scholarly journals Mass Cultivation of Entomopathogenic Nematode in Artificial Media

2016 ◽  
Vol 8 (1) ◽  
pp. 111 ◽  
Author(s):  
Dyah Rini Indriyanti ◽  
Nur Lailatul Muharromah
Keyword(s):  
Entomopathogenic Nematodes ◽  
Entomopathogenic Nematode ◽  
Biology Education ◽  
Chicken Liver ◽  
Dog Food ◽  
Mass Cultivation ◽  
Soybean Powder ◽  
Food Media

<p>Entomopathogenic nematodes Entomopathogenic nematodes (EPN) of the genera Heterorhabditis and Steinernema are commercially used to control pest insect. EPN is widely cultivated through in-vivo and in vitro methods. This research aims to discover the abundance of EPN cultivated in various artificial media. Seven types of media composition were tested in this research: media A (yeast + soybean powder), media B (yeast + chicken liver), media C (yeast + dog food), media D (yolk + soybean powder), media E (yolk + chicken liver), media F (yolk + dog food), and media G (yeast + yolk + dog food). Each media was inoculated in 1.2x103JI/mL. The growth of EPN was observed weekly in 4 weeks. Results showed that EPN could be cultivated using various media; media D, E, F, and G. Highest abundance of EPN is found in the second week of media D for 28164 JI/ml.Cell harvesting is suggested to be conducted during the first and second week to obtain maximum abundance of EPN.</p><p><strong>How to Cite</strong></p><p>Indriyanti, D., &amp; Muharromah, N. (2016). Mass Cultivation of Entomopathogenic Nematode In Artificial Media.<em> Biosaintifika: Journal of Biology &amp; Biology Education</em>, 8(1), 111-118.</p>

In vivo production of entomopathogenic nematodes suppressed by asymptomatic bacterial contaminants

Nematology ◽  
2020 ◽  
Vol 22 (8) ◽  
pp. 917-925
Author(s):  
Akanksha Upadhyay ◽  
Sharad Mohan
Keyword(s):  
Biological Control ◽  
Entomopathogenic Nematodes ◽  
Photorhabdus Luminescens ◽  
Ochrobactrum Anthropi ◽  
Progeny Production ◽  
Bacterial Contaminants ◽  
Significant Difference ◽  
Biological Control Of Insects

Summary Entomopathogenic nematodes (EPN) are excellent biological control agents possessing recycling ability as one of their major attributes. We report the presence of asymptomatic bacteria that can lead to disrupted or low progeny production in Heterorhabditis indica. In a one-to-one in vitro competitive bioassay with contaminants associated with H. indica cuticle, there was a significant suppression in the growth of Sphingomonas koreensis when stressed with the nematode symbiont Photorhabdus luminescens; however, P. luminescens was suppressed when sandwiched between Ochrobactrum anthropi. Bacillus bombysepticus associated with laboratory-reared Galleria when stressed by P. luminescens was significantly suppressed, but not so in the reverse assay. Both O. anthropi and B. bombysepticus were found to be insecticidal to Galleria larvae when fed orally. Tripartite interactive studies on the growth and multiplication of H. indica-P. luminescens symbionts in Galleria larvae, predisposed to S. koreensis, revealed no significant difference initially in the hermaphrodite formation, but subsequently there was a significant decline in the formation of amphimictic females and the final production of infective juveniles. In in vitro studies, none of the contaminants supported the growth and development of axenic H. indica. Adequate precautions should be taken to maintain proper hygiene to eliminate such contaminants while culturing the Galleria and EPN for use in the biological control of insects.

Influence of temperature on the infectivity of entomopathogenic nematodes (Steinernema and Heterorhabditis spp.) to larvae and pupae of the vine weevil Otiorhynchus sulcatus (Coleoptera: Curculionidae)

Nematology ◽  
2000 ◽  
Vol 2 (3) ◽  
pp. 309-317 ◽  
Author(s):  
Steve Long ◽  
John Fenlon ◽  
Paul Richardson
Keyword(s):  
Competing Risks ◽  
Entomopathogenic Nematodes ◽  
Galleria Mellonella ◽  
Commercial Product ◽  
Effect Of Temperature ◽  
Heterorhabditis Megidis ◽  
Otiorhynchus Sulcatus ◽  
The Uk

AbstractThe susceptibilities of early and late instar vine weevil larvae and pupae to three species of entomopathogenic nematodes, indigenous to the UK, were tested in a series of bioassays. Steinernema kraussei (isolates L017 and L137), S. feltiae (the commercial product Nemasys®) and Heterorhabditis megidis (the commercial product Nemasys® H, reared both in vivo in Galleria mellonella larvae and in vitro), were tested at 6, 10 and 18°C for 2 weeks (early instars of O. sulcatus) or 3 weeks (late instars and pupae of O. sulcatus). Nematodes were applied to over 3800 larvae or pupae and there were over 400 untreated controls. Each insect was examined subsequently to determine mortality, and parasitised specimens were dissected to establish whether adult nematodes had developed. Differences in pathogenicity between H. megidis reared in vitro and in vivo were demonstrated. S. kraussei (L137) was consistently the most virulent nematode isolate at low temperatures. The results revealed a significant (P < 0.001) effect of temperature on small larvae of O. sulcatus, but also showed differential levels of mortality, not due to nematodes, for both small larvae and pupae. The use of Abbott's correction for control mortality is challenged and the validity of competing risks theory examined.In einer Reihe von Biotests wurde die Anfälligkeit von frühen und späten Larvenstadien sowie Puppen des Rüsselkäfers Otiorhynchus sulcatus gegenüber drei in UK einheimischen entomopathogenen Nematoden untersucht. Steinernema kraussei (isolate L017 und L137), S. feltiae (Handelsprodukt Nemasys®) und Heterorhabditis megidis (Handelsprodukt Nemasys® H, beide in vivo an Larven von Galleria mellonella und in vitro gezüchtet) wurden bei 6, 10 and 18°C für zwei Wochen (frühe Stadien von O. sulcatus) oder drei Wochen (späte Stadien und Puppen von O. sulcatus) geprüft. Über 3800 Larven oder Puppen wurden mit Nematoden behandelt, daneben gab es über 400 unbehandelte Kontrollen. Anschliessend wurde jedes Insekt untersucht, um die Mortalität zu bestimmen. Parasitierte Exemplare wurden aufpräpariert um festzustellen, ob sich adulte Nematoden entwickelt hatten. Zwischen in vitro und in vivo kultivierten H. megidis konnten Unterschiede in der Pathogenität festgestellt werden. S. kraussei (L137) war bei niedrigen Temperaturen durchgehend das virulenteste Isolat. Die Ergebnisse zeigten eine signifikante (P < 0.001) Wirkung der Temperatur auf kleine Larven von O. sulcatus. Sie zeigten für kleine Larven und Puppen aber auch unterschiedliche Mortlitätsgrade an, die nicht auf Nematoden zurückgingen. Die Anwendung von Abbott's Korrektur zur Prüfung der Mortalität wird kritisch hinterfragt, die Gültigkeit der Theorie der “competing risks” wird geprüft.

scholarly journals Effects of “fresh mechanically deboned meat” inclusion on nutritional value, palatability, shelf-life microbiological risk and digestibility in dry dog food

PLoS ONE ◽  
2021 ◽  
Vol 16 (4) ◽  
pp. e0250351
Author(s):  
Giorgia Meineri ◽  
Alessia Candellone ◽  
Sonia Tassone ◽  
Pier Giorgio Peiretti ◽  
Erica Longato ◽  
...  
Keyword(s):  
Consumer Choice ◽  
Nutritional Value ◽  
Bacterial Count ◽  
In Vitro Digestibility ◽  
Pet Food ◽  
Dog Food ◽  
Fatty Acids Profile ◽  
Valid Instrument

Fresh mechanically deboned meat (MDM) is usually claimed as high-quality ingredient on dry pet food recipes and this aspect may positively influence consumer choice. It is important to determine the scientifically sustainability of this claim and to assess the microbiological safety of MDM inclusion in dry pet food. Objectives were: 1) to evaluate the effect of inclusion of MDM in dry dog food on fatty acid profile and in vivo and in vitro digestibility, proposing a new system (DaisyII Incubator) to measure the in vitro digestibility for dogs; 2) to compare palatability of dry dog food containing MDM with dry dog food in which meat by-products (MBP) are the only animal protein sources; 3) to determine, whether or not, the inclusion of that ingredient changes the microbiology and the storage quality. Results indicated that MDM product was characterized by significant higher nutritional value in terms of fatty acids profile, in vitro digestibility (HV-IVD method) and lower palatability than the MBP product. Microbiological risk assessment showed no microbiological hazards for either product. After 6-months storage, the total mesophilic bacterial count ranged between 1.77 and 2.09 log CFU/g feed, while polyamine values were higher in the MDM (0.37 g/kg) than in the MBP (0.27 g/kg). The DaisyII Incubator was found to be a valid instrument for studying in vitro digestibility also for dogs, providing data simply, quickly, with less variability and costs than in vivo trials. In conclusion, MDM inclusion in dry dog food is microbiologically safe and it can improve its nutritional quality, at the expense of a reduced palatability. The higher polyamine levels fount in MDM-enriched petfood after 6-months storage, however, may represent a possible hazard, and further studies are still warranted.

scholarly journals The entomopathogenic nematode Steinernema hermaphroditum is a self-fertilizing hermaphrodite and a genetically tractable system for the study of parasitic and mutualistic symbiosis

2021 ◽  
Author(s):  
Mengyi Cao ◽  
Hillel T. Schwartz ◽  
Chieh-Hsiang Tan ◽  
Paul W. Sternberg
Keyword(s):  
Entomopathogenic Nematodes ◽  
Genetic Model ◽  
Entomopathogenic Nematode ◽  
Model Development ◽  
Symbiotic Bacteria ◽  
Mutualistic Symbiosis ◽  
Self Fertilization ◽  
Rapid Generation ◽  
Mutant Lines

AbstractEntomopathogenic nematodes, including Heterorhabditis and Steinernema, are parasitic to insects and contain mutualistically symbiotic bacteria in their intestines (Photorhabdus and Xenorhabdus, respectively) and therefore offer opportunities to study both mutualistic and parasitic symbiosis. The establishment of genetic tools in entomopathogenic nematodes has been impeded by limited genetic tractability, inconsistent growth in vitro, variable cryopreservation, and low mating efficiency. We obtained the recently described Steinernema hermaphroditum strain CS34 and optimized its in vitro growth, with a rapid generation time on a lawn of its native symbiotic bacteria Xenorhabdus griffiniae. We developed a simple and efficient cryopreservation method. Previously, S. hermaphroditum isolated from insect hosts was described as first-generation hermaphroditic and second-generation gonochoristic. We discovered that CS34, when grown in vitro, produced consecutive generations of autonomously reproducing hermaphrodites accompanied by rare males. We performed mutagenesis screens in S. hermaphroditum that produced mutant lines with visible and heritable phenotypes. Genetic analysis of the mutants demonstrated that this species reproduces by self-fertilization rather than parthenogenesis and that its sex is determined chromosomally. Genetic mapping has thus far identified markers on the X chromosome and three of four autosomes. We report that S. hermaphroditum CS34 is the first consistently hermaphroditic entomopathogenic nematode and is suitable for genetic model development to study naturally occurring mutualistic symbiosis and insect parasitism.

Comparison of metabolites produced in vitro and in vivo by Photorhabdus luminescens, a bacterial symbiont of the entomopathogenic nematode Heterorhabditis megidis

1998 ◽  
Vol 44 (11) ◽  
pp. 1072-1077 ◽  
Author(s):  
Kaiji Hu ◽  
Jianxiong Li ◽  
Wenjie Wang ◽  
Houming Wu ◽  
Hai Lin ◽  
...  
Keyword(s):  
Galleria Mellonella ◽  
Entomopathogenic Nematode ◽  
Culture Broth ◽  
Bacterial Symbiont ◽  
Natural Source ◽  
Photorhabdus Luminescens ◽  
Heterorhabditis Megidis ◽  
Organic Extracts

The types of metabolites produced by Photorhabdus luminescens C9 when it is introduced by Heterorhabditis megidis 90 into Galleria mellonella larvae are different from those produced in tryptic soy broth. Only 3,5-dihydroxy-4-isopropylstilbene 1 was identified from the organic extracts of P. luminescens culture broth, but both 3,5-dihydroxy-4-isopropylstilbene 1 and 3,5-dihydroxy-4-ethylstilbene 3 were isolated from the organic extracts of nematode-bacterium infected G. mellonella larvae. In addition to two pigments, both of which had been previously reported from P. luminescens C9 culture broth, three pigments, 1,8-dihydroxy-3-methoxy-9,10-anthraquinone 2, 1-hydroxy-2,6,8-trimethoxy-9,10-anthraquinone 6, and 1,4-dihydroxy-2,5-dimethoxy-9,10-anthraquinone 7 were isolated from the organic extracts of G. mellonella larvae infected by the nematode-bacterium complex. Among these, compounds 6 and 7 are novel and isolated from a natural source for the first time.Key words: Photorhabdus luminescens, Heterorhabditis megidis, 1-hydroxy-2,6,8-trimethoxy-9,10-anthraquinone, 1,4-dihydroxy-2,5-dimethoxy-9,10-anthraquinone, pigment.

Nutritional Value of Extruded Dog Food with Mechanically Separated Chicken Meat or Meat By-products.

2019 ◽  
Author(s):  
Giorgia Meineri ◽  
Pier Giorgio Peiretti ◽  
Sonia Tassone ◽  
Alessia Candellone ◽  
Erica Longato ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Chicken Meat ◽  
In Vitro Digestibility ◽  
Pet Food ◽  
Dog Food ◽  
Food Digestibility ◽  
Valid Instrument ◽  
By Products

The objectives of this study were: (a) to evaluate the effect of inclusion of mechanically separated chicken meat (MSCM) in dry dog food on fatty acid profile, in vivo and in vitro digestibility, and palatability as compared with dry dog food containing meat by-products (MBP); (b) to determine, whether or not, the inclusion of the one or the other ingredient changes the microbiology and the storage quality of the two food products; (c) to propose a new system (DaisyII Incubator) to measure the in vitro digestibility of the two products. Their similar chemical composition notwithstanding, the MSCM product had lower palatability but better nutritional quality (with higher polyunsaturated fatty acid [PUFA] content and lower saturated fatty acid [SFA] content) than the MBP product. Microbiological risk assessment showed no microbiological hazards for either product. After 6 months storage, polyamine values were found to be higher in the MSCM than in the MBP. Finally, the DaisyII Incubator proved a valid instrument for the study of in vitro digestibility also for dogs; since it provided data simply, quickly with less variability and cost than obtained with in vivo trials, it could represent the future for pet food digestibility studies. Our results indicate that inclusion of MSCM or MBP as the main protein ingredient in extruded pet food may be used advantageously in product formulations.

Effect of inoculum age and physical parameters on in vitro culture of the entomopathogenic nematode Steinernema feltiae

2016 ◽  
Vol 91 (6) ◽  
pp. 686-695 ◽  
Author(s):  
L.G. Leite ◽  
D.I. Shapiro-Ilan ◽  
S. Hazir ◽  
M.A. Jackson
Keyword(s):  
Liquid Medium ◽  
Solid Medium ◽  
Entomopathogenic Nematode ◽  
Steinernema Feltiae ◽  
Physical Parameters ◽  
Symbiotic Association ◽  
Liquid Cultures ◽  
Inoculum Age

AbstractEntomopathogenic nematodes (EPNs) of the families Steinernematidae and Heterorhabditidae have a symbiotic association with bacteria which makes them virulent against insects. EPNs have been mass produced using in vivo and in vitro methods, including both solid and liquid fermentation. This study assessed the effect of nematode inoculum age on the production of Steinernema feltiae in liquid, solid and biphasic processes. Several physical parameters were also assessed: the effect of medium viscosity, flask size and aeration speed on the recovery and yield of infective juveniles (IJs). Inoculum age treatments included inoculum liquid cultures that were 7, 14, 21 and 28 days old. Nematodes from the same inoculum were added to one liquid medium (liquid culture), one solid medium with bacteria previously grown in sponge (solid culture) and a variation of the solid medium (a biphasic culture), in which the bacteria were first grown in liquid and, then, soaked into the sponges, with the purpose of providing a more homogeneous bacterial culture before nematode inoculation. Experiments were conducted in Erlenmeyer flasks. Eight treatments were established involving combinations of three variables: two media (with and without 0.2% agar), two flask sizes (250 and 150 ml) and two agitation speeds (180 and 280 rpm). The study showed increases in nematode yield for liquid cultures, but not for solid or biphasic cultures, with the advance of the inoculum age up to 28 days of growth. Furthermore, the addition of 0.2% agar to the liquid medium and increasing the aeration rate by using larger flasks with higher agitation speed may increase nematode recovery and final yield. The experiments were conducted using shake flasks but the results may also be applicable for bioreactors.

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