Fall Decay Deceleration in Northern Latitudes: Merely a Matter of Cold?

The decomposition of cadavers and large vertebrate carcasses is the result of complex processes primarily influenced by ambient temperatures. Thus, low temperatures can alter decomposition by curtailing tissue autolysis and bacterial decomposition, and by limiting insect activity contributing to necromass removal. In this study, we tested whether carcass decomposition rate is modulated not only directly by temperature and insect occurrence, but also indirectly by the mediation of interactions among insects by ambient temperature. To test this, a comparative analysis of the decomposition of domestic pig carcasses in summer and fall was conducted in Atlantic Canada. The results indicated that carcass decomposition standardized to account for seasonal differences was significantly decelerated in the fall as opposed to the summer during the later decomposition stages and was sometimes incomplete. Moreover, the arrival, presence, and departure of insects from carcasses during ecological succession differed between summer and fall. Necrodes surinamensis (Fabricius) (Coleoptera: Silphidae) and Creophilus maxillosus (Linnaeus) (Coleoptera: Staphylinidae) maintained higher abundances late during succession in the fall than in the summer and their abundance was related to a decline in decomposition rates, probably because these species feed on dipteran larvae promoting necromass removal. These results demonstrate the variability in response to environmental parameters of insects of forensic importance and support the idea that slowed decomposition in the fall may be exacerbated by changes in interspecific interactions among insects. Furthermore, these results suggest that successional studies of insects carried out in the summer have little forensic utility for cadavers found in cold weather conditions.

Facilitation may not be an adequate mechanism of community succession on carrion.

The facilitation model of ecological succession was advanced by plant ecologists in the late 1970s and was then introduced to carrion ecology in the late 1980s, without empirical evidence of its applicability. Ecologists in both disciplines proposed removing early colonists, in this case fly eggs and larvae, from the substrate to determine whether other species could still colonize, which to our knowledge has never been attempted. Here, we tested the facilitation model in a carrion system by removing fly eggs and larvae from carcasses that were exposed in agricultural fields and assigned to one of the following treatment levels of removal intensity: 0, <5, 50, and 100%. Subsequent patterns of colonisation did not provide support for the applicability of the facilitation model in carrion systems. Although results showed, in part, that the removal of fly eggs and larvae decreased the decomposition rate of carcasses, the removal did not prevent colonization by secondary colonizers. Finally, we discuss future studies and make recommendations as to how the facilitation model could be improved, firstly by being more specific about the scale where facilitation is believed to be occurring, secondly by clearly stating what environmental modification is believed to be involved, and thirdly by disentangling facilitation from priority effects.

Bone marrow-derived macrophages and the CNS: An update on the use of experimental chimeric mouse models and bone marrow transplantation in neurological disorders.

The central nervous system (CNS) is a very unique system with multiple features that differentiate it from systemic tissues. One of the most captivating aspects of its distinctive nature is the presence of the blood brain barrier (BBB), which seals it from the periphery. Therefore, to preserve tissue homeostasis, the CNS has to rely heavily on resident cells such as microglia. These pivotal cells of the mononuclear lineage have important and dichotomous roles according to various neurological disorders. However, certain insults can overwhelm microglia as well as compromising the integrity of the BBB, thus allowing the infiltration of bone marrow-derived macrophages (BMDMs). The use of myeloablation and bone marrow transplantation allowed the generation of chimeric mice to study resident microglia and infiltrated BMDM separately. This breakthrough completely revolutionized the way we captured these 2 types of mononuclear phagocytic cells. We now realize that microglia and BMDM exhibit distinct features and appear to perform different tasks. Since these cells are central in several pathologies, it is crucial to use chimeric mice to analyze their functions and mechanisms to possibly harness them for therapeutic purpose. This review will shed light on the advent of this methodology and how it allowed deciphering the ontology of microglia and its maintenance during adulthood. We will also compare the different strategies used to perform myeloablation. Finally, we will discuss the landmark studies that used chimeric mice to characterize the roles of microglia and BMDM in several neurological disorders. This article is part of a Special Issue entitled: Neuro Inflammation edited by Helga E. de Vries and Markus Schwaninger.

REWRITING ECOLOGICAL SUCCESSION HISTORY: DID CARRION ECOLOGISTS GET THERE FIRST?

Ecological succession is arguably the most enduring contribution of plant ecologists and its origins have never been contested. However, we show that French entomologist Pierre Mégnin, while collaborating with medical examiners in the late 1800s, advanced the first formal definition and testable mechanism of ecological succession. This discovery gave birth to the twin disciplines of carrion ecology and forensic entomology. As a novel case of multiple independent discovery, we chronicle how the disciplines of plant and carrion ecology (including forensic entomology) accumulated strikingly similar parallel histories and contributions. In the 1900s, the two groups diverged in methodology and purpose, with carrion ecologists and forensic entomologists focusing mostly on case reports and observational studies instead of hypothesis testing. Momentum is currently growing, however, to develop the ecological framework of forensic entomology and advance carrion ecology theory. Researchers are recognizing the potential of carcasses as subjects for testing not only succession mechanisms (without assuming space-for-time substitution), but also aggregation and coexistence models, diversity-ecosystem function relationships, and the dynamics of pulsed resources. By comparing the contributions of plant and carrion ecologists, we hope to stimulate future crossover research that leads to a general theory of ecological succession.

C-C chemokine receptor type 2 (CCR2) signaling protects neonatal male mice with hypoxic-ischemic hippocampal damage from developing spatial learning deficits.

Chemokines are a family of cytokines involved in the chemotaxis of leukocytes and other target cells by binding to specific G-protein-coupled receptors on their membranes. As such, the activation of C-C chemokine receptor type 2 (CCR2) is involved in the mobilization of "inflammatory" monocytes from bone marrow and in their recruitment to the brain under inflammatory/pathological conditions. In this study, we investigated whether CCR2 signaling could affect the progression of learning deficits and hippocampal damage in a model of neonatal hypoxic-ischemic (HI) brain injury. Postnatal day 3 wild-type (WT) and CCR2 knockout (KO) mice of both sexes were subjected to the Rice-Vannucci model of neonatal hypoxia-ischemia and were followed for up to 14 weeks. HI CCR2 KO male mice were the only animals to exhibit long-term spatial learning deficits in the T-water maze task, compared to their corresponding sham-operated controls. CCR2 KO mouse pups of both sexes had a lower number of circulating monocytes, although only HI CCR2 KO male mice exhibited reduced numbers of activated macrophages/microglia in the damaged hippocampus, compared to WT mice. However, no differences were observed in hippocampal atrophy between HI CCR2 KO and HI WT mice. These results suggest that CCR2 signaling can protect neonatal mice from developing spatial learning deficits after a HI insult, in a sex-specific fashion.

Anti-inflammatory signaling in microglia exacerbates Alzheimer's disease-related pathology.

In this issue of Neuron, Chakrabarty et al. (2015) and Guillot-Sestier et al. (2015) reveal that the anti-inflammatory cytokine IL-10 inhibits Aß clearance by microglia, worsening cognitive decline in mouse models of Alzheimer's disease (AD). These studies provide further support that pro-inflammatory signaling is an innate immune defense mechanism in AD.

The impact of Ly6Clow monocytes after cerebral hypoxia-ischemia in adult mice.

After an ischemic stroke, mononuclear phagocytic cells such as microglia, macrophages, and monocytes migrate to the lesion site and coordinate an immune response. Monocytes, which are recruited from the bloodstream after ischemic brain injury, can be categorized into two subsets in mice: inflammatory and patrolling monocytes. Although inflammatory monocytes (Ly6C(hi)) seem to have a protective role in stroke progression, the impact of patrolling monocytes (Ly6C(low)) is unknown. To address the role of Ly6C(low) monocytes in stroke, we generated bone marrow chimeric mice in which their hematopoietic system was replaced by Nr4a1(-/-) cells, allowing the complete and permanent ablation of Ly6C(low) monocytes without affecting the Ly6C(hi) subset. We then subjected adult mice to cerebral hypoxia-ischemia using the Levine/Vannucci model. Functional outcomes after stroke such as body weight change, neurologic score, motor functions and spatial learning were not affected. Moreover, depletion in Ly6C(low) monocytes did not change significantly the total infarct size, cell loss, atrophy, the number, or the activation state of microglia/macrophages at the lesion site. These data suggest that Ly6C(low) patrolling monocytes are redundant in the progression and recovery of ischemic stroke.

Real-time in vivo imaging reveals the ability of monocytes to clear vascular amyloid beta.

Alzheimer's disease (AD) is characterized by the accumulation of amyloid beta (Aß) that is assumed to result from impaired elimination of this neurotoxic peptide. Most patients with AD also exhibit cerebral amyloid angiopathy, which consists of Aß deposition within the cerebral vasculature. The contribution of monocytes in AD has so far been limited to macrophage precursors. In this study, we aimed to investigate whether circulating monocytes could play a role in the elimination of Aß. With live intravital two-photon microscopy, we demonstrate that patrolling monocytes are attracted to and crawl onto the luminal walls of Aß-positive veins, but not on Aß-positive arteries or Aß-free blood vessels. Additionally, we report the presence of crawling monocytes carrying Aß in veins and their ability to circulate back into the bloodstream. Selective removal of Ly6C(lo) monocytes in APP/PS1 mice induced a significant increase of Aß load in the cortex and hippocampus. These data uncover the ability of Ly6C(lo) monocytes to naturally target and eliminate Aß within the lumen of veins and constitute a potential therapeutic target in AD.

Effect of variable rates of daily sampling of fly larvae on decomposition and carrion insect community assembly: implications for forensic entomology field study protocols.

Experimental protocols in forensic entomology successional field studies generally involve daily sampling of insects to document temporal changes in species composition on animal carcasses. One challenge with that method has been to adjust the sampling intensity to obtain the best representation of the community present without affecting the said community. To this date, little is known about how such investigator perturbations affect decomposition-related processes. Here, we investigated how different levels of daily sampling of fly eggs and fly larvae affected, over time, carcass decomposition rate and the carrion insect community. Results indicated that a daily sampling of <5% of the egg and larvae volumes present on a carcass, a sampling intensity believed to be consistent with current accepted practices in successional field studies, had little effect overall. Higher sampling intensities, however, slowed down carcass decomposition, affected the abundance of certain carrion insects, and caused an increase in the volume of eggs laid by dipterans. This study suggests that the carrion insect community not only has a limited resilience to recurrent perturbations but that a daily sampling intensity equal to or <5% of the egg and larvae volumes appears adequate to ensure that the system is representative of unsampled conditions. Hence we propose that this threshold be accepted as best practice in future forensic entomology successional field studies.

Evidence for a gender-specific protective role of innate immune receptors in a model of perinatal brain injury.

Hypoxia-ischemia is a common cause of neurological impairments in newborns, but little is known about how neuroinflammation contributes to the long-term outcome after a perinatal brain injury. In this study, we investigated the role of the fractalkine receptor chemokine CX3C motif receptor 1 (CX3CR1) and of toll-like receptor (TLR) signaling after a neonatal hypoxic-ischemic brain injury. Mice deficient in the TLR adaptor proteins Toll/interleukin-1 receptor-domain-containing adaptor protein inducing interferon ß (TRIF) or myeloid differentiation factor-88 (MyD88) and CX3CR1 knock-out (KO) mice were subjected to hypoxia-ischemia at postnatal day 3. In situ hybridization was used to evaluate the expression of TLRs during brain development and after hypoxic-ischemic insults. Behavioral deficits, hippocampal damage, reactive microgliosis, and subplate injury were compared among the groups. Although MyD88 KO mice exhibited no differences from wild-type animals in long-term structural and functional outcomes, TRIF KO mice presented a worse outcome, as evidenced by increased hippocampal CA3 atrophy in males and by the development of learning and motor deficits in females. CX3CR1-deficient female mice showed a marked increase in brain damage and long-lasting learning deficits, whereas CX3CR1 KO male animals did not exhibit more brain injury than wild-type mice. These data reveal a novel, gender-specific protective role of TRIF and CX3CR1 signaling in a mouse model of neonatal hypoxic-ischemic brain injury. These findings suggest that future studies seeking immunomodulatory therapies for preterm infants should consider gender as a critical variable and should be cautious not to abrogate the protective role of neuroinflammation.

Toll-like receptor 4 stimulation with the detoxified ligand monophosphoryl lipid A improves Alzheimer's disease-related pathology.

Alzheimer's disease (AD) is the most common cause of dementia worldwide. The pathogenesis of this neurodegenerative disease, currently without curative treatment, is associated with the accumulation of amyloid ß (Aß) in brain parenchyma and cerebral vasculature. AD patients are unable to clear this toxic peptide, leading to Aß accumulation in their brains and, presumably, the pathology associated with this devastating disease. Compounds that stimulate the immune system to clear Aß may therefore have great therapeutic potential in AD patients. Monophosphoryl lipid A (MPL) is an LPS-derived Toll-like receptor 4 agonist that exhibits unique immunomodulatory properties at doses that are nonpyrogenic. We show here that repeated systemic injections of MPL, but not LPS, significantly improved AD-related pathology in APP(swe)/PS1 mice. MPL treatment led to a significant reduction in Aß load in the brain of these mice, as well as enhanced cognitive function. MPL induced a potent phagocytic response by microglia while triggering a moderate inflammatory reaction. Our data suggest that the Toll-like receptor 4 agonist MPL may be a treatment for AD.

Effects of mild chronic cerebral hypoperfusion and early amyloid pathology on spatial learning and the cellular innate immune response in mice.

Understanding the contribution of cerebrovascular factors in the progression of cognitive decline in Alzheimer's disease (AD) is a key step for the development of preventive therapies. Among these factors, chronic cerebral hypoperfusion is an early component of AD pathogenesis that can predict the progression from mild cognitive impairment to AD. Here, we investigated the effects of a protocol of mild chronic cerebral hypoperfusion in the APPswe/PS1 transgenic mouse model of AD. We observed that the permanent occlusion of the right common carotid artery induced spatial learning impairments in young APPswe/PS1 mice, but not in their wild type littermates. Furthermore, the extent of learning deficits strongly correlated with the number of cortical ß-amyloid plaques, with the mobilization of monocytes into the blood and with the number of bone marrow-derived microglia in the brain. These results indicate that a mild reduction of cerebral blood flow can selectively induce cognitive deficits at an early stage of amyloid pathology, eliciting a cellular innate immune response, even without causing neuronal death.

Hematopoietic MyD88-adaptor protein acts as a natural defense mechanism for cognitive deficits in Alzheimer's disease.

Accumulating evidence supports a critical role of Toll-like receptors in the clearance of Amyloid beta (Aß) by microglial cells. Myeloid differentiation factor 88 (MyD88) is an adaptor protein that bridges the intracellular signal to nucleus for most of these innate immune receptors. We investigated here the role of competent MyD88 hematopoietic stem cells on the cognitive decline of a mouse model of Alzheimer's disease (AD). We generated classical chimeric mouse models using irradiation and transplantation of wild type GFP cells and MyD88-deficient cells. Transplantation of GFP cells essentially rescued the cognitive impairment, whereas MyD88-deficient cells significantly accelerated memory deficits of APP(swe)/PS1 mice. Moreover, we found that monocytes and microglia deficient for MyD88 exhibit a functionally impaired phagocytic reaction to Aß.

MyD88-adaptor protein acts as a preventive mechanism for memory deficits in a mouse model of Alzheimer's disease.

BACKGROUND: Alzheimer's disease (AD) is an age-related neurodegenerative disorder associated with brain innate immune activation mainly mediated by microglia. These cells are known to be activated in the brain of AD patients and to produce inflammatory cytokines and neurotoxic molecules in response to Amyloid beta (Aß). Activation of microglia can also promote Aß clearance via Toll-like receptors (TLRs). Myeloid differentiation factor 88 (MyD88) is the adaptor molecule for most of these innate immune receptors, transducing the intracellular signal from TLRs to nucleus. RESULTS: Here, we report that more than 50% reduction in MyD88 expression in a mouse model of AD accelerated spatial learning and memory deficits. Brain of APPswe/PS1-MyD88+/- mice was characterized by a delay in accumulation of Aß plaques and increased soluble levels of Aß oligomers. Furthermore, inflammatory monocyte subset and brain IL-1ß gene expression were significantly reduced in APPswe/PS1 mice with impaired MyD88 signaling. CONCLUSIONS: These data indicate that activation of MyD88 intracellular signaling pathway, likely by TLRs, acts as a natural innate immune mechanism to restrict disease progression of APPswe/PS1 mice.

A statistical approach based on accumulated degree-days to predict decomposition-related processes in forensic studies.

Using pig carcasses exposed over 3 years in rural fields during spring, summer, and fall, we studied the relationship between decomposition stages and degree-day accumulation (i) to verify the predictability of the decomposition stages used in forensic entomology to document carcass decomposition and (ii) to build a degree-day accumulation model applicable to various decomposition-related processes. Results indicate that the decomposition stages can be predicted with accuracy from temperature records and that a reliable degree-day index can be developed to study decomposition-related processes. The development of degree-day indices opens new doors for researchers and allows for the application of inferential tools unaffected by climatic variability, as well as for the inclusion of statistics in a science that is primarily descriptive and in need of validation methods in courtroom proceedings.

Natural and anthropogenic changes in the insect fauna associated with carcasses in the North American Maritime lowlands.

The insect pool available for carrion visitation and colonisation varies with geographical areas, hence the need to build a comprehensive database wherever such data could be used in forensic investigations. However, most of the geographic records on carrion-related insects are from short-term seasonal studies. Here, we provide the year-round taxonomic composition for the dominant ecosystem of the Maritime lowland ecological region that borders the Gulf of St. Lawrence, Canada, and we examine how this composition is subjected to natural (seasonal) as well as anthropogenic changes. Fresh pig carcasses, used here as human surrogates, were exposed recurrently throughout the whole annual period when carrion-related insects are active in forests and adjacent areas subjected to human-induced land cover changes from agricultural practices. A total of 130 necrophageous and predacious insect species representing 2 orders, 18 families and 75 genera were recovered from carcasses. Abundant fly species were able to visit and/or colonise carcasses exposed in both forests and agricultural fields but the species involved varied throughout the year. Conversely, the complex of abundant coleopterans found on carcasses remained stable throughout the year but differed between forests and agricultural fields. Considering the seasonal and anthropogenic changes that were observed in the complex of carrion-related insects, we stressed that inference on the taxonomic composition in relation to minimum postmortem interval should be restricted to a specific habitat and time of the year. These results also have methodological implications, suggesting that the experimental designs of forensic studies in temperate areas require adjustments to permit robust estimations of minimum postmortem intervals from the insect fauna associated with carcasses.

Predicting the visitation of carcasses by carrion-related insects under different rates of degree-day accumulation.

Common assumptions in forensic entomology are that insects visit and colonize carcasses following a predictable sequence, and that this succession varies among seasons. However, currently available evidence for insect succession on decomposing bodies is essentially descriptive and, to our knowledge, the fine-scale predictability of insect succession with respect to seasons has never been confirmed statistically. In this study, we test these assumptions through the sampling of carrion-related insects attracted to pig carcasses. The study was carried out during the summer and fall of 2006 in rural fields of New Brunswick, Canada. Of the five species of carrion-related insects with high enough occurrence on carcasses to allow modelling, three showed predictable occurrence with respect to degree-day accumulation and seasonal effects. This demonstrates that the occurrence probability of some carrion-related insects on carcasses can be estimated from meteorological records even across seasons with different rates of degree-day accumulation. As opposed to the prevailing idea that adult insects are not reliable for post-mortem interval estimation, the adults of some species exhibited a specific pattern of visitation that could be determined and used in forensic investigations. It is stressed, however, that the statistical predictability of species occurrence must be assessed before any species is considered as a post-mortem interval indicator.