Effects of three antibiotics (cefotaxime, rifampicin and gentamicin) were tested on in vitro shoots cultures of tansy (Tunacetum vulgare L.). These antibiotics were selected because endophytic bacteria isolated from the in vitro shoot cultures of tansy showed that all bacteria were Gram-negative and their growth was reduced by these three antibiotics. Five isolates were Enterobacteriaceae, three were fluorescent Pseudomonas, and two were aerobic bacteria. Increased concentrations of antibiotics caused usually linear or quadratic changes on the initiation of shoot growth, shoot number, growth rate, and shoot height. These changes and changes in pH of the culture media were tansy genotype-dependent following treatments with gentamicin. Also, the treatment with rifampicin or cefotaxime showed a genotype-dependent effect, because they resulted in significantly higher percentage of rooted plants in one of the three tansy genotypes tested. The growth rate and length of shoots were reduced in the media containing both gentamicin and rifampicin, but less so than in media containing both gentamicin and cefotaxime.

Bone fragments are essential structures for the detection of processed animal proteins (PAPs) in feed by light microscopy for official controls according to Annex VI of European Union Regulation EC/152/2009. The preparation of samples submitted for analysis requires a grinding step to make them suitable for microscopic slide preparation and observation. However, there are no technical guidelines set down for this step despite the fact that it can lead to an increase in bone numbers due to fragmentation. This was demonstrated by an in-house study carried out by the Irish National Reference Laboratory (NRL) for animal protein detection. The present collaborative study investigated the possible effects of three different grinding conditions on the final result for a feed adulterated with 0.05 and 0.01% (w/w) of PAP. The microscopic analysis either combined or not with an Alizarin Red staining was carried out by 10 different laboratories. The results demonstrated that although a large variation in the numbers of bone fragments was noted, five of the six different grinding/staining combinations applied at two levels of PAP adulteration did not significantly (at p = 0.05) differ from one another. The only exception occurred when grinding the feed containing 0.05% of PAP with a rotor mill equipped with a 0.5-mm sieve and combined with a staining which resulted in a greater number of bone fragments by forced fragmentation. Overall, the impact of the grinding/staining combinations on the final results was shown to be negligible when considering the regulatory limit of detection (LOD) requirement for the method and the current rules of implementation of the light microscopic method. From a total of 180 analyses carried out on the feed matrix containing 0.05% of PAP no false-negative result was observed, and at a level of 0.01% PAP only 10 false-negative results occurred.

Ten bacteria were isolated from the in vitro shoot cultures of different tansy (Tanacetum vulgare) genotypes. All isolates were Gram-negative. Five isolates belonged to Enterobacteriaceae, three isolates were strains of fluorescent Pseudomonas, and two isolates were strains of other aerobic bacteria. The combined treatment with gentamicin (50 mg·L–1) and rifampicin (25 mg·L–1) prevented the growth of all bacteria, whereas none of the antibiotics (ampicillin, cefotaxime, rifampicin, gentamicin, or streptomycin) controlled all bacteria when used alone. The antibiotics ranged based on their increasing adverse effects on shoot cultures as follows: rifampicin, cefotaxime, and gentamicin. Increased antibiotic concentration reduced the initiation and growth rates of shoots, roots and calli with some exceptions. Low concentrations of gentamicin and cefotaxime both increased the growth rate and shoot number per plant. Rifampicin stimulated the root growth of some of the biotypes tested. The growth rate of calli increased significantly in the presence of both gentamicin (25 mg·L–1) and rifampicin (25 mg·L–1) in the growth media, whereas the use of either antibiotic alone reduced the calli growth relative to the control. Viability of tansy protoplasts was enhanced by rifampicin and to a lesser extend by gentamicin and cefotaxime. The growth of bacteria isolated from tansy tissue culture can be prevented with a combined gentamicin and rifampicin treatment. The growth rate of plants decreased slightly when grown in the media supplemented with the same antibiotic combination. However, the growth retardation was not permanent, and the plants recovered and grew vigorously when transferred to antibiotic free medium.

A survey for the possible occurrence of red‐core disease (Phytophthora fragariae var. fragariae) was carried out on 250 strawberry production sites in Finland. The fields were inspected visually. A total of 1080 samples of strawberry roots were taken in spring and autumn 1995 and examined visually, microscopically and by isolation in the laboratory. Root‐tip bait‐plant tests were performed in the glasshouse to look for the latent presence of the fungus. No red‐core disease was detected in any of the inspected fields or in the examined samples.