On-line gas chromatography combustion/pyrolysis isotope ratio mass spectrometry (HRGC-C/P-IRMS) of pineapple (Ananas comosus L. Merr.) volatiles

By use of extracts prepared by liquid-liquid separation of the volatiles from self-prepared juices of pineapple fruits (Ananas comosus) (n = 14) as well as commercial pineapple recovery aromas/water phases (n = 3), on-line capillary gas chromatography-isotope ratio mass spectrometry was employed in the combustion (C) and the pyrolysis (P) modes (HRGC-C/P-IRMS) to determine the delta(13)C(VPDB) and delta(2)H(VSMOW) values of selected pineapple flavor constituents. In addition to methyl 2-methylbutanoate 1, ethyl 2-methylbutanoate 2, methyl hexanoate 3, ethyl hexanoate 4, and 2,5-dimethyl-4-methoxy-3[2H]-furanone 5, each originating from the fruit, the delta(13)C(VPDB) and delta(2)H(VSMOW) data of commercial synthetic 1-5 and "natural" (biotechnologically derived) 1-4 were determined. With delta(13)C(VPDB) data of pineapple volatiles 1-4 varying from -12.8 to -24.4 per thousand, the range expected for CAM metabolism was observed. Compound 5 showed higher depletion from -20.9 to -28.6 per thousand. A similar situation was given for the delta(2)H(VSMOW) values of 3-5 from pineapple ranging from -118 to -191 per thousand, whereas 1 and 2 showed higher depleted values from -184 to -263 per thousand. In nearly all cases, analytical differentiation of 1-5 from pineapple and natural as well as synthetic origin was possible. In general, natural and synthetic 1-5 exhibited delta(13)C(VPDB) data ranging from -11.8 to -32.2 per thousand and -22.7 to -35.9 per thousand, respectively. Their delta(2)H(VSMOW) data were in the range from -242 to -323 per thousand and -49 to -163 per thousand, respectively.     

Control of Fusarium Wilt of Radish by Combining Pseudomonas putida Strains that have Different Disease-Suppressive Mechanisms

ABSTRACT Biological control of soilborne plant pathogens in the field has given variable results. By combining specific strains of microorganisms, multiple traits antagonizing the pathogen can be combined and this may result in a higher level of protection. Pseudomonas putida WCS358 suppresses Fusarium wilt of radish by effectively competing for iron through the production of its pseudobactin siderophore. However, in some bioassays pseudobactin-negative mutants of WCS358 also suppressed disease to the same extent as WCS358, suggesting that an, as yet unknown, additional mechanism may be operative in this strain. P. putida strain RE8 induced systemic resistance against fusarium wilt. When WCS358 and RE8 were mixed through soil together, disease suppression was significantly enhanced to approximately 50% as compared to the 30% reduction for the single strain treatments. Moreover, when one strain failed to suppress disease in the single application, the combination still resulted in disease control. The enhanced disease suppression by the combination of P. putida strains WCS358 and RE8 is most likely the result of the combination of their different disease-suppressive mechanisms. These results demonstrate that combining biocontrol strains can lead to more effective, or at least, more reliable biocontrol of fusarium wilt of radish.     

Tank-mix adjuvants and pesticide residues: some regulatory and quantitative aspects

Although it is well known that judicious use of adjuvants can increase the performance of foliage-applied sprays of many agrochemicals, little information is available in the public domain about their ultimate effects on pesticide residues in treated crops. In the present work, the influence of Agral (polyoxyethylene nonylphenols), Toil (methyl esters of rapeseed fatty acids) and Bond (styrene-butadiene copolymers) on surface and crop residues of diclofop-methyl/diclofop and propiconazole in wheat and field beans was investigated using a model system simulating field practice. Pesticides were applied as commercial formulations, diclofop-methyl 378 g litre(-1) EC (Hoegrass) and propiconazole 250 g litre(-1) EC (Tilt), at their maximum approved rates, 1135 g AI ha(-1) and 125 g AI ha(-1), respectively, both in the presence or absence of the maximum rate recommended for each candidate adjuvant. No detectable residues of diclofop-methyl or propiconazole were found in wheat 35 days after any of the four applications. However, residues of diclofop were present in this crop, and those from applications containing Agral (0.07 mg kg(-1) fresh weight (FW)) or Bond (0.08 mg kg(-1) FW) were significantly lower than those with no adjuvant (0.14 mg kg(-1) FW) or Toil (0.16 mg kg(-1) FW). Unlike wheat, residues of both diclofop and propiconazole were detected in field beans after harvest. Significantly higher residues of the former were recorded from the applications with Agral or Bond (ca 0.32 mg kg(-1) FW) than with those with no adjuvant or Toil (ca 0.15mg kg(-1) FW). All the propiconazole applications containing adjuvants showed a similar significant increase in residues (0.10-0.16 mg AI kg(-1) FW) over the no-adjuvant treatment (0.05 mg kg(-1) FW) in this crop. There appeared to be little agreement between the apparent amounts of uptake, as indicated by the rates of decline of surface residues up to 5 days after application, and final residues in either target species. On wheat, surface residues of diclofop-methyl decreased from initially ca 20 to as little as 0.02 mg kg(-1) FW using adjuvants; the corresponding values for propiconazole were ca 2 to ca 0.03 mg kg(-1) FW. Recoveries of diclofop-methyl from the surfaces of field beans were much higher than those from wheat, declining from ca 30 to only ca 6 mg kg(-1) FW during the course of 5 days; the corresponding residues for propiconazole were ca 2 mg to 0.15 mg kg(-1) FW. These findings are discussed in relation to uptake results obtained with radiolabelled pesticides and adjuvants in the laboratory, and to the mandatory requirements for pesticide residue data for the authorised use of adjuvants in the UK.     

Use of volumetric capnography to identify pulmonary dysfunction in horses with and without clinically apparent recurrent airway obstruction

OBJECTIVE: To investigate whether volumetric capnography indices could be used to differentiate between horses without recurrent airway obstruction (RAO) and horses with RAO that were in clinical remission or that had clinically apparent RAO. ANIMALS: 70 adult Swiss Warmblood horses (20 used for pleasure riding and 50 used for dressage or show jumping). PROCEDURE: Horses were allocated to 4 groups on the basis of history, clinical signs, results of endoscopy, and cytologic findings (group 1, 21 healthy horses; group 2, 22 horses with RAO that were in remission; group 3, 16 horses with mild RAO; group 4, 11 horses with exacerbated RAO). Expiratory volume and CO2 curves were recorded by use of a computerized ultrasonic spirometer. Volumetric capnograms were plotted, and derived indices were calculated. RESULTS: Dead-space volume (VD) was calculated by use of the Bohr equation (VD(Bohr)) and for physiologic VD (VD(phys)). Ratios for VD(Bohr) to expiratory tidal volume (VT) and VD(phys) to V(T) as well as an index of effective CO2 elimination were significantly different among groups of horses. Age and use of the horses also significantly affected volumetric capnography indices. CONCLUSIONS AND CLINICAL RELEVANCE: Ratios of VD(Bohr) to VT and VD(phys) to VT as well as an index of effective CO2 elimination were sufficiently sensitive measures to distinguish between healthy horses and horses with RAO in remission. To optimize the ability of volumetric capnography indices to differentiate among horses in heterogeneous populations, it is important to account for effects of age and specific use of the horses.     

Effect of prior serial in vivo passage on the frequency of Salmonella enteritidis contamination in eggs from experimentally infected laying hens

Experimental infection models are valuable tools for understanding and preventing the deposition of Salmonella enteritidis inside eggs. Oral inoculation is believed to closely simulate naturally occurring S. enteritidis infections of chickens, but oral infection studies have often generated relatively low frequencies of egg contamination. The present study assessed whether repeated in vivo passage of an S. enteritidis strain could affect its ability to cause egg contamination in experimentally infected hens. The incidence of egg contamination was determined in groups of hens inoculated orally with either a phage type 13a S. enteritidis strain or derivatives of this parent strain that were obtained by three successive rounds of passage and reisolation from tissues of infected hens. Passaged S. enteritidis isolates recovered from ovaries and oviducts induced a significantly higher incidence of egg contamination (16.97%) than was attributed to the parent strain (8.27%). However, passaged S. enteritidis isolates recovered from livers and spleens were not associated with a significantly increased frequency of deposition in eggs. By either inducing or selecting for the expression of relevant microbial properties, passage of S. enteritidis through reproductive tissues of chickens may be useful for improving the efficiency at which experimental infection models produce egg contamination.     

Prevalence of herd specific factors and limb disorders,and their associations in intensive swine production

A longitudinal observational study in 180 pig breeding herds was performed to calculate prevalences of herd specific factors as well as typical limb disorders and to estimate their associations in a 2-step regression analysis. Regarding herd size, genetics, feeding and weight gain herds were distributed almost equal. The population density and the hygiene status were considered proper in most herds. In the farrowing units partially slatted floors of metal or plastic with slats > 9 mm, in the weaning units fully slatted floors of plastic, and in the rearing units fully slatted floors of concrete were most common. Less than 6% of the farms housed their pigs on solid concrete with straw bedding. Herd prevalences of fault floors varied between 18 and 43%. As a herd health problem (morbidity > 25%) claw hematomas and limb abrasions in just 1-week old piglets, overgrown claws and bursa swellings in weaned pigs, and bursa swellings in rearing pigs were wide spread. Leg deformations by osteopathy or arthritis occurred only sporadically. In the risk analysis claw hematomas of piglets were associated with slatted floors, particulary with slats < 10 mm. Abrasions were associated with concrete and rough floor surfaces at all. Overgrown claws and bursa swellings in weaned and in rearing pigs were associated with damaged, slippery or rough floor surfaces. Other associations were not detected. The quality of floor might be more important than the type of housing.     

Molecular biological method cascade for detection, specific isolation and characterization of shiga toxin-producing Escherichia coli (STEC) in food

Shigatoxin-producing E. coli (STEC) and their subgroup, the enterohaemorrhagic E. coli (EHEC) are known since about 25 years. Only EHEC can cause diseases in humans. A molecularbiological method cascade is described in this article. It contains the following steps: preenrichment, enrichment, preparation of samples (DNA extraction), screening PCR, specific STEC isolation by using DNA DNA hybridization, verification of isolates as STEC by using PCR, characterization of STEC isolates. Pulsed field gel electrophoresis and DNA sequencing are additional methods for the detection of clonal correlations. They should be performed in special labs, only. Furthermore, some information about an interlaboratory ring trial within Germany and some conclusions are given.     

Canine brucellosis--a case report

Brucella (B.) canis was isolated from ejaculate of a 4-year old Korthals-Griffon male dog after occurrence of epididymitis and orchitis. Despite several trials of therapy with different antibiotics relapes occurred, with B. canis being isolated from ejaculate, blood and urine samples, respectively. Bacteriological examinations were added by serological testing over a period of about 1.5 years. During the study SAT serum titre steadily dropped from 1:200 to 1:50. By CFT, B. canis antibodies were detectable at the beginning with a titre of 1:320 and to the end of the study with titres between 1:80 and 1:160.