Effect of cytolethal distending toxin of Campylobacter jejuni on adhesion and internalization in cultured cells and in colonization of the chicken gut

Campylobacter jejuni produces cytolethal distending toxin (CDT) that causes host cells to arrest during their cell cycle and that is involved in the pathogenesis of inflammatory diarrhea in humans. To assess the role of CDT in adherence and invasion of different cultured host cells (HeLa and HD-11) and in colonization of the chicken intestine, the genes of C jejuni NCTC11168 encoding the toxin subunits (cdtA, cdtB, and cdtC) were inactivated by insertional mutagenesis. No significant difference was found in adhesion of the wild-type C. jejuni and the isogenic mutants to HeLa and HD-11 cells. All of the mutants exhibited a decrease (>10-fold) in the ability to invade HeLa cells, but no significant difference was noticed for HD-11 cells. The ability of mutants to colonize birds either directly or by horizontal transfer was unchanged. These data indicated that although the production of cytotoxin does not play a role in the adherence to either human or avian cells, it may play a role in the invasion, survival, or both of C. jejuni in human cells, which are more susceptible to C. jejuni internalization. The CDT also does not seem to play a role in the colonization of poultry.     

An assessment of the duration of Mycoplasma hyopneumoniae infection in an experimentally infected population of pigs

Mycoplasma hyopneumoniae (M. hyopneumoniae) is the primary pathogen of enzootic pneumonia (EP), a highly prevalent respiratory disease that affects pigs worldwide. Previous studies have demonstrated that M. hyopneumoniae infection can be longer than 185 days; however, the total duration of infection has not been determined yet. Therefore, the objective of this study was to determine the duration of M. hyopneumoniae infection in asymptomatic carriers. To achieve our goal, 60 pigs were inoculated with M. hyopneumoniae strain 232 and the persistence of M. hyopneumoniae in the respiratory tract was assessed by detection of the bacterial DNA in bronchial swabs and the ability of the infected pigs to transmit the pathogen to sentinels. Infection of the inoculated animals was demonstrated by the detection of M. hyopneumoniae DNA in nasal swabs, seroconversion to the bacteria and the onset of dry coughing. Experimentally infected pigs shed M. hyopneumoniae prior to and after the cough was observed. M. hyopneumoniae DNA was detected in 100% of experimentally infected pigs at 94 days post infection (dpi), 61% at 214dpi and 0% at 254dpi. Experimentally infected pigs transmitted the bacteria to sentinels at 80 and 200dpi. Results of this study have demonstrated that M. hyopneumoniae infected pigs can be incubatory as well as convalescent carriers of the pathogen and that convalescent carriers can remain infectious for up to 200 days. Total clearance of M. hyopneumoniae in the group was evidenced, demonstrating that duration of M. hyopneumoniae infection lasts less than 254 days.     

Alteration of soil water content consequent to root-pruning at a windbreak/crop interface in Nebraska, USA

Root-pruning is generally recommended as an appropriate treatment to reduce competition for soil water and/or nutrients and suppression of crop yield in areas adjacent to windbreaks. Several recent studies suggest, however, that factors other than soil water might be causing yield reduction at the interface. For two consecutive years, we evaluated root-pruning effects on soil water at the windbreak/crop interface under both cropped (soybean [Glycine max (L) Merr.] variety Iroquois, 1997) and non-cropped (1998) conditions in Mead, Nebraska, USA. Volumetric soil water content near the windbreaks was systematically measured at various soil depths, distances from the windbreak, and windbreak exposures using Time Domain Reflectometry (TDR). Overall differences in soil water content between root-pruned and non-pruned plots in soybean were smaller in magnitude at all distances in both the west (windbreak on the east side) and the east (windbreak on the west side) exposures in 1997, compared with the non-cropped condition in the south exposure in 1998. With a soybean crop in 1997, volumetric soil water content in the east exposure averaged 2.3% greater in the top 30 cm of the soil profile at a distance of 0.75H (H = windbreak height) into the field from the windbreak when compared to the non-pruned treatment. In the west exposure, however, the differences were undetectable at corresponding distance and depth. The increase in soybean yield in root-pruned plots corresponded well with the observed differences in soil water content at various distances, especially in the east exposure. Under a non-cropped condition in 1998, soil water content in the root-pruned plots was significantly greater than the non-pruned plots in the top 45-cm profile, averaging 3.3% at 0.75H and 2.2% at 1.0H. Beyond 1.0H, the increase was not significant. These results agree with the previously reported range of crop yield suppression near windbreaks, indicating that soil water competition between the crop and windbreak is highly related to, and probably plays a leading role in yield suppression within the competition zone.This revised version was published online in November 2005 with corrections to the Cover Date.     

Methods of Concentrating Stallion Semen

Removal of excess seminal plasma is sometimes necessary to increase the quality and the longevity of cooled equine semen; moreover, this procedure is an indispensable step aiming to concentrate the sperm cells before freezing equine semen. Typically, the removal of seminal plasma is achieved by centrifugation; however, studies have shown that the force and duration of centrifugation can damage sperm cells and reduce the sperm recovery rate. Recently, new methodologies, such as cushion and filtration, have been described that aim to decrease the mechanical damage of centrifugation to sperm cells. This study aims to compare different methods for concentrating stallion semen.     

Temporal dynamics of root and foliar severity of soybean sudden death syndrome at different inoculum densities

Temporal dynamics of soybean sudden death syndrome (SDS) root and foliar disease severity were studied in growth chamber experiments on susceptible plants exposed to different inoculum densities (0, 10⁰, 10¹, 10², and 10³ conidia g⁻¹ soil) of Fusarium virguliforme. The monomolecular model provided the best fit to describe the progress of root and foliar disease severity over time. Disease severity and area under disease progress curve (AUDPC) both increased in response to increasing inoculum density (P < 0.01), particularly for foliar symptoms. Rate of disease progress increased as inoculum densities increased for both root and foliar disease severities. The incubation period for root and foliar disease severity ranged from 9 to 18 and 15 to 25 days, respectively. Significant differences in root rot severity were most easily detected during the early stages of infection, whereas root rot and foliar severities were only weakly correlated when both were assessed simultaneously at later stages of disease development. Root rot severity assessments performed 15 to 20 days after inoculation (DAI) were most highly correlated (r > 0.9, P < 0.01) with foliar disease severity assessments performed 30 to 50 DAI. Root biomass was reduced by up to 67% at the three highest inoculum densities, indicating the aggressiveness that F. virguliforme possesses as a root rot pathogen on soybeans.     

Finite element modeling for development and optimization of a bone plate for mandibular fracture in dogs

This study aimed to develop a plate to treat fractures of the mandibular body in dogs and to validate the project using finite elements and biomechanical essays. Mandible prototypes were produced with 10 oblique ventrorostral fractures (favorable) and 10 oblique ventrocaudal fractures (unfavorable). Three groups were established for each fracture type. Osteosynthesis with a pure titanium plate of double-arch geometry and blocked monocortical screws of free angulation were used. The mechanical resistance of the prototype with unfavorable fracture was lower than that of the favorable fracture. In both fractures, the deflection increased and the relative stiffness decreased proportionally to the diminishing screw number The finite element analysis validated this plate study, since the maximum tension concentration observed on the plate was lower than the resistance limit tension admitted by the titanium. In conclusion, the double-arch geometry plate fixed with blocked monocortical screws has sufficient resistance to stabilize oblique fractures, without compromising mandibular dental or neurovascular structures.     

Can spleen aspirations be safely used for the parasitological diagnosis of canine visceral leishmaniosis? A study on assymptomatic and polysymptomatic animals

The purpose of this study was to evaluate the safety of spleen aspiration as a sampling technique for the parasitological detection by culture and microscopy of Leishmania (chagasi) infantum. Two hundred and nine domiciled dogs from an endemic area for visceral leishmaniasis in Bahia State, Brazil, were studied. Most dogs (87%) were seropositive for anti-L. chagasi antibodies by ELISA. Clinical signs of disease were recorded and the animals monitored during and after spleen puncture in order to detect possible complications associated with the procedure. From a total of 257 splenic punctures in the 209 animals, only three minor events occurred, with no significant consequence for the animals and no association with risk factors. Leishmania was isolated from 149/180 (83%) seropositive dogs, and from 6/26 (23%) seronegative animals. The procedure did not cause adverse side effects or unnecessary suffering and confirmed the diagnosis in a large percentage of dogs. We conclude that spleen aspiration can be considered an effective and safe procedure for the definitive diagnosis of canine visceral leishmaniosis.     

Electrophysiological and behavioural responses of Pityophthorus pubescens (Coleoptera: Scolytinae) to (E,E)‐α‐farnesene, (R)‐(+)‐limonene and (S)‐(−)‐verbenone in Pinus radiata (Pinaceae) stands in northern Spain

BACKGROUND: Some twig beetles in the genus Pityophthorus (Coleoptera: Scolytinae) may vector pitch canker disease Fusarium circinatum (Niremberg & O'Donnell) of Pinus spp. (Pinaceae). Because Pityophthorus pubescens (Marsh.) has been found to be associated with F. circinatum in the Basque Country (northern Spain), various experiments were conducted to assess the beetle's behavioural responses to (E, E)‐α‐farnesene, (R)‐(+)‐limonene and (S)‐(?)‐verbenone to develop a potential inhibitor to host attraction. These experiments comprise electroantennographic and double‐choice olfactometer tests, as well as field assays in Pinus radiata D. Don stands. RESULTS: Both sexes of P. pubescens showed similar electroantennographic responses to different doses (from 1 ng to 1 µg in decadic steps) of each individual compound, with depolarisations to (S)‐(‐)‐verbenone (100 ng) being similar to those of the aggregation pheromone (+)‐trans‐pityol. In olfactometer assays, both sexes were significantly attracted to (+)‐trans‐pityol, but the attraction was reduced when increasing amounts of the chemicals were added to the pheromone. Particularly relevant was the repellent effect induced by (S)‐(?)‐verbenone at 1 ng dose and higher. In the field, (E, E)‐α‐farnesene, (R)‐(+)‐limonene and (S)‐(?)‐verbenone reduced significantly the number of beetles attracted to (+)‐trans‐pityol and racemic trans‐pityol, with (S)‐(?)‐verbenone being the most effective. CONCLUSIONS: (S)‐(?)‐Verbenone showed an interesting potential for use in the protection of P. radiata stands. A potentially effective strategy, which could be implemented in further, more in‐depth studies, could involve the use of this semiochemical as repellent and (+)‐trans‐pityol‐baited traps as attractant in a ‘push‐pull’ strategy. Copyright © 2012 Society of Chemical Industry     

Demography of <Emphasis Type="Italic">Conyza bonariensis</Emphasis> (Asteraceae) in a ruderal Mediterranean habitat

Conyza bonariensis is an invasive weed of increasing importance in subtropical and warm-temperate regions worldwide, both in non-agricultural habitats and in annual and perennial crops, especially under no-till management. To gain insigths on basic life cycle processes determining the demographic success of C. bonariensis, we studied for this species during two seasons seedling emergence patterns, survival to the adult stage and fecundity in a ruderal Mediterranean habitat in which C. bonariensis was a component of the plant community. The influence of emergence date on survival and fecundity was studied using four successive sowing dates, i.e. cohorts, encompassing the favorable season for plant establishment. The mean rate of seedling emergence was 61%. Emergence patterns were characterized by high initial emergence rates, which were highly dependent on rainfall. The mean rate of survival to the adult stage was 33%. Fecundity reached a mean value of 86,066 achenes and presented density-dependent regulation.. Plant survival and fecundity were positively related to cohort earliness and thus earlier cohorts should preferably be targeted for an effective management of C. bonariensis.     

Genetic Structure of Field Populations of Begomoviruses and of Their Vector Bemisia tabaci in Pakistan

ABSTRACT The genetic structure of field populations of begomoviruses and their whitefly vector Bemisia tabaci in Pakistan was analyzed. Begomoviruses and B. tabaci populations were sampled from different crops and weeds in different locations in Punjab and Sindh provinces, in areas where cotton leaf curl disease (CLCuD) occurs or does not occur. Phylogenetic analysis based on nucleotide sequences of the intergenic region in the viral DNA-A provided evidence of two clusters of isolates: viruses isolated from species in the family Malvaceae, and viruses isolated from other dicotyledon families. Analysis of the capsid protein (CP) open reading frame grouped isolates into three geographical clusters, corresponding to isolates collected in Punjab, Sindh, or both provinces. Random amplified polymorphic DNA analyses of the B. tabaci population showed that intrapopulation diversity was high at both the local and regional scales. Sequence analysis of the mitocondrial cytochrome oxydase I (mt COI) gene showed that the B. tabaci population was structured into at least three genetic lineages corresponding to the previously described Indian, Southeast Asian, and Mediterranean-African clades. The Indian clade was present only in Punjab, the Mediterranean-African only in Sindh, and the Southeast Asian in both provinces. B. tabaci haplotypes of the Indian clade were found only in the Punjab, where CLCuD occurs. Hence, the geographical distribution of virus and vector genotypes may be correlated, because similar phylogenetic relationships were detected for the viral CP and the vector mt COI genes.