[Etiology and molecular-epidemiological characteristics of viral meningo-encephalitis in Zhejiang province, 2002-2018].

Objective: To understand the pathogens and molecular-epidemiologic characteristics of viral meningo-encephalitis in Zhejiang province during 2002 to 2018. Methods: All the samples were collected from suspected patients admitted to the hospitals under the monitoring program. Of the total samples, 2 173 were cerebrospinal fluids while the other 455 were stool specimens. Cerebrospinal fluid (CSF) samples were subject to real-time qPCR for the detection of Human enterovirus (HEV), Mumps virus (MuV), Herpes simplex virus (HSV), Cytomegalovirus (CMV) and Japanese encephalitis virus (JEV). Stool sample were subject to real-time qPCR for HEV. ELISA was used to detect the IgM antibodies in CSF, in the 5 kinds of virus mentioned above. VP1 genes from all RNA-positive specimen were amplified, sequenced, for typing and for evolution analysis. Results: 871 (40.1%) of the 2 173 samples were detected as HEV nucleic acid positive during 2002 to 2018. 654 (38.1%) of the 1 718 CSF sample were HEV nucleic acid positive while 217 (47.7%) of the 455 stool sample were HEV nucleic acid positive. Among the total positive nucleic acid sample, 670 of them were VP1 sequence positive, including 5 HEV-A and 665 HEV-B. There were 23 HEV serotypes, including Coxsackievirus (CV) CVA4, CVA6, CVA9, CVA10, CVB1-5, Echovirus (EchoV; E) E3, E4, E6,E7, E9, E11, E14, E16, E18, E21, E25, E30, E33 and EV-71. The top three serotypes went to E30, E6 and CVB5. These three serotypes presented enhanced viral activity in every several years. 795 CSF samples were detected as virus nucleic acid positive, including 374 HEV, 6 MuV, 5 HSV and 5 CMV, from 2012 to 2015 and in 2018. 5 kinds of IgM antibodies were detected simultaneously in 368 CSF samples, including 2 HEV positive, 6 JEV positive and 1 MuV positive for 5 viruses, respectively. Except for EV-71, there were 517 EchoV and 152 CV viruses presented, with the ratio of 3.4∶1. These two kinds of viruses alternately changed for each predominant epidemic strains in every 3-5 years. Based on VP1, results from the phylogenetic tree showed that HEV from Zhejiang province clustered into HEV-A and HEV-B clades respectively. E30 developed both h and i sub-genotypes. Conclusions: HEV-B seemed the main pathogen for viral meningo-encephalitis in Zhejiang province. Ratio of positive detection on EchoV was significantly higher than that on CV. These two kinds of virus alternately presented changing tendency in every several years. Predominant epidemic strains E30, CVB5 and E6 were presenting enhanced viral activity, also in every several years. High correlation was found in both HEV viral activity from the surveillance sites and in time line of the viral meningo-encephalitis outbreaks.

[Percentile curves on growth among breastfed 1-4 year olds in 8 urban areas].

Objective: To construct the growth percentile curves of weight-, length/height-, head circumference and BMI for 1 to 4 year-olds who had been breastfed in urban areas. Methods: Data was from the longitudinal study on 1 025 breastfed children aged 1 to 4 years, in 8 urban areas during 2008-2012. MLwiN2.25 was selected to construct the multi-level models of weight-for-age,length for-age, head circumference-for-age and BMI-for-age. The models included many growth relevant factors including gender, age, family and social demographic characteristics, perinatal factors, parent biological characteristics, dietary patterns and diseases of childen. Based on these models, predicted values (P(3), P(15), P(50), P(85), P(97)) were estimated to fit the percentiles reference curves. Results: The percentiles reference curves of weight-, length/height, head circumferenceand BMI-for-age for the 1-4 year-olds who had been breastfed in the urban areas were developed. Differences of all the indicators between boys and girls were statistically significant (P<0.001). Weight, length/height, head circumference and BMI were higher in boys than those in girls, with an average differences as 0.56-0.76 kg, 0.89-1.12 cm, 0.64-0.91 cm and 0.31-0.52 kg/m(2). Conclusion: The percentiles reference curves on growth, constructed by the longitudinal observational data and scientific method, were important in reflecting the development of breastfed children in urban areas.

Effects of graded levels of montmorillonite on performance, hematological parameters and bone mineralization in weaned pigs.

The aim of this study was to investigate the effects of graded levels of montmorillonite, a constituent of clay, on performance, hematological parameters and bone mineralization in weaned pigs. One hundred and twenty, 35-d-old crossbred pigs (Duroc×Large White×Landrace, 10.50±1.20 kg) were used in a 28-d experiment and fed either an unsupplemented corn-soybean meal basal diet or similar diets supplemented with 0.5, 1.0, 2.5 or 5.0% montmorillonite added at the expense of wheat bran. Each treatment was replicated six times with four pigs (two barrows and two gilts) per replicate. Feed intake declined (linear and quadratic effect, p< 0.01) with increasing level of montmorillonite while feed conversion was improved (linear and quadratic effect, p<0.01). Daily gain was unaffected by dietary treatment. Plasma myeloperoxidase declined linearly (p = 0.03) with increasing dietary level of montmorillonite. Plasma malondialdehyde and nitric oxide levels were quadratically affected (p<0.01) by montmorillonite with increases observed for pigs fed the 0.5 and 1.0% levels which then declined for pigs fed the 2.5 and 5.0% treatments. In bone, the content of potassium, sodium, copper, iron, manganese and magnesium were decreased (linear and quadratic effect, p<0.01) in response to an increase of dietary montmorillonite. These results suggest that dietary inclusion of montmorillonite at levels as high as 5.0% does not result in overt toxicity but could induce potential oxidative damage and reduce bone mineralization in pigs.

Effects of nano-selenium on performance, meat quality, immune function, oxidation resistance, and tissue selenium content in broilers.

This study was conducted to investigate the effect of nano-selenium (nano-Se) on performance, meat quality, immune function, oxidation resistance, and tissue selenium content in broilers. A total of five hundred forty 1-d-old male Arbor Acres broilers were randomly allotted to 1 of 5 treatments with each treatment being applied to 6 replicates of 18 chicks. The 5 treatments consisted of corn-soybean meal-based diets supplemented with 0.0, 0.3, 0.5, 1.0, or 2.0 mg/kg of nano-Se. The selenium content of the unsupplemented control diet was 0.09 mg/kg for the starter phase (0 to 21 d) and 0.08 mg/kg for the grower phase (22 to 42 d). There were no significant differences (P > 0.05) in performance, meat color, or immune organ index (thymus, bursa, and spleen) due to supplementation with nano-Se. On d 42, a significant quadratic effect of nano-Se was observed on glutathione peroxidase activity, free radical inhibition, contents of IgM, glutathione, and malondialdehyde in serum, on glutathione peroxidase activity, free radical inhibition in liver, and on glutathione peroxidase activity in muscle, with birds fed 0.30 mg/kg of nano-Se exhibiting the best effect and birds fed 2.0 mg/kg of nano-Se showing the worst effect on these parameters. Liver and muscle selenium content increased linearly and quadratically as the dietary nano-Se level increased (P < 0.01), and reached the highest value when 2.0 mg/kg of nano-Se was fed. Based on a consideration of all experiment indexes, 0.3 to 0.5 mg/kg is suggested to be the optimum level of supplementation of nano-Se, and the maximum supplementation of nano-Se could not be more than 1.0 mg/kg in broilers.

Energy and standardized ileal amino Acid digestibilities of chinese distillers dried grains, produced from different regions and grains fed to growing pigs.

Two experiments were conducted to determine the digestibility of crude protein (CP), amino acids and energy in three Chinese corn distillers dried grains with solubles (DDGS), one rice DDGS, one American corn DDGS and one American high protein distillers dried grains (HP-DDG). In Exp. 1, the apparent ileal digestibility (AID) and standardized ileal digestibility (SID) of CP and amino acids in the six samples were determined using cannulated barrows (initial BW: 43.3±1.7 kg). In Exp. 2, the digestible energy (DE) and metabolizable energy (ME) content of these six samples were determined using crossbred barrows (initial BW: 46.0±2.5 kg). The results of the two experiments indicated that Chinese corn DDGS is generally similar to American DDGS in chemical composition, digestibility of amino acids, DE and ME. However, Chinese DDGS had a lower Lys concentration (0.50% vs. 0.74%) and SID Lys (52.3% vs. 57.0%, p<0.01). The DE and ME values in Chinese corn DDGS were 3,427 and 3,306 kcal/kg, respectively. Rice DDGS had a similar DE and ME (3,363 and 3,228 kcal/kg) but higher Lys concentration (0.64% vs. 0.50%) to corn DDGS, while the SID of Lys was quite low (61.8%, p<0.01). HP-DDG had high value of SID of Lys, DE and ME (79.8%, 3,899 and 3,746 kcal/kg). In conclusion, except for a lower Lys concentrations and availability, the chemical composition, digestibility of amino acids, DE and ME values in Chinese corn DDGS are similar to American corn DDGS. Additionally, the rice DDGS had lower Lys content and digestible Lys values than that in corn DDGS. Thirdly, HP-DDG has higher levels of digestible amino acids and energy than DDGS.

Effects of the free fatty acid content in yellow grease on performance, carcass characteristics, and serum lipids in broilers.

This study was conducted to investigate whether the free fatty acid (FFA) content of yellow grease would influence the performance and carcass characteristics of broiler chicks. A total of 432 one-day-old, male Arbor Acres broilers were randomly allotted to 1 of 4 treatments, with each treatment being applied to 6 pens of 18 chicks. The dietary treatments were based on corn and soybean meal and were supplemented with either soybean oil (2.86% FFA) or yellow grease with a low (2.74%), medium (12.59%), or high (19.05%) FFA content. The fat sources were supplemented at 1.5% of the diet during the starter phase (0 to 21 d) and at 3.0% of the diet during the grower phase (22 to 42 d). From d 0 to 42, the BW gains of chicks fed diets containing soybean oil and yellow grease with 2.74% FFA were similar. As the FFA level in the yellow grease increased, the BW gain of chicks decreased (P < 0.01). The reduction in BW gain appeared to be mediated by a reduction in feed intake. The dressing percentage and the percentage of breast muscle in the carcass were significantly (P < 0.01) lower for broilers fed any yellow grease diet compared with birds fed soybean oil. In contrast, abdominal fat was significantly increased in diets containing yellow grease. These results demonstrate that the performance of birds fed yellow grease with a low content of FFA was essentially equal to that of birds fed soybean oil. However, because yellow grease samples containing FFA levels greater than 2.74% negatively affected bird performance, producers should exercise caution with regard to feeding broilers yellow grease with an elevated FFA content. In 42-d-old broilers, serum total cholesterol and low-density lipoprotein cholesterol levels were elevated in birds fed yellow grease, regardless of the dietary level. In contrast, serum high-density lipoprotein cholesterol and triglyceride levels were unaffected by dietary treatment. Although dietary FFA may influence triglyceride-rich lipoprotein metabolism in broilers, an explanation for the observed effects remains elusive.

Effect of stachyose supplementation on growth performance, nutrient digestibility and caecal fermentation characteristics in broilers.

1. The objective of this study was to evaluate whether the oligosaccharide stachyose enhances gastrointestinal tract health by fermentation and proliferation of desirable bacteria species and thus affects growth performance and nutrient digestibility in broilers. 2. A total of 432 1-d-old male Arbor Acres (AA) broilers were randomly allocated to one of 6 treatments, with 12 replicate pens per treatment and 6 birds per pen. Chicks were fed a maize-hamlet protein 300 (HP300) basal diet with 0, 4.0, 8.0, 12.0 or 16.0 g/kg stachyose. A sixth diet contained no HP300 but soybean meal (SBM) and provided 8.7 g/kg stachyose and 3.1 g/kg raffinose. The duration of the study was 42 d. 3. Stachyose contents above 12.0 g/kg depressed group body weights, average daily gain and feed/gain but not feed intake during the whole experimental period. Broiler growth decreased linearly and quadratically with increasing stachyose content. No differences were detected between diets supplemented with 12.0 g/kg stachyose and SBM. 4. Nutrient digestibility tended to decrease but not significantly with increasing stachyose. 5. Stachyose content had no significant positive effects on caecal pH, microflora population and the resulting short-chain fatty acid (SCFA) metabolites during the 42 d experiment, with only butyrate differing significantly in the initial period.

Effects of fish oil supplementation on the performance and the immunological, adrenal, and somatotropic responses of weaned pigs after an Escherichia coli lipopolysaccharide challenge.

Seventy-two crossbred pigs (7.58 +/- 0.30 kg BW) weaned at 28 +/- 3 d of age were used to investigate the effects of fish oil supplementation on pig performance and on immunological, adrenal, and somatotropic responses following an Escherichia coli lipopolysaccharide (LPS) challenge in a 2 x 2 factorial design. The main factors consisted of diet (7% corn oil [CO] or 7% fish oil [FO]) and immunological challenge (LPS or saline). On d 14 and 21, pigs were injected intraperitoneally with either 200 microg/kg BW of LPS or an equivalent amount of sterile saline. Blood samples were collected 3 h after injection for analysis of interleukin-1beta (IL-1beta), prostaglandin E2 (PGE2), cortisol, growth hormone (GH), and insulin-like growth factor (IGF)-I. On d 2 after LPS challenge, peripheral blood lymphocyte proliferation (PBLP) was determined. Lipopolysaccharide challenge decreased ADG (487 vs. 586 g; P < 0.05) and ADFI (as-fed, 776 vs. 920 g; P < 0.05) from d 14 to 21 and ADG (587 vs. 652 g; P < 0.10) from d 21 to 28. Fish oil improved ADG (554 vs. 520 g; P < 0.10) and ADFI (891 vs. 805 g; P < 0.10) from d 14 to 21. On d 14, LPS challenge x diet interactions were observed for IL-1beta (P < 0.10), PGE2 (P < 0.001), and cortisol (P < 0.05) such that these measurements responded to the LPS challenge to a lesser extent (IL-1beta: 93 vs. 114 pg/mL, P < 0.05; PGE2: 536 vs. 1,285 pg/mL, P < 0.001; cortisol: 143 vs. 206 ng/mL, P < 0.05) in pigs receiving the FO diet than in pigs fed the CO diet. In contrast, among LPS-treated pigs, pigs fed the FO diet had higher IGF-I (155 vs. 101 ng/mL; P < 0.10) than those fed the CO diet. On d 21 among LPS-treated pigs, pigs fed FO had lower IL-1beta (70 vs. 84 pg/mL; P < 0.10) and cortisol (153 vs. 205 ng/mL; P < 0.05) than those fed CO. Pigs fed FO had lower PGE2 (331 vs. 444 pg/mL; P < 0.05) and higher IGF-I (202 vs. 171 ng/mL; P < 0.10) compared with those fed CO. Lipopolysaccharide challenge decreased GH (0.27 vs. 0.33 ng/mL; P < 0.05) on d 14, whereas it had no effect on GH on d 21. During both LPS challenge periods, the challenge increased PBLP when these cells were incubated with 8 (1.46 vs. 1.32; P < 0.10) or 16 microg/mL (1.46 vs. 1.30; P < 0.05) of concanavalin A. Fish oil had no effect on PBLP. These results suggest that FO alters the release of proinflammatory cytokines, which might lead to improved pig performance during an immunological challenge.

Effects of lentinan on broiler splenocyte proliferation, interleukin-2 production, and signal transduction.

Lentinan (LEN) is an intensively studied beta-glucan that has been shown to have immunostimulating and antitumor functions. However, it currently is not used in veterinary practice and animal production. This study was conducted to investigate the impacts of LEN on broiler splenocyte proliferation, interleukin-2 (IL-2) production, and to explore its effects on receptor signal transduction by determining signaling molecules, including nitric oxide (NO), cytosolic-free Ca2+, cytosolic cyclic adenosine monophosphate (cAMP), and cyclic guanosine monophosphate (cGMP). The results showed that compared to the control all three measured doses, including 40, 80, and 160 microg/mL LEN, increased splenocyte proliferation and IL-2 production (P < 0.05). Lentinan increased splenocyte NO production and cytosolic-free calcium concentration in a dose-dependent manner (P < 0.001). After incubating splenocytes for 20 min, LEN increased intracellular cAMP and cGMP concentrations at a dose of 80 and 160 microg/mL, respectively. However, after incubating the splenocytes for 60 min, LEN had no effect on cytosolic cAMP and cGMP concentration irrespective of the LEN dosage. Those results indicated that LEN had immunostimulatory effects on splenocytes by increasing splenocyte proliferation and IL-2 production and by activating splenocyte receptor signal transduction.

Effects of Chinese herbal polysaccharides on the immunity and growth performance of young broilers.

Two trials were conducted to study the effects of two Chinese herbal polysaccharides [achyranthan (ACH), a low-molecular-weight polysaccharide, and astragalan (APS), a high-molecular-weight polysaccaride] on the immunity and growth performance of young broilers. Trial 1 was a 28-d growth assay, in which 7-d-old broilers (n = 240) were randomly allotted to one of three dietary treatments, with eight replicate pens per treatment and ten chickens per pen. Dietary treatments included a control corn-soy-fishmeal (Treatment 1), a diet with 200 mg/kg APS (Treatment 2), and a diet with 200 mg/kg ACH (Treatment 3). Blood samples were collected by cardiac puncture on Days 7, 14, 21, and 28 for determination of serum parameters, and chickens were killed on Day 28 to measure immune organ indexes. Trial 2 was an in vitro trial to study the effects of different concentrations of polysaccharides on broiler splenocyte functions. In Trial 1, feeding either APS or ACH had no significant effects on growth performance of broilers relative to the control. However, compared to the control, feeding ACH significantly increased microhemagglutination inhibition (HI) antibody titers, bursa of Fabricius index, serum albumin, serum calcium, and nitric oxide (NO) concentrations at Day 28 (P < or = 0.05). In Trial 2, both polysaccharides showed significant immunostimulating effects. They increased NO and interleukin-2 (IL-2) production of splenocytes and enhanced splenocyte proliferation in a dose-dependent manner (P < 0.05). Those results indicate that the immunostimulating effects of APS are not as pronounced as those of ACH. Achyranthan showed immunostimulating effects in both the growth assay and in vitro studies. Therefore, ACH may be a Chinese herbal polysaccharide that has the potential to be used as a feed additive to improve broilers' immunity.

Optimum ratio of histidine in the piglet ideal protein model and its effects on body metabolism. I. Basal diet formulation based on digestible amino acids according to the ideal protein model for 10 to 20 kg piglets.

A 4 x 4 Latin square design was used to determine ileal apparent digestibility of amino acids (AAs) in corn, soybean meal, feather meal and dried whey in young pigs. The data were then to be used in formulating a basal diet for studies on AA metabolism in young pigs. Eight castrates T-cannulated at terminal ileum (average initial body weight 12.5 +/- 0.62 kg) were divided into 4 groups on the basis of body weight and transferred to individual metabolism crates. They were then fed four experimental diets containing the four feedstuffs to be tested (corn, soybean meal, feather meal and dried whey). The trial lasted 20 days, which included 4 five-day periods for ileal digesta collection. It was found that the digestibility of the AAs was similar to that reported in literature. Based on the findings a basal diet for this research was formulated according to an ideal protein model for the 10 to 20 kg piglet, on the basis of digestible AAs and containing 14.13 MJ/kg digestible energy, 18.22% crude protein, 1.04% digestible lysine and 0.23% digestible histidine.

Optimum ratio of histidine in the piglet ideal protein model and its effects on the body metabolism. II. Optimum ratio of histidine in 10-20 KG piglet ideal protein and its effects on blood parameters.

Two growth trails were conducted to determine the optimum ratio of histidine in 10-20 kg piglet ideal protein model. Four diets containing 0.23%, 0.31%, 0.39% and 0.47% digestible histidine (0, 0.08%, 0.16%, 0.24% crystalline histidine supplemented into the basal diet) were fed to 96 piglets of mean initial body weight 10.3 +/- 1.08 kg for 18 d in Experiment 1. Average daily gain, average daily feed intake and feed conversion efficiency were inhibited (P < 0.05) with the diet containing 0.23% digestible histidine. Performance was maximized with 0.31% digestible histidine. As the dietary histidine increased, blood urea nitrogen and serum cholesterol concentration were influenced significantly. The concentrations of serum histamine and free histidine did not change with increase in digestible histidine from 0.23 to 0.31%, but higher supplementation resulted in a significant linear increase in both serum parameters. It was concluded that the dietary level of 0.23% digestible histidine does not meet the requirement of 10-20 kg piglets. Based on the results from Experiment 1, Experiment 2 was designed to determine the optimum ratio of lysine:histidine in the ideal protein model of 10-20 kg piglet. Ninety-six Large White x Landrace piglets weighing 10.2 +/- 0.88 kg were divided into 4 groups. They were fed four diets containing 0.26, 0.29, 0.32 or 0.35% digestible histidine, formulated by adding 0.03, 0.06, 0.09 or 0.12% crystalline histidine to the basal diet. The trial lasted for 21 days. Results showed that performance was significantly improved with 0.32 and 0.35% digestible histidine. As dietary histidine increased, blood urea nitrogen tended to decrease but not significant at P < 0.05. Serum cholesterol concentration increased with an increase in dietary histidine level and reached a maximum at 0.35%. Serum histamine increased with increasing dietary histidine. Free serum histidine increased linearly with increased dietary histidine. From both experiments it was concluded that the digestible histidine requirement for 10-20 kg piglets was 0.31% and that the optimum ratio of dietary lysine to histidine should be 100:30. The concentrations of cholesterol, histamine and free histidine in serum were sensitive parameters to measure changes in dietary histidine levels.