Lepidopteran species have a variety of defence strategies against bacterial infections.

The insect immune system has versatile ways of coping with microbial insults. Currently, innate immune priming has been described in several invertebrates, and the first insights into its mechanistic basis have been described. Here we studied infections with two different strains of Serratia marcescens bacteria in two different Lepidopteran hosts. The results reveal fundamental differences between the two hosts, a well-known model organism Galleria mellonella and a non-model species Arctia plantaginis. They differ in their strategies for resisting oral infections; priming their defences against a recurring sepsis; and upregulating immunity related genes as a response to the specific pathogen strains. The two bacterial strains (an environmental isolate and an entomopathogenic isolate) differ in their virulence, use of extracellular proteases, survival in the larval gut, and in the immune response they evoke in the hosts. This study explores the potential mechanistic explanations for both host and pathogen specific characters that significantly affect the outcome of Gram-negative bacterial infection in Lepidopteran larvae. The results highlight the need to pay greater attention to the differences between model and non-model hosts, and closely related pathogen strains, in immunological studies.

Long-Term Prophylactic Antibiotic Treatment: Effects on Survival, Immunocompetence and Reproduction Success of Parasemia plantaginis (Lepidoptera: Erebidae).

Hundreds of insect species are nowadays reared under laboratory conditions. Rearing of insects always implicates the risk of diseases, among which microbial infections are the most frequent and difficult problems. Although there are effective prophylactic treatments, the side effects of applied antibiotics are not well understood. We examined the effect of prophylactic antibiotic treatment on the overwintering success of wood tiger moth (Parasemia plantaginis) larvae, and the postdiapause effect on their life-history traits. Four weeks before hibernation larvae were treated with a widely used antibiotic (fumagillin). We monitored moths' survival and life-history traits during the following 10 mo, and compared them to those of untreated control larvae. Prophylactic antibiotic treatment had no effect on survival but we show effects on some life-history traits by decreasing the developmental time of treated larvae. However, we also revealed relevant negative effects, as antibiotic treated individuals show a decreased number of laid eggs and also furthermore a suppressed immunocompetence. These results implicate, that a prophylactic medication can also lead to negative effects on life-history traits and reproductive success, which should be seriously taken in consideration when applying a prophylactic treatment to laboratory reared insect populations.

The oral vaccination with Paenibacillus larvae bacterin can decrease susceptibility to American Foulbrood infection in honey bees-A safety and efficacy study.

Pollination services to increase crop production are becoming more and more important, as we are facing both climate change and a growing world population. Both are predicted to impact food security worldwide. High-density, commercial beekeeping has become a key link in the food supply chain, and diseases have become a central issue in hive losses around the world. American Foulbrood (AFB) disease is a highly contagious bacterial brood disease in honey bees (Apis mellifera), leading to hive losses worldwide. The causative agent is the Gram+ bacterium Paenibacillus larvae, which is able to infect honey bee larvae during the first 3 days of their lives. It can be found in hives around the world with viable spores for decades. Antibiotics are largely ineffective in treating the disease as they are only efficient against the vegetative state. Once a hive shows the clinical manifestation of the disease, the only effective way to eradicate it and prevent the spread of the disease is by burning the hive, the equipment, and the colony. Because of its virulent nature and detrimental effects on honey bee colonies, AFB is classified as a notifiable disease worldwide. Effective, safe, and sustainable methods are needed to ensure the wellbeing of honey bee colonies. Even though insects lack antibodies, which are the main requisites for trans-generational immune priming (TGIP), they can prime their offspring against persisting pathogens. Here, we demonstrate an increased survival of infected honey bee larvae after their queen was vaccinated, compared to offspring of control queens (placebo vaccinated). These results indicate that TGIP in insects can be used to majorly enhance colony health, protect commercial pollinators from deadly diseases, and reduce high financial and material losses to beekeepers. Classification: biological sciences, applied biological sciences.

Increased survival of honeybees in the laboratory after simultaneous exposure to low doses of pesticides and bacteria.

Recent studies of honeybees and bumblebees have examined combinatory effects of different stressors, as insect pollinators are naturally exposed to multiple stressors. At the same time the potential influences of simultaneously occurring agricultural agents on insect pollinator health remain largely unknown. Due to different farming methods, and the drift of applied agents and manure, pollinators are most probably exposed to insecticides but also bacteria from organic fertilizers at the same time. We orally exposed honeybee workers to sub-lethal doses of the insecticide thiacloprid and two strains of the bacterium Enterococcus faecalis, which can occur in manure from farming animals. Our results show that under laboratory conditions the bees simultaneously exposed to the a bacterium and the pesticide thiacloprid thiacloprid had significant higher survival rates 11 days post exposure than the controls, which surprisingly showed the lowest survival. Bees that were exposed to diet containing thiacloprid showed decreased food intake. General antibacterial activity is increased by the insecticide and the bacteria, resulting in a higher immune response observed in treated individuals compared to control individuals. We thus propose that caloric restriction through behavioural and physiological adaptations may have mediated an improved survival and stress resistance in our tests. However, the decreased food consumption could in long-term also result in possible negative effects at colony level. Our study does not show an additive negative impact of sub-lethal insecticide and bacteria doses, when tested under laboratory conditions. In contrast, we report seemingly beneficial effects of simultaneous exposure of bees to agricultural agents, which might demonstrate a surprising biological capacity for coping with stressors, possibly through hormetic regulation.

The maternal transfer of bacteria can mediate trans-generational immune priming in insects.

Parents invest in their offspring by preparing them for defense against pathogens and parasites that only the parents have encountered, a phenomenon known as trans-generational immune priming. We investigated the underlying mechanism using the established lepidopteran model host Galleria mellonella. When larvae were fed with non-pathogenic bacteria, or the entomopathogenic species Pseudomonas entomophila and Serratia entomophila, the activity of lysozyme and phenoloxidase increased in the hemolymph, and immunity-related genes encoding antibacterial proteins such as gloverin were induced. Remarkably, the ingestion of bacteria by female larvae resulted in the differential expression of immunity-related genes in the eggs subsequently laid by the same females, providing evidence for trans-generational immune priming in G. mellonella. To determine the fate of these ingested microbes, the larval diet was supplemented with bacteria carrying a fluorescent label. We observed these bacteria crossing the midgut epithelium, their entrapment within nodules in the hemocoel, their accumulation within the ovary, and ultimately their deposition in the eggs. Therefore, we propose that trans-generational immune priming in Lepidoptera can be mediated by the maternal transfer of bacteria or bacterial fragments to the developing eggs.