The <Emphasis Type="Italic">Rg1</Emphasis> allele as a valuable tool for genetic transformation of the tomato 'Micro-Tom' model system

Background  

The cultivar Micro-Tom (MT) is regarded as a model system for tomato genetics due to its short life cycle and miniature size. However, efforts to improve tomato genetic transformation have led to protocols dependent on the costly hormone zeatin, combined with an excessive number of steps.     

The Effects of Loose Group versus Individual Stall Transport on Glucocorticosteriods and Dehydroepiandrosterone in Yearling Horses

The European Union recently published regulations regarding the welfare of horses during transport, requiring that horses be transported in individual stalls. The objective of this study was to determine whether concentrations of cortisol, corticosterone, or dehydroepiandrosterone (DHEA) differed among horses transported in individual stalls versus in loose groups. A total of 20 yearlings that were regularly handled and accustomed to being tied, but were naïve to transport, were assigned to be transported for 6 hours in either individual stalls or a loose group. The experiment was replicated with a second trial 35 days later following a switchback design. Jugular blood samples were analyzed for plasma cortisol, corticosterone, and DHEA concentrations at pretransport, after 2, 4, and 6 hours of transport, and at 2 and 4 hours after unloading. The data were analyzed using a mixed model repeated measures analysis of variance for treatment effects, whereas differences between sample times within each trial, and pretransport concentrations between trials, were analyzed using paired T-tests. No significant differences were found between treatment groups in concentrations of cortisol (P = .713), corticosterone (P = .370), or DHEA (P = .416). Cortisol and corticosterone concentrations increased significantly during transport, and returned to pretransport concentrations by 2 hours post-transport (P < .01). Changes in concentrations of cortisol and corticosterone indicated that transportation was a significant stressor; however, being transported in a loose group versus individual stalls was not different for these horses.     

A high-throughput method for early screening of coffee (<Emphasis Type="Italic">Coffea</Emphasis> spp.) genotypes for resistance to root-knot nematodes (<Emphasis Type="Italic">Meloidogyne</Emphasis> spp.)

Root-knot nematodes (Meloidogyne spp.) threaten the livelihood of millions of farmers producing coffee worldwide. The use of resistant plants either as cultivars or rootstocks appears to be the single most effective method of control. A screening method was developed to evaluate large populations of plants for resistance to root-knot nematodes. Two coffee cultivars, one susceptible and the other resistant to Meloidogyne paranaensis, were grown under controlled conditions in two substrates: a commercial sieved potting compost and an inert substrate containing sand with a water-absorbent synthetic polymer. Plant growth and development and nematode multiplication were compared for two inoculation dates (2 and 8 weeks after planting) and two evaluation dates (eight and 13 weeks after inoculation). Root growth, but not nematode multiplication, was influenced by the choice of substrate. Evaluation of the differences in root weight and nematode numbers between the different cultivars, substrates and dates of inoculation suggested that an optimal condition could be defined. The best discrimination between susceptible and resistant plants was found in the experiment where inoculation occurred at 2 weeks after planting and evaluation occurred at 8 weeks after inoculation. Because the total duration of this experiment was only 3 months, high-throughput evaluation was possible, opening up new possibilities for screening large germplasm collections and studying the genetic control of root-knot nematode resistance in coffee.     

Prevalence and first genetic identification of Cryptosporidium spp. in cattle in central Viet Nam

We investigated the prevalence of Cryptosporidium infection in relation to age and clinical status in cattle in the central region of Viet Nam. A total of 266 fecal samples from diarrheic and non-diarrheic cattle were examined by the modified Ziehl-Neelsen staining method. Prevalence of Cryptosporidium parvum type infections, those of the Cryptosporidium andersoni type, and mixed infection of both types was 33.5% (89/266), 5.6% (15/266), and 3.4% (9/266), respectively. The infection rate of 44.3% (35/79) of C. parvum in calves less than 6 months old was significantly higher than that of 28.9% (54/187) in cattle greater than 6 months old (P < 0.01). Although no C. andersoni oocysts were detected in calves less than 3 months old, no significant difference was observed between the age groups in the prevalence of C. andersoni infection and mixed infection. The percentage of diarrheic and non-diarrheic cattle identified to be shedding C. parvum oocysts was 46.5% (74/159) and 14.0% (15/107), respectively (P < 0.0001). The risk of diarrhea was 1.7 times greater in C. parvum-infected calves than in their non-infected counterparts. DNA sequences of 18S rRNA genes of C. parvum type and C. andersoni type indicated that they were C. parvum bovine genotype and C. andersoni, respectively. This is the first genetic identification of C. parvum bovine genotype and C. andersoni from cattle in Viet Nam.     

Radiographic, magnetic resonance imaging, computed tomographic, and rhinoscopic features of nasal aspergillosis in dogs

OBJECTIVE: To determine radiographic, magnetic resonance imaging (MRI), computed tomography (CT), and rhinoscopic features of nasal aspergillosis in dogs. DESIGN: Prospective study. ANIMALS: 15 client-owned dogs. PROCEDURE: All dogs had clinical signs of chronic nasal disease; the diagnosis of nasal aspergillosis was made on the basis of positive results for at least 2 diagnostic tests (serology, cytology, histology, or fungal culture) and detection of typical intrasinusal and intranasal fungal colonies and turbinate destruction via rhinoscopy. Radiography, MRI, and CT were performed under general anesthesia. Rhinoscopy was repeated to evaluate lesions and initiate treatment. Findings of radiography, MRI, CT, and rhinoscopy were compared. RESULTS: MRI and CT revealed lesions suggestive of nasal aspergillosis more frequently than did radiography. Computed tomography was the best technique for detection of cortical bone lesions; the nature of abnormal soft tissue, however, could not be identified. Magnetic resonance imaging allowed evaluation of lesions of the frontal bone and was especially useful for differentiating between a thickened mucosa and secretions or fungal colonies; however, fungal colonies could not be differentiated from secretions. Rhinoscopy allowed identification of the nature of intranasal and intrasinusal soft tissue but was not as useful as CT and MRI for defining the extent of lesions and provided no information regarding bone lesions. CONCLUSIONS AND CLINICAL RELEVANCE: The value of CT and MRI for diagnosis of nasal aspergillosis was similar and greater than that of radiography. Rhinoscopy is necessary because it is the only technique that allows direct visualization of fungal colonies.     

Associations between anti-Fasciola hepatica antibody levels in bulk-tank milk samples and production parameters in dairy herds

Our primary objective was to determine the relationships between Fasciola-specific antibody levels in bulk-tank milk and measures of productivity to estimate economic losses that are associated with Fasciola infections. A bulk-tank milk sample was collected in March 2004 from 1105 dairy herds in Flanders and the antibody levels against Fasciola hepatica (ODRf) and Ostertagia ostertagi (ODRo) were determined. The association of ODRf with four production parameters (milk yield, milk-protein %, milk-fat % and inter-calving interval) was assessed by multivariable linear-regression models. Production data were available for 463 out of the 1105 herds sampled. An increase in ODRf from the 25% quantile (0.428) to the 75% quantile (1.064) was associated with a decrease in the annual average milk yield of 0.7kg/(cowday) (P=0.002), with a decrease in the average milk-fat % of 0.06% (P<0.001) and with an increase of the mean inter-calving interval of 4.7 days (P=0.03). No significant relationship was found with the average milk-protein %. When the relationships of ODRf and ODRo with milk yield were tested simultaneously, we saw an additive rather than synergistic effect of concurrent infections.     

Comparison of Multistage Versus One-Stage Destabilization of a Type II External Fixator Used to Stabilize an Oblique Tibial Osteotomy in Dogs

OBJECTIVE: To compare the biomechanical effects of multistage versus one-stage destabilization of a type II external skeletal fixator (ESF) used to stabilize an oblique unstable tibial osteotomy in dogs. STUDY DESIGN: In vitro, in vivo, and ex vivo experimental study. ANIMAL POPULATION: Twelve healthy adult dogs. METHODS: The biomechanical characteristics of the type II ESF used in this study were determined. This fixator was applied to both tibiae of two groups of 6 dogs to stabilize a 2-mm-wide oblique osteotomy. One fixator on each dog remained unchanged throughout the 11-week study (control group). The fixator on the opposite limb was destabilized late and acutely in one group of dogs (single-stage) and early and progressively in the other (multistage). Clinical examination, radiographic examination, and force-plate analysis were used to evaluate the results. All dogs were euthanatized at 11 weeks. All tibiae were scanned to determine the cross-sectional area of the callus in the center of the osteotomy and subjected to biomechanical tests to determine mean pull-out strength of pins and callus strength and stiffness. RESULTS: Stiffness of the type II ESF used in this study was 578 N/mm in axial compression, 0.767 Nm/deg in torsion, 261 N/mm in medio-lateral bending, and 25 N/mm in cranio-caudal bending. Peak vertical forces of the hindlimbs were significantly lower at 2.5 and 5 weeks than before surgery. Peak vertical forces of the hindlimbs did not change before and after destabilization. No significant differences could be detected between the two destabilization sequences or between all control tibiae and pooled destabilized tibiae with regards to radiographic evaluation of the healing osteotomy, cross-sectional periosteal callus area, mean pull-out strength of transfixation pins, callus strength, and callus stiffness. CONCLUSIONS AND CLINICAL RELEVANCE: Bone healing of unstable osteotomies stabilized with a type II ESF is not significantly enhanced by staged destabilization of the fixation as performed in this study.     

High milk neutrophil chemiluminescence limits the severity of bovine coliform mastitis

Polymorphonuclear neutrophil (PMN) function changes during mastitis. To investigate the contribution of milk PMN to the severity of Escherichia coli (E. coli) mastitis, chemiluminescence (CL) of blood and milk PMN and their efficiency to destroy coliform bacteria in the mammary gland were examined following the induction of E. coli mastitis in early lactating cows. To better assess and define the degree of mastitis severity, cows were classified as moderate and severe responders according to milk production loss in the non-infected quarters at post-infection hour (PIH) 48. There was an inverse relationship between pre-infection milk PMN CL and colony-forming units at PIH 6. In moderate cows, the pre-infection blood and milk PMN CL was approximately 2-fold higher than that of severe cows. The probability of severe response increased with decreasing pre-infection PMN CL. At the beginning of the infection blood and milk PMN CL was consistently higher, and milk PMN CL increased faster after infection in moderate cows. At PIH > 48 milk PMN CL in severe cows exceeded that of moderate cows. The somatic cell count (SCC) in moderate cows increased faster than colony-forming units, whereas in severe cows the results were reversed. The kinetics of CL activity for blood and milk PMN before and during the early phase of infection confirmed an impairment in PMN CL activity for severe responding cows. High pre-infection blood and milk PMN CL and the immediate increase of milk PMN CL and SCC after infection limited bacterial growth thereby facilitating the recovery of E. coli mastitis in moderate cows. Our study strengthens the idea that pre-existing milk PMN (a static part of the udder's immune defense) functions as a "cellular antibiotic" before and during infection, and low milk PMN CL is a risk factor for bovine coliform mastitis.     

Antimicrobial resistance and resistance genes in Staphylococcus aureus strains from rabbits

Fifty-six Staphylococcus aureus isolates recovered between 1998 and 2003 from 31 rabbit farms with and without problems of chronic staphylococcosis, were screened for resistance to enrofloxacin, erythromycin, gentamicin, lincomycin, neomycin, penicillin and tetracyclines using the agar dilution test. For penicillin, a disk diffusion test was also performed. The detection of tetP(B), tet(K), tet(L), tet(M), tet(O), tet(T), tet(W), erm(A), erm(B), erm(C) and mec(A) genes was done via a PCR assay. Four isolates showed resistance to erythromycin and lincomycin. These isolates were positive for the erm(C) gene in the PCR. Eleven strains were resistant to tetracyclines and all harboured the tet(K) gene. In the agar dilution test, five isolates showed resistance to penicillin, whereas in the disk diffusion test 12 isolates showed resistance. None of these 12 resistant isolates carried the mec(A) gene. Only one strain showed resistance to gentamicin, and all strains were susceptible to enrofloxacin and neomycin. This study demonstrates that resistance to antimicrobial agents in S. aureus isolates originating from rabbits is relatively rare compared to resistance in S. aureus isolates originating from other animals and humans.