Screening and selection of eubiotic compounds possessing immunomodulatory and anti-Clostridium perfringens properties.

Eubiotics are water and/or feed additives used in poultry to promote gut health and control enteric burden of pathogens, including Clostridium perfringens. While several eubiotic compounds (ECs) are being introduced commercially, it is essential to devise an in vitro model to screen these compounds to assess their immunomodulatory and antimicrobial properties prior to their testing in vivo. A chicken macrophage cell-line (MQ-NCSU) was used to develop an in vitro model to screen the immunological and anti-C. perfringens properties of 10 ECs: monobutyrin, monolaurin, calcium butyrate, tributyrin, carvacrol, curcumin, green tea extract, rosemary extract, monomyristate, and tartaric acid. An optimal concentration for each EC was selected by measuring the effect on viability of MQ-NCSU cells. Cells were then treated with ECs for 6, 12, and 24 h. and expression of interferon-gamma (IFNγ), interleukin (IL)-1ß, IL-6, IL-10, transforming growth factor-beta (TGFß) and cluster of differentiation (CD40) genes, as well as major histocompatibility complex (MHC)-II protein were evaluated. At 6 h post-stimulation, monobutyrin, calcium butyrate, and green tea extract treatments induced a significant downregulation of IFNγ, IL-6, or IL-1ß gene transcription and MHC-II expression, while the IL-10 or TGFß gene expression in these treatments as well as those receiving rosemary extract and tartaric acid was significantly upregulated, when compared to control, suggesting immunomodulatory properties of these ECs. Finally, pretreatment of macrophages with these selected 5 ECs for 24 h followed by C. perfringens infection showed that monobutyrin, green tea extract, rosemary extract, and calcium butyrate treatments can inhibit bacterial growth significantly at 12 and/or 24 h post-infection, when compared to the control. Collectively, our findings show that ECs possessing immunomodulatory and anti-C. perfringens properties can be selected using an in vitro avian macrophage cell-based model so that such ECs can further be tested in vivo for their disease prevention efficacy.

Deoxynivalenol and fumonisin predispose broilers to bacterial chondronecrosis with osteomyelitis lameness.

Bacterial chondronecrosis with osteomyelitis (BCO) lameness is the most critical animal health and welfare issue facing the broiler industry worldwide. It is estimated that 1 to 2% of bird condemnation at marketing age is caused by BCO lameness, resulting in tens of millions of dollars in annual losses. Fast-growing broilers are prone to mechanical stress that triggers bacterial translocation across epithelial barriers into the bloodstream, followed by bacterial colonization in the growth plate of long bones, and eventually, bone necrosis and lameness. Mycotoxins (MTX) are secondary metabolites produced naturally by microfungi, of which deoxynivalenol (DON), fumonisin (FUM), and zearalenone are the most prevalent in corn and soybean-meal-based diets. The presence of these mycotoxins in feed has been proven to reduce the barrier strength of the intestinal tracts and trigger immunosuppressive effects. In this study, we investigated the effects of the DON and FUM-contaminated feeds on the incidence of BCO lameness in broilers reared in both wire- and litter-floors. 720 one-day-old broiler chicks were assigned to the 2 × 2 factorial design: 2 MTX diets containing DON and FUM on wire flooring (MTX-W) and litter flooring (MTX-L), and 2 diets without MTX contamination on control wire flooring (CW) and control litter flooring (CL). Throughout the trial, the cumulative incidence of lameness per treatment was assessed by necropsying the lame birds. Birds in the MTX-W group had a higher incidence of lameness compared to those in CW (73.3% vs. 62.0%) (P < 0.05), and birds in the MTX-L group had a higher incidence of lameness compared to birds in CL (54.0% vs. 34.0%) (P < 0.05). MTX elicited net increases in BCO to a greater degree on litter (+20%) than on wire flooring (+12%). The increased incidence of BCO lameness in the MTX-W coincided with increased intestinal permeability supporting a correlation between intestinal barrier integrity and BCO lameness. To conclude, DON and FUM are predisposing factors for increasing BCO. However, no significant interaction exists between the diet and floor types in inducing lameness in broilers.

Impact of social networks on fertilizer use efficiency in China: Evidence from kiwifruit production in Shaanxi province.

Improving fertilizer use efficiency (FUE) is an effective way to reduce the fertilizer use and non-point source pollution in agriculture production in China. Existing research related to fertilizer use efficiency ignores the impact of famers' social networks, which can significantly affect the fertilizer use and its efficiency. This paper analyzed the impact of social networks on fertilizer use efficiency and the mediator effect of green fertilization technology adoption, based on the calculation of fertilizer use efficiency of 569 kiwifruit farmers in Shaanxi province of China surveyed in 2021 by using stochastic frontier method. The results show that the FUE of kiwifruit production ranged within 0.1027-0.9945, with an average value of 0.333, which showed great variability, and fertilizer input had an average 66.7% reduction potential without reducing output. Social networks and its four dimensions all had a significant positive impact on FUE, in which network trust and network learning had a greater impact on FUE. Farmers' adoption of green fertilization technology played a positive mediator effect in the process of social networks influencing their fertilizer use efficiency, which was mainly promoted by network trust and learning. Farmers' experience in agricultural production, joining the cooperative, the quantity of village agricultural supply points had a significant positive impact on fertilizer use efficiency, while age and gentle of the head of household had a significant negative impact of fertilizer use efficiency. Planting scale had a U-shape impact on FUE. Policies aimed at improving FUE and reducing non-point source pollution should, therefore, focus on improving the social networks of farmers, especially young farmers, strengthening network trust, promoting network learning, network reciprocity and network interaction, strengthening the research and development and extension of green fertilization technology, encouraging farmers to join agricultural cooperative, improving village production conditions, guiding sustainable agricultural production.

The Zonulin-transgenic mouse displays behavioral alterations ameliorated via depletion of the gut microbiota.

The gut-brain axis hypothesis suggests that interactions in the intestinal milieu are critically involved in regulating brain function. Several studies point to a gut-microbiota-brain connection linking an impaired intestinal barrier and altered gut microbiota composition to neurological disorders involving neuroinflammation. Increased gut permeability allows luminal antigens to cross the gut epithelium, and via the blood stream and an impaired blood-brain barrier (BBB) enters the brain impacting its function. Pre-haptoglobin 2 (pHP2), the precursor protein to mature HP2, is the first characterized member of the zonulin family of structurally related proteins. pHP 2 has been identified in humans as the thus far only endogenous regulator of epithelial and endothelial tight junctions (TJs). We have leveraged the Zonulin-transgenic mouse (Ztm) that expresses a murine pHP2 (zonulin) to determine the role of increased gut permeability and its synergy with a dysbiotic intestinal microbiota on brain function and behavior. Here we show that Ztm mice display sex-dependent behavioral abnormalities accompanied by altered gene expression of BBB TJs and increased expression of brain inflammatory genes. Antibiotic depletion of the gut microbiota in Ztm mice downregulated brain inflammatory markers ameliorating some anxiety-like behavior. Overall, we show that zonulin-dependent alterations in gut permeability and dysbiosis of the gut microbiota are associated with an altered BBB integrity, neuroinflammation, and behavioral changes that are partially ameliorated by microbiota depletion. Our results suggest the Ztm model as a tool for the study of the cross-talk between the microbiome/gut and the brain in the context of neurobehavioral/neuroinflammatory disorders.

Assay considerations for fluorescein isothiocyanate-dextran (FITC-d): an indicator of intestinal permeability in broiler chickens.

Fluorescein isothiocyanate-dextran (FITC-d) is being used as an indicator of intestinal paracellular permeability in poultry research. Especially with the industry moving toward antibiotic-free production, intestinal function and integrity issues have been a research focus. An increasing number of scientific conference abstracts and peer-reviewed journal publications have shown that 4-kDa FITC-d is an efficient marker candidate for measurement of intestinal permeability and can be applied in broiler research. However, experimental protocols vary by personnel, instruments used, and research institution, and potential concerns related to this assay have yet to receive the same amount of attention. Understanding protocol consistency within and across laboratories is vital for obtaining accurate, consistent, and comparable experimental results. This review is aimed to 1) summarize different FITC-d assays in broiler research from peer-reviewed publications during the past 6 yr and 2) discuss factors that can potentially affect intestinal permeability results when conducting the FITC-d assay. In summary, it is essential to pay attention to details, including gavage dose, fasting period, sample handling and lab analysis details when conducting the assay in broiler research. Differences in birds (breed/strain, age, and gender) and experimental design (diet, health status/challenge model, and sampling age) need to be considered when comparing serum FITC-d concentration results between different in vivo animal trials.

Caregivers: the potential infection resources for the sustaining epidemic of hand, foot, and mouth disease/herpangina in Guangdong, China?

BACKGROUND: Although several measures have been taken to control hand foot and mouth disease (HFMD) and herpangina (HA), these two diseases have been prevalent in China for 10 years with high incidence. We suspected that adults' inapparent infection might be the cause of the continued prevalence of HFMD/HA infection in mainland China. METHODS: To explore the role of adults (especially caregivers) in the transmission process of HFMD/HA among children, 330 HFMD/HA cases and 330 healthy children (controls) were selected for a case-control study. Then, data were analyzed by logistic regression. RESULTS: Single-variable analyses revealed that caregivers who tested positive for enterovirus was a significant risk factor of HFMD/HA transmission to children (adjusted odds ratio (OR) = 9.22; 95% CI, 1.16 to 73.23). In the final multivariable model, caregiver behavior, such as cooling children's food with mouth (OR = 1.85; 95% CI, 1.11 to 3.08) and feeding children with their own tableware (OR = 2.19; 95% CI, 1.07 to 4.45), significantly increased the risk of transmitting HFMD/HA to children. On the contrary, washing hands before feeding children reduced such risk. CONCLUSIONS: These results implied that the caregivers might be the infectious source or carriers of enterovirus. Therefore, preventing or treating the caregivers' enterovirus infection and improving their hygiene habits, especially when they are in contact with children, could provide a breakthrough for the effective control of HFMD/HA.

Retrospective study identifies infection related risk factors in close contacts during COVID-19 epidemic.

OBJECTIVES: This study aimed to compare the risk of infection of children with that of adults and to explore risk factors of infection with severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) by following up close contacts of COVID-19 patients. METHOD: The retrospective cohort study was performed among close contacts of index cases diagnosed with COVID-19 in Guangzhou, China. Demographic characteristics, clinical symptoms and exposure information were extracted. Logistic regression analysis was employed to explore the risk factors. The restricted cubic spline was conducted to examine to the dose-response relationship between age and SARS-CoV-2 infection. RESULTS: The secondary attack rate (SAR) was 4.4% in 1,344 close contacts. The group of household contacts (17.2%) had the highest SAR. The rare-frequency contact (p < 0.001) and moderate-frequency contact (p < 0.001) were associated with lower risk of infection. Exposure to index cases with dry cough symptoms was associated with infection in close contacts (p = 0.004). Compared with children, adults had a significantly increased risk of infection (p = 0.014). There is a linear positive correlation between age and infection (p = 0.001). CONCLUSIONS: Children are probably less susceptible to COVID-19. Close contacts with frequent contact with patients and those exposed to patients with cough symptoms are associated with an increased risk of infection.

Lack of effective home quarantine: The cause of the continuing prevalence of hand, foot, and mouth disease in China?

BACKGROUND: Home quarantine as a non-pharmaceutical measure is a key strategy for preventing the spread of hand, foot, and mouth disease (HFMD) and herpangina (HA). However, the status of home quarantine remains unclear. This study aimed to provide a theoretical basis for the prevention and control of the HFMD and HA by investigating the situation and determining the factors influencing home quarantine in children with HFMD or HA. METHODS: The convenience sampling method was used in this cross-sectional study involving parents whose children suffered from HFMD or HA in 2019. An online questionnaire tool was used to collect demographic information, socioeconomic status, clinical factors, sources of advice, and details of home quarantine. Logistic regression analysis was used to explore the factors influencing of home quarantine. RESULTS: A total of 3145 parents answered the electronic questionnaire; among them, 317 had children diagnosed with HFMD or HA in 2019. After quality control, 298 questionnaires were included in the analysis. The whole compliance rate of home quarantine for HFMD and HA was 59.1%. Results of multivariate logistic regression analysis indicated that having grandparents as the primary caregiver (OR=2.53, 95% CI: 1.20-5.43) was a risk factor of home-quarantine noncompliance, whereas doctor's notification (OR=0.35, 95% CI: 0.18-0.67) and advice from parents of other infected children (OR=0.53, 95% CI: 0.31-0.91) can increase the compliance with home quarantine. CONCLUSIONS: Home-quarantine measures for HFMD and HA were not well implemented. The caregivers, doctor's notification, and advice from parents of other infected children were associated with home-quarantine compliance. Corresponding measures should be taken to regulate home quarantine.

Zearalenone (ZEN) in Livestock and Poultry: Dose, Toxicokinetics, Toxicity and Estrogenicity.

One of the concerns when using grain ingredients in feed formulation for livestock and poultry diets is mycotoxin contamination. Aflatoxin, fumonisin, ochratoxin, trichothecene (deoxynivalenol, T-2 and HT-2) and zearalenone (ZEN) are mycotoxins that have been frequently reported in animal feed. ZEN, which has raised additional concern due to its estrogenic response in animals, is mainly produced by Fusariumgraminearum (F.graminearum), F.culmorum, F.cerealis, F.equiseti, F.crookwellense and F.semitectums, and often co-occurs with deoxynivalenol in grains. The commonly elaborated derivatives of ZEN are -zearalenol, -zearalenol, zearalanone, -zearalanol, and -zearalanol. Other modified and masked forms of ZEN (including the extractable conjugated and non-extractable bound derivatives of ZEN) have also been quantified. In this review, common dose of ZEN in animal feed was summarized. The absorption rate, distribution ("carry-over"), major metabolites, toxicity and estrogenicity of ZEN related to poultry, swine and ruminants are discussed.

Risk Factors Associated with Dengue Virus Infection in Guangdong Province: A Community-Based Case-Control Study.

Dengue fever (DF) is a mosquito-borne infectious disease that is now an epidemic in China, Guangdong Province, in particular and presents high incidence rates of DF. Effective preventive measures are critical for controlling DF in China given the absence of a licensed vaccination program in the country. This study aimed to explore the individual risk factors for the dengue virus infection in Guangdong Province and to provide a scientific basis for the future prevention and control of DF. A case-control study including 237 cases and 237 controls was performed. Cases were defined for samples who were IgG-antibody positive or IgM-antibody positive, and willing to participate in the questionnaire survey. Additionally, the controls were selected through frequency matching by age, gender and community information from individuals who tested negative for IgG and IgM and volunteered to become part of the samples. Data were collected from epidemiological questionnaires. Univariate analysis was performed for the preliminary screening of 28 variables that were potentially related to dengue virus infection, and multivariate analysis was performed through unconditioned logistic regression analysis to analyze statistically significant variables. Multivariate analysis revealed two independent risk factors: Participation in outdoor sports (odds ratio (OR) = 1.80, 95% confidence interval (CI) 1.17 to 2.78), and poor indoor daylight quality (OR = 2.27, 95% CI 1.03 to 5.03). Two protective factors were identified through multivariate analysis: 2 occupants per room (OR = 0.43, 95% CI 0.28 to 0.65) or ≥3 occupants per room (OR = 0.45, 95% CI 0.23 to 0.89) and air-conditioner use (OR = 0.46, 95% CI 0.22 to 0.97). The results of this study were conducive for investigating the risk factors for dengue virus infection in Guangdong Province. Effective and efficient strategies for improving environmental protection and anti-mosquito measures must be provided. In addition, additional systematic studies are needed to explore other potential risk factors for DF.

Predicting interstate motor carrier crash rate level using classification models.

Ensuring safe operations of large commercial vehicles (motor carriers) remains an important challenge, particularly in the United States. While the federal regulatory agency has instituted a compliance review-based rating method to encourage carriers to improve their safety levels, concerns have been expressed regarding the effectiveness of the current ratings. In this paper, we consider a crash rate level (high, medium, and low) rather than a compliance review-based rating (satisfactory, conditional satisfactory, and unsatisfactory). We demonstrate an automated way of predicting the crash rate levels for each carrier using three different classification models (Artificial Neural Network, Classification and Regression Tree (CART), and Support Vector Machine) and three separate variable selection methods (Empirical Evidence, Multiple Factor Analysis, Garson's algorithm). The predicted crash rate levels (high, low) are compared to the assigned levels based on the current safety rating method. The results indicate the feasibility of crash rate level as an effective measure of carrier safety, with CART having the best performance.

Dengue Infection Spectrum in Guangzhou: A Cross-Sectional Seroepidemiology Study among Community Residents between 2013 and 2015.

The majority of dengue virus infections are asymptomatic, which could potentially facilitate the transmission of dengue fever and increase the percentage of sever dengue fever manifestations. This cross-sectional study explored the sero-prevalence of dengue virus infection in Guangzhou to clarify the infection spectrum. In total, 2085 serum samples were collected from residents of 34 communities. All samples were selected from a 200,000-sample database holding serum collected from community residents living in Liwan and Yuexiu districts of Guangzhou between September 2013 and August 2015, and 17 to 28 individuals of each age group were chosen per month. Dengue immunoglobulin G (IgG) and immunoglobulin M (IgM) antibodies were tested by enzyme-linked immunosorbent assay. Symptomatic infected individuals were identified via follow-up questionnaires. Among 2085 serum samples, anti-dengue IgG and IgM positive rates were 11.80% and 3.98%, respectively. The IgG antibody positive rate increased with age and was higher in poorly educated people than in highly educated people and in married individuals than in single individuals. Approximately 96.71% of dengue virus infections and an estimated 13.68% of the whole population were asymptomatic. Such high asymptomatic-infection rates have an impact on the local spread of dengue fever. Stricter surveillance, such as a network of rapid diagnostic laboratories, screening of residents in the epidemic season, and other integrated control measures are necessary.

Dynamic Changes of Pectin Epitopes in Cell Walls during the Development of the Procambium-Cambium Continuum in Poplar.

The change of pectin epitopes during procambium-cambium continuum development was investigated by immunolocalization in poplar. The monoclonal antibody JIM5 labels homogalacturonan (HGA) with a low degree of esterification, and the monoclonal antibody JIM7 labels HGA with a high degree of methyl-esterification. Arabinan, rather than galactan, and HGA with low degree of esterification were located in the cell walls of procambial, while HGA with a low degree of esterification was located in the tangential walls, and galactan was located in both the tangential and radial walls of procambial, yet nearly no arabinan was located in the tangential walls of the cambial cells. The changes in pectin distribution took place when periclinal divisions appeared within a procambial trace. The distribution difference of pectin epitopes was also present in procambium-cambium derivatives. The arabinan existed in all cell walls of primary xylem, but was absent from the tangential walls of secondary xylem cells. The galactan existed only in mature primary phloem. Furthermore, 19 pectin methylesterases (PMEs) genes were identified by RNA sequencing, six genes presented highly differentially and were supposed to be involved in the cell wall esterification process. The results provide direct evidence of the dynamic changes of pectin epitopes during the development of the procambium-cambium continuum in poplar.

Effects of essential oil supplementation of a low-energy diet on performance, intestinal morphology and microflora, immune properties and antioxidant activities in weaned pigs.

A total of 144 weaned piglets were used to evaluate the effects of essential oil (EO) supplementation of a low-energy diet on performance, apparent nutrient digestibility, small intestinal morphology, intestinal microflora, immune properties and antioxidant activities in weaned pigs. Pigs received a low-energy diet (negative control, NC, digestible energy = 3250 kcal/kg), NC plus 0.025% EO or a positive control diet (PC, digestible energy = 3400 kcal/kg) for 28 days. Growth performance was similar between the EO group and PC group. However, EO supplementation increased (P < 0.05) average daily gain and the apparent digestibility of dry matter, crude protein and energy compared with pigs fed the NC diet. Greater (P < 0.05) villus height and lower (P < 0.05) counts of Escherichia coli and total anaerobes in the rectum in the EO group were observed compared with NC or PC groups. Pigs fed EO diet had higher (P < 0.05) concentrations of albumin, immunoglobulin A (IgA), IgG and total antioxidant capacity and lower fecal score than pigs fed the PC and NC diets. Above all, this study indicates that supplementation of EO to a low-energy pig diet has beneficial results and obtains similar performance compared with normal energy (PC) diet.

Determination and prediction of the energy content and amino acid digestibility of peanut meals fed to growing pigs.

Two experiments were conducted to determine the content of digestible energy (DE) and metabolisable energy (ME) as well as the apparent ileal digestibility (AID) and standardised ileal digestibility (SID) of amino acids in peanut meal (PNM) for growing pigs. In Experiment 1, 78 growing pigs (46.8 ± 2.6 kg) were randomly allotted to 1 of 13 diets, including a corn-soya bean meal basal diet and 12 PNM test diets. In Experiment 2, 12 growing barrows (48.7 ± 2.8 kg) were allotted to one of two 6 × 6 Latin squares. The treatments include a N-free diet and 10 PNM test diets. The results of Experiment 1 showed that the DE and ME differed (p < 0.05) among the 12 PNM samples. On a dry matter basis, the DE and ME content ranged from 14.5 to 16.4 MJ/kg (mean 15.6 MJ/kg) and from 12.7 to 15.5 MJ/kg (mean 13.9 MJ/kg), respectively. The apparent total tract digestibility (ATTD) of gross energy (GE) was 82.2%. The DE of PNM could be precisely predicted by equations including NDF combined with GE or crude protein (CP) with an R(2) value of 0.91 and 0.92, respectively. For the AID and SID for lysine, the results of Experiment 2 indicated variations among PNM sources ranging from 59.6% to 76.7% and 64.8% to 80.9%, respectively. However, for CP, variations for AID and SID were lower and ranged from 70.2% to 81.9% and 75.7% to 85.6%, respectively. The results indicate that the concentration of lysine was the best single predictor to estimate the digestibility of amino acids. However, further work is needed to investigate the reason for the variation in the digestibility of lysine and avoid processing procedures that are detrimental to lysine digestibility.