Inter-generational Consequences for Growing C.elegans in Liquid

AbstractIn recent years, studies in Caenorhabditis elegans nematodes have shown that different stresses can generate multigenerational changes. Here we show that worms that grow in liquid media, and also their plate-grown progeny, are different from worms whose ancestors were grown on plates. It has been suggested that C.elegans might encounter liquid environments in nature, although actual observations in the wild are few and far between. In contrast, in the lab, growing worms in liquid is commonplace, and often used as an alternative to growing worms on agar plates, to control the composition of the worms’ diet, to starve (and synchronize) worms, or to grow large populations for biochemical assays. We found that plate-grown descendants of M9 liquid media-grown worms were longer than control worms, and the heritable effects were apparent already very early in development. We tested for the involvement of different known epigenetic inheritance mechanisms, but could not find a single mutant in which these intergenerational effects are canceled. While we found that growing in liquid always leads to inter-generational changes in the worms’ size, trans-generational effects were found to be variable, and in some cases the effects were gone after 1 -2 generations. These results demonstrate that standard cultivation conditions in early life can dramatically change the worms’ physiology in adulthood, and can also affect the next generations.

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Inter-generational consequences for growing Caenorhabditis elegans in liquid

Philosophical Transactions of the Royal Society B Biological Sciences ◽

10.1098/rstb.2018.0125 ◽

2019 ◽

Vol 374 (1770) ◽

pp. 20180125 ◽

Cited By ~ 6

Author(s):

Itamar Lev ◽

Roberta Bril ◽

Yunan Liu ◽

Lucila Inés Ceré ◽

Oded Rechavi

Keyword(s):

Caenorhabditis Elegans ◽

Early Life ◽

Epigenetic Inheritance ◽

Evolutionary Medicine ◽

C Elegans ◽

Generational Effects ◽

Large Populations ◽

Two Generations ◽

Generational Changes ◽

In The Wild

In recent years, studies in Caenorhabditis elegans nematodes have shown that different stresses can generate multigenerational changes. Here, we show that worms that grow in liquid media, and also their plate-grown progeny, are different from worms whose ancestors were grown on plates. It has been suggested that C. elegans might encounter liquid environments in nature, although actual observations in the wild are few and far between. By contrast, in the laboratory, growing worms in liquid is commonplace, and often used as an alternative to growing worms on agar plates, to control the composition of the worms' diet, to starve (and synchronize) worms or to grow large populations for biochemical assays. We found that plate-grown descendants of M9 liquid medium-grown worms were longer than control worms, and the heritable effects were already apparent very early in development. We tested for the involvement of different known epigenetic inheritance mechanisms, but could not find a single mutant in which these inter-generational effects are cancelled. While we found that growing in liquid always leads to inter-generational changes in the worms’ size, trans-generational effects were found to be variable, and in some cases, the effects were gone after one to two generations. These results demonstrate that standard cultivation conditions in early life can dramatically change the worms' physiology in adulthood, and can also affect the next generations. This article is part of the theme issue ‘Developing differences: early-life effects and evolutionary medicine’.

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Antifungal activity of extracts from endophytic fungi associated with Smallanthus maintained in vitro as autotrophic cultures and as pot plants in the greenhouse

Canadian Journal of Microbiology ◽

10.1139/w2012-088 ◽

2012 ◽

Vol 58 (10) ◽

pp. 1202-1211 ◽

Cited By ~ 14

Author(s):

Luiz H. Rosa ◽

Nurhayat Tabanca ◽

Natascha Techen ◽

Zhiqiang Pan ◽

David E. Wedge ◽

...

Keyword(s):

Antifungal Activity ◽

Endophytic Fungi ◽

Plant Pathogens ◽

Cultivation Conditions ◽

Smallanthus Sonchifolius ◽

Fungal Plant Pathogens ◽

In The Wild ◽

Fungal Assemblages ◽

Pot Plants

The endophytic fungal assemblages associated with Smallanthus sonchifolius (Poepp.) H. Rob. and Smallanthus uvedalius (L.) Mack. ex Small growing in vitro autotrophic cultures and in the greenhouse were identified and evaluated for their ability to produce bioactive compounds. A total of 25 isolates were recovered that were genetically closely related to species of the genera Bionectria , Cladosporium , Colletotrichum , Fusarium , Gibberella , Hypocrea , Lecythophora , Nigrospora , Plectosphaerella , and Trichoderma . The endophytic assemblages of S. sonchifolius presented a greater diversity than the group isolated from S. uvedalius and demonstrated the presence of dominant generalist fungi. Extracts of all fungi were screened against the fungal plant pathogens. Ten extracts (41.6%) displayed antifungal activities; some of them had a broad antifungal activity. The phylotypes Lecythophora sp. 1, Lecythophora sp. 2, and Fusarium oxysporum were isolated from in vitro autotrophic cultures and displayed antifungal activity. The presence of bioactive endophytic fungi within S. sonchifolius and S. uvedalius suggests an ecological advantage against pathogenic attacks. This study revealed reduced numbers of endophytes in association with both Smallanthus species in controlled cultivation conditions compared with the endophytic communities of hosts collected in the wild environments. Even as reduced endophytic communities, these fungi continue to provide chemical protection for the host.

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Testing for a role of Dnmt2 in paternal trans-generational immune priming

10.1101/422063 ◽

2018 ◽

Cited By ~ 2

Author(s):

Nora K E Schulz ◽

Maike F Diddens-de Buhr ◽

Joachim Kurtz

Keyword(s):

Potential Role ◽

Epigenetic Inheritance ◽

Red Flour Beetle ◽

Adult Offspring ◽

Paternal Effects ◽

Immune Priming ◽

Transgenerational Epigenetic Inheritance ◽

Generational Effects ◽

Underlying Mechanisms

AbstractTrans-generational effects from fathers to offspring are increasingly reported from diverse organisms, but the underlying mechanisms are often unknown. Paternal trans-generational immune priming (TGIP) was demonstrated in the red flour beetle Tribolium castaneum: non-infectious bacterial exposure (priming) of fathers protects their offspring against an infectious challenge. Here we studied a potential role of the Dnmt2 (now also called Trdnmt1) gene, which encodes a highly conserved enzyme that provides CpG methylation to a set of tRNAs and has previously been reported to be involved in transgenerational epigenetic inheritance in mice. We first studied gene expression and found that Dnmt2 was expressed throughout life, with high expression in testes. Knockdown of Dnmt2 in fathers slowed down offspring larval development and increased mortality of the adult offspring upon bacterial infection. However, the observed effects were independent of the paternal priming treatment. In conclusion, our results point towards a role of Dnmt2 for paternal effects, while elucidation of the mechanisms behind paternal TGIP needs further studies.

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Intergenerational effects of a paternal Western diet during adolescence on offspring gut microbiota, stress reactivity and social behavior

10.1101/2021.09.16.460599 ◽

2021 ◽

Author(s):

Carina Bodden ◽

Terence Y. Pang ◽

Yingshi Feng ◽

Faria Mridha ◽

Geraldine Kong ◽

...

Keyword(s):

Gut Microbiota ◽

Disease Susceptibility ◽

Stress Reactivity ◽

Control Diet ◽

Epigenetic Inheritance ◽

Overweight And Obesity ◽

Male Dominance ◽

Metabolic Dysfunction ◽

Negative Consequences ◽

Intergenerational Effects

The global consumption of highly processed, calorie-dense foods has contributed to an epidemic of overweight and obesity, along with negative consequences for metabolic dysfunction and disease susceptibility. As it becomes apparent that overweight and obesity have ripple effects through generations, understanding of the processes involved is required, in both maternal and paternal epigenetic inheritance. We focused on the patrilineal effects of a Western-style high-fat (21%) and high-sugar (34%) diet (WD) compared to control diet (CD) during adolescence and investigated F0 and F1 mice for physiological and behavioral changes. F0 males (fathers) showed increased body weight, impaired glycemic control, and decreased attractiveness to females. Paternal WD caused significant phenotypic changes in F1 offspring, including higher body weights of pups, increased Actinobacteria abundance in the gut microbiota (ascertained using 16S microbiome profiling), a food preference for WD pellets, increased male dominance and attractiveness to females, as well as decreased behavioral despair. These results collectively demonstrate the long-term intergenerational effects of a Western-style diet during paternal adolescence. The behavioral and physiological alterations in F1 offspring provide evidence of adaptive paternal programming via epigenetic inheritance. These findings have important implications for understanding paternally mediated intergenerational inheritance, and its relevance to offspring health and disease susceptibility.

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Effects of early life adversity on maternal effort and glucocorticoids in wild olive baboons

10.1101/2020.12.11.418095 ◽

2020 ◽

Author(s):

Sam K Patterson ◽

Katie Hinde ◽

Angela B Bond ◽

Benjamin C Trumble ◽

Shirley C Strum ◽

...

Keyword(s):

Early Life ◽

Life Experiences ◽

Social Rank ◽

Epigenetic Inheritance ◽

Papio Cynocephalus ◽

Papio Anubis ◽

Intergenerational Effects ◽

Early Life Adversity ◽

Olive Baboons ◽

Maternal Effort

Adverse experiences during early life exert important effects on development, health, reproduction, and social bonds, with consequences often persisting across generations. A mother's early life experiences can impact her offspring's development through a number of pathways, such as maternal care, physiological signaling through glucocorticoids, or even intergenerational effects like epigenetic inheritance. Early life adversity in female yellow baboons (Papio cynocephalus) predicts elevated glucocorticoids, reduced sociality, shortened lifespan, and higher offspring mortality. If baboon mothers with more early life adversity, experience poorer condition and struggle to provide for their offspring, this could contribute to the persisting transgenerational effects of adversity. Here, we examined the effects of mothers' early life adversity on their maternal effort, physiology, and offspring survivability in a population of olive baboons, Papio anubis. Mothers who experienced more adversity in their own early development exerted greater maternal effort (i.e., spent more time nursing and carrying) and had higher glucocorticoid metabolites than mothers with less early life adversity. Offspring of mothers with more early life adversity had reduced survivability compared to offspring of mothers with less early life adversity. There was no evidence that high maternal social rank buffered against the effects of early life adversity. Our data suggest early life experiences can have lasting consequences on maternal effort and physiology, which may function as proximate mechanisms for intergenerational effects of maternal experience.

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Starvation Responses Throughout the Caenorhabditiselegans Life Cycle

Genetics ◽

10.1534/genetics.120.303565 ◽

2020 ◽

Vol 216 (4) ◽

pp. 837-878

Author(s):

L. Ryan Baugh ◽

Patrick J. Hu

Keyword(s):

Life Cycle ◽

Epigenetic Inheritance ◽

Egg Laying ◽

Food Sources ◽

Adult Health ◽

Adaptive Responses ◽

Comprehensive Overview ◽

C Elegans ◽

Maternal Provisioning ◽

In The Wild

Caenorhabditis elegans survives on ephemeral food sources in the wild, and the species has a variety of adaptive responses to starvation. These features of its life history make the worm a powerful model for studying developmental, behavioral, and metabolic starvation responses. Starvation resistance is fundamental to life in the wild, and it is relevant to aging and common diseases such as cancer and diabetes. Worms respond to acute starvation at different times in the life cycle by arresting development and altering gene expression and metabolism. They also anticipate starvation during early larval development, engaging an alternative developmental program resulting in dauer diapause. By arresting development, these responses postpone growth and reproduction until feeding resumes. A common set of signaling pathways mediates systemic regulation of development in each context but with important distinctions. Several aspects of behavior, including feeding, foraging, taxis, egg laying, sleep, and associative learning, are also affected by starvation. A variety of conserved signaling, gene regulatory, and metabolic mechanisms support adaptation to starvation. Early life starvation can have persistent effects on adults and their descendants. With its short generation time, C. elegans is an ideal model for studying maternal provisioning, transgenerational epigenetic inheritance, and developmental origins of adult health and disease in humans. This review provides a comprehensive overview of starvation responses throughout the C. elegans life cycle.

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Grandmother’s chemical legacy of pesticide exposure: bi-generational effects and acclimation in a model invertebrate

10.1101/2020.11.25.397554 ◽

2020 ◽

Author(s):

Rikke Poulsen ◽

Henrik H. De Fine Licht ◽

Martin Hansen ◽

Nina Cedergreen

Keyword(s):

First Generation ◽

Pesticide Exposure ◽

Continuous Exposure ◽

Biological Organization ◽

Intergenerational Effects ◽

Single Generation ◽

Generational Effects ◽

Direct Association ◽

Chemical Legacy ◽

Different Levels

AbstractMan-made chemicals are a significant contributor to the ongoing deterioration of ecosystems. Currently, risk assessment of these chemicals is based on observations in a single generation of animals, despite potential adverse intergenerational effects. Here, we investigate the effect of the fungicide prochloraz across three generations of Daphnia magna. We studied both the effects of continuous exposure over all generations and the effects of first-generation (F0) exposure on two subsequent, non-exposed, generations. Effects at different levels of biological organization were monitored. Acclimation to prochloraz was found after continuous exposure. Following F0-exposure, non-exposed F1-offspring showed no significant effects. However, in the F2 animals, several parameters differed significantly from controls. A direct association between grandmaternal effects and toxic mode of action of prochloraz was found, showing that chemicals can be harmful not only to the exposed generation, but also to subsequent generations and that effects may even skip a generation.

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Characterization of differentially detectable Mycobacterium tuberculosis in the sputum of subjects with drug sensitive or drug resistant tuberculosis before and after two months of therapy

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00608-21 ◽

2021 ◽

Author(s):

Kayvan Zainabadi ◽

Kathleen Frances Walsh ◽

Stalz Charles Vilbrun ◽

Laurent Daniel Mathurin ◽

Myung Hee Lee ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Drug Resistant ◽

Second Line ◽

Liquid Media ◽

Treatment Regimens ◽

Resistant Tuberculosis ◽

Solid Media ◽

Large Populations ◽

Before And After

Standard methods for enumerating Mycobacterium tuberculosis (Mtb) in patient sputa can miss large populations of viable Mtb that are unable to grow either on solid media or in liquid media if not extensively diluted. Because these bacteria can be detected in liquid media after limiting dilution, they have been termed differentially culturable or differentially detectable Mtb (DD Mtb). Treatment with isoniazid, rifampin, pyrazinamide and ethambutol (HRZE) for 1-2 weeks has been shown to increase the representation of DD Mtb in the sputum of drug sensitive (DS) tuberculosis (TB) patients. However, little is known about DD Mtb after longer periods of treatment with HRZE, or in patients with drug resistant (DR) TB who receive second-line therapies. Here we measured the proportion of DD Mtb in the sputum of 47 subjects, 29 with DS TB and 18 with DR TB, before initiation of their treatment regimens and at 2 weeks and 2 months thereafter. Prior to treatment, DD Mtb represented the majority of Mtb in the sputum of 21% of subjects with DS TB and this proportion rose to 65% after 2 weeks of treatment with first-line drugs. In subjects with DR TB, DD Mtb was found in the sputum of 29% of subjects prior to treatment initiation, and this proportion remained steady at 31% after 2 weeks of treatment with second-line drugs. By 2 months, DD Mtb was detected in the sputum of only 2/15 (13.3%) subjects with DS TB and 0/15 of subjects with DR TB. One of the DS subjects whose sputum was positive for DD Mtb at month 2 later experienced treatment failure.

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