Identification of the Chinese Box Orange (Severinia Buxifolia) as an Alternative Host of the Bacterium Causing Citrus Huanglongbing

The Chinese box orange (Severinia buxifolia) was shown by graft-inoculation and psyllid-transmission tests to be an alternative host of the bacterium causing citrus Huanglongbing (HLB). A PCR-based assay for detection of the HLB bacterium (HLBB) was used to monitor HLBB. In graft-inoculation tests, the Chinese box orange (CBO) grafted with HLBB-infected scions of Luchen sweet orange (LSO) were positive for HLBB, 2–3 months after grafting. The back-grafting test demonstrated that HLBB-infected CBO scions could transmit HLBB back to LSO hosts via grafting. In psyllid-transmission tests, psyllids (insect vectors) transmitted HLBB to CBO plants, in which HLBB could be detected 3–4 months after inoculation. Acquisition-access tests of psyllids revealed that HLBB-free psyllids can acquire HLBB from diseased CBO hosts and can transmit HLBB back to the LSO plants. A field survey verified the presence of HLBB-infected CBO plants in the vicinity of citrus orchards. In this paper, CBO is shown to be a susceptible host plant in which HLBB can exist and replicate. It is also a donor plant from which HLBB can be transmitted to citrus hosts by grafting or by psyllid vectors.     

Characterization of monoclonal antibodies against avian reovirus S1133 protein sigmaA synthesized in Escherichia coli

Monoclonal antibodies (MAbs) were prepared against avian reovirus S1133 protein sigmaA (esigmaA) synthesized in Escherichia coli. MAbs were characterized and used to develop a diagnostic test. Ten MAbs were selected for competitive binding assay following coupling with horseradish peroxidase. The results indicated that these MAbs delineated two epitopes I and II of esigmaA. An immuno-dot binding assay was used to detect the effect of denaturation on antibody recognition of the epitopes. All MAbs bound to esigmaA in its native form. After denaturation by boiling in SDS and 2-mercaptoethanol, the binding of MAbs recognizing epitope I was fully abolished. However, the reactivity of MAbs recognizing epitope II was not affected. MAbs 31 and 32, recognizing epitopes I and II, respectively, were selected for the cross-reactivity to heterologous reovirus strains. The results suggest that the two epitopes are highly conserved among these virus strains. A MAb capture enzyme-linked immunosorbent assay (ELISA) procedure was developed using MAbs 32 and 31 to detect reovirus protein sigmaA in samples from tendon tissues of infected bird and chicken embryo fibroblast (CEF) cell cultures. Avian reovirus sigmaA antigens in tendon specimens were detected from the inoculated birds as early as 2 days post-inoculation (PI), approximated a peak at 7 days PI, and maintained this until 16 days PI, then decreased gradually. A clear difference in absorbance values between the tendon samples of the avian reovirus- and mock-infected birds is obtained. Positive results were also obtained from avian reovirus-infected CEF and from the tendon tissues of naturally infected broilers. These results indicated that the MAb capture ELISA is a useful methods for the detection of avian reovirus from chickens suspected to have avian reovirus infections.     

Effect of tumor infiltrating lymphocytes on the expression of MHC molecules in canine transmissible venereal tumor cells

Canine transmissible venereal tumor (CTVT) can be allo-transplanted across major histocompatibility complex barriers. The expression of MHC molecules is usually low in the progression (P) stage and then greatly increases during tumor regression (R). We investigated the effects of tumor infiltrating lymphocytes (TIL) on the expression of MHC molecules of CTVT cells. Isolated, viable CTVT cells were inoculated at each of 12 sites (1 x 10(8) CTVT cells per site) on the back of six, mixed-breed dogs. Tumor masses were collected every 2-3 weeks and prepared for histopathologic, immunocytochemistry, flow cytometry and immunoblotting studies. The level of MHC expression on tumor cells from different stages of growth was measured. Initially, expression of MHC I and II molecules in P phase CTVT was low. Twelve weeks post-inoculation (PI), expression increased dramatically and it continued to increase during R phase. Tumor growth slowed after 12 weeks PI and tumors entered R phase around 17 weeks PI. We hypothesize that CTVT evades host immunosurveillance and grows progressively for 12 weeks, when it becomes vulnerable and subject to the host's anti-tumor immune responses. We further demonstrated that R phase, but not P phase, TIL were closely associated with the over-expression of MHC I and II molecules by CTVT cells. The number and proportion of TIL were higher in R phase tumors. Supernatants, from R phase co-cultures (CTVT+TIL) and TIL only, promoted MHC I and II expression on P phase CTVT cells. After culturing alone for 1 month, expression of MHC classes I and II molecules in R phase CTVT cells decreased to the level of P phase CTVT cells. However, the above-mentioned supernatants restored their expression of MHC I and II molecules. In contrast, supernatants from P phase TIL or CTVT cells increased expression slightly or had no effect. Therefore, TIL, not CTVT cells, produce the effective substance (s) to promote the expression of MHC molecules by the tumor cells. Heat treated supernatant was unable to promote the expression of MHC I and II molecules by CTVT cells. In conclusion, TIL isolated from R phase CTVT secreted a heat-sensitive, soluble substance(s) that triggered over-expression of MHC I and II after 12 weeks PI. This caused the tumor to enter R phase and helped stop CTVT growth. Our findings will facilitate the understanding and further investigation of the mechanisms that initiate host immune surveillance against tumors.     

Inoculation of cucumber roots with zoospores of mycoparasitic and plant pathogenic Pythium species: Differential zoospore accumulation, colonization ability and plant growth response

Hydroponically grown cucumber (Cucumis sativus) seedlings were inoculated with zoospores of 1 mycoparasitic (Pythium oligandrum) and 2 pathogenic (Pythium aphanidermatum and Pythium group F) Pythium spp. During the first 2 days after inoculation, all the Pythium spp. caused reduction in the root length. However, roots treated with Pythium oligandrum quickly reached the length of the control and on the 8th day, and for the rest of the experimental period, stimulation of root elongation was noted. Pythium oligandrum was not pathogenic on cucumber and no differences in the fresh weights of control and Pythium oligandrum inoculated plants were observed in the course of the experiment. Pythium group F and Pythium aphanidermatum were pathogenic on cucumber seedlings, but their pathogenicities differed. Thus, while Pythium group F had a constant, negative influence on root length and plant growth, measured as fresh weight, Pythium aphanidermatum caused generalized necroses of the root system, inhibiting consistently root elongation and plant growth and finally causing plant death. Moreover, the zoospores of 2 mycoparasitic species, Pythium oligandrum and Pythium periplocum, were not attracted to roots of cucumber and accumulated on the roots in very low numbers compared to those of the pathogenic species, Pythium aphanidermatum, which were strongly attracted and accumulated in large numbers. Finally, it was also found that Pythium oligandrum colonized the roots very poorly, while Pythium group F and Pythium aphanidermatum were significantly better root colonizers. The significance of these findings is discussed in relation to the ecology of Pythium species and biocontrol.     

A new ent-kaurane diterpenoid from Croton tonkinensis leaves

A new ent-kaurane diterpenoid (1) was isolated from the leaves of Croton tonkinensis. The structure of 1 was determined as ent-7beta-hydroxy-15-oxokaur-16-en-18-ol from spectroscopic methods.     

Detection of Novel Reassortant Influenza A (H3N2) and H1N1 2009 Pandemic Viruses in Swine in Hanoi,Vietnam

From May to September 2013, monthly samples were collected from swine in a Vietnamese slaughterhouse for influenza virus isolation and serological testing. A(H1N1)pdm09 viruses and a novel H3N2 originating from reassortment between A(H1N1)pdm09 and novel viruses of the North American triple reassortant lineage were isolated. Serological results showed low seroprevalence for the novel H3N2 virus and higher seroprevalence for A(H1N1)pdm09 viruses. In addition, serology suggested that other swine influenza viruses are also circulating in Vietnamese swine.     

Red mold dioscorea has greater hypolipidemic and antiatherosclerotic effect than traditional red mold rice and unfermented dioscorea in hamsters

Monascus-fermented red mold dioscorea (RMD) was proven to produce higher monacolin K levels than red mold rice (RMR) in our previous study. The goal of this study is to investigate whether the novel RMD had more hypolipidemic and antiatherosclerotic effect than traditional red mold rice. The daily dose of RMR for adults was recommended as 1 g, which corresponded to 96 mg/kg/day for hamsters. Therefore, high cholesterol diet-induced hyperlipidemic hamsters were daily administrated with a 0.5-fold (48 mg/kg/day), a 1-fold (96 mg/kg/day), or a 5-fold dose (480 mg/kg/day) of RMD for 8 weeks. Furthermore, a 1-fold dose of RMR (96 mg/kg/day) and unfermented dioscorea (96 mg/kg/day) were also respectively used to evaluate the effect of hypolipidemic and antiarteriosclerosis. The results indicated that only needing a 0.5-fold dose of RMD was able to significantly lower total cholesterol (by 13.78%, p<0.001), triglyceride (by 38.74%, p<0.01), and low-density lipoprotein cholesterol levels (by 43.11%, p<0.05) as well as maintain a high-density lipoprotein cholesterol level, as compared to the hyperlipidemic group. RMD including a higher monacolin K level and a dioscorea substrate was able to exhibit a more significant difference in the hypolipidemic effect than RMR or unfermented dioscorea. Both RMR and dioscorea exhibited potent in vitro antioxidative ability and in vivo protection against hypolipidemia-induced oxidative stress. Therefore, the antioxidative ability of RMD provided by Monascus metabolites (dimerumic acid, tannin, phenol, etc.) as well as dioscorea was able to perform more antiatherosclerotic effects on increasing total antioxidant status, catalase, and superoxide dismutase activity and repressing lipid peroxidation and atherosclerotic plaque than RMR and dioscorea.     

Improving the ratio of monacolin K to citrinin production of Monascus purpureus NTU 568 under dioscorea medium through the mediation of pH value and ethanol addition

Using dioscorea root as substrate of Monascus species was found to stimulate monacolin K (cholesterol-lowering agent) formation in our previous study, but the mycotoxin-citrinin has never been studied. This study used dioscorea root as the liquid medium to culture Monascus purpureus NTU 568 using a 6.6 L jar fermentor. Culture pH value, dioscorea concentration, and ethanol concentration were used as the factors of response surface methodology (RSM) to investigate the optimal culture condition for high monacolin K production and low citrinin formation. Monacolin K and citrinin formation of M. purpureus NTU 568 under submerged dioscorea medium were respectively found to be significantly formed by 148% and 147%, as compared to that under submerged rice medium. The reason is due to the pH value (3.5) of dioscorea medium involved in the formation of Monascus cell amount and secondary metabolite. RSM results further indicated that lowering the pH value to 2.5 would result in high monacolin K and citrinin concentrations as well as high biomass in fixed dioscorea amount, implying that pH value may stimulate the formation of monacolin K and citrinin through increasing Monascus cell amount. Lowering dioscorea and ethanol concentration was able to increase the ratio of monacolin K level to citrinin level. The optimal culture condition (pH 5.7, 1% dioscorea concentration, and 0.5% ethanol concentration) would increase monacolin K levels to 27.9 mg/g (by 47%) and decrease citrinin level to 2.15 microg/g (by 54%), as compared to control conditions (pH 3.5, 5% dioscorea, and ethanol free).     

Dietary nano-chromium tripicolinate increases feed intake and decreases plasma cortisol in finisher gilts during summer

Chromium (Cr) is an essential mineral element and has been used in pig diets to improve growth performance, insulin sensitivity, immune response and carcase traits and to reduce heat or other stress responses. The aims of thiss study were to determine the impact of nano-sized chromium tripicolinate (nCrPic) on growth performance, feed efficiency and carcase characteristics of finisher gilts during the summer period. A total of 60 finisher Large White x Landrace gilts were stratified on initial weight and then within strata randomly allocated into two treatment groups in three replicates during mid-summer for 28 days. All pigs were housed in individual pens and had ad libitum access to feed and water. Pigs were fed either a control finisher diet (wheat-based diet containing 13.8 MJ digestible energy (DE) per kilogram and 0.56 g available lysine/MJ DE) or a control diet containing 400 ppb Cr as nCrPic. Dietary nCrPic supplementation increased feed intake by 6 % over the entire study (P?=?0.05). In particular, dietary nCrPic increased average daily feed intake (ADFI) by 8 % (P?=?0.02) during the final 2 weeks of the study. Moreover, dietary nCrPic tended to improve average daily feed (ADFI) over the entire study (P?=?0.09). However, there were no significant effects of nCrPic on feed conversion ratio (FCR), final weight, hot standard carcase weight (HCWT), P2 depth or dressing percentage. Plasma cortisol was decreased by 25 % (P?=?0.06) by dietary nCrPic supplementation. However, there were no effects of nCrPic on plasma glucose, insulin and nonesterified fatty acids (NEFA), might because of the higher feed intake. In conclusion, this study demonstrates that dietary nCrPic supplementation at 400 ppb can increase feed intake in finisher gilts during mid-summer, suggesting that nCrPic can ameliorate some of the negative effects of heat stress in pigs, possibly via decreased of circulating cortisol.     

Histological and histochemical changes in the digestive tract of white sturgeon larvae during ontogeny

Ontogenetic changes in digestive tract histology and digestive enzyme histochemistry were investigated 11 to 36 days post-hatch in white sturgeon Acipenser transmontanus larvae. From initiation of exogenous feeding (12 days post-hatch), larvae were fed a commercial salmonid diet for the ensuing 24 days. The digestive system of white sturgeon displayed a high degree of morphologic organization and functionality at the onset of exogenous feeding. An enhancement of digestive capacities occurred with transition to active feeding. On day 2 of feeding, there was a clear increase of alkaline phosphatase, aminopeptidase M, dipeptidyl peptidase IV, and -glutamyl transpeptidase activity in the brush border of the spiral intestine. This strong activity is an apparent confirmation of the importance of this segment of the intestine for protein digestion and nutrient absorption. The functional development of the pyloric intestine occurred on day 4 and was concomitant with an increase in the activity of brush border and cytoplasmic enzymes such as acetylcholinesterase, dipeptidyl peptidase II, - and -galactosidases. The absence of acetylcholinesterase, lactase, nonspecific esterase, and weak activity of exopeptidases and alkaline phosphatase in the anterior intestine suggests that this segment of the intestine may be less important in nutrient absorption than the pyloric and spiral intestines. The observed quantitative and qualitative differences in enzyme activity along the intestine indicate a high degree of specialization of each segment for specific digestive and absorptive processes.