Characterization of infectious bursal disease viruses isolated in 2007 from Delmarva commercial broiler chickens

A study was performed in 2007 to isolate and characterize infectious bursal disease viruses (IBDVs) in commercial broilers grown in the Delmarva (DMV) Peninsula region of the United States. Bursae of Fabricius were collected weekly from 1 to 4 wk of age from broilers on 10 farms with a history of poor performance. Microscopic pathology was used to determine the infectious bursal disease (IBD) status of the broilers. Bursae from 1- and 2-wk-old broilers did not show IBD microscopic lesions. Moreover, broilers on 1 of the 10 farms were IBD lesion free at 3 and 4 wk of age. However, 3 of 9 and 9 of 9 farms yielded broilers with IBD-affected bursae from 3- and 4-wk-old commercial broilers, respectively. Ten IBDV isolates were recovered from 3 of 3 lesion-positive bursal pools at 3 wk of age and 7 of 9 lesion-positive bursal pools at 4 wk of age. Analysis of the viral protein (VP) 2 genes identified all isolates as serotype 1 Delaware (Del) variant viruses. Five field isolates, each representing different molecular clades of the Delaware variant viruses, were selected for further study. Experimental infection of specific-pathogen-free white leghorn chickens with isolates DMV/4813/07, DMV/4947/07, DMV/4955/07, DMV/5038/07, and DMV/5041/07 produced gross and microscopic pathology of the bursa consistent with Delaware variant infection. Monoclonal antibody testing showed DMV/4813/07, DMV/4947/07, DMV/ 4955/07, and DMV/5041/07 to be similar to previous recognized variant viruses. However, DMV/5038/07 was found to be unreactive with the monoclonal antibodies that typically recognize reference strains STC, Del E, GLS, RS593, and AL2. In a challenge of immunity study, 10-day-old progeny from breeders immunized with a commercially available inactivated IBDV vaccine containing the Del E and classic strains were protected to a lesser degree against isolate DMV/5038/07 compared to Del E challenge based on microscopic lesion scores (P < 0.01) of the bursa. This result suggests the virus is antigenically different from the Del E strain contained in the vaccine. Collectively, the monoclonal antibody and progeny challenge of immunity findings suggest DMV/5038/07 is antigenically different from the Del E strain contained in the vaccine.     

Bleeding canker of horse chestnut (Aesculus hippocastanum) in Ireland: incidence,severity and characterization using DNA sequences and real‐time PCR

A survey of bleeding canker disease, caused by Pseudomonas syringae pv. aesculi, was undertaken across Ireland. Incidence has become severe and can be considered epidemic, as 61% of the 1587 horse chestnut trees surveyed showed symptoms of the disease. Bacteria were isolated from a sample of trees and characterized using gyrBDNA sequencing. DNA was also extracted directly from wound tissue. The Irish P. syringae pv. aesculi genotype was identical to genotypes previously sequenced with gyrB from the UK and some other locations in Europe. Real‐time PCR, using existing primers and a newly designed, more pathovar‐specific primer set, was assessed for use in disease screening. With molecular screening, a total of 11 trees from a sample of 55 tested positive for P. syringae pv. aesculi in Ireland. It was more efficient to extract DNA directly from wound tissue, especially fresh bark, for disease detection than to undertake bacterial isolation with subsequent molecular analysis. A further set of sequencing primers was developed for the amplification of the gyrB gene from P. syringae pv. aesculi and their specificity was shown using a diverse sample of bacterial isolate DNAs. The study also isolated and identified other bacterial species from diseased material; some of these are known pathogens (Brenneria nigrifluens, P. marginalis and P. syringae) or have previously been identified as potentially beneficial endophytes of host trees (Erwinia billingiae, E. tolentana, P. fluorescens, P. putida and Raoultella).     

Effects of cultural methods and physical weed control on intrarow weed numbers, manual weeding and marketable yield in direct-sown leek and bulb onion

Manual weeding of intrarow weeds in direct-sown leek and bulb onion crops grown organically can be very labour-intensive. Four field experiments, two in direct-sown leek and two in direct-sown bulb onion were made in this investigation to study the effects of physical and cultural methods on intrarow weed numbers, time consumption for hand-weeding and marketable yield parameters. The physical methods considered were: pre-emergence flaming and harrowing, and post-emergence hoeing close to the row (only leek) and vertical brush weeding. The cultural methods were: seed priming, slurry placement and cultivar choice (only leek). Generally, flaming plus brush weeding gave the highest intrarow weed control at 92% and 87%, respectively, in the two leek experiments and 39% (only brush weeding was effective) and 74%, respectively, in the two onion experiments. Time consumption for hand-weeding after the different treatments was linearly related to the remaining numbers of intrarow weeds, with no significant influences of the experimental factors on the general relationship. Generally, the cultural methods had no significant influence on the effects of physical weeding in terms of their effect against intrarow weeds. They did not affect the tolerance or robustness of the crop plants against negative impact from the physical control methods. However, generally, seed priming and cultivar choice did improve yield in the leek experiments and seed priming also did so in the one experiment with onion.     

Parasites of domestic and wild animals in South Africa. XXXVIII. Ixodid ticks collected from 23 wild carnivore species

Ixodid ticks were collected from 104 wild carnivores belonging to 23 species in various nature reserves and on several farms in all nine provinces of South Africa. Seven feral cats in a nature reserve were also examined. Twenty-four tick species belonging to seven genera were recovered and identified. Amongst these ticks we consider the adults of Haemaphysalis leachi, Haemaphysalis spinulosa, Haemaphysalis zumpti, Ixodes rubicundus, Rhipicentor nuttalli, Rhipicephalus simus and Rhipicephalus turanicus to be true parasites of wild carnivores. Although numerous adult Rhipicephalus appendiculatus and Rhipicephalus zambeziensis were collected from some lions these were either sick or old animals. The immature stages of seven species regularly utilized wild carnivores as hosts on an opportunistic basis.     

Effects of treadmill exercise on cortical bone in the third metacarpus of young horses.

The effects of exercise and relative inactivity on cortical bone were compared in young horses. Two groups were used; one was given a 14-week programme of exercise (n = 6) and the other kept as unexercised controls (n = 6). The first nine weeks of exercise involved trotting and cantering (2 to 4 km d-1 at speeds up to 12 m s-1) on a treadmill set at an incline of 3 degrees. Over the next five weeks the horses were trained at near maximal speeds (that is, up to 14.5 m s-1) with no incline of the treadmill. At the end of the programme marked differences in cortical porosity and distribution of subperiosteal osteogenesis at the mid-shaft of the third metacarpal bone were found between the groups. Histomorphometrical examination of the dorsal cortex showed minimal bone remodelling in the exercised horses, but extensive modelling as evidenced by the large amount of subperiosteal bone formation. In contrast, the unexercised horses had significantly more bone remodelling and less formation of subperiosteal bone. The histomorphometric and microradiographic findings provided an explanation for changes in the non-invasive bone measurements that occurred during training. Bone mineral content of the mid-metacarpus was found to increase more in the exercised than the unexercised horses despite a lower overall growth in bodyweight. In those horses that completed the full training programme, ultrasound speed increased significantly by the end of the training programme. It remained unchanged in the horse that did not complete the full exercise programme and decreased slightly in the unexercised horses. The difference in ultrasound speed between the groups was considered to reflect differences in intracortical bone porosity, endosteal bone formation and alterations in skin thickness. The stiffness of cortical bone increased significantly in the exercised horses but remained unaltered in the unexercised horses.     

CO2-stunning of slaughter pigs: effects on EEG, catecholamines and clinical reflexes

Investigations were carried out on the response of slaughter pigs to CO2 stunning with two different gas concentrations (80 vol%, 90 vol%, 73 s) under practical conditions in a one gondola-dip-lift system. EEG measurements were performed and blood constituents such as catecholamines (adrenaline, nor-adrenaline) and lactate as well as clinical reactions (nasal septum and corneal reflex, heart beats) investigated. Special EEG-electrodes were adapted for the measurement on pigs and a mobile data logger was prepared for the use in the lairage of the slaughter house and in the stunning unit. The CO2 concentrations were measured continuously close to the head of the pigs when transported up and down in the stunning gondola. The results show that the technique is suited to monitor the effects of different CO2 gas concentrations on the EEG of the pigs under practical conditions. There is strong evidence that CO2 concentrations of 80 vol% applied over 70 s as required by law are not sufficient to stun pigs properly. A large part of the animals still showed typical reflexes when leaving the stunning pit. When an atmosphere of 90 vol% CO2 is applied, most animals are already dead before bleeding commences. This may create problems in respect to meat hygiene. The blood analysis revealed very high concentrations of catecholamines after stunning. The values for adrenaline and nor-adrenaline in the sticking blood rose by a factor of about 1000 as compared to the concentrations in blood samples taken in the lairage before stunning. It seems necessary to revise the current legislation on gas stunning and to look in greater detail in the effects of CO2 stunning on the welfare of slaughter pigs.     

Copper resistance genes from different xanthomonads and citrus epiphytic bacteria confer resistance to Xanthomonas citri subsp. citri

Genetic exchange is considered to be an important process in the selective adaptation of microorganisms to shifting and challenging environmental conditions. As a consequence of the copious use of copper bactericides, many species of plant pathogenic bacteria, including Xanthomonas citri subsp. citri (Xcc), have developed resistance to copper. This study assesses whether copper resistant (Cu^R) strains of other Xanthomonas species and citrus epiphytic bacteria pose a risk for the development of copper resistance in Xcc. Cu^R epiphytic bacteria were isolated on MGY agar from citrus leaves collected in two citrus groves treated with copper bactericides in Florida. Horizontal gene transfer of copper resistance genes was investigated within different Xanthomonas species and from citrus epiphytic bacteria to Xanthomonas. Cu^R epiphytic bacteria from citrus were screened for the presence of copper resistance genes homologous to copL, copA and copB genes from Xcc and characterized regarding tolerance to copper. Copper resistance determinants from a citrus epiphytic strain of Stenotrophomonas maltophilia (Stm) were cloned and expressed in Xcc and other Xanthomonas strains. Copper resistance genes in Xcc were determined to be present on a large (~300?kb) conjugative plasmid. Cu resistance was transferred via conjugation from two copper resistant citrus strains, Xcc and X. alfalfae subsp. citrumelonis (Xac), and two tomato pathogens, X. euvesicatoria (Xe) and X. perforans (Xp), to Xcc. PCR analysis revealed that two Cu^R strains from citrus, an epiphytic Xanthomonas ssp. and a strain of Stm, harboured homologs of the copper resistance genes found in Cu^R Xcc. The introduction of copLAB gene cluster from Stm into different xanthomonads conferred copper resistance to sensitive strains of Xcc, Xac, Xe and Xp. Based on these results there is a low, but significant, likelihood of horizontal gene transfer of copper resistance genes from other xanthomonads or epiphytic bacteria to Xcc in nature.