Neurotransmitter acetylcholine mediates the mummification of Ophiocordyceps sinensis-infected Thitarodes xiaojinensis larvae.

Parasites can manipulate host behavior to facilitate parasite transmission. One such host-pathogen interaction occurs between the fungus Ophiocordyceps sinensis and the ghost moth Thitarodes xiaojinensis. O. sinensis is involved in the mummification process of infected host larvae. However, the underlying molecular and chemical mechanism for this phenomenon is unknown. We characterized the small molecules regulating host behaviors and the altered metabolites in infected and mummified host larvae. Lipid-related metabolites, such as phosphatidylcholine, were identified in infected and mummified larvae. Decreased levels of the neurotransmitter acetylcholine (ACh) and elevated choline levels were observed in the brains of both the infected and mummified larvae. The aberrant activity of acetylcholinesterase (AChE) and relative mRNA expression of ACE2 (acetylcholinesterase) may mediate the altered transformation between ACh and choline, leading to the brain dysfunction of mummified larvae. Caspofungin treatment inhibited the mummification of infected larvae and the activity of AChE. These findings indicate the importance of ACh in the mummification of host larvae after O. sinensis infection.IMPORTANCEOphiocordyceps sinensis-infected ghost moth larvae are manipulated to move to the soil surface with their heads up in death. A fruiting body then grows from the caterpillar's head, eventually producing conidia for dispersal. However, the underlying molecular and chemical mechanism has not been characterized. In this study, we describe the metabolic profile of Thitarodes xiaojinensis host larvae after O. sinensis infection. Altered metabolites, particularly lipid-related metabolites, were identified in infected and mummified larvae, suggesting that lipids are important in O. sinensis-mediated behavioral manipulation of host larvae. Decreased levels of the neurotransmitter acetylcholine were observed in both infected and mummified larvae brains. This suggests that altered or reduced acetylcholine can mediate brain dysfunction and lead to aberrant behavior. These results reveal the critical role of acetylcholine in the mummification process of infected host larvae.

Characteristics of unilateral and bilateral bovine twin foetuses.

Twin pregnancy in cattle is undesirable for a number of reasons, including a higher abortion risk compared to pregnancies with a single foetus. Yet, the abortion risk is significantly influenced by the intrauterine location of the foetuses, that is, the abortion risk is several times higher if they are implanted in the same uterine horn (unilateral twin pregnancy) than if they are implanted with one foetus in each uterine horn (bilateral twin pregnancy). The reason for the higher abortion risk in unilateral twin pregnancies is unknown, but it may be related to malnutrition of the outermost foetus due to a limited placental capacity, as is the case for equine twin foetuses. A slaughterhouse study was performed and the foetuses of cattle pregnant with twins were measured. We identified 65 cases of twin pregnancies, of which 35 were unilateral twin pregnancies and 30 were bilateral twin pregnancies. There was no significant difference between the outermost and the more centrally located foetus in unilateral twin pregnancies in terms of body weight and length of the metacarpal diaphysis. Growth retardation of the outermost foetus could therefore not be confirmed as the cause of the higher abortion risk in unilateral bovine twin pregnancies. Four cases of pre-slaughter foetal mortality were identified. In three of these cases, both twins were dead, of equal size and at a comparable level of degradation. In the fourth case, with approximately 40-day-old twin foetuses of equal size, only one of the foetuses showed signs of pre-slaughter death.

Mummification in a forensic context: an observational study of taphonomic changes and the post-mortem interval in an indoor setting.

The objective of this study was to evaluate the presence of mummification in an indoor setting, with an emphasis on the forensic perspective. A dataset of 102 forensic autopsy cases was assessed for distribution of desiccation of skin and soft tissue (i.e., subcutaneous fat and musculature) and for moist decompositional (i.e., putrefactive) changes. Further, possible correlation with the post-mortem interval (PMI) was evaluated, as well as the effects of clothing coverage of the body. The results indicated that yellow to orange parchment-like desiccated skin was found at significantly shorter PMIs than reddish brown to black leathery desiccated skin, even when soft tissue desiccation was included in the comparative analysis. Clothing appeared to have a significant decelerating effect on the extent of desiccation on the legs, but findings in regard to whole body or torso/arms were inconclusive. A large variation in PMIs was evident as regards fully desiccated skin (PMI 18-217 days), indicating difficulties in PMI estimation due to a variable repressive effect on the decompositional process per se in an indoor setting. For the specific case in forensic practice, no definite conclusion can be drawn from the observed desiccation changes to the PMI. One way forward might be creating a systematic and standardized method for describing different desiccation types, as well as other cooccurring decompositional changes and how they relate to the PMI, as a foundation for a future quantification model.

"Salt mummification" - atypical method of embalming a corpse.

It is extremely rare for table salt to be used to preserve a dead body in criminal cases. In the case presented here, after the death of his 85-year-old mother, a son kept her body preserved in table salt for about 2 years to extort social benefits (pension). Before her death, the woman had been hospitalised twice due to chronic diseases. The case has been examined by the multi-disciplinary team. The unusual conditions in which the corpse was stored influenced its good condition (close to mummification), with limited colonisation of the corpse by necrophagous insects and insects involved in soft tissue biolysis (i.e. selected Diptera or Coleoptera). The use of table salt inhibited the growth of most fungi which would normally be present on a corpse stored in ambient conditions, and the corpse's surface was colonised by halophilic fungus (Scopulariopsis brevicaulis).

Technical note: A comparison between rehydrating solutions in the pretreatment of mummified and corified skin for forensic microscopic examination.

Microscopic examination of mummified or corified skin may be of extreme importance for forensic purposes. However, standard histological samples in these cases are low-end, and preparation is burdened by several problems and so are diagnostic results: an improvement of these types of specimens is therefore advantageous. This study aims to identify the best performing rehydration solution among a fabric softener, a body lotion, and Sandison's rehydrating solution. Samples of skin undergoing mummification or corification were collected from 25 corpses and each sample was divided into 4 fragments: one of these fragments was directly fixated in 4% formalin, one was previously treated with a tissue softener, another one was previously treated with a body lotion, and the last one was treated with Sandison's solution. After 72 h, the pretreated samples were post-fixated in 4% formalin and then prepared for standard histological examination staining the histological slides with hematoxylin-eosin and Masson's trichrome. At the microscopic examination, samples directly fixated in formalin were characterized by usual marked structural alterations and altered stainability, typical of such dry tissues. Vice versa, those previously treated appeared to be better-preserved even though with different improvement levels: body lotion made a medium-low-grade restoration of the tissues, and fabric softener a high-grade restoration, while Sandison's rehydrating solution produced an optimal grade restoration. Sandison's rehydrating solution was confirmed to be the best rehydrating substance for mummified and corified skin. Fabric softener could be, however, considered a valid substitute, being productive of high-grade microscopic yield.

Rethinking the Process of Animal Mummification in Ancient Egypt: Molecular Characterization of Embalming Material and the Use of Brassicaceae Seed Oil in the Mummification of Gazelle Mummies from Kom Mereh, Egypt.

The study of animal mummification in ancient Egypt has recently received increasing attention from a number of modern scholars given the fact that this part of ancient Egyptian funerary and religious history is a practice yet to be fully understood. In this study, nine samples of embalming matter were extracted from six gazelle mummies from the archaeological site of Kom Mereh (modern village of Komir), dated to the Roman period of dominance in ancient Egypt. All samples were analyzed for the presence of inorganic and organic matter applying a multi-analytical approach based on Fourier transform infrared spectroscopy (FT-IR) and gas chromatography-mass spectrometry (GC-MS). Furthermore, in order to identify more specific compounds such as bitumen and beeswax in studied balms, each sample was subjected to a solid phase extraction (SPE) and saponification separation process, respectively. The results of this study revealed that the majority of the analyzed embalming substances sampled from six gazelle mummies from Kom Mereh were complex mixtures of plant oils, animal fats, conifer resin, and beeswax. In this regard, this study was able to report a practice until now unmentioned in the scientific literature, namely, the use of cruciferous oil, derived from seeds of Brassicaceae plants, in animal mummification.

Transcriptome analysis reveals key genes and pathways associated with piglet fetal mummification.

China has the largest pork consumption worldwide. However, the high incidence of piglet fetal mummification (3%-5%) is an important factor that causes the slow improvement of pig reproductive capacity, and the genetic mechanism is still unclear. This study aimed to identify candidate genes associated with piglet fetal mummification. RNA-seq technology was used to compare transcriptome profiling of blood from healthy and mummified piglets at different stages of pregnancy (35, 56, 77, and 98 days). A total of 137-420 differentially expressed genes (DEGs) were detected at each stage. Seven DEGs were significantly differentially expressed at various stages. IL-9R, TLR8, ABLIM3, FSH-α, ASCC1, PRKCZ, and GCK may play important roles in the course of piglet fetal mummification. The differential genes we identified between the groups were mainly enriched in immune and inflammation regulation, while others were mainly enriched in reproduction. Considering the function of candidate genes, IL-9R and TLR8 were suggested as the most promising candidate genes involved in mummified piglet traits. We speculate that during pregnancy, it may be the combined effects of the above-mentioned inflammation, immune response, and reproduction-related signaling pathways that affect the occurrence of mummified piglets and further affect pig reproduction.

Technical note-stabilization of cadaveric corified and mummified skin thanks to prolonged temperature.

Microscopic study and interpretation of skin samples obtained from mummified/corified cadavers request the adoption of special laboratory techniques in order to avoid that the histological samples may detach from the slide during staining processes. We have assessed an experimental and extended drying of corified and mummified skin samples in order to evaluate an eventual improvement after staining procedures. Samples of skin undergoing different post-mortem putrefactive phenomena-from parchment-like to mummification-were collected from 16 corpses and divided in halves: one half was fixated in buffered formalin while the other half was previously treated with Sandison's rehydrating solution. The so-obtained sections were subsequently dried for one night at 37 °C and stained with Hematoxylin & Eosin staining and Masson's Trichrome staining according to Goldner. At microscope observation, 69% of the dyed sections stained with H&E and 53% of the sections stained with Masson's Trichrome was lost during different steps of the coloring procedures. We then proceeded with a new cutting of the slides and decided to experimentally dry them for 5 days at a mean temperature of 37 °C: purpose of this procedure was to gradually remove the possible liquid component of the samples probably culpable of the detachment. With this modified procedure, all the sections resulted to be preserved in the following staining steps and showed, moreover, an improvement of the morphological structure, especially in those previously treated with Sandison's rehydrating solution. In those cases where a histological examination is requested on skin samples collected from corified or mummified cadavers, this technique is highly recommended in order to obtain microscopic slide of high quality.

An octavalent vaccine provides pregnant gilts protection against a highly virulent porcine parvovirus strain.

BACKGROUND: Porcilis® Ery+Parvo+Lepto is an octavalent inactivated ready-to-use vaccine that contains Erysipelothrix rhusiopathiae (Ery), porcine parvovirus (PPV), and six serogroups of Leptospira (Lepto). The efficacy of Porcilis® Ery + Parvo+Lepto against reproductive problems associated with porcine parvovirus (PPV) infection was evaluated in pregnant gilts. For this, a group of ninegilts was vaccinated twice (at 5 and 6 months old) with Porcilis® Ery + Parvo+Lepto (Group 1), while a group of eight gilts was included as unvaccinated controls (Group 2). All pigs were artificially inseminated 4 weeks after the second vaccination. They were challenged during early gestation with PPV-27a, a virulent cluster D strain, and euthanized to collect their offspring by hysterectomy around day 90 in pregnancy. Antibody responses against PPV in gilts were measured, and the presence of PPV in progeny was also determined. RESULTS: No clinical signs were observed after vaccination. After PPV challenge, all foetuses from the vaccinated gilts were alive (132/132), while in the unvaccinated group only 41% were alive (46/112), 19.6% were dead and 39.4% of the offspring (44/112) were mummified. PPV could be detected by qPCR in 14% of the progeny from vaccinated gilts at an average of 4.7 log10/ml, whereas this was significantly higher in the control group, where 90% of the progeny were PPV positive, with titres of 9.8 log10/ml on average. CONCLUSIONS: The present study demonstrates that vaccination of gilts with Porcilis® Ery + Parvo+Lepto was safe and induced an immune response sufficient to protect progeny against PPV by reducing transplacental infection.

Report on fetal mummification in the scalloped hammerhead shark Sphyrna lewini.

This study describes the fetal mummification process in two embryos of a 310 cm total length scalloped hammerhead shark Sphyrna lewini caught in southeastern Brazil, in December 2017. Fourteen embryos were observed in total, in which two males in the left uterus presented different stages of mummification. Both mummified embryos were covered by an exudate (i.e., a mucous substance), indicating a hematic mummification process. All embryos were at the placentotrophic stage of development, indicating that they were close to parturition. An intrinsic characteristic is suggested as possible etiology for this condition, such as umbilical torsion, because both embryos were at different sizes and, therefore, at different development stages. In addition, the sample size did not allow the authors to presume any pollution effect once only one female was observed. Finally, fetal mummification and other embryonic development disorders might have populational impacts due to reduction in embryo survival and, consequently, recruitment. For this reason and considering that S. lewini is categorized as a "critically endangered species," this study's results have conservational relevance.

The environmental variables that impact human decomposition in terrestrially exposed contexts within Canada.

Little is known about the nature and trajectory of human decomposition in Canada. This study involved the examination of 96 retrospective police death investigation cases selected using the Canadian ViCLAS (Violent Crime Linkage Analysis System) and sudden death police databases. A classification system was designed and applied based on the latest visible stages of autolysis (stages 1-2), putrefaction (3-5) and skeletonisation (6-8) observed. The analysis of the progression of decomposition using time (Post Mortem Interval (PMI) in days) and temperature accumulated-degree-days (ADD) score found considerable variability during the putrefaction and skeletonisation phases, with poor predictability noted after stage 5 (post bloat). The visible progression of decomposition outdoors was characterized by a brown to black discolouration at stage 5 and remnant desiccated black tissue at stage 7. No bodies were totally skeletonised in under one year. Mummification of tissue was rare with earlier onset in winter as opposed to summer, considered likely due to lower seasonal humidity. It was found that neither ADD nor the PMI were significant dependent variables for the decomposition score with correlations of 53% for temperature and 41% for time. It took almost twice as much time and 1.5 times more temperature (ADD) for the set of cases exposed to cold and freezing temperatures (4°C or less) to reach putrefaction compared to the warm group. The amount of precipitation and/or clothing had a negligible impact on the advancement of decomposition, whereas the lack of sun exposure (full shade) had a small positive effect. This study found that the poor predictability of onset and the duration of late stage decomposition, combined with our limited understanding of the full range of variables which influence the speed of decomposition, makes PMI estimations for exposed terrestrial cases in Canada unreliable, but also calls in question PMI estimations elsewhere.

Mannitol mediates the mummification behavior of Thitarodes xiaojinensis larvae infected with Ophiocordyceps sinensis.

Introduction: Parasites can facilitate their own spread and reproduction by manipulating insect hosts behavior, as seen in the interaction between Thitarodes xiaojinensis and Ophiocordyceps sinensis. Infection by O. sinensis leads to the mummification of T. xiaojinensis larvae, but the underlying mechanisms remain mysterious. Methods: The morphology of O. sinensis infected larvae and fungal growth were first observed. Subsequently, the metabolite changes in the larvae before and after infection with the fungus were analyzed by LC/MS and targeted metabolomics. The expression of mannitol-related genes was detected using RT-qPCR, and morphological changes in larvae were observed after injection of different concentrations of mannitol into the O. sinensis-infected larvae. Results: Significant changes were found in phenotype, fungal morphology in hemocoel, larval hardness, and mannitol metabolites in infected, mummified 0 h larvae and larvae 5 days after mummification behavior. Surprisingly, the occurrence of mummification behavior was accompanied by fungal dimorphism, as well as the absence of mannitol in both infected and non-infected larvae, until the initial accumulation of mannitol and the expression of mannitol-associated genes occurred at the time of mummification behavior. The presence of mannitol may promote fungal dimorphism to mediate changes in fungal toxicity or resistance, leading to the end of the fungus-insect coexistence period and the incidence of mummification behavior. Furthermore, mannitol injections increase the mummification rate of the infected larvae without significant difference from the normal mummification phenotype. Discussion: This finding suggests the importance of mannitol in the mummification of host larvae infected with O. sinensis.

Still waters run deep: a unique case of mummification under extreme conditions.

In rural Crete, Greece, a mummified body was discovered inside a screw-top plastic barrel next to a dry riverbed. Local authorities conducted a field investigation along with a forensic pathologist, and the body was transferred inside the barrel to the Forensic Medicine Unit of the University of Crete for further investigation. Forensic examination included radiographs taken using a portable X-ray machine in situ followed by extraction of the remains, a full-body postmortem computed tomography scan, autopsy, DNA extraction, tissue and biofluid sampling, and histological and toxicological analyses. Two butane camping gases were recovered from inside the barrel. The remains were determined to belong to a middle-aged adult male. The postmortem interval was estimated to be over 4 months. DNA comparison revealed that the body belonged to a 58-year-old man who was reported missing 28 months prior to discovery. Examinations showed no evidence of skeletal or other trauma, and death was attributed to asphyxia due to oxygen deprivation. Careful examination of the barrel, which bore several peculiar modifications, in conjunction with contextual information pertaining to the deceased's personal life, led to successful case resolution. Key points: Advanced decomposition and taphonomic alterations present challenges for forensic practitioners.A case of a set of mummified remains discovered inside a plastic screw-top barrel is reported.A multidisciplinary approach, including examination of the barrel and contextual information regarding the deceased, led to case resolution.

The Belowground-Aboveground Interactions of Zucchini: The Effects of Trichoderma afroharzianum Strain T22 on the Population and Behavior of the Aphid Aphis gossypii Glover and Its Endoparasitoid Aphidius colemani Viereck.

Fungi belonging to the genus Trichoderma have received high consideration in agriculture due to their beneficial effects on crops from their plant promotion effects and protection from disease. A role of Trichoderma fungi in triggering plant defense mechanisms against insect pests, either directly or by natural enemy attraction, has been proposed, even if the results in different studies are controversial. In this present study, using zucchini plants as a model species, we investigated the effects of Trichoderma afroharzianum strain T22 plant inoculation on the cotton aphid Aphis gossypii and its endoparasitoid Aphidius colemani. Our results showed that the inoculation with T. afroharzianum T22 promotes A. gossypii population growth and makes zucchini more attractive to the aphid. The higher abundance of aphids on Trichoderma-inoculated zucchini was compensated for by a higher presence of the mummies of Aphidius colemani. In this present study, we recorded a higher zucchini biomass, thereby confirming that Trichoderma can act as a plant growth inducer.

A Rare Phenomenon of Natural Precocious Mummification.

Immediately after death, specific changes occur in the human body, leading to the total dissolution of the soft tissues and internal organs. In some cases, when in suitable conditions, the decomposition process could stop and be displaced by mummification. The last one is time-consuming and needs several weeks to months to set in completely. We present a case of a 34-year-old man found dead 16 days after being last seen alive in a stage of complete mummification. Natural mummification occurring in less than one month is termed precocious mummification and is rarely observed in temperate regions. With only a few cases reported globally, this case is essential for the forensic community. It will help better know the mummification processes and estimate the time since death.

Toxicological Analysis of Drugs in Human Mummified Bodies and Proposed Guidelines.

From palaeopathology to forensic taphonomy, mummified human bodies constitute biological archives of paramount importance. Toxicology analysis of endobiotics and xenobiotics has already shown value to archaeological mummies research with detecting heavy metals, sedative-hypnotic drugs, and stimulants. Thanks to the large window of drug detection in hair and nails, the information from such studies has increased the scientific community's knowledge regarding past populations' lifestyles. Still, few bibliographic references exist regarding toxicology reports in mummified bodies from forensic settings. Here, the authors aim to draw attention to the valuable contribution of toxicology analysis, taking into account previously conducted studies and their findings. Given that sample collection on mummified bodies from forensic scenarios may not always happen in laboratories or autopsy rooms, the authors also suggest guidelines for in situ sampling of forensic mummies. It is expected that the present technical note will encourage experts to perform toxicology analysis in mummified bodies and publish their case reports more often.

Examination on the Occurrence of Coinfections in Diagnostic Transmittals in Cases of Stillbirth, Mummification, Embryonic Death, and Infertility (SMEDI) Syndrome in Germany.

The stillbirth, mummification, embryonic death, and infertility (SMEDI) syndrome is most commonly associated with porcine parvovirus 1 (PPV1) infections. Little is known about the occurrence of coinfections with SMEDI-associated pathogens and the associations among these pathogens. In our study, we included 40 SMEDI-affected litters from 18 different farms. In total, 158 out of 358 available fetuses from diagnostic transmittals were selected by systematic random sampling and examined for PCV2, PCV3, PPV1, and Leptospira spp. by q-PCR. Results from diagnostic materials showed the following results: in eleven farms, PCV2 was present; in nine farms, PPV1 was present; in five farms, PCV3 was present; and in two farms, Leptospira spp. was present. The detection of Leptospira spp. was significantly associated with a PCV2 coinfection (OR: 26.3; p < 0.001). PCV3 positivity resulted in a reduced probability of detecting PCV2 in the corresponding fetus (OR: 0.078; p = 0.008). Fetal maceration was associated with Leptospira spp. detection (OR: 8.6; p = 0.003), whereas mummification (p = 0.047), reduced crown-rump length (p < 0.001), and bodyweight (p = 0.001) of fetuses were significantly associated with PPV1 and PCV2 coinfection and thus, presumably, a shorter time to death after infection, indicating an enhanced negative effect on the development of fetuses with PCV2 + PPV1 coinfection.

Neuropathologic findings in a young woman 4 years following declaration of brain death: case analysis and literature review.

Brain death (death by neurologic criteria) is declared in 2% of all in-hospital deaths in the United States. Published neuropathology studies of individuals maintained on cardiorespiratory support are generally decades old, and notably include only 3 cases with long intervals between brain and "somatic" death (68 days, 101 days, 20 years). Here, we share our observations in a young woman supported for nearly 4½ years following declaration of brain death after oropharyngeal surgery. While limited by tissue availability and condition, we found evidence of at least partial perfusion of the superficial cerebral and cerebellar cortices by external carotid and vertebral arteries (via meningeal and posterior pharyngeal branches), characterized by focal cellular reaction and organization. Dural venous sinuses had thrombosis and recanalization, as well as iron deposition. In nonperfused brain areas, tissue "mummification," akin to that seen in certain postmortem conditions, including macerated stillbirths and saponification (adipocere formation), was identified, and are reviewed herein. Unfortunately, correlation with years-earlier clinical and radiographic observations was not possible. Nevertheless, we feel that our careful neuropathologic inspection of this case expands the understanding of the spectrum of human brain tissue alterations possible in a very rarely seen set of conditions.

Dermestes (s.str.) haemorrhoidalis (Coleoptera: Dermestidae)-The Most Frequent Species on Mummified Human Corpses in Indoor Conditions? Three Cases from Southwestern Poland.

Beetles of the family Dermestidae, especially of the genus Dermestes Linnaeus, 1758, are often identified on corpses in late stages of decomposition. They usually feed on remains devoid of organs and soft tissues or when the corpses undergo mummification. In Europe, eight species from two subgenera Dermestes and Dermestinus Zhantiev, 1967, have so far been identified on human corpses. Despite the relatively frequent presence of Dermestes sp. in experimental studies conducted in Poland, no reports concerning Dermestes directly collected from human corpses have been published to date. This article again describes observations of Dermestidae collected from human corpses found in indoor conditions in Wroclaw, the capital of the Dolnoslaskie Voivodeship. For the second time, there is evidence of the presence of Dermestes (s.str.) haemorrhoidalis on human corpses-a species considered to be relatively rare, as evidenced by faunistic data published from Poland, as well as the results of ongoing experiments of forensic interest.

Blunt head trauma in a peculiar case of partial mummification. Murder or accidental event?

The finding of a partially mummified body presenting signs of trauma requires the forensic pathologist to conduct a careful and complex examination; multidisciplinary analysis is often necessary.We report a case where the partially mummified corpse of an elderly man was found in his own home more than seven years after death. Complete post-mortem investigation revealed a cranial fracture and an acute subdural haematoma.An in-depth multidisciplinary analysis provided important information on the modality and cause of death but it was not possible to establish whether the trauma and death resulted from an accidental event or from an assault.