During the early pre and postnatal life, host and environmental factors can impart a major influence on immune development, thus shaping lifelong disease resistance. Two major factors known to influence immune function and mortality in animals and people are early life stress and biological sex. How these two factors interact to shape long-term immune development and later life disease risk is poorly understood. Here we investigated how early weaning, a common early life stressor in pigs, and biological sex impacts long-term systemic inflammatory responses and hypothalamic–pituitary–adrenal axis (HPA axis) activation later in life. Ten-week-old female (F), intact-male (IM) and castrated-male (CM) pigs that were randomly assigned to early weaning (EW) and later weaning (LW) (at 15 or 28 days of age, respectively) were intramuscularly injected with either saline vehicle or lipopolysaccharide (LPS) to induce a systemic inflammatory response. Complete blood counts (CBC), proinflammatory cytokines, cortisol, testosterone, estradiol, and rectal temp were measured at 0 h, 2 h, and 4 h post-LPS challenge. At 4 h post-LPS, peritoneal fluid (PF) and white blood cells (WBC) were collected for differential analysis. LPS challenge significantly increased rectal temp and plasma cortisol level in all treatment groups. Together, the CBC results and immune cell counts in peritoneal cavity indicated that EW-F exhibited greater systemic immune response characterized by increased neutrophils to lymphocytes ratio (NLR) and enhanced neutrophil trafficking to the peritoneal cavity. Early weaning had an opposite effect on IM and CM pigs, which exhibited a suppressed LPS-induced neutrophil migration. Early weaning induced significantly greater cortisol responses only in IM pigs indicating a heightened HPA axis responses in EW-IM. how early weaning and biological sex affect immune and stress responses in pigs. Together, these results demonstrate that early weaning and biological sex and castration shape later life immune responses in pigs and provides insight into potential mechanisms driving sex differences in later life inflammatory disease risk and mortality.

Cockroaches are important global urban pests from aesthetic and health perspectives. Insecticides represent the most cost-effective way to control cockroaches and limit their impacts on human health. However, cockroaches readily develop insecticide resistance, which can quickly limit efficacy of even the newest and most effective insecticide products. The goal of this research was to understand whole-body physiological responses in German cockroaches, at the metatranscriptome level, to defined insecticide selection pressures. We used the insecticide indoxacarb as the selecting insecticide, which is an important bait active ingredient for cockroach control. Six generations of selection with indoxacarb bait produced a strain with substantial (>20×) resistance relative to inbred control lines originating from the same parental stock. Metatranscriptome sequencing revealed 1,123 significantly differentially expressed (DE) genes in ≥two of three statistical models (81 upregulated and 1,042 downregulated; FDR P < 0.001; log2FC of ±1). Upregulated DE genes represented many detoxification enzyme families including cytochrome-P450 oxidative enzymes, hydrolases and glutathione-S-transferases. Interestingly, the majority of downregulated DE genes were from microbial and viral origins, indicating that selection for resistance is also associated with elimination of commensal, pathogenic and/or parasitic microbes. These microbial impacts could result from: (i) direct effects of indoxacarb, (ii) indirect effects of antimicrobial preservatives included in the selecting bait matrix, or (iii) selection for general stress response mechanisms that confer both xenobiotic resistance and immunity. These results provide novel physiological insights into insecticide resistance evolution and mechanisms, as well as novel insights into parallel fitness benefits associated with selection for insecticide resistance.

Biological sex is a major host factor influencing risk for infectious disease-associated mortality, and chronic inflammatory and metabolic diseases. Research in human and rodent models -has revealed sex differences that exist across organ systems during health and disease that may contribute to sex biases in disease risk. Despite the robust and growing literature on the role of sex as a risk factor in human disease, comparatively little attention has been focused on investigating the role of biological sex in disease susceptibility in agriculturally important animal populations such as the pig. To date, comparisons between sexes have focused on carcass composition, growth rate, and feed efficiency in pigs. However, there is a large gap in the literature regarding the effects of biological sex on other integral aspects of health and disease. The objective of this review is to highlight the available literature reporting sex differences in pig health and biology with an emphasis on sex differences in mortality, immunity, and gastrointestinal (GI) physiology and to address biological sex as a significant biological variable in disease risk and research study design. A basic overview of the biology of sex differences including the major hormonal and genetic/chromosomal mechanisms of sexual differentiation and the developmental periods in which sex differences emerge will be covered. This review will also discuss how production-relevant management and environmental factors (e.g., wean age, castration, stress, and nutrition) interact with biological sex to shape host immune and GI development and function. Perceived gaps in knowledge and areas of future research will also be discussed.

Early-life adversity (ELA) is linked with the increased risk for inflammatory and metabolic diseases in later life, but the mechanisms remain poorly understood. Intestinal epithelial glucose transporters sodium-glucose-linked transporter 1 (SGLT1) and glucose transporter 2 (GLUT2) are the major route for intestinal glucose uptake but have also received increased attention as modulators of inflammatory and metabolic diseases. Here, we tested the hypothesis that early weaning (EW) in pigs, an established model of ELA, alters the development of epithelial glucose transporters and coincides with elevated markers of metabolic inflammation. The jejunum and ileum of 90-day-old pigs previously exposed to EW (16 days wean age), exhibited reduced SGLT1 activity (by ∼ 30%, P < 0.05) than late weaned (LW, 28 days wean age) controls. In contrast, GLUT2-mediated glucose transport was increased (P = 0.003) in EW pigs than in LW pigs. Reciprocal changes in SGLT1- and GLUT2-mediated transport coincided with transporter protein expression in the intestinal brush-border membranes (BBMs) that were observed at 90 days and 150 days of age. Ileal SGLT1-mediated glucose transport and BBM expression were inhibited by the β-adrenergic receptor (βAR) blocker propranolol in EW and LW pigs. In contrast, propranolol enhanced ileal GLUT2-mediated glucose transport (P = 0.015) and brush-border membrane vesicle (BBMV) abundance (P = 0.035) in LW pigs, but not in EW pigs. Early-weaned pigs exhibited chronically elevated blood glucose and C-reactive protein (CRP) levels, and adipocyte hypertrophy and upregulated adipogenesis-related gene expression in visceral adipose tissue. Altered development of intestinal glucose transporters by EW could underlie the increased risk for later life inflammatory and metabolic diseases.NEW & NOTEWORTHY These studies reveal that early-life adversity in the form of early weaning in pigs causes a developmental shift in intestinal glucose transport from SGLT1 toward GLUT2-mediated transport. Early weaning also induced markers of metabolic inflammation including persistent elevations in blood glucose and the inflammatory marker CRP, along with increased visceral adiposity. Altered intestinal glucose transport might contribute to increased risk for inflammatory and metabolic diseases associated with early-life adversity.

A sex bias exists in mast cell (MC)-associated inflammatory diseases with females at increased risk. Our previous studies demonstrated that MCs exhibit a sexually dimorphic phenotype with male MCs exhibiting significantly reduced concentration and release of MC granule mediators (histamine, serotonin, and proteases) which was associated with reduced severity of anaphylaxis in males. Further, we showed that perinatal androgens are important in organizing male MC sex differences. The mechanisms by which androgens organize MC sex differences remain poorly understood. Utilizing male mice with reduced androgen receptor (AR) expression induced by Cre-LoxP technology (induced TFM; iTfm), we tested the hypothesis that ARs mediate sex differences in MC histamine storage and release, and severity of anaphylaxis. Isolated peritoneal MCs (pMCs) from iTFm male mice exhibited a feminized MC phenotype characterized by a greater level of histamine compared with control male pMCs, but similar to female controls. Following induction of IgE-mediate anaphylaxis in vivo, iTFM male and female pMCs contained lower (n=5–6; P<0.01) histamine contents compared with control males, suggesting enhanced release of histamine. Preliminary results also indicate that iTFm male mice exhibit greater IgE-mediated histamine releases and hypothermia. Experiments with bone marrow derived MCs (BMMCs) showed that IgE-mediated histamine release from iTFm male and female BMMCs was greater (n=5–6; P<0.01) relative to control male BMMCs. Together, these studies demonstrate that sex differences in MC phenotype and function are mediated via Ars and that iTFm mice are a valid model to explore AR-mediated sex differences in MC function and disease pathogenesis. Supported by NIH R01 HD072968, NIH R21 AI140413

Early postnatal life represents a critical window of GI immune development when lifelong immune function is shaped. Yet, little is known on how early life events impact long-term immune development in animals and people. Based on our previous research, we investigated how biological sex and early weaning (EW) stress interact to shape long-term GI immune development in the pig. Female (F), male intact (MI) and male castrated (MC) piglets (Yorkshire-cross) were weaned at 17 d (EW) or at 26 d of age (Later weaning; LW). At 77 d of age, tissue samples were harvested for RNA-seq, qPCR, and immunoassays. Serum cytokine and ileal mucosal IgG responses were measured following oral and intramuscular vaccination for Lawsonia intracellularis (LI) and PCV2, respectively. Compared with LW pigs, EW MC and MI pigs had reduced body weight gain (P<0.05). The impact of biological sex and EW were observed in vaccine responses. Serum from F showed higher PCV2-specific IgG titers and LI-antibody levels compared with MC and MI pigs. EW F had higher IgG titers in response to vaccination compared with LW F. In contrast, EW MC had suppressed antibody titers compared with LW MC. Pathway analysis of RNA-seq data from ileal mucosa revealed enrichment of genes associated with increased in immune-related pathways (e.g. inflammation) in EW F. In contrast, EW MC exhibited enrichment for genes associated with a down-regulation of immune response (e.g. immune cell migration). Together, these studies showed that biological sex and EW have a major impact on GI immune development in the pig. An understanding of the role and underlying sex-specific mechanisms driving altered immune development in EW pigs could reveal new targets for enhancing early and long-term immune development.

BACKGROUND:Ozone gas is commercially used for deodorization and microbial control. Its efficacy against stored product insect pests is well documented. In the midst of the common bed bug (Cimex lectularius L.) outbreak, claims were made that ozone gas was effective for their control. This study was conducted to determine baseline ozone concentrations and exposure times required for the control of an insecticide-susceptible C. lectularius strain under laboratory conditions. Dichlorvos (DDVP), an organophosphate class fumigant insecticide was used as a positive control. RESULTS:Nymphs and adults were more susceptible to ozone than eggs. Complete (100%) nymph and adult mortality was achieved at an ozone concentration (C) of 1500 ppm and exposure time (T) of 180 min, or concentration × time product (CT) of 270 000 ppm-min, whereas eggs required an eightfold higher CT (2 040 000 ppm-min). DDVP vapor was 2070-, 2542- and 450-fold more potent than ozone, against nymphs, adults and eggs, respectively. CONCLUSIONS:Baseline ozone toxicity data provide insights on the practicality of using this gas for the management of common bed bugs. High ozone CT products required for C. lectularius control, particularly eggs, suggest that its use for treating infested human dwellings is not feasible due to logistic, safety and monetary concerns. © 2020 Society of Chemical Industry.

Mast cell (MC)-associated diseases, including allergy/anaphylaxis and neuroinflammatory pain disorders, exhibit a sex bias, with females at increase risk. While much attention has been directed toward adult sex hormones as drivers of sex differences, that female sex bias in MC-associated diseases is evident in prepubertal children, suggesting early-life origins of sex differences which have yet to be explored. Utilizing rodent models of MC-mediated anaphylaxis, our data here reveal that, 1) compared with females, males exhibit significantly reduced severity of MC-mediated anaphylactic responses that emerge prior to puberty and persist into adulthood, 2) reduced severity of MC-mediated anaphylaxis in males is linked with the naturally high level of perinatal androgens and can be recapitulated in females by perinatal exposure to testosterone proprionate, 3) perinatal androgen exposure guides bone marrow MC progenitors toward a masculinized tissue MC phenotype characterized by decreased concentration of prestored MC granule mediators (e.g., histamine, serotonin, and proteases) and reduced mediator release upon degranulation, and 4) engraftment of MC-deficient Kit(W-sh/W-sh) mice with adult male, female, or perinatally androgenized female MCs results in MC-mediated anaphylaxis response that reflects the MC sex and not host sex. Together, these data present evidence that sex differences in MC phenotype and resulting disease severity are established in early life by perinatal androgens. Thus, factors affecting levels of perinatal androgens could have a significant impact on MC development and MC-associated disease risk across the life span.

Dried Distillers Grains with Solubles (DDGS) has been used as a grain-based supplement in animal feed. The focus of this study was to investigate the effects of chemical and physical characteristics that may affect DDGS susceptibility to Tribolium castaneum (Herbst) infestation. Multiple regression-based models were built to determine if the diet's chemical and physical characteristics affect T castaneum development. Based on the regression models, diet particle size was the most influential factor in larval development. Larval weight increased more rapidly on T. castaneum normal laboratory diet, 90% flour/10% brewer's yeast (F/Y), followed by ground DDGS, compared to raw DDGS with different particle sizes. To determine the effect of diet particle size on larval development, experiments were conducted to increase particle size by pelletizing F/Y using a rotary drum granulation apparatus. T. castaneum larval stage lasted significantly longer on F/Y granules with a particle size of 1.4 mm or larger. Although the chemical properties of DDGS (protein, fat, fiber and ash content) were not significant predictors of T. castaneum larval development, larval developmental time and weight gain on ground DDGS was significantly slower than on F/Y, highlighting the potential importance of nutritional factors not accounted for in our analysis. Our results conclude that storing DDGS as raw ingredient and in the granule or pellet form with large particle size may help limit T. castaneum infestations. Although granulation or pelletization may not be economically feasible for small manufacturing plants, larger producers may benefit from the reduced risk of insect contamination associated with secondary feeders such as T. castaneum. (C) 2018 Elsevier Ltd. All rights reserved.

The German cockroach (Blattella germanica L.) is a worldwide pest that lives exclusively in human environments. B. germanica threatens human health by producing asthma-triggering allergens, vectoring pathogenic/antibiotic-resistant microbes, and by contributing to unhealthy indoor environments. While insecticides are essential for reducing cockroach populations and improving health outcomes, insecticide resistance has been a consistent barrier to cockroach control since the 1950s. We conducted seminal field studies to compare three insecticide resistance intervention strategies for cockroaches and evaluated resistance evolution across multiple generations. Using pre-treatment resistance assessment to drive decisions, we found that single active ingredient (AI) treatments can successfully eliminate cockroaches if starting resistance levels are low. We further established that rotation treatments intuitively reduce selection pressure, and are effective when insecticides with no/low resistance are used. We also found that mixture products containing thiamethoxam + λ-cyhalothrin AIs were universally ineffective and highly repellent; and finally, evolution of cross-resistance among AIs is a significant, previously unrealized challenge.

Insecticide resistance in German cockroaches (Blattella germanica (L.)) has been a barrier to effective control since its first documentation in the 1950s. A necessary first step toward managing resistance is to understand insecticide susceptibility profiles in field-collected strains so that active ingredients (AIs) with lowest resistance levels can be identified. As a first step in this study, diagnostic concentrations (DCs) were determined for 14 insecticide AIs based on lethal concentrations that killed 99% or 90% of the individuals from a susceptible lab strain (JWax-S). Next, cockroaches were collected from two low-income multifamily housing complexes in Danville, IL, and Indianapolis, IN, and used to establish laboratory strains. These strains were screened against the 14 AI-DCs in vial bioassays, and susceptibility profiles were determined by comparing percent mortalities between the field strains relative to the JWax-S strain. Results revealed lowest resistance of field strains to boric acid, abamectin, dinotefuran, clothianidin, thiamethoxam, and chlorfenapyr. For the AIs hydramethylnon and imidacloprid, field strains did not display survivorship different than the lab strain, but > 90% mortality was never achieved. Lastly, both field strains displayed resistance to indoxacarb, fipronil, acetamiprid, beta-cyfluthrin, bifenthrin, and lambda-cyhalothrin, but at varying levels. These results satisfy two objectives. First, baseline monitoring DCs were established for 14 insecticides presently registered for use against cockroaches, which represents a useful resource. Second, our findings reveal insecticide AIs with lowest resistance levels for use in forthcoming field studies that will investigate impacts of different insecticide deployment strategies on resistance management and evolution in cockroach field populations.

German cockroach males possess tergal glands that secrete a combination of oligosaccharides, lipids and proteins. Four major proteins occur in the secretion, with one being the 63 kDa alpha‐amylase Blattella germanica Tergal Gland protein‐1 (BGTG‐1). Denaturing and starch gel electrophoresis coupled with peptide sequencing verified amylase activity for the BGTG‐1 protein. BGTG‐1 gene expression profiles were determined by using quantitative real‐time PCR to compare messenger RNA abundance among isolated tissues of males, females and gravid females. Differences in BGTG‐1 gene expression occurred among male tissues, with tergal gland tissue showing the highest expression. Tissues of nongravid and gravid females had significantly lower expression in comparison with male tergal glands (gravid females lowest). RNA interference (RNAi) was used to silence BGTG‐1 gene expression by injecting BGTG‐1 homologous double‐stranded RNA (dsRNA) into male cockroaches. Groups injected with BGTG‐1 dsRNA showed ∼90% lower BGTG‐1 gene and protein expression compared to controls, which correlated with lower amylase activity in colorimetric assays. However, behavioural assays comparing precopulatory behaviour and mating success between RNAi and control males did not reveal differences. These results connect amylase gene expression and activity in tergal gland tissue but suggest other factors, such as other tergal gland components, may contribute more strongly to mating success.

Understanding the susceptibility of commercially available animal feeds which contain Dried Distiller's Grains with Solubles (DDGS) to insect infestation is the first step in developing a pest management strategy for a stored product. This study is a nascent analysis of animal feed containing DDGS susceptibility to Tribolium castaneum (Herbst) infestation, one of the most important post-harvest insect pests world-wide. Three types of animal feed containing 5-10% DDGS in their formula mixtures were obtained from two feed manufacturers in Indiana, USA including: crumbled poultry feed, pelletized juvenile frog feed, and pelletized adult frog feed. Besides commercially available animal feed, laboratory manufactured feed were simulated by mixing Flour/Yeast (F/Y), T. castaneum normal laboratory diet, with 10, 20, 30, 40, and 80% DDGS (% based on weight). Larval developmental time was used to evaluate feed susceptibility to T castaneum infestation. The T. castaneum larval period was significantly increased when fed on pelletized adult frog feed, pelletized juvenile frog feed, and crumbled poultry feed (29.8 +/- 0.5, 26.0 +/- 0.3, and 21.9 +/- 0.3 d respectively), compared with F/Y (16.1 +/- 0.3 d). The larval period was significantly decreased however, when fed on ground poultry feed (15.5 +/- 0.2 d). Insect development on laboratory manufactured feeds was similar to F/Y regardless of DDGS ratios added in this study (15.9-16.9 d). These results suggested that first; particle size (PS) was one of the influential factors on susceptibility of commercially available animal feed to T castaneum apart from feed chemical composition, and second, adding 10-80% DDGS to the laboratory manufactured feed did not change feed susceptibility to T castaneum infestation. (C) 2017 Elsevier Ltd. All rights reserved.

Demand for Dried Distiller's Grains with Solubles (DDGS) in international markets and the United States has increased during the past few years. Knowledge of DDGS supplemented animal feed vulnerability to insect infestation is critical for safe feed storage. To assess this vulnerability, it is necessary to know how DDGS is susceptible to insect infestation, while stored as raw ingredient. This research focused on the susceptibility of different types of DDGS (raw and ground) to red flour beetle, T. castaneum, infestation under 30% and 50% relative humidity (r.h.) regimes. Larval period at 30% r.h. increased 2-3 fold on raw DDGS diets with larger particle sizes (PSs) compared with their normal laboratory diet, a mixture of flour and yeast (9:1) (F/Y). However, grinding DDGS samples and increasing the r.h. to 50% decreased the amount of time required for insect development thus increasing DDGS vulnerability to T castaneum infestation compared with raw DDGS at r.h. of 30%. As was expected, T. castaneum egg and pupal development were not affected by diet or humidity. The results suggested that DDGS as a raw ingredient at 30% r.h. was not a suitable food source for T. castaneum and given a choice, the majority of T. castaneum adults prefer laboratory diet over DDGS. Additionally, fecundity was significantly lower on DDGS compared with the control diets (F/Y and ground corn (GCORN)). These results indicated that these types of DDGS were not suitable developmental diets compared with the F/Y diet if stored at 30% r.h. with larger PSs. (C) 2015 Elsevier Ltd. All rights reserved.

The influence of temperature in combination with gender, age, and mating status was examined to determine minimum temperature and percent of cigarette beetle, Lasioderma serricorne (F.) that initiate flight in a laboratory flight chamber. Tests were conducted on temperatures between 17.5 degrees C and 37.5 degrees C at intervals of 2.5 degrees C. We found that flight initiation of L serricorne is highly dependent on environmental temperature, gender, age and mating status of individuals. The minimum detectable temperature at which all categories of males (virgin young, virgin old, mated young and mated old) initiated flight was 22.5 degrees C, while virgin young, virgin old, mated young and mated old females were observed to initiate flight at 25, 22.5, 25, and 27.5 degrees C, respectively. Young virgin males and older virgin females had the highest percentage of flight initiation among all categories of their gender and thus the greatest probability of being detected. No flight initiation is predicted for males at temperatures below 19.7 degrees C, thus, environmental temperature should be strongly considered when interpreting pheromone monitoring programs in food storage. (C) 2012 Elsevier Ltd. All rights reserved.

Pheromone traps have been used successfully for monitoring of stored-product insects in the facilities but factors that could influence accuracy have not been fully examined. In this study, we examined the influence of lure (food for females and sex pheromone for males) on minimum flight initiation temperature (MFIT1) and percentage of 6-9 d-old mated cigarette beetles, Lasioderma serricorne (F.) that initiate flight. Temperature had a greater influence on minimum flight initiation temperature compared to food lure for young mated females or sex pheromone lure for young mated males. There was no significant effect of sex pheromone lure on the MFIT of 6-9 d-old mated male cigarette beetles nor was there a significant effect of food lure on the MFIT of 6-9 d-old mated females. (C) 2013 Elsevier Ltd. All rights reserved.