Synthesis and characterization of MnO2/Eggplant carbon composite for enhanced supercapacitors.

In this paper, the eggplant carbon (EC)was derived from eggplant skin by one-step carbonization method. Subsequently, the MnO2/eggplant carbon (MnO2/EC) composite was prepared viain-situ hydrothermal method. The morphology and structure as well as electrochemical performance were investigated through a series of characterization and tests. The results showed that the urchin shaped structures of MnO2 was successfully loaded on the surface of EC. The electrochemical studies indicated that the specific capacitance of the MnO2/ECcomposite could reach 652.5F/g at 0.5 A/g in 1 M Na2SO4 aqueous electrolyte. In addition, the MnO2/EC composite exhibits excellent cyclic stability after 10000 cycles, which might be ascribed to the synergistic effect of MnO2 and EC for the improvement of electrochemical performance. Taken together, this work demonstrated that MnO2/EC composite can be used in the aspect of energy storage for high-performance supercapacitors.

[Application of limiting antigen avidity enzyme immunoassay for estimating HIV-1 incidence in men who have sex with men].

Objective: To estimate the incidence of HIV-1 infection in men who have sex with men (MSM) in key areas of China through HIV-1 limiting antigen avidity enzyme immunoassay (LAg-Avidity EIA), analyze the deviation from the actual results and identify influencing factors, and provided reference for improving the accuracy of estimation results. Methods: Based on the principle of the cohort randomized study design, 20 cities were selected in China based on population size and the number of HIV-positive MSM. The sample size was estimated to be 700 according to the HIV-1 infection rate in MSM. MSM mobile phone app. was used to establish a detection appointment and questionnaire system, and the baseline cross-sectional survey was conducted from April to November 2019. LAg-Avidity EIA was used to identify the recent infected samples. The incidence of HIV-1 infection was calculated and then adjusted based on the estimation formula designed by WHO. The influencing factors were identified by analyzing the sample collection and detection processes. Results: Among the 10 650 blood samples from the participants, 799 were HIV-positive in initial screening, in which 198 samples (24.78%) missed during confirmation test. Only 621 samples were received by the laboratory. After excluding misreported samples, 520 samples were qualified for testing. A total of 155 samples were eventually determined as recent infection through LAg-Avidity EIA; Based on the estimation formula , the incidence of HIV-1 infection in MSM in 20 cities was 4.06% (95%CI:3.27%-4.85%), it increased to 5.53% (95%CI: 4.45%-6.60%)after the adjusting for sample missing rate. When the sample missing rate and misreporting rate were both adjusted, the incidence of HIV-1 infection in the MSM increased to 5.66% (95%CI:4.67%-6.65%). The actual incidence of HIV-1 infection in MSM in the 20 cities might be between 4.06% and 5.66%. Conclusions: Sample missing and misreporting might cause the deviation of the estimation of HIV-1 infection incidence. It is important to ensure the sample source and the quality of sample collection and detection to reduce the deviation in the estimation of HIV-1 infection incidence.

[Effects of different crystalloid fluids resuscitation on renal structure and function in sepsis rats].

To evaluate the effects of resuscitation with normal saline and sodium potassium magnesium calcium and glucose injection on renal structure and function in septic rats. Rat model of sepsis was established by ligation and perforation of cecum. Male SD rats were divided into four groups: sham operation group, sepsis group, saline resuscitation group, sodium potassium magnesium calcium and glucose injection resuscitation group. Blood gas analysis was performed at the end of resuscitation. The rats were sacrificed 72 hours after resuscitation. Blood samples were taken to measure the plasma levels of blood urea nitrogen (BUN), creatinine, interleukin-1ß (IL-1ß), interleukin-6 (IL-6) and tumor necrosis factorα (TNFα). Caspase-3 expression was detected by immunohistochemistry in kidney sections. The degree of renal injury was evaluated by regular HE staining and electron microscope. Compared with normal saline resuscitation, sodium potassium calcium magnesium glucose injection resuscitation could decrease the levels of BUN, serum creatinine, IL-1ß, IL-6 and TNFα (P<0.05) , reduce the expression of caspase-3 (P<0.05) , and improve the renal injury score (P<0.05) . Sodium potassium calcium magnesium glucose injection resuscitation can significantly improve the renal function of sepsis rats with less pathological damage of the kidney.

Effect of propofol on myocardial ischemia-reperfusion injury through MAPK/ERK pathway.

OBJECTIVE: The aim of this study was to investigate the effects of propofol on myocardial ischemia-reperfusion injury (MIRI) and the mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) pathway. MATERIALS AND METHODS: Primary cells were first isolated from rats. The effects of propofol on the apoptosis of primary myocardial cells and the expression of apoptosis-related proteins were detected via flow cytometry and Western blotting, respectively. Meanwhile, the effect of propofol on MIRI model after ischemia for 2 h and reperfusion for 24 h was detected as well. Subsequently, the effect of propofol on the activity of proteins in myocardial tissues was detected using transcriptome sequencing and VIPER method. The effect of propofol on myocardial tissues was detected via 2, 3, 5- triphenyl tetrazolium chloride (TTC) staining, hematoxylin-eosin (HE) staining, and Masson staining. Besides, the effect of propofol on myocardial function was detected using the BL-420F hemodynamic system. Propofol effect on the MAPK/ERK signaling pathway was determined via Western blotting in vivo. Finally, the effects of propofol on the content of serum lactate dehydrogenase (LDH), creatine kinase (CK), total superoxide dismutase (T-SOD), and malondialdehyde (MDA) were detected using relevant kits. RESULTS: Propofol activated the MAPK/ERK signaling pathway in a dose-dependent manner in primary myocardial cells, which also reduced the apoptotic rate of myocardial cells. The results of TTC staining, HE staining, and Masson staining showed that propofol significantly reduced MIRI in a dose-dependent manner in vivo. ERK inhibitor PD-98059 could significantly reduce the cardioprotective effect of propofol. Propofol significantly decreased the content of serum LDH, CK, and MDA (p<0.05), while it increased the content of T-SOD (p<0.05). According to the hemodynamic study, statistically significant differences were observed in left ventricular systolic pressure (LVSP), maximal rate of the decrease of left ventricular pressure (-dp/dtmax), and left ventricular end-diastolic pressure (LVEDP) between propofol group and model group (p<0.01). The results of Western blotting revealed that propofol increased the protein expression level of p-ERK1/2 in a dose-and time-dependent manner in vivo. Furthermore, the expression level of p-ERK1/2 remarkably increased at 8 h after ischemia-reperfusion. CONCLUSIONS: Propofol exerts a cardio-protective effect on MIRI through the MAPK/ERK pathway.

Dual functions of eicosapentaenoic acid-rich microalgae: enrichment of yolk with n-3 polyunsaturated fatty acids and partial replacement for soybean meal in diet of laying hens.

Microalgae (Nannochloropsis sp., NS), with high contents of eicosapentaenoic acid (EPA) and crude protein, may be one of the important n-3 polyunsaturated fatty acid (PUFA) sources and potential protein feed ingredient. The purposes of this study were to enrich yolk with n-3 PUFA by dietary EPA-rich NS supplementation and to evaluate whether it is feasible to partly substitute for soybean meal in laying hens diet. A total of 360 37-wk-old healthy Lohmann Brown laying hens, with similar laying rate and body weight, were randomly allotted to 5 groups (6 replicates, 12 birds/replicate) and fed 5 experimental diets (0, 1, 2, 4, and 8% NS) for 4 wk. The hen performance and egg quality (except yolk color) were not affected (P > 0.05) by the NS supplemental diets. Yolk color score was increased as NS supplementation in diets (P < 0.001), and peaked on about the seventh day in all NS supplemental groups. The concentration of total n-3 PUFA was increased (P < 0.001), while total n-6 PUFA and n-6/n-3 ratio were decreased (P < 0.001) in yolk with increasing NS levels in diets. The 8% NS group had highest docosahexaenoic acid (DHA) and total n-3 PUFA levels, reaching 111.6 mg DHA and 148.6 mg total n-3 PUFA per egg. Maximum DHA, total n-3 PUFA, very long-chain (LC-) n-3 PUFA, and LC-PUFA levels were all observed at day 13 of NS supplementation. In conclusion, dietary NS supplementation enriched yolk with n-3 PUFA (especially DHA) and enhanced yolk color score without adverse effects on performance and egg quality, and indicated the practical feasibility of partial replacement for soybean meal in laying hens diet.

Effect of gut stress induced by oxidized wheat gluten on the growth performance, gut morphology and oxidative states of broilers.

This study was to investigate the effect of oxidized wheat gluten (OG) on growth performance, gut morphology and its oxidative states of broilers. One hundred and eighty-day-old male broilers (10 chicks/pen) were randomly allocated into three dietary treatments: control diet (CON), diet with 8% wheat gluten (WG) and diet with 8% OG with six pens/treatment. Body weight (BW) (21 and 35 days) and average daily gain (ADG) (1-21 days and 22-35 days) decreased (p < .05) and feed conversion ratio (FCR) (1-21 days and 22-35 days) increased (p < .05) in OG treatment. Feed intake (FI) decreased (p < .05) in WG and OG treatments during 22-35 days. However, FI was not influenced by dietary treatments during 1-21 days (p > .05). The OG-fed broilers had a lower faecal pH value (p < .05) and higher faecal moisture content (p < 05) at 14, 21, 28 and 35 days. Villus height, crypt depth and V/C value were not different (p > .05) among treatments at 21 and 35 days. Lipid peroxidation (LPO) (21 and 35 days) and malondialdehyde (MDA) (35 days) content in crop of OG treatment increased (p < .05). Oxidized glutathione (GSSG) (21 days), LPO (21 and 35 days) and MDA (21 and 35 days) content in ileum of OG treatment increased (p < .05). The reduced glutathione/oxidized glutathione (GSH/GSSG) (21 days) and (GSH) (35 days) in ileum of OG treatment decreased (p < .05). The present findings indicate that OG might be a stressor for broiler gut, which could induce oxidative stress both in crop and in ileum, and the diarrhoea as well. The growth performance of broiler was consequently depressed.

[The advance of detection technology of HIV self-testing].

At present, China's AIDS testing increased rapidly, but there are still many people living with HIV do not recognize their status, thus postponing the antiviral treatment time. HIV self-testing (HST) is an effective method to expand the testing, not only simple operation, easy to get a result, effectively protect the detection privacy, expand the selection of testers, suit to the entire population, but also the premise and basis of other AIDS comprehensive prevention measures, all over the world are promoting it. Because the HST has controversies in the window period, price and before and after controversial, and our country is in the initial stage of HST, so it is not to develop related policies, but more and more countries are in accordance with their own situations are modified or developed to allow to use rapid detection of AIDS policy to regulate the field. This paper analyzed and summarized the advantage and influence factors of HST promotion, HST believes that in the long term, the advantages outweigh the disadvantages, we need to formulate relevant policies, and improve the sensitivity of the kit, shorten the window period of time, production and promotion of operation standard of video, specification and testing the operating practices, preventing and reporting the possible social harm, investigation and understanding of the needs of the people of the crowd, to maximize the advantages of HST, find more infection, so as to curb the epidemic of AIDS.

Apparent metabolizable and net energy values of corn and soybean meal for broiler breeding cocks.

The AME and net energy (NE) values of 4 corn varieties, including 2 normal corn varieties (Zheng Dan 958 and Xian Yu 335), and one each of waxy corn and sweet corn, and 2 soybean meal samples including regular (RSBM) and dehulled soybean meal (DSBM), were determined in 2 experiments for broiler breeding cocks using the indirect calorimetry method. The 4 test diets in Experiment 1 consisted of each test corn, which replaced 40% of the corn-soybean meal basal diet, and the test diets in Experiment 2 contained 25% RSBM or DSBM, which was used to replace the corn basal diet. Thirty (Experiment 1) or 18 (Experiment 2) 50-week-old Arbor Acre (AA) broiler breeding cocks were used in a completely randomized design. After a 7 d dietary adaptation period, 6 birds as replicates from each treatment were assigned to individual respiration chambers for energy measurement via gaseous exchange and total excreta collection for 10 d. In Experiment 1, the AME, ME intake (MEI), retained energy (RE), NE, and NE:AME ratio values were higher (P < 0.001) in the test diets as compared with the corn-soybean meal basal diet. The AME and NE values in the sweet corn diet were higher (P < 0.05) than those values in the other 3 test diets. The heat production (HP), fasting heat production (FHP), and respiration quotient (RQ) were not influenced by the various experimental diets. The respective AME and NE values were 3,785, 3,775, 3,738, and 3,997 kcal/kg (DM basis), and 2,982, 3,006, 2,959, and 3,146 kcal/kg (DM basis) for Zheng Dan 958, Xian Yu 335, waxy corn, and sweet corn. Birds fed a corn basal diet in Experiment 2 had higher AME, MEI, RE, NE, and NE:AME ratio values (P < 0.001). Soybean meal substitution had no effect on HP, FHP, or RQ. The average AME and NE content was 2,492 and 1,581 kcal/kg (DM basis) for RSBM, and 2,580 and 1,654 kcal/kg (DM basis) for DSBM, respectively.

Improved thermal lattice Boltzmann model for simulation of liquid-vapor phase change.

In this paper, an improved thermal lattice Boltzmann (LB) model is proposed for simulating liquid-vapor phase change, which is aimed at improving an existing thermal LB model for liquid-vapor phase change [S. Gong and P. Cheng, Int. J. Heat Mass Transfer 55, 4923 (2012)10.1016/j.ijheatmasstransfer.2012.04.037]. First, we emphasize that the replacement of ∇·(λ∇T)/∇·(λ∇T)ρc_{V}ρc_{V} with ∇·(χ∇T) is an inappropriate treatment for diffuse interface modeling of liquid-vapor phase change. Furthermore, the error terms ∂_{t_{0}}(Tv)+∇·(Tvv), which exist in the macroscopic temperature equation recovered from the previous model, are eliminated in the present model through a way that is consistent with the philosophy of the LB method. Moreover, the discrete effect of the source term is also eliminated in the present model. Numerical simulations are performed for droplet evaporation and bubble nucleation to validate the capability of the model for simulating liquid-vapor phase change. It is shown that the numerical results of the improved model agree well with those of a finite-difference scheme. Meanwhile, it is found that the replacement of ∇·(λ∇T)/∇·(λ∇T)ρc_{V}ρc_{V} with ∇·(χ∇T) leads to significant numerical errors and the error terms in the recovered macroscopic temperature equation also result in considerable errors.

Revised Chapman-Enskog analysis for a class of forcing schemes in the lattice Boltzmann method.

In the lattice Boltzmann (LB) method, the forcing scheme, which is used to incorporate an external or internal force into the LB equation, plays an important role. It determines whether the force of the system is correctly implemented in an LB model and affects the numerical accuracy. In this paper we aim to clarify a critical issue about the Chapman-Enskog analysis for a class of forcing schemes in the LB method in which the velocity in the equilibrium density distribution function is given by u=∑_{α}e_{α}f_{α}/ρ, while the actual fluid velocity is defined as u[over ̂]=u+δ_{t}F/(2ρ). It is shown that the usual Chapman-Enskog analysis for this class of forcing schemes should be revised so as to derive the actual macroscopic equations recovered from these forcing schemes. Three forcing schemes belonging to the above class are analyzed, among which Wagner's forcing scheme [A. J. Wagner, Phys. Rev. E 74, 056703 (2006)10.1103/PhysRevE.74.056703] is shown to be capable of reproducing the correct macroscopic equations. The theoretical analyses are examined and demonstrated with two numerical tests, including the simulation of Womersley flow and the modeling of flat and circular interfaces by the pseudopotential multiphase LB model.

Pinning-Depinning Mechanism of the Contact Line during Evaporation on Chemically Patterned Surfaces: A Lattice Boltzmann Study.

In this paper, the pinning and depinning mechanism of the contact line during droplet evaporation on chemically stripe-patterned surfaces is numerically investigated using a thermal multiphase lattice Boltzmann (LB) model with liquid-vapor phase change. A local force balance in the context of diffuse interfaces is introduced to explain the equilibrium states of droplets on chemically patterned surfaces. It is shown that when the contact line is pinned on a hydrophobic-hydrophilic boundary, different contact angles can be interpreted as the variation of the length of the contact line occupied by each component. The stick-slip-jump behavior of evaporating droplets on chemically patterned surfaces is well captured by the LB simulations. Particularly, a slow movement of the contact line is clearly observed during the stick (pinning) mode, which shows that the pinning of the contact line during droplet evaporation on chemically stripe-patterned surfaces is actually a dynamic pinning process and the dynamic equilibrium is achieved by the self-adjustment of the contact lines occupied by each component. Moreover, it is shown that when the surface tension varies with the temperature, the Marangoni effect has an important influence on the depinning of the contact line, which occurs when the horizontal component (toward the center of the droplet) of the force caused by the Marangoni stress overcomes the unbalanced Young's force toward the outside.

First Report of Mucor Rot on Stored 'Gala' Apple Fruit Caused by Mucor piriformis in Pennsylvania.

Mucor piriformis E. Fischer causes Mucor rot of pome and stone fruits during storage and has been reported in Australia, Canada, Germany, Northern Ireland, South Africa, and portions of the United States (1,2). Currently, there is no fungicide in the United States labeled to control this wound pathogen on apple. Cultural practices of orchard sanitation, placing dry fruit in storage, and chlorine treatment of dump tanks and flumes are critical for decay management (3,4). Cultivars like 'Gala' that are prone to cracking are particularly vulnerable as the openings provide ingress for the fungus. Mucor rot was observed in February 2013 at a commercial packing facility in Pennsylvania. Decay incidence was ~15% on 'Gala' apples from bins removed directly from controlled atmosphere storage. Rot was evident mainly at the stem end and was light brown, watery, soft, and covered with fuzzy mycelia. Salt-and-pepper colored sporangiophores bearing terminal sporangiospores protruded through the skin. Five infected apple fruit were collected, placed in an 80-count apple box on trays, and temporarily stored at 4°C. Isolates were obtained aseptically from decayed tissue, placed on potato dextrose agar (PDA) petri plates, and incubated at 25°C with natural light. Five single sporangiospore isolates were identified as Mucor piriformis based on cultural characteristics according to Michailides and Spotts (1). The isolates produced columellate sporangia attached terminally on short and tall, branched and unbranched sporangiophores. Sporangiospores were ellipsoidal, subspherical, and smooth. Chlamydospore-like resting structures (gemmae), isogametangia, and zygospores were not evident in culture. Mycelial growth was examined on PDA, apple agar (AA), and V8 agar (V8) at 25°C with natural light. Isolates grew best on PDA at rates that ranged from 38.4 ± 5.3 to 34.5 ± 2.41 mm/day, followed by AA from 30.5 ± 1.22 to 28.5 ± 2.51 mm/day, and V8 from 29.2 ± 3.0 to 26.7 ± 2.17 mm/day. Species-level identification was conducted by isolating genomic DNA, amplifying a portion of the 28S rDNA gene, and directly sequencing the products. MegaBLAST analysis of the 2X consensus sequences revealed that all five isolates were 99% identical to M. piriformis (GenBank Accession No. JN2064761) with E values of 0.0, which confirms the morphological identification. Koch's postulates were conducted using organic 'Gala' apples that were surface sanitized with soap and water, then sprayed with 70% ethanol and allowed to air dry. Wounds 3 mm deep were created using the point of a finishing nail and then inoculated with 50 µl of a sporangiospore suspension (1 × 105 sporangiospores/ml) for each isolate. Ten fruit were inoculated with each isolate, and the experiment was repeated. The fruit were stored at 25°C in 80-count boxes on paper trays for 14 days. Decay observed on inoculated 'Gala' fruit was similar to symptoms originally observed on 'Gala' apples from storage and the pathogen was re-isolated from inoculated fruit. This is the first report of M. piriformis causing postharvest decay on stored apples in Pennsylvania and reinforces the need for the development of additional tools to manage this economically important pathogen. References: (1) T. J. Michailides, and R. A. Spotts. Plant Dis. 74:537, 1990. (2) P. L. Sholberg and T. J. Michailides. Plant Dis. 81:550, 1997. (3) W. L. Smith et al. Phytopathology 69:865, 1979. (4) R. A. Spotts. Compendium of Apple and Pear Diseases and Pests: Second Edition. APS Press, St. Paul, MN, 2014.

First Report of Pyrimethanil Resistance in Botrytis cinerea from Stored Apples in Pennsylvania.

Botrytis cinerea Pers.: Fr. (teleomorph Botryotinia fuckeliana [de Bary] Whetzel) causes gray mold on apple fruit. A survey of commercial packinghouses in Washington State revealed that it accounted for 28% of the decay in storage (1). Fungicide application coupled with cultural practices are the primary method of control as all commercial apple cultivars are susceptible to gray mold. In February 2013, gray mold was observed at ~5% incidence for commercially packed 'Gala' apple fruit that had been treated with Penbotec (active ingredient: pyrimethanil, Shield-Bright, Pace International) prior to controlled atmosphere storage in Pennsylvania. Eight infected apple fruit were collected, placed in 80 count boxes on cardboard trays, and stored at 4°C. One isolate was obtained from each decayed apple, placed on potato dextrose agar (PDA) petri plates, and incubated at 20°C with natural light. Eight single-spore isolates were identified as B. cinerea based on cultural characteristics. Species level identification was executed by obtaining mycelial genomic DNA, amplifying the ITS rDNA, and sequencing the ~550-bp amplicon directly (2). MegaBLAST analysis of the 2X consensus for the 8 isolates revealed 100% identity to B. cinerea ITS sequences in GenBank (KF156296.1 and JX867227.1) with E values of 0.0, thus confirming the morphological identification. Minimum inhibitory concentration (MIC) was determined using conidial suspensions obtained from ~14-day-old plates (104 spores/ml) and a range (0 to 500 µg/ml) of technical grade pyrimethanil on three replicated 96-well microtiter plates containing a defined medium for each experiment. Conidial proliferation was inhibited at 250 µg/ml for all eight isolates and the experiment was conducted four times. To further define the resistance levels between the isolates, mycelial growth analysis using a plug of actively growing mycelium from the margin of ~3-day-old plates was conducted with a defined medium three times with technical grade pyrimethanil with three plates per experiment. Five isolates grew at 250 µg/ml (highly resistant), while three did not (moderately resistant). To assess resistance in vivo, organic 'Gala' apples were rinsed with soap and water, sprayed with 70% ethanol, placed on trays, and allowed to air dry. Apples were wounded with a sterile finishing nail, inoculated with 20 µl of a conidial suspension (104 spores/ml) of either a moderately or a highly resistant isolate, and dipped in the labeled application rate of Penbotec at 500 µg/ml or sterile water for 30 s. Fruit were stored in 100 count boxes at 22°C for 5 days and decay incidence and severity were recorded. Ten fruit composed a replicate per treatment and the experiment was repeated. Water inoculated controls were symptomless and water-dipped inoculated fruit had 100% decay. Penbotec-treated fruit had 100% decay incidence and mean lesion diameters of 37.6 (±13.1 mm) for the highly, and 35.7 (±9.0 mm) for the moderately resistant isolate. This is the first report of pyrimethanil resistance in B. cinerea from decayed apples collected from a commercial packinghouse in Pennsylvania. The results indicate that pyrimethanil resistance has developed in B. cinerea, which can result in control failures on Penbotec-treated fruit during storage. Furthermore, it emphasizes the need for additional tools to manage gray mold on apple fruit and may pose issues for export concerning the spread of fungicide-resistant inoculum. References: (1) Y.-K. Kim and C. L. Xiao. Plant Dis. 92:940, 2008. (2) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

First Report of Penicillium expansum Isolates Resistant to Pyrimethanil from Stored Apple Fruit in Pennsylvania.

Apples in the United States are stored in low-temperature controlled atmospheres for 9 to 12 months and are highly susceptible to blue mold decay. Penicillium spp. cause significant economic losses worldwide and produce mycotoxins that contaminate processed apple products. Blue mold is managed by a combination of cultural practices and the application of fungicides. In 2004, a new postharvest fungicide, pyrimethanil (Penbotec 400 SC, Janseen PMP, Beerse, Belgium) was registered for use in the United States to control blue mold on pome fruits (1). In this study, 10 blue mold symptomatic 'Red Delicious' apples were collected in May 2011 from wooden bins at a commercial facility located in Pennsylvania. These fruit had been treated with Penbotec prior to controlled atmosphere storage. Ten single-spore Penicillium spp. isolates were analyzed for growth using 96-well microtiter plates containing Richards minimal medium amended with a range of technical grade pyrimethanil from 0 to 500 µg/ml. Conidial suspensions adjusted to 1 × 105 conidia/ml were added to three 96-well plates for each experiment; all experiments were repeated three times. Nine resistant isolates had prolific mycelial growth at 500 µg/ml, which is 1,000 times the discriminatory dose that inhibited baseline sensitive P. expansum isolates from Washington State (1). However, one isolate (R13) had limited conidial germination and no mycelial proliferation at 0.5 µg/ml and was categorized as sensitive. One resistant (R22) and one sensitive (R13) isolate were selected on the basis of their different sensitivities to pyrimethanil. Both isolates were identified as P. expansum via conventional PCR using ß-tubulin gene-specific primers according to Sholberg et al. (2). Analysis of the 2X consensus amplicon sequences from R13 and R22 matched perfectly (100% identity and 0.0 E value) with other P. expansum accessions in GenBank including JN872743.1, which was isolated from decayed apple fruit from Washington State. To determine if pyrimethanil applied at the labeled rate of 500 µg/ml would control R13 or R22 in vivo, organic 'Gala' apple fruit were wounded, inoculated with 50 µl of a conidial suspension (1 × 104 conidia/ml) of either isolate, dipped in Penbotec fungicide or sterile water, and stored at 25°C for 7 days. Twenty fruit composed a replicate within a treatment and the experiment was performed twice. Non-inoculated water-only controls were symptomless, while water-dipped inoculated fruit had 100% decay with mean lesion diameters of 36.8 ± 2.68 mm for R22 and 38.5 ± 2.61 mm for R13. The R22 isolate caused 30% decay with 21.6 ± 5.44 mm lesions when inoculated onto Penbotec-treated apples, while the R13 isolate had 7.5% decay incidence with mean lesion diameters of 23.1 ± 3.41 mm. The results from this study demonstrate that P. expansum pyrimethanil-resistant strains are virulent on Penbotec-treated apple fruit and have the potential to manifest in decay during storage. To the best of our knowledge, this is the first report of pyrimethanil resistance in P. expansum from Pennsylvania, a major apple growing region for the United States. Moreover, these results illuminate the need to develop additional chemical, cultural, and biological methods to control this fungus. References: (1) H. X. Li and C. L. Xiao. Phytopathology 98:427, 2008. (2) P. L. Sholberg et al. Postharvest Biol. Technol. 36:41, 2005.

Analysis of antigen epitopes and molecular pathogenic characteristics of the 2009 H1N1 pandemic influenza A virus in China.

In order to further predict the epidemic trend and develop vaccines for 2009 H1N1 virus, we monitored its epitopes and molecular pathogenic characteristics during the epidemic process. We also analyzed the similarity of antigenic and genetic characteristics among the novel 2009 H1N1, representative seasonal H1N1 strains, and vaccine strains. 2009 H1N1 isolates had high similarity of hemagglutinin (HA) antigenic sites with H1N1 viruses isolated before 1940 and up to 80.0% similarity with 1918 H1N1. The elderly people born before 1940 have relatively low 2009 H1N1 infection rate, which might be responsible for their previous infection with either 1918 H1N1 virus or an early progeny. Compared to seasonal H1N1 vaccine strains from 1999 to 2010, the HA, neuraminidase (NA), and nucleoprotein (NP) proteins of the isolates had highly conserved CTL epitopes (60.5-65.8%, 69.6-82.6%, and 76.7%, respectively). The seriousness and mortality rate of 2009 H1N1 infections were similar to seasonal influenza, which may be related to the molecular characteristics of low toxicity of 2009 H1N1 and cross-T-cell immunity, due to vaccination or exposure to seasonal H1N1 virus. Some strains of 2009 H1N1 acquired mutations at antigenic and glycosylation sites. It is of particular interest that Haishu/SWL110/10 and Beijing/SE2649/09, isolated after November 2009, gained a new glycosylation site at the position 179 of HA protein, near the RBD. Thus, in the future, vaccination with glycosylated 2009 H1N1 virus may prevent the seasonal epidemic caused by strains with glycosylation site mutation near the receptor binding domain (RBD).

Transport stress in broilers. II. Superoxide production, adenosine phosphate concentrations, and mRNA levels of avian uncoupling protein, avian adenine nucleotide translocator, and avian peroxisome proliferator-activated receptor-gamma coactivator-1alpha in skeletal muscles.

The effect of transport stress on superoxide production and adenosine phosphate concentration in addition to avian uncoupling protein (avUCP), avian adenine nucleotide translocator, and avian peroxisome proliferator-activated receptor-gamma coactivator-1alpha mRNA levels of skeletal muscles in broilers was investigated. Arbor Acres chicks (n = 360, 46 d old, males) were randomly allotted to 1 of 5 treatments: unstressed control, 45-min (short-term) transport with 45-min (short-term) recovery, 45-min transport with 3-h (long-term) recovery, 3-h (long-term) transport with 45-min recovery, and 3-h transport with 3-h recovery. Each treatment consisted of 6 replicates with 12 birds each. All birds (except control group) were transported according to a designed protocol. Transport time affected reactive oxygen species production in the thigh muscle (P < 0.05), adenosine triphosphate (ATP) content and energy charge (EC) in both breast and thigh muscles (P < 0.05 for all 4 comparisons), ATP:adenosine diphosphate (ADP) ratio in the breast muscle (P < 0.05), and avUCP mRNA levels in the thigh muscle (P < 0.05). Long-term transport increased (P < 0.05) reactive oxygen species production, ATP content, ATP:ADP ratio, and EC in the thigh muscle, but it decreased ATP content, ATP:ADP ratio, and EC in the breast muscle. Long-term transport increased avUCP mRNA in the thigh muscle (P < 0.05). Long-term recovery increased the ATP (P < 0.05) and ADP (P < 0.05) concentrations, avian adenine nucleotide translocator mRNA (P < 0.05), and avian peroxisome proliferator-activated receptor-gamma coactivator-1alpha mRNA (P < 0.05) in the thigh muscle, whereas EC decreased (P < 0.05) in the breast muscle. There were interactions between transport and recovery time on ATP (P < 0.05), EC (P < 0.05), and avUCP mRNA level (P < 0.05) in the thigh muscle. This study suggests that long-term transport accelerates muscle energy metabolism and lipid peroxidation. A long-term recovery may help alleviate cellular damage and maintain meat quality by reducing the rate of energy metabolism and scavenging of free radicals formed.

Transport stress in broilers: I. Blood metabolism, glycolytic potential, and meat quality.

The effect of transport stress on blood metabolism, glycolytic potential, and meat quality in broilers was investigated. Arbor Acres chicks (n = 360, 1 d old, males) were randomly allotted to 1 of 5 treatments: unstressed control, 45-min (short-term) transport with 45-min (short-term) recovery, 45-min transport with 3-h (long-term) recovery; 3 h (long-term) transport with 45-min recovery, and 3-h transport with 3-h recovery. Each treatment consisted of 6 replicates with 12 birds each. On d 46, all birds (except the control group) were transported according to a designed protocol. Transport time affected plasma glucose level (P<0.05) and glycogen level (P=0.06) in breast muscle as well as the area (P<0.01) and density (P<0.01) of IIa fibers. Glucose concentration increased slightly during the first 45 min of transport and then decreased dramatically in the long-term transported broilers (P<0.05). Long-term transport decreased the concentration of breast glycogen (P=0.06) and affected the size of IIa fibers in tibialis anterior by decreasing the area (P<0.01) with an increase in density (P<0.01). However, a long-term recovery after transport contributed to the homeostasis of blood corticosterone (CORT, P=0.05) and low levels of glycogen (P<0.05), lactate (P<0.01), and glycolytic potential (P<0.01) in thigh muscles. Interactions existed between transport and recovery time on area (P<0.05) and density (P<0.01) of IIa fibers. Furthermore, plasma nonesterified fatty acids increased significantly in the 3-h transport with 3-h recovery group (P<0.05) in comparison with the control. These results suggested that transport induced the release of plasma CORT and glycopenia, which affected the contractive status of muscle fibers by changing their area and density, and enhanced glycolysis and even lipolysis. A long-term recovery after transport was beneficial in lowering plasma CORT levels and reducing muscle glycolysis, which might improve broiler meat quality.

Effect of Saccharomyces cerevisiae fermentation product on immune functions of broilers challenged with Eimeria tenella.

Three hundred sixty 1-d-old male Arbor Acres broilers were randomly allotted to 6 groups with a 2x3 factorial arrangement of treatments. Three supplemental levels (0, 0.25, and 0.50%) of Saccharomyces cerevisiae fermentation product (XP) were fed to control and Eimeria tenella-infected broilers. Growth performance and immune response criteria were measured after coccidian infection. Broiler ADG was lowered (P<0.01) by coccidian infection and improved by XP supplementation (P=0.04). Supplementation of XP increased CD3+, CD4+, and CD8+ T-lymphocyte counts (P<0.05) and the ratio CD4+:CD8+ in blood (P=0.06) and spleen (P=0.04) as well as ileum intraepithelial lymphocyte count, cecal tonsil secretory IgA counts, serum lysozyme content (P<0.01), and albumin:globulin ratio (P=0.02). These results suggest that dietary XP supplementation could improve immune function and growth performance in coccidia-infected broilers.

Molecular characterization of the surface glycoprotein genes of an H5N1 influenza virus isolated from a human in Guangdong, China.

In March 2006, a human H5N1-infected case was found in Guangdong province, China. Here, we molecularly characterized the hemagglutinin (HA) and neuraminidase (NA) genes of the A/China/GD01/06 (GD01) strain causing the infection. The phylogenetic analyses suggested that the HA and NA genes of GD01 and recent human H5N1 viruses from different provinces of China were probably derived from a common ancestor and the H5N1 human infection was acquired directly from affected poultry. At the cleavage site of HA, GD01 contained multiple basic amino acids, a feature characteristic of highly pathogenic avian influenza A viruses. The virus possessed Gln222, Gly224, Ser223, Asn182, Gln192 residues adjacent to the receptor-binding site, preferential for recognizing SAalpha2, 3Gal. In addition, the GD01 NA amino acid sequence possessed Asn344 and Phe466, which might be related to the low-pH stability of the sialidase activity and gastrointestinal symptoms of the patient.