Pathogenic Effects of Cloned Genomic DNA of Porcine Circovirus-like Virus P1 on Neonatal Mice via Different Inoculation Routes

[Objective] The paper was to explore the pathogenicity of cloned genomic DNA of porcine circovirus-like virus Pl to neonatal mice via different inoculation routes （brain, liver and muscle）. [Method] Cloned genomic DNA of P1 was inoculated to neonatal mice via different routes of brain, liver and muscle. Tissues of heart, liver, spleen, lung, kidney and brain were taken from neonatal mice at 7, 14 and 21 d post inoculation, re- spectively. Pl in various tissues were qualitatively and quantitatively detected by using ordinary PCR and quantitative real-time PCR. Meanwhile, histopathological changes were analyzed. [Result] Pl was detected in neonatal mice inoculated through three different routes. The viral load of tis- sues at 7 d post inoculation was significantly higher than those at 14 and 21 d post inoculation. Moreover, muscle inoculation led to the highest viral load in all tissues of neonatal mice. [Conclusion] Pl infection caused different degrees of pathological damage to heart, liver, lung, kidney and brain in neonatal mice.

Poisoning-free effect of calcium on grain refinement of Mg-3%Al alloy containing trace Fe by carbon inoculation

Mg-3%Al alloy was modified by combining Ca addition with carbon inoculation. The effects of Fe addition and addition sequence on the grain refinement were investigated. A higher grain refining efficiency could be obtained for the Mg-Al alloy modified by combining Ca addition with carbon inoculation. Fe addition and addition sequence had no obvious effect on the grain refinement. Ca addition could effectively avoid grain-coarsening resulting from Fe in the carbon-inoculated Mg-Al alloy. The Al-C-O particles, actually being Al4C3, should act as potent substrates for a-Mg grains in the sample treated by combining Ca addition with carbon inoculation. However, the duplex-phase particles of AI4C3 coated on Al-Fe or Al-C-Fe should be the potent substrates for a-Mg grains if Fe existed in the Mg-Al melt. Ca addition can contribute to the formation of the particles of Al4C3 coated on Al-Fe or Al-C-Fe, regardless of the Fe addition sequence. The poisoning effect of Fe was effectively inhibited in the carbon-inoculated of Mg-Al alloy due to Ca addition, namely, Ca has a poisoning-free effect.

Grain refinement of pure aluminum by direct current pulsed magnetic field and inoculation

The combined effects of direct current pulsed magnetic field （DC-PMF） and inoculation on pure aluminum were investigated, the grain refinement behavior of DC-PMF and inoculation was discussed. The experimental results indicate that the solidification micro structure of pure aluminum can be greatly refined under DC-PMF. Refinement of pure aluminum is attributed to electromagnetic undercooling and forced convection caused by DC-PMF. With single DC-PMF, the grain size in the equiaxed zone is uneven. However, under DC-PMF, by adding 0.05% （mass fraction） Al5Ti-B, the grain size of the sample is smaller, and the size distribution is more uniform than that of single DC-PMF. Furthermore, under the combination of DC-PMF and inoculation, with the increase of output current, the grain size is further reduced. When the output current increases to 100 A, the average grain size can decrease to 113 μn.

Fungal inoculation induces agarwood in young Aquilaria malaccensis trees in the nursery

Fungi are often used to induce agarwood inAquilariatrees. This study was conducted to evaluate the effect of several fungi on agarwood formation over time in youngAquilaria malaccensis （Lam.） trees. Typical changes in the length and light intensity of the resulting discoloration were observed after three and six month periods following inoculation. Wood samples were observed microscopically and classified into several light intensity groups. The discoloration length was meas-ured longitudinally. The duration after inoculation affected the mean of discoloration length： the 6-month old sample （1.70 cm） had a wider discoloration zone when compared to the 3-month old sample （1.17 cm）. When measuring the discoloration intensity, a positive relationship with time was perceived. Digital images, captured using a camera-equipped microscope, revealed that wood samples collected after six months appeared to be 1.8-times darker than after three months. We concluded that time, not the species of any of the tested fungi, had significant effect on discoloration length and intensity. Gas chromatography/mass spec-trometry （GCMS） analysis of the 6-month old sample yielded some important agarwood compounds such as benzylacetone, anisylacetone, guaiene and palustrol. This demonstrates that the tested fungi have the ability to induce agarwood formation in nurseryA. malaccensis trees.

Synergistic refining mechanism of Mg-3%Al alloy refining by carbon inoculation combining with Ca addition

Mg-3%Al alloy was refined by carbon inoculation combining with 0.2%Ca addition.High grain refining efficiency was obtained and the synergistic refining mechanism was deeply discussed in the present study.Al-C-O particles,actually Al4C3 particles,were formed in the carbon-inoculated Mg-3%A1 alloy acting as nuclei forα-Mg grains.Ca addition had no obvious effect on size distribution of the nucleating particles.Ca segregation was proved on Al4C3 particles,which should reduce the interfacial energy of nuclei/Mg.The constitutional undercooling in front of nucleus/liquid was increased from 0.12℃to 0.15℃induced by 0.2%Ca addition.The synergistic grain refining efficiency can be attributed to the higher constitutional undercooling and lower the interface energy of nucleus/Mg induced by Ca addition.More nucleating particles with small size could be activated acting as potent nuclei ofα-Mg grains.Consequently,Mg-3%Al alloy could be effectively refined due to the synergistic effect induced by carbon inoculation combining with Ca.

Early inoculation with caecal fermentation broth alters small intestine morphology,gene expression of tight junction proteins in the ileum,and the caecal metabolomic profiling of broilers

Background:The establishment of stable microbiota in early life is beneficial to the individual.Changes in the intestinal environment during early life play a crucial role in modulating the gut microbiota.Therefore,early intervention to change the intestinal environment can be regarded as a new regulation strategy for the growth and health of poultry.However,the effects of intestinal environmental changes on host physiology and metabolism are rarely reported.This study was conducted to investigate the effects of early inoculation with caecal fermentation broth on small intestine morphology,gene expression of tight junction proteins in the ileum,and cecum microbial metabolism of broilers.Results:Our data showed that early inoculation with caecal fermentation broth could improve intestine morphology.The small intestine villus height was significantly increased(P<0.05)in the intervened broilers compared to the control group,especially on day 28.A similar result was observed in the ratio of villus height to crypt depth(P<0.05).Meanwhile,we found early inoculation significantly increased(P<0.05)the expression levels of zonula occludens-1(ZO1)on days 14 and 28,claudin-1(CLDN1)on day 28,whereas the gene expression of claudin-2(CLDN2)was significantly decreased(P<0.05)on days 14 and 28.Gas chromatography time-of-flight/mass spectrometry(GC-TOF/MS)technology was further implemented to systematically evaluate the microbial metabolite profiles.Principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA)displayed a distinct trend towards separation between the fermentation broth group(F group)and the control group(C group).The differentially expressed metabolites were identified,and they were mainly functionally enriched in beta-alanine metabolism and biosynthesis of unsaturated fatty acids.In addition,1,3-diaminopropane was selected as a key biomarker that responded to early inoculation with caecal fermentation broth.Conclusions:These results provide insight into intestinal metabolomics and confirm that early inoculation with caecal fermentation broth can be used as a potential strategy to improve intestinal health of broilers.

Effect of Inoculation with Effective Microorganisms and Leachate Recycle on Degradation of Municipal Refuse

Biodegradation of waste in landfill is a slow process requiring decades for completion. Accelerated degradation of municipal refuse in modulated landfill environments may alleviate or eliminate pollution to the land, water and air. In this work, nineteen effective microorganisms (EMs) were isolated from old landfill refuse by enrichment culturing techniques and used for the inoculum of municipal refuse. The preliminary experiments demonstrate that a combination of EMs inoculation in landfill with leachate recycle resulted in increased rates of decomposition and faster process stability. The concentrations of COD, VFA and SO4^2- in digester with EMs inoculation and leachate recycle decreased more rapidly than others. Gas production from digester with EMs inoculation and leachate recycle commenced around 32 days, which is a week shorter than with leachate recycle only. And peak cumulative gas production was obtained much earlier in digester with EMs inoculation and leachate recycle (150 days) compared to 180 days with leachate recycle only. Moreover, in the first two months, the rate of settlement in digester with EMs inoculation and leachate recycle was more rapid than others.

Seedling Petri-dish inoculation method:A robust,easy-to-use and reliable assay for studying plant-Ralstonia solanacearum interactions

Ralstonia solanacearum causes a lethal bacterial wilt disease in many crops,leading to huge losses in crop production every year.Understanding of plant-R.solanacearum interactions will aid to develop efficient strategies to control the disease.As a soilborne pathogen,R.solanacearum naturally infects plants via roots.A huge limitation in studying plant-R.solanacearum interactions is the large variation of R.solanacearum infection assay due to the variable soil conditions and uneven inoculum exposure.Here,we developed a robust and reliable Petri-dish inoculation method which allows consistent and stable infection in young plant seedlings.This method is easy to use,takes about only 10 days from seed germination to the completion of inoculation assay,and requires less inoculum of bacteria as well as growth chamber space.We proved the efficacy of the seedling Petri-dish inoculation method by analyzing plant defense primed by molecular patterns,resistance of defense-related plant mutants,and virulence of R.solanacearum mutants.Furthermore,we demonstrated that the seedling Petri-dish inoculation method can be applied to other host plants such as tobacco and has great potential for high-throughput screening of resistant plant germplasms to bacterial wilt in the future.

Turning, Microbial Inoculation and Use of Eisenia foetida in Household Organic Waste Composting

On site composting of organic household wastes is an economical and environmentally tiiendly way to manage municipal wastes. In this manuscript authors evaluated the importance of turning the wastes and of inoculating microorganisms and worms in order to unprove the composting process at domestic scale. Four treatments （Control without turning- C-, Control with Turning- CT- inoculated Mountain Microorganism with turning- MM- and Worms without turning- W-） were tested in a random experiment with tour replicates. Sixteen composting bins were fed with 300 kg of organic wastes from a local street market. The process of compostnig lasted 13 weeks after which all the composts were sifted and submitted to a range of chemical, physical and biological analysis. According to the results MM slightly increased the initial temperature and enzymatic activiry. This investigation outcome don＇t provide of sufficient grounds for a precise recommendation about worms inoculation in compost bins. A decrease between 75% and 80% in the fresh weight of the organic waste was found in the first thirteen weeks after starting the composting process in all the treatments. The quality level of the compost was acceptable, with very low heavy metal content. Turn over helps to keep the product hygienic especially after the inoculation with MM microbial starts. In conclusion and according to the results of this investigation, the use of 320 L compost bins for the organic waste management is strongly advisable.

Research of Synthetic Rules with Mechanical Properties vs Inoculation Degree on Gray Cast Iron

To fit the special needs of gray cast iron as a new engineering material,i.e.,higher σ b and lower HB,or lower σ b and higher HB,using the method of heat analyzing and polyparameters analytic figures,the synthetic rules of carbon saturation degree,inoculation degree and mechanical properties can be exposed.The result shows that,according to the needs of the part′s mechanical properties,referring to the synthetic rules of gray cast iron,the corresponding technological measure and composition proportion can be defined,by which we will get the exact mechanical properties that could achieve the result expected completely.Thus,the casting for gray cast iron might be 'designed'from'selected'.

Indoor Inoculation and Identification Technology of Sweet (Hot) Pepper Virus

[Objective] The paper was to establish an affordable indoor virus inoculation and identification technology. [Method] Taking Tobacco Mosaic Virus （TMV） and Cucumber Mosaic Vires （CMV） of sweet （hot） pepper as the sources of virus, an affordable indoor virus inoculation and identification technology was developed in the paper. [ Result] The suitable inoculation concentration of CMV was five to ten times, and the best seedling age for inoculation was five to six leav- es. Suitable inoculation concentration of TMV was 20 to 30 times, and the best seedling age for inoculation was three to six leaves. Single inoculation technology was mainly used for indoor virus inoculation and identification of sweet （hot） pepper, and complex inoculation technology could also be adopted with first, st inoculation of CMV and late inoculation of TMV. For mixed inoculation technology, CMV： TMV should be 1： 1. Complex inoculation and mix inoculation should base on the tech- nology of single inoculation. Disease resistant materials, AID1-W22-dg176, ABgl-W22-48123, AB91-DL-6428, HY031-2-8-1-6, BYT-4-1-3-6-8, JFG-2-1-2-6, JF8S-1-1-5-4-8 and I＇502-1-1-3-5, were identified by this method. [ Conclusion] This research provided scientific basis for standardization of indoor inoculation of sweet （hot） pepper virus.

Preparation and Application on the Inactivated Bacterins of Yellow and White Scour of Piglets

Pathogenic Escherichia coli strains were isolated from yellow and white scour samples in pig farms. The strains were used to make autologous inactivated bacterins,and the bacterins were applied to multiple infected farms. The results showed that in the group of inactivated bacterins,the infected piglet number was less,the course of disease was shorter and the symptoms was milder. The piglet E. coli inactivated bacterins could significantly reduced morbidity of yellow and white scour of piglets,the preventive effects of E. coli autologous inactivated bacterins against yellow and white scour of piglets were much better than that of genetically engineered vaccines.

Mycorrhizal Inoculation Effect on the Forage Cowpea Biomass Production in Burkina Faso

In Burkina Faso, breeding occupies an important place in the country’s economy, but is based essentially on the exploitation of the natural resources (pasture) which remains limited in extent due to the extension of land cultivated by agricultural practices extensive. Food is one of the most important limiting factors in animal production, hence the need to develop mechanisms to improve the production of food resources. This study is a contribution to fodder production improvement. In this study, cowpea was grown in a greenhouse for 60 days and inoculated with two inocula of arbuscular mycorrhizal fungi. The growth parameters were measured at 30 and 60 days after sowing. Shoot, root and total biomass were evaluated 60 days after sowing. The results show Yac 2 mix inoculum improves height growth (95.5%), height relative growth rate (525%), collar diameter (138%), collar diameter relative growth rate (328.57%), shoot biomass (396.3%), root biomass by (205.66%), total biomass by (320%), total nitrogen of aerial parts (92.39%) and total phosphorus of aerial parts (143.48%) compared to control. Yac 2 mix appears the most effective of all inoculum used in this study.

Phosphorus Fertilizer Rating and Rhizobia Inoculation for Improved Productivity of Cowpea in Northern Uganda

Cowpea (Vigna unguiculata) is an important legume crop in the tropics and subtropical regions of the world. It is mainly grown for its leaves and grains, and to a lesser extent as a fodder crop. Cowpea is considered as the most important food grain legume in the dry savannas of tropical Africa. This study compared the yield of local (Agondire) and improved (SECOW 2W) cowpea varieties grown on an Oxisol. Inorganic P at levels of 0, 10, 20, and 40 kg·ha-1 was tested on each variety with or without rhizobia inoculation. The experiments were set up in a randomized complete block design and replicated thrice during the short and long rains of the 2015/2016 seasons on fifteen fields in Arua district, northern Uganda. Agondire responded significantly (P < 0.05) better than SECOW 2W when high rates of inorganic phosphorus (40 kg P ha-1) were applied. A significant increase of 26.4% and 28.4% in grain yield of Agondire and SECOW 2W, respectively was obtained after inoculation with rhizobia. We concluded that inoculation and P fertilizer application increased the yield of both varieties, but with inoculation, SECOW 2W performs much better at lower P fertilizer rates than Agondire. Therefore, we recommend growing of SECOW 2W under inoculation with 20 kg P ha-1 and an application of 40 kg P ha-1 for Agondire local cowpea variety in northern Uganda.

Influence of different inoculation methods on the evaluation of the resistance to rice sheath blight(Rhizoctonia solani Kuhn)

To establish a standard system for geneticstudies on sheath blight resistance, a field testwas conducted at the experimental farm ofYangzhou University to compare several pro-cedures for inoculating rice plants with R.solani Kuhn (RH9). The varieties used wereJasmine 85, Teqing (resistant or moderatelyresistant), and Lemont (susceptible). They

In-vitro vs in-vivo Inoculation: Screening for Resistance of Australian Rice Genotypes Against Blast Fungus

To assist with rapid screening for rice blast resistance as a precursor in a breeding program, the susceptibility to rice blast of 13 rice genotypes from Australia was evaluated in May to June 2013 using three distinct inoculation methods（spot, filter paper and standard methods） at seedling, vegetative and reproductive stages. The results revealed that the spot and filter paper inoculation methods were successful in discerning susceptibility to the rice blast disease（P ≤ 0.05）. Disease susceptibility declined significantly from the vegetative to reproductive stages. The standard method was conducted at three different stages for pot plants grown inside the mist house. However, low temperatures did not produce disease symptoms except in a few genotypes. Among the 13 rice genotypes screened, AAT9 expressed a highly resistant response, and AAT4, AAT6, AAT10, AAT11, AAT13, AAT17 and AAT18 expressed resistance at various stages. The results will be useful for selecting elite genotypes for disease tolerance where rice blast is prevalent. In addition, the resistant genotypes can serve as a gene pool used in breeding programmes to develop new resistant genotypes.

Drought adaptation of Bauhinia faberi var.Microphylla seedlings with dual inoculation of arbuscular mycorrhizal fungi

Bauhinia faberi via.Microphylla(BFM)is an important tree species for vegetation restoration in the dry valley of southwestern China.However,there were few studies on the application of arbuscular mycorrhizal fungi(AMF)in improving the drought adaptation of BFM.In order to investigate the response of BFM to water stress(WS),we tested four inoculation treatments((no AMF,Control),Glomus mosseae(GM),Glomus intraradices(GI),Glomus mosseae+Glomus intraradices(GMI))in pots,experimented under three field water holding capacity(WHC)of 70%,50%and 30%.The changes of seedling survival rate(SR),AMF relative root length colonization rate(Col),growth,photosynthetic parameters,water status and leaf nutrients were examined.The results showed that under 30%WHC drought conditions,SR with dual inoculation of AMF was not higher than with single inoculation of GM,suggesting that increasing the diversity of AMF did not definitely improve plant SR,and that the species of inoculated AMF might have an important impact on SR.The sensitivity of dual inoculated Col to water stress was lower than that of single inoculation,which was more favorable for dual inoculated BFM seedlings adapting to drought environment.The overall drought resistance ability(D)also showed that dual inoculation of AMF improved plant drought adaptation compared with single inoculation,which was related to the higher Col of dual inoculated AMF.This study is of practical importance to promote vegetation restoration in arid areas in a cost-effective and environmentally friendly manner.

Isolation,identification and artificial inoculation of Ustilago esculenta on Zizania latifolia

The smut Ustilago esculenta is an endophytic fungus that invades Zizania latifolia(Jiaobai)and plays an important role in inducing the host stem to form a swollen gall.The aims of this study were to investigate isolation,identification and inoculation methods of U.esculenta from Jiaobai.The spore suspension culturing method was the best method of isolating U.esculenta,and yeast-like fungal strains and septate hyphal strains were obtained.The universal primers ITS1 and ITS4 amplified a ribosomal DNA spacer element from all tested strains,while specific primers for Lam16A gene generated a 442 bp product only with septate hyphae strains.De novo transcriptome assembly results showed that the yeast-like fungal strains did not match Ustilago,and genomic analysis suggested the septate hyphae strains were highly similar to known smut fungi.Therefore,the septate hyphae strains were identified as U.esculenta,which indicated that the ITS primers were not species-specific but that Lam16A-specific primers can distinguish U.esculenta accurately and easily.In vitro,U.esculenta grew quickly on PDA medium at 28°C in darkness.Furthermore,an inoculation method was established to produce swollen stems by injecting a spore suspension of U.esculenta at OD600=2.5 into the basal culm internode of male Jiaobai.

A STUDY OF THE FORMATION OF LIVER METASTASIS AND ITS MECHANISM USING THE INTRASPLENIC INOCULATION OF CANCER CELLS

In order to establish an animal model with hepatic metastasis intrasplenic inoculation of carcinoma cells from murine uterine cervical carcinoma (U14) was employed. Results showed a high incidence of hepatic metastasis could be obtained through the intrasplenic inoculation of 1 × 106 carcinoma cells. Removal of the primary carcinoma through splenec-tomy at different intervals after intrasplenic inoculation proved that the hepatic metastatic mechanism was not due to mechanical pressure but occurred spontaneously. This experimental model provides a useful means for studying the mechanism and prevention of hepatic metastasis.