Say is one of the most widely used biological controllers in biological plant protection against a number of harmful lepidopterans, including especially dangerous pests of corn, soy, fruit and vegetable crops. As a result of research conducted in 2017, food specialization and parasitic activity of three different populations of H.hebetor were studied. Two races have been identified for mass rearing and application: pyralid and leaf roller (against corn moth, bean pod borer, apple and plum moths), and pyralid owl-moth (against cotton moth, corn borer, bean pod borer and boxwood moth). As a result of studies of biological features and trophic needs, it has been determined that caterpillars of mill moth (Ephestia cuhniellia Zella) should be used as a host insect for laboratory cultivation of the stock population of the Habrobracon pyralid and leaf roller race (race No. 1). For the introduced from South Kazakhstan the H.hebetor pyralid and noctuid race the most productive rearing is on the caterpillars of large bee moth (Galleria mellonela L.). Optimal temperature for rearing of both races is 26-28 ° C, relative air humidity is 70% and photoperiod is not less than 16 hours. It has been noticed that before laying eggs on the host’s caterpillars, the Habrobracon female preliminarily paralyzes the victim, piercing the sheath with ovipositor. As a result, the caterpillar stops eating and is immobilized. In 3-4 days larvae hatch out of the laid on the caterpillar eggs. The larvae feed on the contents of the caterpillars for 4-5 days, then pupate and after 6-8 days an adult insect leaves the cocoon. The development of one generation lasts 13-16 days, one cocoon includes one parasite. 1,000 large bee moth caterpillars used for infection provide on average 5.8-6.0 thousand cocoons, of which an average of 4.5-4.7 thousand parasites fly out.


Author(s):  
Yi Chieh Lim ◽  
Kathleen S Ensbey ◽  
Carolin Offenhäuser ◽  
Rochelle C J D’souza ◽  
Jason K Cullen ◽  
...  

Abstract Background Despite significant endeavor having been applied to identify effective therapies to treat glioblastoma (GBM), survival outcomes remain intractable. The greatest nonsurgical benefit arises from radiotherapy, though tumors typically recur due to robust DNA repair. Patients could therefore benefit from therapies with the potential to prevent DNA repair and synergize with radiotherapy. In this work, we investigated the potential of salinomycin to enhance radiotherapy and further uncover novel dual functions of this ionophore to induce DNA damage and prevent repair. Methods In vitro primary GBM models and ex vivo GBM patient explants were used to determine the mechanism of action of salinomycin by immunoblot, flow cytometry, immunofluorescence, immunohistochemistry, and mass spectrometry. In vivo efficacy studies were performed using orthotopic GBM animal xenograft models. Salinomycin derivatives were synthesized to increase drug efficacy and explore structure-activity relationships. Results Here we report novel dual functions of salinomycin. Salinomycin induces toxic DNA lesions and prevents subsequent recovery by targeting homologous recombination (HR) repair. Salinomycin appears to target the more radioresistant GBM stem cell–like population and synergizes with radiotherapy to significantly delay tumor formation in vivo. We further developed salinomycin derivatives which display greater efficacy in vivo while retaining the same beneficial mechanisms of action. Conclusion Our findings highlight the potential of salinomycin to induce DNA lesions and inhibit HR to greatly enhance the effect of radiotherapy. Importantly, first-generation salinomycin derivatives display greater efficacy and may pave the way for clinical testing of these agents.


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

2018 ◽  
Vol 13 (5) ◽  
pp. 161
Author(s):  
Evan Purnama Ramdan ◽  
Efi Toding Tondok ◽  
Suryo Wiyono ◽  
Sri Hendrastuti Hidayat ◽  
Widodo Widodo

Stem rot disease caused by Phytophthora capsici is an important disease on chilli. Eight endophytic fungi that had been isolated and screened based on pathogenecity test were further tested for their potential as the biological control agent of the stem rot disease of chilli. The endophytic fungi suspension was applied twice during the trial. The first application was on 100 seed lot, by soaking them in 100 mL of suspension. The second application was on the 3 weeks-old chili seedlings by drenching them with 10 mL suspension per plant. The concentration of endophytic fungi  in the suspension was 2.8 × 106 cfu mL-1. The disease intensity and AUDPC value were measured for 4 weeks after the pathogen inoculation. The growth inhibition test of P. capsici  was performed in vitro and the colonization abilities of endophytic fungi were observed at 4 weeks-old chilli seedlings. Eight endophytic fungi  inhibited the growth of the P. capsici, and two of those isolates namely Penicillium strain MAG1 and Penicillium strain PAB2 showed antibiosis mechanism. Endophytic fungi has the ability more to colonize at the root (26–60%) than in the stem (20–40%). Fusarium strain MAGR1 has the highest level of endophytic colonization i.e. 60% compared to others.  Based on in vivo assay, six endophytic fungi isolates, i.e. Fusarium strain MAGR1, Penicillium strain MAG1, Penicillium strain PAB2, sterile hyphae HAJ1, sterile hyphae HAJ2, and  sterile hyphae PBG7, showed the potency to control stem rot disease with inhibition level of 25.5–35.5%


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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