Titers of Anti-Brucella Antibodies by Enzyme Linked Immunosorbant Assay in Vaccinated and Unvaccinated Brucellosis Infected Cattle

Brucellosis is an important re-emerging zoonotic disease caused by Brucella organisms. In the absence of a Differentiation of Infected from Vaccinated Animal (DIVA) assay for bovine Brucellosis, it becomes difficult to assess whether the anti-Brucella antibody response in an animal is due to vaccination or infection. We compared the anti-Brucella antibody titers of naturally Brucellosis affected unvaccinated cows, previously vaccinated infected cows, normal healthy vaccinated cows and healthy unvaccinated calves. The titers of anti-Brucella antibodies were estimated by indirect ELISA. The mean titer (log10) was found to be 1.518 ± 0.005 in case of naturally Brucellosis affected cattle which had been vaccinated during calf hood. The mean titer in case of naturally infected cattle which had never been vaccinated was 1.5441 ± 0.005. The mean titer in healthy unaffected cattle vaccinated during calf hood was 1.504 ± 0.002 and that of unvaccinated healthy calves was 0.560 ± 0.016. It was interesting to find that the antibody titers in naturally affected cattle which had never been vaccinated were very significantly (p < 0.01) higher than those of Brucellosis affected cows which had been vaccinated during calf hood. The titer in vaccinated infected cattle was very significantly (p < 0.01) higher than that of uninfected vaccinated cows.

Analysis of sex pheromone production and field trapping of the Asian corn borer(Ostrinia furnacalis Guenée)in Xinjiang,China

Identifying the sex pheromone systems of local pest populations facilitates their management,especially for moth species that show significant geographic variation in sex pheromone communication.We investigated the pheromone production and behavioral responses of the Asian corn borer(Ostrinia furnacalis Guenée;ACB)in Xinjiang,China.The ACB produces three compounds:(Z)-12-tetradecenyl acetate(Z12-14:Ac)and(E)-12-tetradecenyl acetate(E12-14:Ac)which are two sex pheromone compounds,and n-tetradecyl acetate(14:Ac)which has variable roles in mediating behavioral responses.The ratios of these three compounds produced in female gland are geographically distinct among different populations.Quantitative analysis of pheromone production showed that the proportions of Z12-14:Ac in the E/Z isomers(i.e.,Z and E12-14:Ac)and the proportions of 14:Ac in the ternary blend respectively averaged 60.46%(SD=5.26)and 25.00%(SD=7.37),with their probabilities normally or near-normally distributed.Trapping experiments in a cornfield indicated that deploying the E/Z isomers and the three compounds in rubber septa close to their gland ratios yielded the most captured males,while other ratios that deviated from the gland ratios showed reduced field captures.The ternary blend was significantly more attractive to males than the E/Z isomers in the field,indicating a functional role of 14:Ac as the third pheromone component used by the local population.Additionally,the dose-response test demonstrated that the application of the three compounds at dosages between 200 and 350μg attracted significantly more males compared to other dosages.Therefore,the characterization of this local ACB pheromone system provides additional information about its geographic variation and serves as a basis for optimizing the pheromone-mediated control of this pest in Xinjiang.

Phylogenetic and epidemiological characteristics of H9N2 avian influenza viruses in Shandong Province, China from 2019 to 2021

H9N2 avian influenza virus(AIV) has widely circulated in poultry worldwide and sporadic infections in humans and mammals. During our surveillance of chicken from 2019 to 2021 in Shandong Province, China, we isolated 11 H9N2AIVs. Phylogenetic analyses showed that the eight gene segments of the 11 isolates were closely related to several sublineages of Eurasian lineage: BJ/94-like clades(HA and NA genes), G1-like clades(PB2 and M genes), and SH/F/98-like clades(PB1, PA, NP and NS genes). The isolates showed mutation sites that preferentially bind to humanlike receptors(HA) and mammalian fitness sites(PB2, PB1 and PA), as well as mutations in antigen and drug resistance sites. Moreover, studies with mice revealed four isolates with varying levels of pathogenicity. The average antibody titer of the H9N2 AIVs was 8.60 log2. Based on our results, the epidemiological surveillance of H9N2 AIVs should be strengthened.

Effects of Three Kinds of Chinese Medicine Polysaccharides on the Immune Effect of Newcastle Disease Vaccine

240 14-day-old healthy and non-immune Roman chicken were randomly divided into 8 groups, including blank control group （BC group）, immune control group （IC group）, and immunity groups of each Chinese medicine. On the day of the first immunity, 3 d before the second immunity, the day of the second immunity and 3 d after the second immunity, high-dose concentration and low-dose concen- tration of Astragalus polysaccharide （ASP）, Epimedium polysaccharide （EPP） and Isatis roots polysaccharide （IRPS） were used for the immunity groups of each Chi- nese medicine using the gavage, and 0.5 ml for each chick, and the equivalent normal saline was used for the blank control group and vaccine control group. On the 7^th, 14^th, 21^st and 28^th day after the first immunity, 10 chicken of each group were randomly got and weighed, and the antibody titer and the changes of the pro- liferation of T lymphocyte were measured. The results showed that 3 kinds of Chi- nese medicine polysaccharides all can increase the weight of chicken, improve HI antibody titer of Newcastle disease, and promote the proliferation of peripheral T lymphocyte, in which the effect of IRPS at low dosage is the best.

Precore/basal core promoter mutants quantification throughout phases of hepatitis B virus infection by Simpleprobe

AIM:To investigate precore/basal core promoter(PC/BCP) mutants throughout hepatitis B virus(HBV) infection and to determine their relationship to hepatitis B early antigen(HBeA g) titers.METHODS:We enrolled 191 patients in various stages of HBV infection at the Huashan Hospital and the Taizhou Municipal Hospital from 2010 to 2012.None of the patients received antiviral therapy.HBV DNA from serum,was quantified by real-time PCR.The HBV genotype was determined by direct sequencing of the S gene.We used the Simpleprobe ultrasensitivequantitative method to detect PC/BCP mutants in each patient.We compared the strain number,percentage,and the changes in PC/BCP mutants in different phases,and analyzed the relationship between PC/BCP mutants and HBe Ag by multiple linear regression and logistic regression.RESULTS:Patients with HBV infection(n = 191) were assigned to groups by phase:Immune tolerance(IT) = 55,Immune clearance(IC) = 67,Low-replicative(LR) = 49,and HBeA g-negative hepatitis(ENH) = 20.Of the patients(male,112; female,79) enrolled,122 were HBe Ag-positive and 69 were HBe Ag-negative.The median age was 33 years(range:18-78 years).PC and BCP mutation detection rates were 84.82%(162/191) and 96.86%(185/191),respectively.In five HBe Ag-negative cases,we detected double mutation G1896A/G1899 A.The logarithm value of PC mutant quantities(log10 PC) significantly differed in IT,IC,and LR phases,as well as in the ENH phase(F = 49.350,P < 0.001).The logarithm value of BCP mutant quantities(log10 BCP) also differed during the four phases(F = 25.530,P < 0.001).Log10 PC and log10 BCP values were high in the IT and IC phases,decreased in the LR phase,and increased in the ENH phase,although the absolute value at this point remained lower than that in the IT and IC phases.PC mutant quantity per total viral load(PC%) and BCP mutant quantity per total viral load(BCP%) differed between phases(F = 20.040,P < 0.001; F = 10.830,P < 0.001),with PC% and BCP% gradually increasing in successive phases.HBeA g titers negatively correlated with PC%(Spearman's rho =-0.354,P < 0.001) and BCP%(Spearman's rho =-0.395,P < 0.001).The negative correlation between PC% and HBeA g status was significant(B =-5.281,P = 0.001),but there was no such correlation between BCP% and HBeA g status(B =-0.523,P = 0.552).CONCLUSION:PC/BCP mutants become predominant in a dynamic and continuous process.Log10 PC,log10 BCP,PC% and BCP% might be combined to evaluate disease progression.PC% determines HBeA g status.

Effect of Se-enriched Lactobacillus on Antibody of NDV of Broilers

This study was conducted to investigate the effect of Se-enriched lactobacillus on antibody of Newcastle Disease Virus(NDV)of broilers.One thousand day-old AA broiler chickens were selected from a chicken farm of Jinghai County,and the chickens were divided randomly into two groups,which respectively had five repeat groups with 100 chickens each,one named basal diet group,and the other named Se-enriched lactobacillus group.Corn soybean meal diet was selected for basal diet group,while 2%se-enriched lactobacillus was added into corn soybean meal diet for the other group.The method of feeding was free diet,drinking,and routine immunization program was adopted.This test period was 42 d.Two chickens were selected randomly in each repeat for slaughtering and collecting serums on the 14th,28th and 42nd d.Assessments on the effect of Se-enriched lactobacillus in antibody titer of NDV of broilers were made by hemagglutination-inhibition test.The results indicated that antibody titer of NDV was increased significantly by Se-enriched lactobacillus.

Lost Immune Markers for Major Affective Disorders: Could They Help Screen Airline Pilots?

In the 1980s it was discovered that quantitative IgG titers for antibodies directed against the hu-man-obligate DNA viruses were reliably and reproducibly correlated negatively with mood in in-dividuals with major affective disorders. This knowledge was lost to confirmation bias and uncontrolled studies that only looked at Epstein-Barr virus antibodies. The latter led to a long American fad of misdiagnosing major affective disorders as a “chronic Epstein-Barr virus syndrome.” This misdiagnosis even became part of the American popular culture during the 1980s. In light of fatal airline crashes intentionally caused by pilots, the antiviral IgG titers should be revisited as a means of screening pilots for current dysphoria or mania/hypomania due to major affective disorders.

Antibacterial activities of selected medicinal plants in traditional treatment of human wounds in Ethiopia

Objective:To evaluate the activity of selected Ethiopian medicinal plants traditionally used for wound treatment against wound-causing bacteria.Methods:Samples of medicinal plants(Achyranthes aspera,Brucea antidysenteriea,Datura stramonium,Croton macrostachyus,Acokanthera xchimperi.,Phytolacca dodecandra,Milhttia ferruginea,and Solanum incanum)were extracted using absolute methanol and water and tested for their antimicrobial activities against clinical isolates and standard strains of wound-causing bacteria using agar well diffusion and micro titer plate methods.Results:Most of the plant extracts had antibacterial activities,among which Acokanthera schimperi and Brucea antidysenteriea inhibited growth of 100%and 35%of the test organisms,respectively.Methanolic extracts had higher activities compared with their corresponding aqueous extracts.The most susceptible organism to the extracts was Streptococcus pyogens while the most resistant were Escherichia coli and Proteus vulgaris.Conclusions:This finding justifies the use of the plants in wound healing and their potential activity against woundcausing bacteria.Their toxicity level and antimicrobial activity with different extraction solvents should further be studied to use them as sources and templates for the synthesis of drugs to control wound and other disease-causing bacteria.

Evaluation of thermotherapy against Huanglongbing (citrus greening) in the greenhouse

Huanglongbing （HLB, or citrus greening） is the most destructive disease of citrus, which is associated with Candidatus Liberibacter asiaticus （Las）. Few management options are available, aside from preventive measures such as removing infected plants, planting disease-free seedlings, and managing the insect vector. In this study, we assessed the efficacy of thermotherapy against HLB under controlled greenhouse conditions. A total of 60 two-year-old, graft-infected Citrus reticulata Blanco plants were used. The plants were randomly divided into three groups （45℃, 48℃, and untreated control）, with five plants/replicate （rep） and four reps/treatment. The treated plants were placed in phytotrons for a 4-h treatment session, repeated once per week for three consecutive weeks. Disease remission was observed eight weeks post-treatment. Real-time PCR assays revealed that Las titers in HLB-affected seedlings were significantly reduced in both 45 and 48℃ treatments four weeks after treatment, with the exception of eight plants. In contrast, Las titers in the untreated control plants increased significantly during the same period, with a maximum increase of 28-fold. Except for seven plants, Las titers in the new flushes of treated plants decreased more than 90% eight weeks after treatment. Las titers in mature leaves of treated plants decreased 56 and 60% in average at 45 and 48℃, respectively, eight weeks after treatment. The HLB symptoms and Las titer of seedings were markedly alleviated eight weeks after treatment in both 45 and 48℃ treatments. Our results laid a good foundation for the further development of citrus free-disease seedling cultivation and Huanglongbing control in the field. The whole plants were replaced for scion or branch in previous as the research object in this study, and the expression of Huanglongbing symptoms combined with real-time polymerase chain reaction （PCR） were used to evaluate the effect of heat treatment in the greenhouse.

Comparison of RFFIT Tests with Different Standard Sera and Testing Procedures

The World Health Organization （WHO） standard assay for determining antibody level is the rapid fluorescent focus inhibition test （RFFIT） and is used to determine the degree of immunity after vaccination against rabies. To compare the difference in RFFIT results between the laboratories of The National Institute of Infectious Disease in Japan （NIID） and the Chinese Centre for Disease Control （CCDC） as well the influence of the choice of standard serum （STD） for the detection, the two laboratories detection methods were simultaneously manipulated by RFFIT. The reference serums used in NIID and the WHO standard serum used in CCDC were compared in the same RFFIT detection to determine the titer of four sera samples C1, Sl, S2 and S4 in parallel, and the titers of the detected sera samples were calculated using the standard formula for neutralizing antibody titer. No significant difference was found in RFFIT methods from the two laboratories and the RFFIT testing procedures of the two laboratories have good consistency. However, different titers were obtained with the tentative internal standard serum （TI-STD） produced by adjusting to 2.0 IU of WHO standard serum in NIID and the WHO STD. The titer determined with the TI-STD was higher than that determined with WHO STD, This difference appears to be significant and requires further investigation

PRESENT STATUS OF RESEARCH ABROAD CONCERNING THE EFFECT OF ACUPUNCTURE AND MOXIBUSTION ON IMMUNOLOGIC FUNCTIONS

Much research has been conductedabroad in recent years concerningacupuncture and moxibustion on theimmunologic functions of the organism.Anoutline of this is presented as follows.EFFECTS OF ACUPUNCTUREAND MOXIBUSTION ON NORMALIMMUNOLOGIC FUNCTIONS

Disease Progression Patterns of SHIV-KB9 in Rhesus Macaques of Chinese Origin in Comparison with Indian Macaques

Objective To develop a model of SHIV-KB9/Chinese origin rhesus （Ch Rh） macaques for vaccine research and to compare the pathogenesis of SHIV-KB9 in Ch Rh macaques with that reported in Indian rhesus （Ind Rh） macaques. Methods Seven mamu-A＊01 negative Ch Rh macaques were inoculated intravenously with 1-10000 MID50 of SHIV-KB9. The monkeys were monitored for viral load, CD4, CDS, SHIV-specific antibody and virus genetic variation. The results were compared with those previously observed in Ind Rh macaques. Results As compared to that observed in Ind Rh macaques, SHIV-KB9 in Ch Rh macaques displayed three identical disease progression patterns. However, the primary pattern was not identical between the two subspecies. The level of plasma viremia differed in SHIV-KB9-infected Ch Rh macaques which exhibited different outcomes from those in Ind Rh macaques. Generally, the values of viral load and the maintenance of CD4^＋ T cells were associated with humoral responses. Otherwise, the viral genetic distances （divergence, diversity） were larger in animals （M419, M425） with their CD4^＋ T cells profoundly depleted. Conclusion The model of SHIV-KB9/Ch Rh macaques displays a relatively slow progression to AIDS compared with Ind Rh macaques, which may more accurately reflect the potential of candidate vaccines in humans.

Effects of Chinese Medicinal Herb on Growth and Immunization of Laying Chicks

In order to evaluate effects of Chinese medicinal herb (CMH) on growth and immunization of laying male chicks,eleven CMHs were used in this study,which are Astragalus membranacens (AM),Schisandra chinensis (SC),Ligustrum Lucidum(LL),Codonopsis,Scutellaria baicalensis (SB),Atractylodes macrocephala(AMA),Haw,Ginger,Acanthopanax scenticosns (AS),Angelica and Lycium,added to basal diet as 1%,respectively,with the basal diet supplemented with 50 mg·kg^(-1) bacitracin zinc(BZ) as the control.The body weight (BW) of birds were recorded at 1,21 and 42 days after birth,individually.The birds were vaccinated 0.50 mL against Newcastle disease (ND) with La sota vaccinel (containing mineral oil as adjuvant) by i.m.inoculation at 21 d of age.All of birds was vaccinated with F48E9 NDV by i.m.inoculation at 49 day of age.Blood samples were taken via wing vein from each bird on the day receiving the vaccination (Day-1) and on 14,21,28,30 and 34 d after vaccination (Days 14,21,28,30 and 34). Results showed that bodyweight (BW) and bodyweight gain(BWG) of the trial groups are similar to the control group.No significant differences of relative weight (RW) of bursa and spleen were observed among trial groups except for AS (P<0.05).Serum antibody titers of SC,LL,Codonopsis,SB,AMA and Lycium groups were significantly increased (P<0.05) in contrast to these control group on day-21 or 28 after immunized ND La sota vaccinel,which suggested that SC,LL,Codonopsis,SB,AMA and Lycium could augment antibody formulation.Furthermore,Compared with the control,antibody titers in SC,Codonopsis,AS,Lycium,SB and AMA group were higher (P<0.05) after vaccinated with F48E9 NDV,which suggested that SC,Codonopsis,AS,Lycium,SB and AMA have activity of antivirus.In conclusion,CMH used in the present study have similar effect on BW and BWG of chicks compared with the control.Moreover,these eleven CMH have little influence on RW of immunized organ except AS.However,SC,Codonopsis,LL,Lycium,SB and AMA could augment antibody formulation.Furthermore,SC,Codonopsis,AS,Lycium,SB and AMA have activity of antivirus compared with the control.

St udy on Hsp90 Expression in Different Tissues and Its Antibody in Serum of Chickens Infected with Marek's Diseases

To investigate the dynamic change of heat shock protein 90 （Hsp90） in the genesis and development of tumor, we successfully established tumor animal model using Marek’s disease and then determined the location of Hsp90 in the tumor tissue using immunohistochemistry method, the antibody titer level of Hsp90 in the serum and the expression level in the tissue using enzyme-linked immunosorbent assay （ELISA） method. Our result showed that Hsp90 location in the tumor tissue was signiifcantly associated with the tumor cell and most in the cytoplasm of the tumor cell, and Hsp90 expression level in the tissue and the antibody titer level in the serum was most signiifcantly increased with the development of tumor. This is the ifrst report to show the presence of Hsp90 in tumor tissues induced by the Marek’s disease, with its expression correlated to the tumoral grading. These data may also be valuable for developing new molecular anti-cancer therapies.

Distribution pattern and titer of Candidatus Liberibacter asiaticus in periwinkle(Catharanthus roseus)

Candidatus Liberibacter asiaticus （CaLas）, an uncultured Gram-negative alphaproteobacterium, is the causal agent of Huanglongbing （HLB） in citrus. CaLas resides in phloem sieve tubes and has been shown to be unequally distributed in different tissues. Although HLB is a disease of citrus plants, it has been demonstrated that periwinkle can serve as an experimental host of CaLas, which can be transmitted from citrus to periwinkle via the parasitic plant dodder （Cuscuta spp.）. To investigate the distribution of CaLas in various periwinkle tissues, the bacteria were transmitted from an infected periwinkle plant to healthy periwinkles by top-grafting. The movement of the inoculum and associated titer changes were observed over time in various tissues. CaLas could be detected in the leaves, main stems, and roots of infected periwinkle by conventional PCR, and in all three tissues a clear time-dependent change in CaLas titer was observed, with titer increasing soon after inoculation and then decreasing as disease symptoms became severe. The highest titer was found at 25, 35 and 35 days after inoculation in leaves, main stems and roots, respectively. The titer in leaves was much higher than in the main stems and roots at the same time point, and the spatial distribution of CaLas in the leaves, main stems and roots of infected periwinkle was uneven, similar to what has been shown in citrus. The results provide guidance for selecting the proper periwinkle tissues and sampling times for early detection of CaLas.

Mortality rates from a Nigerian isolate of the <i>Infectious Bursa Disease Virus</i>and passive haemagglutination antibody titer that protects chicks against challenge with the virus isolate

To determine passive haemagglutination (PHA) antibody titer that would protect chicks against Nigerian isolates of the Infectious Bursa Disease Virus (IBDV), five groups of chicks aged 30 days which had different antibody titers were challenged with a Nigerian isolate of virulent IBDV. Mortality rates of the different groups were plotted against their respective mean PHA antibody titers. A group with zero antibody titer had a mortality rate of 75% while those with PHA antibody titers of 185.6, 243.2, 256 and 307.2 had mortality rates of 40%, zero, zero and zero respectively. Linear equation generated for a line of best fit of the graph of mortality rates of the chicks on their IBD antibody titers gave antibody titer (X) at which mortality (Y) would be zero as 300. A mortality of 75% and the high antibody level needed to protect chicks suggest that the isolate may be a hypervirulent strain.

Enhancement of the Immune Response to Rabbit Hemorrhagic Disease Vaccine in Young Rabbits by Advanced Vaccination and Chinese Herbal Adjuvants

Experiments were conducted to determine the effects of advanced vaccination and Chinese herbal adjuvants （CHA）, containing astragalus polysaccharides （APS） and ginsenosides （GS） on the immune response to rabbit hemorrhagic disease （RHD） vaccine in young rabbits. In experiment 1, 5 New Zealand rabbits of each group at 30, 35, 40, or 45 days of age were injected with 2 mL of inactivated RHD vaccine, respectively. The dynamic changes of antibody titers were tested by the hemagglutination inhibition （HI） method. In experiment 2, 30 New Zealand rabbits at 35 days of age were randomly assigned to 5 treatment groups, representing inoculation with 3 mL of non-adjuvant RHD vaccine, CHA-RHD vaccine, CHA-HA vaccine （half dose antigen）, aluminium adjuvant-RHD vaccine, and PBS, respectively. The dynamic changes of peripheral lymphocyte proliferation and serum antibody titers were tested by the MTT method and the HI method. The results showed that the titer of maternal HI antibody in the 35-day-old rabbits was lower than the protective level of 3 log2, while on days 7 to 49 after the vaccination, the antibody titers were higher than 3 log2. The CHA promoted the lymphocyte proliferation and enhanced the serum antibody titer （P〈 0.05）. These findings from the two experiments suggested that advanced vaccination and Chinese herbal adjuvants significantly enhanced the immune response to vaccine against RHD, and effectively protected the young rabbits against RHD challenge.

Chemometric identification of canonical metabolites linking critical process parameters to monoclonal antibody production during bioprocess development

A deeper understanding of the biological events occurring when bioprocess parameters changed will be of great value in improving the monoclonal antibodies (mAbs) production. Design of experiment (DoE) was applied to investigate the effect of process parameters (pH, temperature shift and dissolve oxygen (DO)) on protein titer. The key metabolites connecting the critical process parameters (CPPs) with monoclonal antibody production were identified by different chemometrics tools. Finally, the biological events of marker metabolites relating with titer improvement were concluded. pH and temperature shift were identified as CPPs that affect the target protein titer. A series of metabolites influenced by the altered CPPs and correlated with protein titer were screened by principal component analysis (PCA) and Pearson' correlation test. The marker metabolites and their pathways linking CPPs to target protein titer in different culture phases were summarized. Metabolomics and chemometrics are promising data-driven tools to shine light into the biological black box between the bioprocess parameters and process performance.

Protective effect of neurotrophin-3 secreting myoblasts against kainic acid-induced exitotoxic lesions in rat hippocampus

Neurotrophic factors are able to promote neuronalsurvival and prevent neuronal death induced by variousinjuries and have potential uses in the treatment ofneuronal degenerative diseases and neuropathy. Due tothe existence of brain blood barrier, neurotrophic factorscan not be ased in CNS diseases directly. Gene therapy isa good candidate to overcome this problem.

Preliminary study of retroviral mediated transfer of multidrug resistance gene into CD34^+ cells originated from human bone marrow

To elucidate the feasibility of retroviral mediatedtransfer of multidrug resistance gene (mdrl) intohematopoietic stem/progenitor cells, the CD34^+ cellswere isolated from bone marrow by using a high-gradient.magnetic cell sorting system, and the efficiency ofretroviral mediated gene transfer was studied.