To investigate the infection patterns of porcine epidemic diarrhea virus (PEDV) in Korean pig farms, a total of 4,768 swine sera samples from 159 pig farms were taken twice, in June (n=82) and October (n=77) in 2007. In each farm selected for the survey, 10 samples from breeding pigs and 4 from each of the 5 age groups (30, 60, 90, 120, and 150 days) were taken, and all serum samples were tested for PEDV by the serum neutralization test. The overall seroprevalence was 62.6% (2,983/4,768), with the highest prevalence in breeding pigs (93.5%, 1,485/1,589). The prevalence showed an increasing trend with increasing age (30.8, 27.2, 44.7, 61.6, and 71.2% respectively in the 30, 60, 90, 120, and 150 days age groups) (p<0.0001 for $x^2$ trend test). The association between age and PEDV prevalence was similar in both surveys, indicating that the infection of PEDV seemed to be occurring repeatedly in the farms surveyed. This inference could also be explained by the fact that prevalence in sows was very high despite low vaccination coverage, as they are continuously exposed to PEDV in potentially infected farms for a longer period. Based on the neutralizing antibody levels in sows and growing pigs, the majority of farms (91.8%, n=146 farms) were endemically infected with PEDV, and most of pigs seemed to be intensively infected with PEDV at around early growth (41.8%) and weaning (31.5%). On the other hand, serum neutralizing antibodies were not detected in pigs older than 30 days of age in farms classified as having no PEDV infection (n=13 farms), indicating the level of maternal antibody against PEDV is decreased on a non-detectable level before the piglet is 60 days old in the field situation. The results indicated that most farms surveyed in 2007 were affected with endemic PEDV infection. Therefore, a national monitoring and control program for the endemic type PEDV infection needs further attention.
Porcine reproductive and respiratory syndrome virus (PRRSV) has plagued pig populations worldwide causing severe economical impacts. In order to establish effective strategies for prevention of PRRS, infection patterns on the herd level are primarily evaluated. In the present study, therefore, serological and virological analyses were conducted in 20 pig farms suffering from PRRS. Seroprevalence levels in each farm were grouped into 3 patterns: SN (Stable sow groups/Not infected piglet groups, SI (Stable sow groups and Infected piglet groups), and UI (Unstable sow groups and Infected piglet groups). The rates of each serological pattern were 15% (n=3), 10% (n=2), and 75% (n=15), respectively. In addition, the pattern analysis was extended to virological monitoring on the same farms that further included suckling pig groups. As a result, the infection pattern was classified into 4 categories: SNI (Stable sow groups/Not infected suckler groups/Infected piglet groups), SII (Stable sow groups/Infected suckler groups/Infected piglet groups), UNI (Unstable sow groups/Not infected suckler groups/Infected piglet groups), and UII (Unstable sow groups/Infected suckler groups/Infected piglet groups). The rates of each viroprevalence were estimated at 50% (n=10), 30% (n=6), 10% (n=2), and 10% (n=2), respectively. PRRSV viroprevalence results of suckling pig groups revealed that 8 farms were considered virus positive. In 2 farms among these farms, PRRSV appeared to be transmitted vertically to suckling piglets from their sows. In contrast, piglet-to-piglet horizontal transmission of PRRSV seemed to occur in sucking herds of the remaining farms. Thus, this virological analysis on suckling piglets will provide useful information to understand PRRSV transmission routes during the suckling period and to improve a PRRS control programs. Our seroprevalence and viroprevalence data found that infection patterns between sow and piglet groups are not always coincident in the same farm. Remarkably, 15 farms belonging to the UI seroprevalence pattern showed four distinct viroprevalence patterns (SNI; 7, SII; 4, UNI; 2 and UII; 2). Among these farms, 11 farms with unstable seroprevalence sow groups were further identified as the stable viroprevalence pattern. These results indicated that despite the absence of typical seroconversion, PRRSV infection was detected in several farms, implying the limitation of serological analysis. Taken together, our data strongly suggests that both seroprevalence and viroprevalence should be determined in parallel so that a PRRS control strategies can be efficiently developed on a farm level.
This study was performed to investigate antigenic expression patterns in the course of HSV-1 infection. In SDS-PAGE analysis, HSV-1 antigens were detected, and among them, antigens in the size of 39, 47, 63, 86, 101, 105, 135, 159, and 181 kDa appear to be expressed in the most dominant forms. BALB/c mice were infected with HSV-1 for 29 days and antigenic expression from HSV-1 was investigated by Western blot analysis using anti-HSV-1 sera collected every two days from BALB/c mice infected with HSV-1. Most of HSV-1 antigens appeared sporadically as the infection progressed. However, antigens in the sizes of 63kDa and 135kDa were expressed from day 1 and 3, respectively, and existed continuously during the course of infection for 29 days, suggesting that they are the most dominant antigens inducing immune response durign HSV-1 infection, and they could be the target antigens for the development of vaccines. The isotype levels of IgA, IgGl, and IgM increased till the 17 th day infection and then started to decrease. During this course. IgGl was the most dominant isotype. In an indirect immunofluorescent assay, antibodies exhibited surface binding to the Vero cell infected with HSV-1, demonstrating that HSV-1 antigens are expressed on the surface of Vero cells.
Trichomonas vaginalis induces proinflammation in cervicovaginal mucosal epithelium. To investigate the signaling pathways in $TNF-{\alpha}$ production in cervical mucosal epithelium after T. vaginalis infection, the phosphorylation of PI3K/AKT and MAPK pathways were evaluated in T. vaginalis-infected SiHa cells in the presence and absence of specific inhibitors. T. vaginalis increased $TNF-{\alpha}$ production in SiHa cells, in a parasite burden-dependent and incubation time-dependent manner. In T. vaginalis-infected SiHa cells, AKT, ERK1/2, p38 MAPK, and JNK were phosphorylated from 1 hr after infection; however, the phosphorylation patterns were different from each other. After pretreatment with inhibitors of the PI3K/AKT and MAPK pathways, $TNF-{\alpha}$ production was significantly decreased compared to the control; however, $TNF-{\alpha}$ reduction patterns were different depending on the type of PI3K/MAPK inhibitors. $TNF-{\alpha}$ production was reduced in a dose-dependent manner by treatment with wortmannin and PD98059, whereas it was increased by SP600125. These data suggested that PI3K/AKT and MAPK signaling pathways are important in regulation of $TNF-{\alpha}$ production in cervical mucosal epithelial SiHa cells. However, activation patterns of each pathway were different from the types of PI3K/MAPK pathways.
Porcine cytomegalovirus (PCMV) is a betaherpesvirus which causes reproductive failure in breeding sows and generalized infection in newborn piglets. It has worldwide distribution including Korea. Serological survey on this virus has been reported in 76.3% of pigs, but virological survey and epidemiological analysis on PCMV distribution have been reported in only a few papers in Korea. In this study, we investigated the virological prevalence and infection status of PCMV on a farm level in selected swine farms with respiratory diseases. A total of 1,938 blood samples taken from groups of pigs of different ages were collected from 31 farms distributed nationwide in 2006 and 2007 and tested by PCR to detect the presence of PCMV. Virological prevalence at farm level and pig level were 96.8% and 17.5%, respectively, suggesting that PCMV has endemically infected Korean pig herds. The prevalence at farm level in gilts, sows and suckling piglet groups were 16.7%, 36.7% and 56.7%, indicating that vertical infections frequently occurred in conception or newborn stage. Thereafter, detection rates of PCMV were slightly increased in pig groups aged 40 and 70 days (70.0% and 73.3%), and then gradually decreased as they aged - 33.3% in 100, 26.7% in 130 and 16.7% in 160 day old pig groups. The prevalence at pig level has similar patterns to that at farm level. With the passage of time, the variation of infection patterns of PCMV was investigated in four PCMV-positive farms. Three blood samples were collected at intervals of 6 months in each farm, and examined for presence of PCMV using PCR. The results revealed that once PCMV was introduced to the pig farms, it continuously circulated between and within groups of sows and piglets in those farms. Taken together, it can be concluded that PCMV has endemically infected Korean pig farms and has the potential risk for emerging pathogen in combination with the known endemic pathogens including porcine reproductive, respiratory syndrome virus and porcine circovirus type 2. Therefore, more research is needed on diagnosis, epidemiology and control strategy for PCMV on the field.
IL-23 and IL-12 are structurally similar and critical for the generation of efficient cellular immune responses. Toxoplasma gondii induces a strong cell-mediated immune response. However, little is known about IL-23 secretion profiles in T. gondii-infected immune cells in connection with IL-12. We compared the patterns of IL-23 and IL-12 production by THP-1 human monocytic cells in response to stimulation with live or heat-killed T. gondii tachyzoites, or with equivalent quantities of either T. gondii excretory/secretory proteins (ESP) or soluble tachyzoite antigen (STAg). IL-23 and IL-12 were significantly increased from 6 hr after stimulation with T. gondii antigens, and their secretions were increased with parasite dose-dependent manner. IL-23 concentrations were significantly higher than those of IL-12 at the same multiplicity of infection. IL-23 secretion induced by live parasites was significantly higher than that by heat-killed parasites, ESP, or STAg, whereas IL-12 secretion by live parasite was similar to those of ESP or STAg. However, the lowest levels of both cytokines were at stimulation with heat-killed parasites. These data indicate that IL-23 secretion patterns by stimulation with various kinds of T. gondii antigens at THP-1 monocytic cells are similar to those of IL-12, even though the levels of IL-23 induction were significantly higher than those of IL-12. The detailed kinetics induced by each T. gondii antigen were different from each other.
Nontuberculous mycobacteria (NTM) are a major cause of opportunistic infections in immunocompromised patients, making the reliable and rapid identification of NTM to the species level very important for the treatment of such patients. Therefore, this study evaluated the usefulness of the novel target genes tuf and tmRNA for the identification of NTM to the species level, using a PCRrestriction fragment length polymorphism analysis (PRA). A total of 44 reference strains and 17 clinical isolates of the genus Mycobacterium were used. The 741 bp or 744 bp tuf genes were amplified, restricted with two restriction enzymes (HaeIII/MboI), and sequenced. The tuf gene-PRA patterns were compared with those for the tmRNA (AvaII), hsp65 (HaeIII/HphI), rpoB (MspI/HaeIII), and 16S rRNA (HaeIII) genes. For the reference strains, the tuf gene-PRA yielded 43 HaeIII patterns, of which 35 (81.4%) showed unique patterns on the species level, whereas the tmRNA, hsp65, rpoB, and 16S rRNA-PRAs only showed 10 (23.3%), 32 (74.4%), 19 (44.2%), and 3 (7%) unique patterns after single digestion, respectively. The tuf gene-PRA produced a clear distinction between closely related NTM species, such as M. abscessus (557-84-58) and M. chelonae (477-84-80-58), and M. kansasii (141-136-80-63-58-54-51) and M. gastri (141-136-117-80-58-51). No difference was observed between the tuf-PRA patterns for the reference strains and clinical isolates. Thus, a diagnostic algorithm using a tuf gene-targeting PRA is a promising tool with more advantages than the previously used hsp65, rpoB, and 16S rRNA genes for the identification of NTM to the species level.
Although the Korean isolate KI-1 of Toxoplasma gondii has been considered to be a virulent type I lineage because of its virulent clinical manifestations, its genotype is unclear. In the present study, genotyping of the KI-1 was performed by multilocus PCR-RFLP and microsatellite sequencing. For 9 genetic markers (c22-8, c29-2, L358, PK1, SAG2, SAG3, GRA6, BTUB, and Apico), the KI-1 and RH strains exhibited typical PCR-RFLP patterns identical to the type I strains. DNA sequencing of tandem repeats in 5 microsatellite markers (B17, B18, TUB2, W35, and TgM-A) of the KI-1 also revealed patterns characteristic of the type I. These results provide strong genetic evidence that KI-1 is a type I lineage of T. gondii.
Pepper and tomato plants infected with two Clavibacter species, C. capsici and C. michiganensis have shown different patterns of disease development depending on their virulence. Here, we investigated how pepper and tomato plants respond to infection by the high-virulent or low-virulent Clavibacter strains. For this, we chose two strains of each Clavibacter species to show different virulence level in the host plants. Although low-virulent strains showed less disease symptoms, they grew almost the same level as the high-virulent strains in both plants. To further examine the response of host plants to Clavibacter infection, we analyzed the expression patterns of plant defense-related genes in the leaves inoculated with different strains of C. capsici and C. michiganensis. Pepper plants infected with high-virulent C. capsici strain highly induced the expression of CaPR1, CaDEF, CaPR4b, CaPR10, and CaLOX1 at 5 days after inoculation (dai), but their expression was much less in low-virulent Clavibacter infection. Expression of CaSAR8.2 was induced at 2 dai, regardless of virulence level. Expression of GluA, Pin2, and PR2 in tomato plants infected with high-virulent C. michiganensis were much higher at 5 dai, compared with mock or low-virulent strain. Expression of PR1a, Osmotin-like, Chitinase, and Chitinase class 2 was increased, regardless of virulence level. Expression of LoxA gene was not affected by Clavibacter inoculation. These results suggested that Clavibacter infection promotes induction of certain defense-related genes in host plants and that differential expression of those genes by low-virulent Clavibacter infection might be affected by their endophytic lifestyle in plants.
Adenoviral vectors are among the most promising vectors available for human gene therapy. However, the use of recombinant adenoviral vectors, including replicationcompetent adenovirus (RCA), raises a variety of safety concerns in relation to the development of new therapies based on gene therapy. To examine how organic compounds change in rat plasma following the injection of adenovirus, $\beta$-galactosidase expressing recombinant adenovirus (designated rAdLacZ) or RCA, we investigated the content of fatty acids (FAs), which are important biochemical indicators in pathological conditions. Pattern recognition analysis on the level of FAs in rat plasma is described for the visual discrimination of adenovirus infection groups from normal controls. Plasma FAs from four control rats (normal group), and from four rats with rAdLacZ infection and six rats with RCA infection (the two abnormal groups), were examined by gas chromatography-mass spectrometry in selected ion monitoring modes as their tert-butyldimethylsilyl derivatives. In total, 20 FAs were positively detected and quantified. The results of the Student's t-test on the normal mean of two abnormal groups, the levels of three FAs (p<0.05) from rAdLacZ group and eleven FAs (p<0.05) from RCA group were significantly different. When star symbol plotting was applied to the group mean values of 20 FAs after normalization to the corresponding normal mean values, the resulting eicosagonal star patterns of the two infected groups were distorted into similar shapes, but were distinguishable from each other. Thus, these approaches will be useful for screening and monitoring of diagnostic markers for the effects of infection following the use of adenoviral vectors in gene therapy.
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