Initial Stages Of Infection Research Articles

How mobility habits influenced the spread of the COVID-19 pandemic: Results from the Italian case study

Starting from December 2019 the world has faced an unprecedented health crisis caused by the new Coronavirus (COVID-19) due to the SARS-CoV-2 pathogen. Within this topic, the aim of the paper was to quantify the effect of mobility habits in the spread of the Coronavirus in Italy through a multiple linear regression model. Estimation results showed that mobility habits represent one of the variables that explains the number of COVID-19 infections jointly with the number of tests/day and some environmental variables (i.e. PM pollution and temperature). Nevertheless, a proximity variable to the first outbreak was also significant, meaning that the areas close to the outbreak had a higher risk of contagion, especially in the initial stage of infection (time-decay phenomena). Furthermore, the number of daily new cases was related to the trips performed three weeks before. This threshold of 21 days could be considered as a sort of positivity detection time, meaning that the mobility restrictions quarantine commonly set at 14 days, defined only according to incubation-based epidemiological considerations, is underestimated (possible delays between contagion and detection) as a containment policy and may not always contribute to effectively slowing down the spread of virus worldwide. This result is original and, if confirmed in other studies, will lay the groundwork for more effective containment of COVID-19 in countries that are still in the health emergency, as well as for possible future returns of the virus.

When Appearance Misleads: The Role of the Entomopathogen Surface in the Relationship with Its Host.

Currently, potentially harmful insects are controlled mainly by chemical synthetic insecticides, but environmental emergencies strongly require less invasive control techniques. The use of biological insecticides in the form of entomopathogenic organisms is undoubtedly a fundamental resource for the biological control of insect pests in the future. These infectious agents and endogenous parasites generally act by profoundly altering the host’s physiology to death, but their success is closely related to the neutralization of the target insect’s immune response. In general, entomopathogen parasites, entomopathogenic bacteria, and fungi can counteract immune processes through the effects of secretion/excretion products that interfere with and damage the cells and molecules typical of innate immunity. However, these effects are observed in the later stages of infection, whereas the risk of being recognized and neutralized occurs very early after penetration and involves the pathogen surface components and molecular architecture; therefore, their role becomes crucial, particularly in the earliest pathogenesis. In this review, we analyze the evasion/interference strategies that entomopathogens such as the bacterium Bacillus thuringiensis, fungi, nematocomplexes, and wasps implement in the initial stages of infection, i.e., the phases during which body or cell surfaces play a key role in the interaction with the host receptors responsible for the immunological discrimination between self and non-self. In this regard, these organisms demonstrate evasive abilities ascribed to their body surface and cell wall; it appears that the key process of these mechanisms is the capability to modify the surface, converting it into an immunocompatible structure, or interaction that is more or less specific to host factors.

Structure, Dynamics and Cellular Insight Into Novel Substrates of the Legionella pneumophila Type II Secretion System.

Legionella pneumophila is a Gram-negative bacterium that is able to replicate within a broad range of aquatic protozoan hosts. L. pneumophila is also an opportunistic human pathogen that can infect macrophages and epithelia in the lung and lead to Legionnaires’ disease. The type II secretion system is a key virulence factor of L. pneumophila and is used to promote bacterial growth at low temperatures, regulate biofilm formation, modulate host responses to infection, facilitate bacterial penetration of mucin gels and is necessary for intracellular growth during the initial stages of infection. The L. pneumophila type II secretion system exports at least 25 substrates out of the bacterium and several of these, including NttA to NttG, contain unique amino acid sequences that are generally not observed outside of the Legionella genus. NttA, NttC, and NttD are required for infection of several amoebal species but it is unclear what influence other novel substrates have within their host. In this study, we show that NttE is required for optimal infection of Acanthamoeba castellanii and Vermamoeba vermiformis amoeba and is essential for the typical colony morphology of L. pneumophila. In addition, we report the atomic structures of NttA, NttC, and NttE and through a combined biophysical and biochemical hypothesis driven approach we propose novel functions for these substrates during infection. This work lays the foundation for future studies into the mechanistic understanding of novel type II substrate functions and how these relate to L. pneumophila ecology and disease.

Analysis of the initiation of viral infection under flow conditions with applications to transmission in feed

While kinetic models are widely used to describe viral infection at various levels, most of them are focused on temporal aspects and understanding of corresponding spatio-temporal aspects remains limited. In this work, our attention is focused on the initial stage of infection of immobile cells by virus particles (“virions”) under flow conditions with diffusion. A practical example of this scenario occurs when humans or animals consume food from virion-containing sources. Mathematically, such situations can be described by using a model constructed in analogy with those employed in chemical engineering for analysis of the function of a plug-flow reactor with dispersion. As in the temporal case, the corresponding spatio-temporal model predicts either the transition to a steady state or exponential growth of the populations of virions and infected cells. The spatial distributions of these species are similar in both of these regimes. In particular, the maximums of the populations are shifted to the upper boundary of the infected region. The results illustrating these conclusions were obtained analytically and by employing numerical calculations without and with the dependence of the kinetic parameters on the coordinate. The model proposed has also been used in order to illustrate the effect of antiviral feed additives on feedborne infection towards curbing disease transmission.

Fundamental Basis of COVID-19 Pathogenesis

Abstract At the end of 2019, a new coronavirus infection occurred in the People’s Republic of China with an epicentre in the city of Wuhan. On February 11th, 2020, the World Health Organization assigned the official name of the infection caused by the new coronavirus – COVID-19. COVID-19 has affected people from all over the world given that the infection was noted in 200 countries resulting in annunciation of the pandemic situation. Human corona viruses cause mild to moderate respiratory infections. At the end of 2002, a new coronavirus appeared (SARS-CoV), the causal agent of atypical pneumonia, which caused acute respiratory distress syndrome (ARDS). The initial stage of COVID-19 infection is the penetration of SARS-CoV-2 into target cells that have angiotensin converting enzyme type II receptors. The virus enters the body through the respiratory tract and interacts primarily with toll-like receptors (TLRs). The events in SARS-Cov-2 induced infection follow the next scenario: epithelial cells via TLRs recognize and identify SARS-Cov-2, and after that the information is transmitted to the transcriptional NF-κB, which causes expression of the corresponding genes. Activated in this way, the epithelial cells begin to synthesize various biologically active molecules. The results obtained on preclinical material indicate that ROS generation increases and the antioxidant protection decreases, which plays a major role in the pathogenesis of SARS-CoV, as well as in the progression and severity of this respiratory disease.

Paracoccidioides brasiliensis infection increases regulatory T cell counts in female C57BL/6 mice infected via two distinct routes.

Studies that show an overview of the peripheral immune response in a model of Paracoccidioides brasiliensis (Pb) infection in females are scarce in the literature. We sought to characterize the innate and adaptive immune responses in female C57BL/6 mice infected with Pb through two distinct routes of administration, intranasal and intravenous. In addition to the lung, P. brasiliensis yeast cells were observed in liver and brain tissues of females infected intravenously. To our knowledge, our study is the first to prove the presence of this pathogenic fungus in the cerebral cortex of female mice. During the initial stages of infection, augmented expression of both MHCII and CD86 was observed on the surface of CD11c+ pulmonary antigen-presenting cells (APCs) in intranasally and intravenously infected females. However, CD40 expression was downregulated in these cells. Concomitantly with increasing serum IL-10 levels, we noted that splenic dendritic cells (DCs) from both intravenously- and intranasally-infected female mice had acquired an immature phenotype. Further, increased T regulatory cell counts were observed in female mice infected via both routes, along with an increase in the infiltration of IL-10-producing CD8+ T cells into the lungs. Moreover, we noted that P. brasiliensis infection resulted in enhanced IL-10 production - by CD11c+ APCs in the lung tissue - and induction of Th17 polarization. Taken together, our results suggest that P. brasiliensis could modulates the immune response in female mice by influencing the balance between regulatory T cells (Tregs) and Th17 polarization.

Computed tomography manifestations in super early stage 2019 novel coronavirus pneumonia.

BackgroundThe recent outbreak of pneumonia cases in Wuhan, PR China, was caused by a novel beta coronavirus, the 2019 novel coronavirus (COVID-19).PurposeTo summarize chest computed tomography (CT) manifestations of the early stage of COVID-19 infection and provide a piece of reliable imaging evidence for initial screening and diagnosis.Material and MethodsFrom 10 January 2020 to 10 February 2020, we continuously observed chest CT imaging of 14 patients with clinically suspected new coronavirus infection in the two weeks after onset of symptoms. Ground-glass opacity (GGO), consolidation, reticular pattern, and ground-glass mimic nodules in each patient’s chest CT image were recorded.ResultsWe enrolled 14 patients, of which nine patients had the infection confirmed by reverse transcription polymerase chain reaction (RT-PCR). Five patients were highly suspected of infection. All cases had epidemiological evidence. GGO was a dominant imaging manifestation in the initial days of infection. GGO performance accounts for 40% in 1– 2 days, 90% in 3– 6 days, and 85% in 7– 10 days. With disease progression, consolidation appeared on follow-up CT. Consolidation performance accounts for 0% in 1– 2 days, 40% in 3– 6 days, and 71% in 7– 10 days. The lesions are mostly near the pleura. The number of lesions and the extent of the lesions increased as the disease progressed.ConclusionPatients with novel coronavirus pneumonia have characteristic CT features in the initial stage of infection, which can be used as an essential supplement for nucleic acid examination.

Dynamics in a simple evolutionary-epidemiological model for the evolution of an initial asymptomatic infection stage

Pathogens exhibit a rich variety of life history strategies, shaped by natural selection. An important pathogen life history characteristic is the propensity to induce an asymptomatic yet productive (transmissive) stage at the beginning of an infection. This characteristic is subject to complex trade-offs, ranging from immunological considerations to population-level social processes. We aim to classify the evolutionary dynamics of such asymptomatic behavior of pathogens (hereafter "latency") in order to unify epidemiology and evolution for this life history strategy. We focus on a simple epidemiological model with two infectious stages, where hosts in the first stage can be partially or fully asymptomatic. Immunologically, there is a trade-off between transmission and progression in this first stage. For arbitrary trade-offs, we derive different conditions that guarantee either at least one evolutionarily stable strategy (ESS) at zero, some, or maximal latency of the first stage or, perhaps surprisingly, at least one unstable evolutionarily singular strategy. In this latter case, there is bistability between zero and nonzero (possibly maximal) latency. We then prove the uniqueness of interior evolutionarily singular strategies for power-law and exponential trade-offs: Thus, bistability is always between zero and maximal latency. Overall, previous multistage infection models can be summarized with a single model that includes evolutionary processes acting on latency. Since small changes in parameter values can lead to abrupt transitions in evolutionary dynamics, appropriate disease control strategies could have a substantial impact on the evolution of first-stage latency.

Structural and Proteomic Characterization of the Initiation of Giant Virus Infection

Since their discovery, giant viruses have expanded our understanding of the principles of virology. Due to their gargantuan size and complexity, little is known about the life cycles of these viruses. To answer outstanding questions regarding giant virus infection mechanisms, we set out to determine biomolecular conditions that promote giant virus genome release. We generated four infection intermediates in Samba virus (Mimivirus genus, lineage A) as visualized by cryoelectron microscopy (cryo-EM), cryoelectron tomography (cryo-ET), and scanning electron microscopy (SEM). Each of these four intermediates reflects similar morphology to a stage that occurs invivo. We show that these genome release stages are conserved in other mimiviruses. Finally, we identified proteins that are released from Samba and newly discovered Tupanvirus through differential mass spectrometry. Our work revealed the molecular forces that trigger infection are conserved among disparate giant viruses. This study is also the first to identify specific proteins released during the initial stages of giant virus infection.

Psychological stress creates an immune-suppressive environment that increases the susceptibility of aged mice to Mycobacterium tuberculosis infection

Abstract Age is a risk factor for chronic infections, including tuberculosis (TB). A second compounding factor of TB susceptibility is psychological stress, which is particularly acute in older individuals. While the influence of psychological stress in TB is well documented, the mechanisms by which stress influences M.tb susceptibility in the elderly are almost completely unknown. To study the effects of chronic psychological stress on M.tb infection in old mice, we employed a mouse social stressor model (called social disruption stress or SDR) which involves repeated social defeat in subordinate mice. Young (3 month) and old (18 month) C57BL/6 mice were subjected to SDR and bronchial fluid (BF) and alveolar macrophages (AMs) were isolated by lung lavage. Our studies showed that SDR created an immune-suppressive lung environment in old mice, based on lower levels of pro-inflammatory cytokines and chemokines in bronchial fluid (BF) or their mRNA expression by AMs. AMs from old SDR mice compared to non-stressed mice were also deficient in their ability to mount an inflammatory cytokine response or induce MHC class II mRNA expression. To determine the effect of SDR on M.tb growth, young and old mice were subjected to SDR and infected with M.tb. At 30 days, SDR in both young and old mice reduced M.tb burden. However, this was transient as at 60 days M.tb burden was higher in old mice compared to young mice and SDR further significantly increased M.tb burden in old mice. Also we found that SDR at 60 days in old mice induced significantly higher levels of IL-10 and IL-17 mRNA in the lung, which favors growth of M.tb. In conclusion, our studies suggest that psychological stress during the initial stage of Mtb infection has long-term impact on the control of the infection.

Antibiotic resistance in Pseudomonas aeruginosa and adaptation to complex dynamic environments.

Antibiotic resistance has become a serious threat to human health (WHO Antibacterial Agents in Clinical Development: an Analysis of the Antibacterial Clinical Development Pipeline, Including Tuberculosis. Geneva: World Health Organization; 2017), and the ability to predict antibiotic resistance from genome sequencing has become a focal point for the medical community. With this genocentric prediction in mind, we were intrigued about two particular findings for a collection of clinical Pseudomonas aeruginosa isolates (Marvig et al. Nature Genetics 2015;47:57–64; Frimodt-Møller et al. Scientific Reports 2018;8:12512; Bartell et al. Nature Communications 2019;10:629): (i) 15 out of 52 genes found to be frequently targeted by adaptive mutations during the initial infection stage of cystic fibrosis airways (‘candidate pathoadaptive genes’) (Marvig et al. Nature Genetics 2015;47:57–64) were associated with antibiotic resistance (López-Causapé et al. Fronters in Microbiology 2018;9:685; López-Causapé et al. Antimicrobal Agents and Chemotherapy 2018;62:e02583-17); (ii) there was a parallel lack of resistance development and linkage to the genetic changes in these antibiotic-resistance-associated genes (Frimodt-Møller et al. Scientific Reports 2018;8:12512; Bartell et al. Nature Communications 2019;10:629). In this review, we highlight alternative selective forces that potentially enhance the infection success of P. aeruginosa and focus on the linkage to the 15 pathoadaptive antibiotic-resistance-associated genes, thereby showing the problems we may face when using only genomic information to predict and inform about relevant antibiotic treatment.

Phage capsid nanoparticles with defined ligand arrangement block influenza virus entry.

Multivalent interactions at biological interfaces occur frequently in nature and mediate recognition and interactions in essential physiological processes such as cell-to-cell adhesion. Multivalency is also a key principle that allows tight binding between pathogens and host cells during the initial stages of infection. One promising approach to prevent infection is the design of synthetic or semisynthetic multivalent binders that interfere with pathogen adhesion1-4. Here, we present a multivalent binder that is based on a spatially defined arrangement of ligands for the viral spike protein haemagglutinin of the influenza A virus. Complementary experimental and theoretical approaches demonstrate that bacteriophage capsids, which carry host cell haemagglutinin ligands in an arrangement matching the geometry of binding sites of the spike protein, can bind to viruses in a defined multivalent mode. These capsids cover the entire virus envelope, thus preventing its binding to the host cell as visualized by cryo-electron tomography. As a consequence, virus infection can be inhibited in vitro, ex vivo and in vivo. Such highly functionalized capsids present an alternative to strategies that target virus entry by spike-inhibiting antibodies5 and peptides6 or that address late steps of the viral replication cycle7.

A Dual Role for the Plasminogen Activator Protease During the Preinflammatory Phase of Primary Pneumonic Plague.

Early after inhalation, Yersinia pestis replicates to high numbers in the airways in the absence of disease symptoms or notable inflammatory responses to cause primary pneumonic plague. The plasminogen activator protease (Pla) is a critical Y. pestis virulence factor that is important for early bacterial growth in the lung via an unknown mechanism. In this article, we define a dual role for Pla in the initial stages of pulmonary infection. We show that Pla functions as an adhesin independent of its proteolytic function to suppress early neutrophil influx into the lungs, and that Pla enzymatic activity contributes to bacterial resistance to neutrophil-mediated bacterial killing. Our results suggest that the fate of Y. pestis infection of the lung is decided extremely early during infection and that Pla plays a dual role to tilt the balance in favor of the pathogen.

Comprehensive analysis of the dermatophyte Trichophyton rubrum transcriptional profile reveals dynamic metabolic modulation.

The environmental challenges imposed onto fungal pathogens require a dynamic metabolic modulation, which relies on activation or repression of critical factors and is essential for the establishment and perpetuation of host infection. Wherefore, to overcome the different host microenvironments, pathogens not only depend on virulence factors but also on metabolic flexibility, which ensures their dynamic response to stress conditions in the host. Here, we evaluate Trichophyton rubrum interaction with keratin from a metabolic perspective. We present information about gene modulation of the dermatophyte during early infection stage after shifting from glucose- to keratin-containing culture media, in relation to its use of glucose as the carbon source. Analyzing T. rubrum transcriptome using high-throughput RNA-sequencing technology, we identified the modulation of essential genes related to nitrogen, fatty acid, ergosterol, and carbohydrate metabolisms, among a myriad of other genes necessary for the growth of T. rubrum in keratinized tissues. Our results provide reliable and critical strategies for adaptation to keratin and confirm that the urea-degrading activity associated with the reduction in disulfide bonds and proteolytic activity facilitated keratin degradation. The global modulation orchestrates the responses that support virulence and the proper adaptation to keratin compared with glucose as the carbon source. The gene expression profiling of the host-pathogen interaction highlights candidate genes involved in fungal adaptation and survival and elucidates the machinery required for the establishment of the initial stages of infection.

Differences in physiological traits at the initial stage of Fusarium head blight infection in wheat

Wheat (Triticum aestivum L.) is leading cereal crop worldwide, but its yield is highly affected due to various diseases, especially Fusarium head blight (FHB), which affects the metabolism of plants. The present study was conducted at the Agricultural Institute Osijek using three winter wheat cultivars (Apache, Bezostaya1, and U1) during 2016/2017. The objectives of our studies were to examine differences in physiological characteristics of FHB resistance among wheat cultivars in the early stage of infection. The FHB incidence and severity was the highest in 'Bezostaya1'. Results suggest that activation of some anti-oxidative enzymes in the first 2 h after Fusarium attack was not efficient to prevent disease. 'Apache', which revealed an average FHB incidence, efficiently activated defence response through phenol metabolism elevation. The most effective defence response trough activation of anti-oxidative enzymes triggered by H2O2 was revealed in 'U1', which resulted in a minimal FHB incidence and disease severity. The obtained results confirm differences in defence strategies of wheat genotypes.

Transcriptomic Analysis of the Brown Planthopper, Nilaparvata lugens, at Different Stages after Metarhizium anisopliae Challenge.

Nilaparvata lugens is one of the major pests of rice and results in substantial yield loss every year. Our previous study found that the entomopathogenic fungus Metarhizium anisopliae showed effective potential for controlling this pest. However, the mechanisms underlying M. anisopliae infection of N. lugens are not well known. In the present study, we further examined the transcriptome of N. lugens at 4 h, 8 h, 16 h, and 24 h after M. anisopliae infection by Illumina deep sequencing. In total, 174.17 Gb of data was collected after sequencing, from which 23,398 unigenes were annotated by various databases, including 3694 newly annotated genes. The results showed that there were 246 vs 75, 275 vs 586, 378 vs 1055, and 638 vs 182 up- and downregulated differentially expressed genes (DEGs) at 4 h, 8 h, 16 h, and 24 h after M. anisopliae infection, respectively. The biological functions and associated metabolic processes of these genes were determined with the Clusters of Orthologous Groups (COG), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. The DEGs data were verified using RT-qPCR. These results indicated that the DEGs during the initial fungal infection appropriately reflected the time course of the response to the fungal infection. Taken together, the results of this study provide new insights into the molecular mechanisms underlying the insect host response to fungal infection, especially during the initial stage of infection, and may improve the potential control strategies for N. lugens.

Genetic Resistance to Johne's Disease in Cattle

Paratuberculosis (Johne's disease) is an economically significant, infectious disease of ruminants caused by Mycobacterium avium subsp paratuberculosis, and characterized by progressive weight loss and nonresponsive diarrhea. Genetic factors have been associated with differences in host susceptibility to bovine paratuberculosis. Research has been aimed at detecting associations between susceptibility and polymorphisms at candidate genes with no definitive results. Several gene products are candidates for this association. Interferon gamma is an inducible cytokine with a crucial role in the innate host response to intracellular bacteria. Toll-like receptors are trans-membrane structures responsible for coordination of innate and adaptive immune responses. The SLC11A1 gene plays an important role in innate immunity, preventing bacterial growth in macrophages during the initial stages of infection. The objective of this candidate gene case-control study was to characterize the distribution of polymorphisms in three candidate genes related to the immune function; interferon gamma (BoIFNG), toll-like receptor 4 (TLR4), and solute carrier family 11 member1 (SLC11A1) genes, and test their association with paratuberculosis infection in cattle.

Public health impact of foodborne exposure to naturally occurring virulence-attenuated Listeria monocytogenes: inference from mouse and mathematical models.

Listeriosis is a clinically severe foodborne disease caused by Listeria monocytogenes (Lm). However, approximately 45% of Lm isolates in food carry a virulence-attenuating single-nucleotide polymorphism in inlA, which normally facilitates crossing the intestinal barrier during the initial stages of infection. We hypothesized that (i) natural exposure to virulence-attenuated (vA) Lm strains through food can confer protective immunity against listeriosis attributable to fully virulent (fV) strains and (ii) current food safety measures to minimize exposure to both Lm strains may have adverse population-level outcomes. To test these hypotheses, we evaluated the host response to Lm in a mouse infection model and through mathematical modelling in a human population. After oral immunization with a murinized vA Lm strain, we demonstrated the elicitation of a CD8+ T-cell response and protection against subsequent challenge with an fV strain. A two-strain compartmental mathematical model of human exposure to Lm with cross-protective immunity was also developed. If food safety testing strategies preferentially identify and remove food contaminated by vA strains (potentially due to their common occurrence in foods and higher concentration in food compared to fV strains), the model predicted minimal public health benefit to potentially adverse effects. For example, reducing vA exposures by half, while maintaining fV exposures results in an approximately 6% rise in annual incidence.

Proteomic and phosphoproteomic analyses reveal several events involved in the early stages of bovine herpesvirus 1 infection.

Herpesviruses are predicted to express more than 80 proteins during their infection cycle. The proteins synthesized by the immediate early genes and early genes target signaling pathways in host cells that are essential for the successful initiation of a productive infection and for latency. In this study, proteomic and phosphoproteomic tools showed the occurrence of changes inMadin-Darby bovine kidney cells at the early stageof the infectionbybovine herpesvirus 1 (BoHV-1). Proteins that had already been described in the early stage of infection for other herpesviruses but not for BoHV-1 were found. For example, stathmin phosphorylation at the initial stage of infection is described for the first time. In addition, two proteins that had not been described yet in the early stages of herpesvirus infections in general were ribonuclease/angiogenin inhibitor and Rab GDP dissociation inhibitor beta. The biological processes involved in these cellular responses were repair and replication of DNA, splicing, microtubule dynamics, and inflammatory responses. These results reveal pathways that might be used as targets for designing antiviral molecules against BoHV-1 infection.

Combined effect of the entomopathogenic fungus Metarhizium robertsii and avermectins on the survival and immune response of Aedes aegypti larvae.

Combination of insect pathogenic fungi and microbial metabolites is a prospective method for mosquito control. The effect of the entomopathogenic fungus Metarhizium robertsii J.F. Bischoff, S.A. Rehner & Humber and avermectins on the survival and physiological parameters of Aedes aegypti (Linnaeus, 1762) larvae (dopamine concentration, glutathione S-transferase (GST), nonspecific esterases (EST), acid proteases, lysozyme-like, phenoloxidase (PO) activities) was studied. It is shown that the combination of these agents leads to a synergistic effect on mosquito mortality. Colonization of Ae. aegypti larvae by hyphal bodies following water inoculation with conidia is shown for the first time. The larvae affected by fungi are characterized by a decrease in PO and dopamine levels. In the initial stages of toxicosis and/or fungal infection (12 h posttreatment), increases in the activity of insect detoxifying enzymes (GST and EST) and acid proteases are observed after monotreatments, and these increases are suppressed after combined treatment with the fungus and avermectins. Lysozyme-like activity is also most strongly suppressed under combined treatment with the fungus and avermectins in the early stages posttreatment (12 h). Forty-eight hours posttreatment, we observe increases in GST, EST, acid proteases, and lysozyme-like activities under the influence of the fungus and/or avermectins. The larvae affected by avermectins accumulate lower levels of conidia than avermectin-free larvae. On the other hand, a burst of bacterial CFUs is observed under treatment with both the fungus and avermectins. We suggest that disturbance of the responses of the immune and detoxifying systems under the combined treatment and the development of opportunistic bacteria may be among the causes of the synergistic effect.