Influence of E1-deleted recombinant adenoviruses on B7.1 and IL-2 expression in C1498 cells

BACKGROUND: Knowing that adenoviral vectors could initiate innate immunity, the ability of E1-deleted recombinant adenovirus (Ad-E1Delta) in induction of B7.1 and IL-2 molecules was studied. METHODS: The expression of green fluorescent protein in C1498 cells following transfection of these cells with adenovirus green fluorescent protein vector confirmed the ability of adenovirus vectors in infecting the cells and inducing the expression of the gene of interest. The expression of B7.1 molecule on the surface of the cells was assayed upon infection with Ad-E1Delta vector. Adenovirus-IL-2/B7.1 vector capable of inducing IL-2 and B7.1 expression in the cells was used as the positive control vector. RESULTS: According to the FACS results, about 4.17% of normal cells expressed B7.1 on their surface, while this level was increased in Ad-E1Delta transduced cells up to 14.43%. These results demonstrate that Ad-E1Delta vector considerably (about 3 folds) increases the expression of B7.1 on the cells. No detectable IL-2 was secreted into the medium of non-transduced and Ad-E1Delta transduced cells. CONCLUSION: Data indicate that the infection of C1498 cells with recombinant adenoviruses stimulates expression of B7.1 on the cell surface rather than secretion of IL-2 into the medium.     

A QTL on chromosome 2DS of ‘Sumai 3’ increases susceptibility to <Emphasis Type="Italic">Fusarium</Emphasis> head blight in wheat

Much effort has been invested in identifying molecular markers in wheat (Triticum aestivum L.) linked to quantitative trait loci (QTL) that confer resistance to Fusarium head blight (FHB) caused by Fusarium graminearum Schwabe [teleomorph Gibberella zeae (Schwein) Petch]. Even after several generations of crossing and selection by many wheat breeding programs, resistance of the Chinese spring wheat cultivar ‘Sumai 3’ (PI 481542) remains among the most effective. It therefore seems that undocumented resistance QTL present in Sumai 3 were not detected in various mapping studies. Using an extremely susceptible Tibetan landrace (‘Y1193-6’; unknown pedigree) in the creation of a mapping population with Sumai 3, the objective of this research was to identify undocumented resistance QTL in Sumai 3. This was accomplished through collecting disease index (DI) and Fusarium damaged kernel (FDK) phenotypic values along with 305 Diversity Array Technology (DArT) and 52 Simple Sequence Repeat (SSR) marker genotypes on 160 F_2:6 recombinant inbred lines (RILs). Disease response evaluations were based on four (two greenhouse and two field) experiments where spray inoculation methods were used. Three QTL were identified on chromosome arms 3BS, 6BL and 2DS explaining 26.1, 10.7 and 18.9% of the phenotypic variation for DI, respectively. The same QTL were also significantly associated with reduced FDK scores and explained 28.0, 11.0 and 23.0% of phenotypic variation. Lines within the mapping population were placed in eight categories with respect to their various QTL combinations. Lines with no QTL were the most susceptible, whereas those with the Sumai 3-derived 3BS and 6BL QTL combined with the 2DS QTL from Y1193-6 were the most resistant. Though the 3BS and 6BL QTL are well-documented, the 2DS resistance QTL, which was contributed by the susceptible parent, confers increased susceptibility when derived from Sumai 3. In this study no new FHB QTL from Sumai 3 was discovered, but results suggest that Sumai 3 contains a QTL for susceptibility on chromosome arm 2DS. Selection against this QTL may potentially increase resistance levels among Sumai 3-derived populations.     

Risk factors for <Emphasis Type="Italic">Mycobacterium avium</Emphasis> subspecies <Emphasis Type="Italic">paratuberculosis</Emphasis> in Fars province (Southern Iran) dairy herds

A cross-sectional study was conducted from March to August 2006 in dairy herds in Fars province, southern Iran to determine the herd-level risk factors for infection with Mycobacterium avium subspecies paratuberculosis (MAP). Statistical analysis using multivariable logistic regression showed that contamination of udders of periparturient cows with manure (OR = 6.4, P = 0.02) and history of having suspected cases of Johne's disease in the herd (OR = 6.7, P = 0.04) were significantly associated with the herd infection status. No relationship between breed, herd size and other management practices with the infection status of the herd were found in this study. Implementing high sanitary measures in the farm, particularly with respect to manure handling and cleaning could be considered as one of the important aspects in controlling disease in the region as well as in the future educational effort.     

Effects of administration of two growth hormone-releasing hormone plasmids to gilts on sow and litter performance for the subsequent three gestations

Objective-To determine whether a novel optimized plasmid carrying the porcine growth hormone-releasing hormone (GHRH) wild-type cDNA administered at a lower dose was as effective at eliciting physiologic responses as a commercial GHRH plasmid approved for use in Australia. Animals-134 gilts. Procedures-Estrus was synchronized and gilts were bred. Pregnant gilts were assigned to 2 treatment groups (40 gilts/group) or 1 untreated control group (24 gilts). Gilts in one of the treatment groups received the commercial GHRH plasmid, whereas gilts in the other treatment group received a novel optimized GHRH plasmid; both plasmids were administered IM in the right hind limb, which was followed by electroporation. Sow and litter performance were monitored for the 3 gestations after treatment. Results-A significant increase in insulin-like growth factor-I concentrations, decrease in perinatal mortality rate, increase in the number of pigs born alive, and increase in the weight and number of pigs weaned were detected for both groups receiving the GHRH-expressing plasmids, compared with values for the control group. Additionally, there was a significant decrease in sow attrition in GHRH-treated females, compared with attrition in the control group, during the 3 gestations after treatment. Conclusions and Clinical Relevance-Both of the GHRH plasmids provided significant benefits for sow performance and baby pig survivability for pregnant and lactating sows and their offspring during the 3 gestations after treatment, compared with results for untreated control gilts. Use of a novel optimized plasmid reduced the effective plasmid dose in these large mammals.     

A novel approach to denitrification processes in a zero-discharge recirculating system for small-scale urban aquaculture

This paper presents an innovative process to solve the nitrate build-up problem in recirculating aquaculture systems (RAS). The novel aspects of the process lie in a denitrification bioreactor system that uses solid cotton wool as the primary carbon source and a unique degassing chamber. In the latter, the water is physically stripped of dissolved gaseous O₂ (by means of a Venturi vacuum tube), and the subsequent denitrification becomes more efficient due to elimination of the problems of oxygen inhibition of denitrification and aerobic consumption of cotton wool. The cotton wool medium also serves as a physical barrier that traps organic particles, which, in turn, act as an additional carbon source for denitrification. Operation in the proposed system gives an extremely low C/N ratio of 0.82 g of cotton wool/g of nitrate N, which contributes to a significant reduction of biofilter volume. The additional advantage of using solid cotton wool as the carbon source is that it does not release organic residuals into the liquid to be recycled. Operation of the system over a long period consistently produced effluents with low nitrate levels (below 10 mg N/l), and there was only a very small need to replace system water. The overall treatment scheme, also incorporating an aerobic nitrification biofilter and a granular filtration device, produced water of excellent quality, i.e., with near-zero levels of nitrite and ammonia, a sufficiently high pH for aquaculture, and low turbidity. The proposed system thus provides a solution for sustainable small-scale, urban aquaculture operation with a very high recovery of water (over 99%) and minimal waste disposal.     

From farm to fork--Mycobacterium avium ssp. paratuberculosis (MAP) as zoonotic agent?

Mycobacterium avium ssp. paratuberculosis (MAP) is the causative agent of the paratuberculosis (Para Tb) in ruminants. In addition, this pathogen has been suspected to be implicated in the pathogenesis of Morbus Crohn disease (MC), causing chronic inflammatory intestine changes of humans. The participation of MAP in this illness is discussed intensively and has very contradictory opinions. On the one hand several times succeeded in proving MAP DNA in changed human tissues as well as, in recent time, the bacteria has been isolated from patient's blood. On the other hand there are many publications which support the opposite opinion. In critical evaluation of already available data, therefore the hypothesis can be formulated that MAP could possibly take part in the MC of humans. The reliable verification of this hypothesis will only be possible, if the diagnostic procedures can be refined upon the substantial deficit concerning the sensitivity and/or specificity of the diagnostic methods. In addition, till now there is lack of optimized statistically case control studies. The conceivable transmission of the bacteria to humans by the direct animal contact has been considered as possible vector, furthermore, MAP has been detected in pasteurised milk and other food of animal origin. The prevalence data, usually estimated by ELISA for milk cattle stock show over 80% prevalence in many counties of the Federal Republic of Germany with an individual case prevalence ranging between 1% and 17% in different stocks. Comparable data are present also from other countries as well as for small ruminants. MAP has been concerned as a global problem, moreover the high spreading rate of MAP in wild animal populations as well as the considerable ability of the bacteria to survive in different stages of the infectious- and contamination-cycle, which might hardly be broken through. Thus it requires intensive research efforts for the development of the methodical diagnostic process as basis for valid epidemiological investigations of animals, humans and food.     

Transmission of sheep scrapie to elk (Cervus elaphus nelsoni) by intracerebral inoculation: final outcome of the experiment.

This is a final report of an experimental transmission of sheep scrapie agent by intracerebral inoculation to Rocky Mountain elk (Cervus elaphus nelsoni). It documents results obtained in experimental (n = 6) and control (n = 2) elk. During the first 2 years postinoculation (PI), 3 animals died or were euthanized because of infection or injuries other than spongiform encephalopathy (SE). In years 3 and 4 PI, 3 other inoculated elk died after brief terminal neurological episodes. Necropsy of these animals revealed moderate weight loss but no other gross lesions. Microscopically, characteristic lesions of SE were seen throughout the brain and spinal cord, and the tissue was positive for proteinase K-resistant prion protein (PrPres) by immunohistochemistry (IHC) and by Western blot. Scrapie-associated fibrils (SAF) were observed by negative-stain electron microscopy in the brain of elk with neurologic signs. PrPres and SAF were not detected in the 3 inoculated elk necropsied during the first 2 years or in the 2 control animals. Retrospective analysis of the gene-encoding cervid PrP revealed a polymorphism at codon 132. The elk with SE were either homozygous (MM) or heterozygous (LM). These findings confirm that intracerebral inoculation of sheep scrapie agent results in SE with accumulations of PrPres in the central nervous system of elk. Based on morphologic and IHC findings, the experimentally induced SE cannot be distinguished from chronic wasting disease of elk with currently available diagnostic techniques.     

Experimental transmission of chronic wasting disease agent from mule deer to cattle by the intracerebral route.

This communication reports final observations on experimental transmission of chronic wasting disease (CWD) from mule deer to cattle by the intracerebral route. Thirteen calves were inoculated intracerebrally with brain suspension from mule deer naturally affected with CWD. Three other calves were kept as uninoculated controls. The experiment was terminated 6 years after inoculation. During that time, abnormal prion protein (PrP(res)) was demonstrated in the central nervous system (CNS) of 5 cattle by both immunohistochemistry and Western blot. However, microscopic lesions suggestive of spongiform encephalopathy (SE) in the brains of these PrP(res)-positive animals were subtle in 3 cases and absent in 2 cases. Analysis of the gene encoding bovine PRNP revealed homozygosity for alleles encoding 6 octapeptide repeats, serine (S) at codon 46, and S at codon 146 in all samples. Findings of this study show that although PrP(res) amplification occurred after direct inoculation into the brain, none of the affected animals had classic histopathologic lesions of SE. Furthermore, only 38% of the inoculated cattle demonstrated amplification of PrP(res). Although intracerebral inoculation is an unnatural route of exposure, this experiment shows that CWD transmission in cattle could have long incubation periods (up to 5 years). This finding suggests that oral exposure of cattle to CWD agent, a more natural potential route of exposure, would require not only a much larger dose of inoculum but also may not result in amplification of PrP(res) within CNS tissues during the normal lifespan of cattle.     

A possible involvement of gibberellin in the mechanism of Avenafatua resistance to triallate and cross-resistance to difenzoquat

Avena fatua L. populations from numerous fields in Alberta have simultaneously developed resistance to recommended rates of two dissimilar herbicides, triallate and difenzoquat. We used exogenous applications of gibberellic acid (GA₃) to investigate the possibility that endogenous gibberellins were involved in the A. fatua 's resistance mechanism. For control plants, without applied GA₃, shoots of the most resistant (R) populations emerged more rapidly and elongated faster than shoots of the susceptible (S) populations. Increases in shoot elongation in response to exogenous GA₃ were significantly lower in R populations compared with S populations. This suggests that R populations may have elevated endogenous gibberellin levels, relative to S populations. Additionally, inhibition of S population shoot elongation and shoot anatomical abnormalities caused by relatively low concentrations of triallate and difenzoquat could be prevented by exogenous application of GA₃. These results suggest that there may be a phytohormonal involvement in the mechanism of triallate/difenzoquat resistance in A. fatua . That is, higher endogenous gibberellin levels in R populations may result in meristematic growth that is rapid enough to preclude phytotoxic levels of these herbicides from reaching the shoot meristem, which is the likely site of action.     

Management of root-knot disease of tomato by the application of fly ash in soil

The effects of fly ash at different concentrations (0, 10, 20, 30 … 100% vol./vol. in soil) on plant growth and yield were investigated in tomato plants infected or noninfected with root-knot nematode, Meloidogyne incognita (2000 juveniles per plant) in clay pots. An increase in fly ash concentration in the soil correspondingly increased the availability of carbonates, bicarbonates, sulphate, chlorides, B, P, K, Ca, Mg, Mn, Cu and Zn in the soil. The porosity, water-holding capacity, pH, conductivity and cation exchange capacity also increased progressively in the fly ash amended soil. Ash application enhanced plant growth, leaf pigment concentrations, fruit production, weight of fruit/plant and mean fruit weight of both nematode-infected and noninfected tomato plants, being maximum in the soil containing 50 or 60% fly ash. The yield enhancements were 93.6 (infected plants) and 84% (noninfected plants) at 50 and 60% fly ash levels, respectively. Fly ash treatments adversely affected root invasion by juveniles, disease intensity and reproduction of the nematodes. A gradual increase in the ash concentration in soil caused a corresponding decrease in the numbers of invading juveniles, galls and egg masses per root system and eggs/egg mass, being lowest at 100% fly ash i.e. 52, 16, 10 and 81 against 289, 137, 131 and 238 (control), respectively. Linear regression suggested 40% fly ash as the most economic level, enhancing yield of infected plants by 96% and suppressing the nematode disease and reproduction by 63 and 76%, respectively.