Implementation of two high through-put techniques in a novel application: detecting point mutations in large EMS mutated plant populations

Background  

The establishment of mutant populations together with the strategies for targeted mutation detection has been applied successfully to a large number of organisms including many species in the plant kingdom. Considerable efforts have been invested into research on tomato as a model for berry-fruit plants. With the progress of the tomato sequencing project, reverse genetics becomes an obvious and achievable goal.     

Hard-setting behaviour of some structurally weak tropical soils

Some soils set hard on drying and may limit crop productivity. The hard-setting behaviour is influenced by soil properties and management. The objective of this study was to relate physical properties of soils sampled from 3 sites in Nigeria to soil strength or degree of hard-setting and to management. Clearing the vegetation and tillage led to a decrease in organic matter content and wet aggregate stability and increased bulk density leading to an increase in soil strength. The soil strength increased as the water content decreased but the increase was much more marked for mechanized cleared or tilled soils than for the less disturbed treatments of forestry or no-till cropping.     

Transendoscopic biopsy of the horse's airway mucosa

Transtracheal wash and bronchoalveolar lavage are diagnostic techniques that have been adopted from human medicine for monitoring inflammatory changes in the airway of the horse. Transendoscopic biopsy has also proven to be a valuable tool for obtaining samples of the airway mucosa in human patients. A transendoscopic technique was developed in this study for obtaining a respiratory mucosal biopsy from standing, sedated horses. Six normal adult horses were sampled at eight-week intervals for a total of three sample periods. Horses were monitored for adverse effects of the technique and none were noted. Sample sites were completely healed after eight weeks with no gross or histologic abnormalities. Biopsy samples were 3 to 4 millimeters in diameter, and 17 of 18 samples provided interpretable histological sections. Methods for handling, staining and evaluating tissue were also developed. The results of this study demonstrated that airway mucosal biopsy is a safe, repeatable technique that can be performed in the sedated, standing horse.     

Prevalence of enteropathogenic Escherichia coli in naturally occurring cases of poult enteritis-mortality syndrome

Enteropathogenic Escherichia coli (EPEC) previously were identified in poult enteritis-mortality syndrome (PEMS)-affected turkeys and associated as a cause of this disease. In the present study, the prevalence of EPEC in PEMS-affected turkeys was examined retrospectively with archived tissues and intestinal contents collected from 12 PEMS-affected turkey flocks in 1998. Formalin-fixed intestinal tissues were examined by light and electron microscopy for attaching and effacing (AE) lesions characteristic of EPEC, and frozen (-75 C) intestinal contents were examined for presence of EPEC. Escherichia coli isolates were characterized on the basis of epithelial cell attachment, fluorescent actin staining (FAS) test, and presence of E. coli attaching/effacing (EAE), shigalike toxin (SLT) type I, SLT II, and bundle-forming pilus (BFP) genes by polymerase chain reaction procedures. EPEC isolates were examined for pathogenicity and ability to induce AE lesions in experimentally inoculated young turkeys. AE lesions were identified by light microscopy in Giemsa-stained intestines from 7 of 12 PEMS-affected turkey flocks. Lesions consisted of bacterial microcolonies attached to epithelial surfaces with epithelial degeneration at sites of attachment and inflammatory infiltration of the lamina propria. Electron microscopy confirmed the identity of AE lesions in six of seven flocks determined to have AE lesions by light microscopy. EPEC were identified in 4 of 12 flocks on the basis of the presence of EAE genes a nd absence of SLT I and SLT II genes; all isolates lacked BFP genes. EPEC isolates produced AE lesions and variable mortality in turkeys coinfected with turkey coronavirus. In total, EPEC were associated with 10 of 12 (83%) naturally occurring PEMS cases on the basis of identification of AE lesions and/or EPEC isolates. These findings provide additional evidence suggesting a possible role for EPEC in the pathogenesis of PEMS.     

Reproductive Performance and Reconception of Nellore Cows According to Their Pure‐ or Cross‐Bred Calves

The objective of the present research was to evaluate the effect of a calf's genetic group on the productive and reproductive efficiency of its Nellore dam. Fixed‐time artificial insemination was applied to 800 cows using semen extracted from Nellore, Simmental and Angus Red bulls. Four hundred eleven cows calved, producing 119 Nellore, 103 ½Simmental–½Nellore and 189 ½Nellore–½Angus Red calves. The second mating period, which paired Nellore cows with Simmental bulls, was initiated 10 days after parturitions began and lasted for 5 months. Based on the two successive parturitions, the cumulative parturition rate for calving periods of 3, 4 and 5 months was calculated. Although no significant difference was observed for birth weight among the genetic groups, cross‐bred calves weighed, on average, 10% more than did pure‐bred calves at the age of 205 days. Nellore dams experienced a gestation period that was 7 days longer than did the cross‐bred dams, and the former showed a higher parturition rate at 90 and 120 days of the calving season, but not at 150 days (calving rates of 80.6, 76.4 and 76.2% for mothers of Nellore, ½Nellore–½Angus Red and ½Nellore–½Simmental, respectively, p > 0.05). At 90 and 120 days, Nellore dams produced more kg of calf per mated dam. In conclusion, in a short breeding season, Nellore dams nursing pure‐bred Nellore calves were found to have a higher biological efficiency compared with Nellore dams nursing cross‐bred calves.     

Hepatic emphysema associated with ultrasound-guided liver biopsy in a dog

An eleven-year-old Chinese Crested Powder Puff dog presented with polydipsia/polyuria, inappetence, diarrhea and vomiting underwent an ultrasound-guided percutaneous liver biopsy. Two days post-biopsy the clinical condition of the dog acutely deteriorated with fever, dyspnea, ataxia and subcutaneous emphysema. Radiographs and ultrasound showed focal severe hepatic emphysema in the region of the previous liver biopsy. Post-mortem examination revealed chronic hepatitis with dissecting fibrosis, acute hepatitis with hemorrhage and in the hindlimb musculature extensive hemorrhage and necrosis. Pure cultures of the gas producing bacteria Clostridium perfringens were isolated in samples from the hind limb musculature. We propose that the hepatic emphysema in the region of the biopsy site was a result of a clostridial infection.     

Physiological and biochemical consequences of electroimobilisation in conscious sheep

An electroimmobilisation device has been developed to facilitate the automated shearing of sheep, but there is little information on its effects on the body. We have studied its effects on the cardiovascular system and on intermediary metabolism in sheep. Electroimmobilisation caused statistically significant increases in mean arterial pressure, heart rate, cardiac output, renal and hepatic and hindquarter glucose and lactate flux, organ and whole body oxygen flux, hindquarter blood flow and core temperature and decreases in arterial and posterior vena cava blood pH, renal and hepatic blood flow and PaCO2. Notably, no change occurred in PaO2. The metabolic changes demonstrated the capacity of sheep to respond to the increased muscular and cardiovascular work induced by electroimmobilisation. Pulmonary function was not compromised during electroimmobilisation as judged from blood gas changes, and the acid/base changes were rapidly reversed after electroimmobilisation. The recovery to control conditions for all perturbations generally took no longer than 30 min, consistent with a rapid and physiologically adequate reversal by the animal's homeostatic mechanisms.     

Experimental infection of turkeys with Mycoplasma gallisepticum of low virulence, transmissibility, and immunogenicity.

Three-week-old turkeys were inoculated intranasally with approximately 10(6) colony-forming units (CFU) of putative variant Mycoplasma gallisepticum (MG) strains M876, M35, or the virulent S6 reference strain. Uninoculated turkeys in each group served as contact sentinels. The hemagglutination-inhibition (HI) test and enzyme-linked immunosorbent assay (ELISA) were used to determine serologic responses. MG was isolated from 100% and 92% of S6- and M876-inoculated turkeys, respectively, on day 7 PI. However, culture-positive rates among M876-inoculated turkeys declined more rapidly, transmission to contact sentinels took longer and occurred at lower rates, and serologic responses measured by HI and ELISA were lower than in S6-infected turkeys. Testing sera from inoculated turkeys for antibodies to MG in homologous and heterologous ELISA systems indicated that strain M876 was significantly (P less than 0.05) less immunogenic than S6 (days 62 and 95 PI), and that the homologous ELISA was more sensitive (P less than 0.005). MG strain M35 failed to infect turkeys in three attempts, even though the inocula used were viable on culture media.