Significant Differences In Pathogenicity Research Articles

Genetic Diversity and Virulence of Phytopathogenic Burkholderia glumae Strains Isolated from Rice Cultivars in Valleys of the High Jungle of Perú.

Burkholderia glumae causes bacterial leaf blight in rice, and its global spread has been exacerbated by climate change. To understand the genetic diversity and virulence of B. glumae strains isolated from rice cultivars in Perú, 47 isolates were obtained from infected rice fields, all belonging to B. glumae, and confirmed by recA and toxB sequences. The BOX-PCR typing group has 38 genomic profiles, and these turn into seven variable number tandem repeats (VNTR) haplotypes. There was no correlation between clustering and geographical origin. Nineteen strains were selected for phenotypic characterization and virulence, using both the maceration level of the onion bulb proxy and inoculation of seeds of two rice cultivars. Several strains produced pigments other than toxoflavin, which correlated with onion bulb maceration. In terms of virulence at the seed level, all strains produced inhibition at the root and coleoptile level, but the severity of symptoms varied significantly between strains, revealing significant differences in pathogenicity. There is no correlation between maceration and virulence scores, probably reflecting different virulence mechanisms depending on the host infection stage. This is the first study to evaluate the VNTR diversity and virulence of Peruvian strains of B. glumae in two commercial cultivars.

Molecular epidemiology of Mycoplasma pneumoniae pneumonia in children, Wuhan, 2020–2022

BackgroundMycoplasma pneumoniae (M. pneumoniae) is an important pathogen of community-acquired pneumonia in children. The factors contributing to the severity of illness caused by M. pneumoniae infection are still under investigation. We aimed to evaluate the sensitivity of common M. pneumoniae detection methods, as well as to analyze the clinical manifestations, genotypes, macrolide resistance, respiratory microenvironment, and their relationship with the severity of illness in children with M. pneumoniae pneumonia in Wuhan.ResultsAmong 1,259 clinical samples, 461 samples were positive for M. pneumoniae via quantitative polymerase chain reaction (qPCR). Furthermore, we found that while serological testing is not highly sensitive in detecting M. pneumoniae infection, but it may serve as an indicator for predicting severe cases. We successfully identified the adhesin P1 (P1) genotypes of 127 samples based on metagenomic and Sanger sequencing, with P1-type 1 (113/127, 88.98%) being the dominant genotype. No significant difference in pathogenicity was observed among different genotypes. The macrolide resistance rate of M. pneumoniae isolates was 96% (48/50) and all mutations were A2063G in domain V of 23S rRNA gene. There was no significant difference between the upper respiratory microbiome of patients with mild and severe symptoms.ConclusionsDuring the period of this study, the main circulating M. pneumoniae was P1-type 1, with a resistance rate of 96%. Key findings include the efficacy of qPCR in detecting M. pneumoniae, the potential of IgM titers exceeding 1:160 as indicators for illness severity, and the lack of a direct correlation between disease severity and genotypic characteristics or respiratory microenvironment. This study is the first to characterize the epidemic and genomic features of M. pneumoniae in Wuhan after the COVID-19 outbreak in 2020, which provides a scientific data basis for monitoring and infection prevention and control of M. pneumoniae in the post-pandemic era.

Phenotype and genotype characterization of Botrytis cinerea isolates from cut roses in Yunnan, China

AbstractBotrytis cinerea is considered to be the second most destructive fungal pathogen worldwide, causing severe pre‐ and postharvest losses in cut roses. However, to date, no systematic research on its characteristics in cut roses has been reported. In our study, a total of 100 isolates from cut roses from Yunnan, China, were analysed. A combination of morphological characteristics and phylogenetic analysis of RPB2 revealed that 100 isolates were of the species B. cinerea. These isolates were pathogenic on unwounded detached rose petals. Pathogenicity was evaluated according to the size of petal spots and categorized into grades 0–5, from weak to strong. The inhibition rate varied greatly following the addition of procymidone (21.9%–100%) and cyprodinil (25.0%–92.3%). Analysis of Bc‐hch sequences revealed all isolates belong to phylogenetic Group II. According to the presence or absence of transposable elements, 87, 5, 4 and 4 isolates were identified as transposa, vacuma, Boty‐only and Flipper‐only types, respectively. Detection of mating type indicated that all isolates were heterothallic with 45% belonging to MAT1‐1 and 55% to MAT1‐2. Isolates were divided into four subpopulations when analysing single‐nucleotide polymorphisms at the genomic level, with a significant difference in pathogenicity between subpopulations 1 and 4. The evolutionary tree indicated that isolates AN‐02, AN‐22 and SM‐C18 were clustered in the root, suggesting an earlier evolutionary time than other isolates; moreover, they all exhibited low pathogenicity (grade 1). This systematic study of the characteristics of B. cinerea will provide significant support for grey mould control and further research.

Characterization and pathogenicity of Pratylenchus vandenbergae stat. nov. (Tylenchina: Pratylenchidae), a highly pathogenic root‐lesion nematode parasitizing crops in Kenya and South Africa

AbstractA highly fecund root‐lesion nematode parasitizing a variety of crops was discovered and found to be widespread in Kenya and South Africa. These populations were molecularly identical to Pratylenchus teres teres and P. teres vandenbergae based on D2‐D3 of 28S rDNA and Hsp90 sequences. However, based on morphological differences with the original description of P. teres and its different geographical distribution, Pratylenchus vandenbergae stat. nov., previously known as subspecies P. teres vandenbergae, is recognized as a separate species. This species is characterized by a slightly offset labial region with three annuli, en face morphology belonging to Group II, lateral field with four incisures with two outer bands areolated as observed under a light microscope and the inner band also partially areolated at the vulva region as observed in scanning electron microscopy view, a robust stylet (14–17 μm) with rounded knobs and subcylindrical tail with annulated tail tip. This important and common species remained largely under the radar, probably due to identifications based only on sequence similarity, including mislabelled Pratylenchus bolivianus sequences in GenBank. Pathogenicity of P. vandenbergae stat. nov. in finger millet roots was confirmed through acid fuchsin staining, and reproduction and pathogenicity tests in maize, soybean, sunflower and tomato plants demonstrated its capacity to affect crop growth. Life cycle comparison with P. penetrans in vitro showed a similar life cycle length but significantly higher fecundity. Finally, significant differences in pathogenicity and reproduction of P. vandenbergae stat. nov. were found both at genotype level (finger millet genotypes OKHALE‐1 & KNE1034) and crop level (maize, soybean, sunflower and tomato plants).

Unraveling the role of type 1 fimbriae in Salmonella pathogenesis: insights from a comparative analysis of Salmonella Enteritidis and Salmonella Gallinarum

Significant differences in pathogenicity between Salmonella Enteritidis and Salmonella Gallinarum exist despite the fact that S. Gallinarum is a direct descendant of S. Enteritidis. It was hypothesized that such various properties may be in part the result of differences in structure and functions of type 1 fimbriae (T1Fs). In S. Enteritidis, T1Fs bind to oligomannosidic structures carried by host cell glycoproteins and are called mannose-sensitive T1Fs (MST1F). In S. Gallinarum, T1Fs lost ability to bind such carbohydrate chains, and were named mannose-resistant MRT1Fs (MRT1F). Therefore, the present study was undertaken to evaluate the role of MST1Fs and MRT1Fs in the adhesion, invasion, intracellular survival and cytotoxicity of S. Enteritidis and S. Gallinarum toward chicken intestinal CHIC8-E11cells and macrophage-like HD11 cells. Using mutant strains: S. Enteritidis fimH::kan and S. Gallinarum fimH::kan devoid of T1Fs and in vitro assays the following observations were made. MST1Fs have a significant impact on the chicken cell invasion by S. Enteritidis as MST1F-mediated adhesion facilitates direct and stable contact of bacteria with host cells, in contrast to MRT1Fs expressed by S. Gallinarum. MST1Fs as well as MRT1Fs did not affected intracellular viability of S. Enteritidis and S. Gallinarum. However, absolute numbers of intracellular viable wild-type S. Enteritidis were significantly higher than S. Enteritidis fimH::kan mutant and wild-type S. Gallinarum and S. Gallinarum fimH::kan mutant. These differences, reflecting the numbers of adherent and invading bacteria, underline the importance of MST1Fs in the pathogenicity of S. Enteritidis infections. The cytotoxicity of wild-type S. Enteritidis and its mutant devoid of MST1Fs to HD11 cells was essentially the same, despite the fact that the number of viable intracellular bacteria was significantly lower in the mutated strain. Using HD11 cells with similar number of intracellular wild-type S. Enteritidis and S. Enteritidis fimH::kan mutant, it was found that the lack of MST1Fs did not affect directly the cytotoxicity, suggesting that the increase in cytotoxicity of S. Enteritidis devoid of MST1Fs may be associated with crosstalk between T1Fs and other virulence factors.

The pathogenicity and immunogenicity of Pasteurella multocida in low and rich iron media

Pasteurella multocida has the highest prevalence among animal populations and may cause hemorrhagic septicemia in cattle and pneumonia in sheep and goats. It has known that the pathogenicity of this organism depends on the levels of iron. Present study attempts to evaluate any changes in blood parameters and visceral organs of mice by cultivation of P.multocida under iron sufficient and insufficient conditions. P.multocida was cultured on three media: BHI alone, supplemented with bipyridyl and iron. They injected into the mice by intra-peritoneal rout. At the same time inactivated antigens were prepared from all medium and mice were immunized two weeks apart and blood samples were collected until weeks 9. Visceral organs were collected from dead mice for pathological studies. In contrast to immunogenicity and antibody titers, there are significant differences in pathogenicity and blood parameters in rich and limit iron condition.

Colletotrichum Species Associated with Anthracnose in Salix babylonica in China.

Salix babylonica L. is a popular ornamental tree species in China and widely cultivated in Asia, Europe, and North America. Anthracnose in S. babylonica poses a serious threat to its growth and reduces its medicinal properties. In 2021, a total of 55 Colletotrichum isolates were isolated from symptomatic leaves in three provinces in China. Phylogenetic analyses using six loci (ITS, ACT, CHS-1, TUB2, CAL, and GAPDH) and a morphological characterization of the 55 isolates showed that they belonged to four species of Colletotrichum, including C. aenigma, C. fructicola, C. gloeosporioides s.s., and C. siamense. Among them, C. siamense was the dominant species, and C. gloeosporioides s.s. was occasionally discovered from the host tissues. Pathogenicity tests revealed that all the isolates of the aforementioned species were pathogenic to the host, and there were significant differences in pathogenicity or virulence among these isolates. The information on the diversity of Colletotrichum spp. that causes S. babylonica anthracnose in China is new.

Effect of Crop Rotation on Fusarium Mycotoxins and Fusarium Species in Cereals in Sichuan Province (China).

The present study was performed to evaluate the effect of crop rotation on Fusarium mycotoxins and species in cereals in Sichuan Province. A total of 311 cereal samples were randomly collected and analyzed from 2018 to 2019 in Sichuan Province. The results of mycotoxin analysis showed that the major trichothecene mycotoxins in Sichuan Province were nivalenol (NIV) and deoxynivalenol (DON), and the mean concentration of total trichothecenes (including NIV, fusarenone X [4ANIV], DON, 3-acetyldeoxynivalenol [3ADON], and 15-acetyldeoxynivalenol [15ADON]) in wheat was significantly higher than that in maize and rice. The concentration of total trichothecenes in the succeeding crops was significantly higher than that in the previous crops. In addition, wheat grown after maize had reduced incidence and concentration of trichothecene mycotoxins compared with that grown after rice, and ratooning rice grown after rice had increased incidence and concentration of trichothecene mycotoxins. Our data indicated that Fusarium asiaticum with the NIV chemotype was predominant in wheat and rice samples, while the number of the NIV chemotypes of F. asiaticum and Fusarium meridionale and the 15ADON chemotype of Fusarium graminearum in maize were almost the same. Although the composition of Fusarium species was affected by crop rotations, there were no differences when comparing the same crop rotation except for the maize-wheat rotation. Moreover, the same species and chemotype of Fusarium strains originated from different crops in various rotations, but there were no significant differences in pathogenicity in wheat and rice. These results contribute to the knowledge of the effect of crop rotation on Fusarium mycotoxins and species affecting cereals in Sichuan Province, which may lead to improved strategies for control of Fusarium mycotoxins and fungal disease in China.

Detection and characterization of difenoconazole resistance in Stagonosporopsis citrulli from watermelon and muskmelon in Zhejiang Province of China

The watermelon and muskmelon productions are important agriculture pillar industries of Zhejiang Province in China. Difenoconazole is an imidazole-class sterol, 14-demethylase inhibitor (DMI), which has been in use for several years to control gummy stem blight (GSB) caused by Stagonosporopsis species. However, the detection and characterization of difenoconazole resistance in GSB have not been clarified. In this study, a total of 112 isolates were collected from samples of GSB on watermelon and muskmelon in five locations of Zhejiang Province, China. All of the isolates were identified as Stagonosporopsis citrulli through morphology and multiplex PCR analysis. The determination of their resistance to difenoconazole via the discriminatory dosage method showed that the total resistance frequency was 89.3%. Among the resistant sub-population, 36.6% had high-level resistance to difenoconazole (DifHR), while 46.4% and 6.3% had low- (DifLR) and moderate-level resistance (DifMR), respectively. Additionally, the difenoconazole showed a positive cross-resistance with four DMIs, i.e., tebuconazole, prochloraz, metconazole, and mefentrifluconazole, but not hexaconazole. The phenotypic analysis found that the difenoconazole resistant (DifR) isolates demonstrated attenuated ability in both the mycelial growth and sporulation compared with the difenoconazole sensitive (DifS) isolates, while there was no significant difference in pathogenicity on watermelon leaves between the DifR and DifS isolates. Further exploration of the mechanism related to difenoconazole resistance of S. citrulli isolates revealed that the resistance to difenoconazole involved four types of mutations in CYP51, i.e., G463S for DifLR, I444M, Y446H, and A464G for DifHR. No over-expression of the cyp51 gene was found in the tested DifR isolates. Furthermore, it was found that 5% of the DifR isolates were significantly more sensitive to difenoconazole after being treated with 20 μg/mL chlorpromazine hydrochloride, indicating that the efflux mechanism may be involved in these difenoconazole-resistant isolates. Together, our study results suggested that S. citrulli had a strong resistance to difenoconazole on watermelon and muskmelon, and the mutations in cyp51and changes in fungicide efflux were responsible for the emergence of difenoconazole resistance in S. citrulli.

Genetic Variations among Different Variants of G1-like Avian Influenza H9N2 Viruses and Their Pathogenicity in Chickens.

Since it was first discovered, the low pathogenic avian influenza (LPAI) H9N2 subtype has established linages infecting the poultry population globally and has become one of the most prevalent influenza subtypes in domestic poultry. Several different variants and genotypes of LPAI H9N2 viruses have been reported in Egypt, but little is known about their pathogenicity and how they have evolved. In this study, four different Egyptian LPAI H9N2 viruses were genetically and antigenically characterized and compared to representative H9N2 viruses from G1 lineage. Furthermore, the pathogenicity of three genetically distinct Egyptian LPAI H9N2 viruses was assessed by experimental infection in chickens. Whole-genome sequencing revealed that the H9N2 virus of the Egy-2 G1-B lineage (pigeon-like) has become the dominant circulating H9N2 genotype in Egypt since 2016. Considerable variation in virus shedding at day 7 post-infections was detected in infected chickens, but no significant difference in pathogenicity was found between the infected groups. The rapid spread and emergence of new genotypes of the influenza viruses pinpoint the importance of continuous surveillance for the detection of novel reassortant viruses, as well as monitoring the viral evolution.

Identification and phylogenetic analysis of the pathogen of jute anthracnose in China

<p id="C3">Anthracnose is a major fungal disease that seriously affects the yield and quality in jute fiber. Identifying the types of anthracnose pathogens and determining their pathogenicity will help jute disease-resistant breeding and develop elite disease-resistant gene resources. The 92 strains were isolated and purified from samples of diseased jute, which were collected from 7 jute production areas including Fuzhou and Zhangzhou in Fujian Province, Xinyang City in Henan Province, Liu’an City in Anhui Province, Hangzhou City in Zhejiang Province, Nanning City in Guangxi Province, and Changsha City in Hunan Province. The sequence analysis of the rDNA-ITS region showed that 11 out of the 92 strains were typical anthracnose pathogens. Further identification of the phylogenetic tree of the LSU region and morphological characteristics revealed that 10 strains, including ZZ4 and GX19, were <italic>Colletotrichum gloeosporioides</italic>, while the strain of CS3 was <italic>Colletotrichum dematium</italic>. The pathogenicity test of artificial inoculation indicated that there was a significant difference in pathogenicity between <italic>Colletotrichum gloeosporioides</italic> and <italic>Colletotrichum dematium</italic>. These results laid a basis for identification of resistance gene loci and effective controlling of anthracnose in jute.

First Report of Root Rot onkiwifruitsCaused byAthelia rolfsiiin Hefei, China.

Kiwifruits (Actinidia ssp.), known as "King of vitamin C", have been wildly cultivated. In August 2020, about 15% of A. deliciosa (cv. Xuxiang) and A. macrosperma (rootstock) plants displayed symptoms typical of root rot at a farm in Hefei (117°25'E, 31°86'N), Anhui Province of China (Fig.1 a-b). Symptoms first appeared at the root and stem junction which were covered by cottony white mycelium during warm and humid summer. Then, the infected tissues were rotted, and subsequently the whole plant withered. Tan to brown sclerotia were observed on the basal stem epidermis and soil surface surrounding the stem (Fig.1 c-d). Infected plant tissues and sclerotia were collected for isolating the fungal pathogen. The samples were surface sterilized in 70% alcohol for 30 s, followed by 2% sodium hypochlorite for 3 min, washed five times with sterile double-distilled water (ddH2O), dried, placed on potato dextrose agar, and incubated at 25 °C in the dark. In total, twelve fungal isolates were obtained. The mycelia of all the isolates were white with a fluffy appearance (Fig.1 e). Sclerotia formed after 7 days were initially white (Fig.1 f) and gradually turned to dark brown (Fig.1 g) measuring 0.67 to 2.03 mm in diameter (mean = 1.367 ± 0.16 mm;n= 30). Hyphae were hyaline, septate. Some cells possessed multiple nuclei (Fig.1 h) and clamp connections (Fig.1 i). No spores were observed. For species-level identification, ITS1/ITS4 and EF1-983F/EF1-2218R primers were used to amplify the internal transcribed spacer regions (ITS) and translation elongation factor-1 alpha regions (TEF-1α), respectively (White et al. 1990; Rehner & Buckley 2005). Based on ITS and TEF-1α sequence analyses, all 12 isolates were categorized into two groups, group one including isolates NC-1 and NC-6~10 and group two containing NC-2~5 and NC-11~12. The length of ITS sequences for NC-1 (MW311079) was 684bp and 99% to 100% similar to Athelia rolfsii (MN610007.1, MN258360.1). Similarly, ITS sequences for NC-2 (MW311080) were 99% to 100% similar to A. rolfsii (MH858139.1; MN872304.1). Also, TEF-1α sequences of NC-1 (MW322687) and NC-2 (MW322688) were 96% to 99% similar to sequences of A. rolfsii (MN702794.1, GU187681.1, MN702789.1). Based on morphology and phylogenetic analyses (Fig.1 j&k), the isolates NC-1 and NC-2 were identified as Athelia rolfsii (anamorph Sclerotium rolfsii) (Mordue. 1974; Punja. 1985). To fulfill Koch's postulates, ten sclerotia of NC-1 were incorporated into the soil near stems of healthy Xuxiang plants (Fig.2 a). Similar treatments were also used for plants of A. macrosperma or A. arguta (Fig.2 g&m). Each control group had the same number of plants (n=3) for inoculating with ddH2O. The plants were kept in an incubator with a relative humidity of 80% and temperature of 28°C with 16/8 hours light/dark photoperiod. After twenty days, the pathogen-inoculated plants developed similar symptoms of root rot observed in the field (Fig.2 b-d, h-j, n-o). Similarly, four days after inoculation with sclerotia, leaves developed water-soaked lesions (Fig.2 e, k&p). No significant difference in pathogenicity was observed between NC-1 and NC-2. Non-inoculated control plants remained disease-free (Fig.2 f, l&q). The pathogenicity experiments were repeated three times. The pathogen was re-isolated from infected tissues and sclerotia, and isolates were confirmed as A. rolfsii by the ITS sequences. A. rolfsii has been reported to cause root rot in kiwifruit in the USA (Raabe. 1988). To our knowledge, this is the first report A. rolfsiicausing root rot on kiwifruits in China.

Analysis of the Fusarium graminearum Species Complex from Gramineous Weeds Near Wheat Fields in Jiangsu Province, China.

Several weed species are known as alternative hosts of the Fusarium graminearum species complex (FGSC), and their epidemiological significance in Fusarium head blight (FHB) has been investigated; however, scant information is available regarding FGSC occurrence in weeds near Chinese wheat fields. To evaluate the potential role of gramineous weeds surrounding wheat fields in FHB, 306 FGSC isolates were obtained from 210 gramineous weed samples in 2018 in Jiangsu Province. Among them, 289 were Fusarium asiaticum, and the remainder were F. graminearum. Trichothecene genotype and mycotoxin analyses revealed that 74.3% of the F. asiaticum isolates were the 3-acetyldeoxynivalenol (3ADON) chemotype, and the remainder were the nivalenol (NIV) chemotype. Additionally, 82.4% of F. graminearum isolates were the 15-acetyldeoxynivalenol (15ADON) chemotype, and the remainder were the NIV chemotype. FHB severity and trichothecene analysis indicated that F. asiaticum isolates with the 3ADON chemotype were more aggressive than those with the NIV chemotype in wheat. 3ADON and NIV chemotypes of F. asiaticum isolated from weeds and wheat showed no significant differences in pathogenicity in wheat. All selected F. asiaticum isolates produced perithecia, with little difference between the 3ADON and NIV chemotypes. These results highlight the epidemiology of the FGSC isolated from weeds near wheat fields, with implications for reducing FHB inoculum in China.

Variations in Pathogenicity of Three Different Forms of Rhizoctonia Inoculum and Assessment of Cultivar Resistance of Sugar Beet

Rhizoctonia solani causes pre-emergence and post-emergence damping-off, as well as crown and root rot of sugar beet (Beta vulgaris L.), which significantly affects the yield returns in the USA and Europe. The pathogen overwinters as sclerotia or melanized mycelium. Traditionally, the resistance of cultivars to R. solani is evaluated by scoring disease reactions at the crowns and roots of older seedlings, thus resistance is not evaluated during seed germination. Moreover, earlier studies evaluated cultivars resistance to R. solani using colonized whole barley or wheat grains which, unlike sclerotia, are artificial inocula of the pathogen that require time, space and technical know-how to produce. Moreover, colonized grains are prone to contamination with other pathogens, consumed by rodents/birds while applied in the field, and are often uneconomic. Considering those limitations, a study was undertaken (1) to develop in vitro methods to generate large-scale sclerotia, (2) to compare pathogenic potentials of sclerotia, mycelia, and colonized barley grains for optimization of dampingoff assays, and (3) to evaluate Rhizoctonia resistance of selected commercial cultivars during the seed germination phase. Comparing six different culture media, we found that R. solani had the highest radial growth (8.9 ± 0.04, cm³) at 8-days and the maximum number of sclerotia produced (203 ± 4.6) at 28-days in CV8 medium. We demonstrated significant differences in pathogenicity of the three different forms of R. solani inocula and susceptibility of cultivars to preand post-emergence damping-off. The highest pre-emergence damping-off and root rot were observed with sclerotia, and the highest post-emergence dampingoff was recorded with both sclerotial and colonized barley inocula. In addition, varietal differences in susceptibility to pre- and post-emergence damping-off were noted. The highest pre-emergence damping-off was recorded on cv Crystal 101RR and lowest in Maribo MA 504. The highest post-emergence damping-off was recorded on BTS 8500 and the lowest in Crystal 467. The maximum mean root rot was observed in BTS 8500, BTS 8606, and Crystal 101R. Our studies demonstrated that sclerotia serve as efficient natural inocula, reemphasized that host-pathogen interactions differ at the early vs. late stages of sugar beet growth, and highlighted the need to reevaluate commercial sugar beet cultivars for resistance at the seed germination stage.

Effect of Antibacterial Root Canal Sealer on Persistent Apical Periodontitis.

The infection of Enterococcus faecalis and its interacting microorganisms in the root canal could cause persistent apical periodontitis (AP). Antibacterial root canal sealer has favorable prospects to inhibit biofilms. The purpose of this study was to investigated the antibacterial effect of root canal sealer containing dimethylaminododecyl methacrylate (DMADDM) on persistent AP in beagle dogs for the first time. Persistent AP was established by a two-step infection with Enterococcus faecalis and multi-bacteria (Enterococcus faecalis, Lactobacillus acidophilus, Actinomyces naeslundii, Streptococcus gordonii). Root canal sealer containing DMADDM (0%, 1.25%, 2.5%) was used to complete root canal filling. The volume of lesions and inflammatory grade in the apical area were evaluated by cone beam computer tomography (CBCT) and hematoxylin-eosin staining. Both Enterococcus-faecalis- and multi-bacteria-induced persistent AP caused severe apical destruction, and there were no significant differences in pathogenicity between them. DMADDM-modified sealer significantly reduced the volume of periapical lesion and inflammatory grade compared with the control group, among them, the therapeutic effect of the 2.5% group was better than the 1.25% group. In addition, E. faecalis-induced reinfection was more sensitive to the 2.5% group than multi-bacteria reinfection. This study shows that root canal sealer containing DMADDM had a remarkable therapeutic effect on persistent AP, especially on E. faecalis-induced reinfection.

Genetic Characterization and Pathogenicity Analysis of Recently Isolated Fowl Adenovirus 8b in Korea.

A Korean field strain of fowl adenovirus (FAdV) 8b was isolated from chickens showing high mortality. Isolated FAdV-8b strains with the hexon and fiber genes were genetically analyzed. The Korean FAdV-8b (K194/19) strain isolated in 2019 showed higher sequence identity with the FAdV-8b strain isolated in China but lower sequence identity with the Korean FAdV-8b (K187/08) strain isolated in 2008. The K194/19 strain formed a distinct subcluster within the FAdV-8b cluster in a phylogenetic tree based on hexon and fiber genes. FAdV can infect day-old chicks through vertical transmission, and so blood samples were obtained from 54-, 60-, and 63-wk-old parent chickens. FAdV-specific antibody levels were investigated with ELISA and virus neutralization (VN) tests with the K194/19 and K187/08 strains as antigens. In VN tests, all sera neutralized the K187/08 strain. However, the K194/19 strain was neutralized by sera collected from 60- and 63-wk-old chickens but not sera obtained from 54-wk-old chickens, indicating natural infection. Finally, to determine the pathogenicity of the K194/19 strain, 1-day-old and 4-wk-old specific-pathogen-free birds were infected with the K194/19 and K187/08 strains. No significant difference in pathogenicity was observed between the two strains. Although the K194/19 strain showed similar pathogenicity with the K187/08 strain, differences in nucleotide and amino acid sequences of the hexon and fiber genes may determine the evasion ability of the K187/08 neutralizing antibody, indicating the need for development of a novel FAdV vaccine.

Characterization of mycotoxigenic Alternaria species isolated from the Tunisian halophyte Cakile maritima

Cakile maritima is a typical halophyte of the Mediterranean coasts. In addition to its ecological and industrial properties, C. maritima has antiscorbutic, diuretic and purgative roles in folk remedies. This plant is infected by different fungal species, mainly belonging to Alternaria genus. Two-hundred Alternaria strains were collected from four different pedo-climatic areas in Tunisia, from C. maritima fresh plant tissues showing symptoms of Alternaria infection. Phylogenetic analyses of 79 representative Alternaria strains, were carried out using multi-locus gene sequencing. All the strains clustered in the Alternaria Section: 47 strains had high homology with A. alternata reference strain, 13 grouped with A. arborescens reference strain, 12 grouped with A. mali reference strain, and seven strains were not well defined with A. mali as their closest species. In vitro production of tenuazonic acid (TA), alternariol (AOH), alternariol-monomethyl ether (AME), and altenuene (ALT) was evaluated. Approx. 68% of strains simultaneously produced AOH, AME and TA. Only two A. alternata and one A. mali strains were ALT producing. Pathogenicity tests on leaves of C. maritima were carried out with 41 representative strains. Alternaria arborescens showed the greatest pathogenicity compared to A. alternata and A. mali, although no statistically significant differences in pathogenicity were observed. This is the first study on Tunisian populations of Alternaria species isolated from the extremophile C. maritima.

The pathogenicity of duck hepatitis A virus types 1 and 3 on ducklings

Duck hepatitis A virus (DHAV) is one of the pathogens that cause fatal duck viral hepatitis (DVH) in ducklings, which is an acute and contagious disease with a high mortality rate. Despite a continuing official duck vaccination program, DHAV infection remains a major threat to the duck industry. Considerable changes were observed in the epidemiology of DHAV-1/-3 in China over time. Therefore, comparing the pathogenicity of different DHAV serotypes can provide a theoretical basis for the diagnosis and prevention of DVH. In this study, we systematically investigated the effects of infection with DHAV-1/-3 field strains on clinical signs, gross lesions, histopathological changes, viral RNA detection, enzymatic systems, and metabolite concentrations. The results demonstrated that the major macroscopic and microscopic lesions in ducks infected with DHAV-1/-3 in the liver, brain, spleen, pancreas, and kidneys exhibited no significant differences. After 24 h of infection, DHAV quickly appeared in blood and major organs. Significant changes in clinical chemical markers together with histopathological lesions and viral RNA detection indicated that the liver is the major target organ for both viruses, resulting in impaired of liver integrity and function. In addition, we found that both viruses were able to invade both central and peripheral immune organs. Also lipase plasma activity was substantially affected by DHAV-1/-3, indicating that the integrity and function of the pancreas was compromised. However, there was no significant difference in pathogenicity between DHAV-1 and -3. The results of this study provide new insights into the pathogenesis of DHAV-1/3, two viruses that cause serious depression, metabolic disorders, and immunosuppression.

Construction and immunological evaluation of recombinant Newcastle disease virus vaccines expressing highly pathogenic porcine reproductive and respiratory syndrome virus GP3/GP5 proteins in pigs

Highly pathogenic porcine reproductive and respiratory syndrome (HP-PRRS) poses a significant threat to the pig industry, for which vaccination is considered to be an effective means of prevention and control. Here, we developed two recombinant Newcastle disease virus (NDV) LaSota-vectored PRRS candidate vaccines, rLaSota-GP5 and rLaSota-GP3-GP5, using reverse genetic techniques. The two recombinant viruses exhibited a high degree of genetic stability after 10 successive generations in chicken embryos. There was no significant difference in pathogenicity compared with the rLaSota parent strain in poultry, mice and pigs. The recombinant viruses could not be detected in the feeding environment of immunized pigs, but could be detected in the organs and tissues of pigs for no more than 10 days after immunization. Importantly, in contrast to rLaSota-GP5, rLaSota-GP3-GP5 elicited both significant humoral and cellular immune responses in pigs. In particular, the neutralizing antibody titer in the rLaSota-GP3-GP5 group was 1.51 times significantly higher than that of the commercial vaccine group at 42 days post-immunization. At the same time, there was significant difference in the level of IFN-γ between the rLaSota-GP3-GP5 group and the commercial vaccine group. Furthermore, the viral load in the organs and tissues of rLaSota-GP3-GP5-immunized pigs was substantially lower than that of unimmunized pigs after being challenged with HP-PRRS virus GD strain. These results suggest that rLaSota-GP3-GP5 is a safe and promising candidate vaccine, and there is potential for further development of a recombinant virus vaccine for PRRS using NDV.

Comparison of Pathogenicity and Transmissibility of Influenza B and D Viruses in Pigs.

Influenza viruses are important pathogens causing respiratory disease in humans and animals. In contrast to influenza A virus (IAV) that can infect a wide range of animal species, other influenza viruses, including influenza B virus (IBV), influenza C virus (ICV), and influenza D virus (IDV) have a limited host range. Swine can be infected with all four different genera of influenza viruses. IAV infection of pigs causes the well-known swine influenza that poses significant threats to human and animal health. However, influenza virus infection of pigs with IBV, ICV, and IDV are not well-characterized. Herein, we compared pathogenicity of IBV and IDV using intratracheal and intranasal infection of pigs, which are IAV seropositive, and commingled naïve pigs with the infected animals to determine their transmissibility. Both viruses caused fever and some lung lesions, replicated in the lungs of infected pigs, but only IDV transmitted to the contact animals. Although IBV and IDV displayed differing levels of replication in the respiratory tract of infected pigs, no significant differences in pathogenicity of both viruses were observed. These results indicate that both IBV and IDV can replicate, and are pathogenic in pigs.