Assessment of toxicity and persistence of Yersinia entomophaga and its Yen-Tc associated toxin.

BACKGROUND: The insect-pathogenic bacterium Yersinia entomophaga MH96 is currently under development as a microbial pesticide active against various pasture and crop pests such as the diamondback moth Plutella xylostella and the cotton bollworm Helicoverpa armigeria. To enable nonrestricted field trials of Y. entomophaga MH96, information on the persistence and nontarget effects of the bacterium and its Yen-Tc proteinaceous toxin are required. RESULTS: The Y. entomophaga Yen-Tc associated toxin was found to have limited persistence on foliage and is inactivated by UV light. The Yen-Tc was rapidly degraded in ovine or bovine rumen fluid or the intestinal fluid of H. armigera. In H. armigera an intestinal protein of >50 kDa was found to cleave the Yen-Tc bond. Assessment of Y. entomophaga persistence on foliage and in soil found that after 42 days the bacterium could not be detected in soil at 20% soil moisture content but persisted for 72 days at 30-40% soil moisture. Nontarget effects of Y. entomophaga towards earthworms found that the bacterium afforded no adverse effects on worm growth or behavior. A summary of historic Yen-Tc and Y. entomophaga persistence and toxicity data is presented. CONCLUSION: The bacterium Y. entomophaga and its Yen-Tc associated toxin have limited persistence in the environment, with the Yen-Tc being susceptible to UV inactivation and proteolytic degradation, and the bacterium persisting longer in soil of a high moisture content. © 2020 Society of Chemical Industry.

The Draft Genome Sequence of the Yersinia entomophaga Entomopathogenic Type Strain MH96T.

Here we report the draft genome of Yersinia entomophaga type strain MH96T. The genome shows 93.8% nucleotide sequence identity to that of Yersinia nurmii type strain APN3a-cT, and comprises a single chromosome of approximately 4,275,531 bp. In silico analysis identified that, in addition to the previously documented Y. entomophaga Yen-TC gene cluster, the genome encodes a diverse array of toxins, including two type III secretion systems, and five rhs-associated gene clusters. As well as these multicomponent systems, several orthologs of known insect toxins, such as VIP2 toxin and the binary toxin PirAB, and distant orthologs of some mammalian toxins, including repeats-in-toxin, a cytolethal distending toxin, hemolysin-like genes and an adenylate cyclase were identified. The genome also contains a large number of hypothetical proteins and orthologs of known effector proteins, such as LopT, as well as genes encoding a wide range of proteolytic determinants, including metalloproteases and pathogen fitness determinants, such as genes involved in iron metabolism. The bioinformatic data derived from the current in silico analysis, along with previous information on the pathobiology of Y. entomophaga against its insect hosts, suggests that a number of these virulence systems are required for survival in the hemocoel and incapacitation of the insect host.

Temperature-Dependent Galleria mellonella Mortality as a Result of Yersinia entomophaga Infection.

The bacterium Yersinia entomophaga is pathogenic to a range of insect species, with death typically occurring within 2 to 5 days of ingestion. Per os challenge of larvae of the greater wax moth (Galleria mellonella) confirmed that Y. entomophaga was virulent when fed to larvae held at 25°C but was avirulent when fed to larvae maintained at 37°C. At 25°C, a dose of ~4 × 10(7) CFU per larva of a Y. entomophaga toxin complex (Yen-TC) deletion derivative, the Y. entomophaga ΔTC variant, resulted in 27% mortality. This low level of activity was restored to near-wild-type levels by augmentation of the diet with a sublethal dose of purified Yen-TC. Intrahemocoelic injection of ~3 Y. entomophaga or Y. entomophaga ΔTC cells per larva gave a 4-day median lethal dose, with similar levels of mortality observed at both 25 and 37°C. Following intrahemocoelic injection of a Yen-TC YenA1 green fluorescent protein fusion strain into larvae maintained at 25°C, the bacteria did not fluoresce until the population density reached 2 × 10(7) CFU ml(-1) of hemolymph. The observed cells also took an irregular form. When the larvae were maintained at 37°C, the cells were small and the observed fluorescence was sporadic and weak, being more consistent at a population density of ~3 × 10(9) CFU ml(-1) of hemolymph. These findings provide further understanding of the pathobiology of Y. entomophaga in insects, showing that the bacterium gains direct access to the hemocoelic cavity, from where it rapidly multiplies to cause disease.