Staphylococci isolated from animals and food with phenotypically reduced susceptibility to beta-lactamase-resistant beta-lactam antibiotics

The antibiotic resistance pattern of 1921 Staphylococcus strains isolated from animals and food within the last two years were examined using diffusion tests. Among them there were only 35 strains of S. aureus having an inhibition zone diameter of 15 mm or less, and 4 strains of coagulase-negative staphylococci (CNS) having a zone diameter of 18 mm or less to 1-microg oxacillin disk. These 39 strains were examined also by E-test to oxacillin and for the detection of the mecA gene by PCR in order to determine whether they might be real methicillin-resistant staphylococci. Among the 39 strains there were only two that were susceptible to penicillin by disk diffusion method; however, further examination by the penicillinase test showed that they produced beta-lactamase. While 19 (15 S. aureus, 4 CNS) strains were resistant and 7 strains were intermediate to oxacillin in disk diffusion test, the E-test gave 8 resistant and 5 intermediate results. Six out of the 8 oxacillin-resistant strains examined by disk diffusion and E-test harboured the mecA gene. Thus only 6 out of the examined 1921 strains proved to be mecA positive. These methicillin-resistant, mecA-positive strains (5 of the S. aureus strains and 1 of the S. epidermidis) originated from two dairy herds. The results prove that methicillin-resistant S. aureus (MRSA) strains in animals are really rare in Hungary. Eighteen strains were chosen and screened for minimal inhibitory concentration (MIC) of oxacillin with or without clavulanic acid or sulbactam, and three of them produced methicillinase enzyme.     

Energy imbalance related predisposition to mastitis in group-fed high-producing postpartum dairy cows

The energy imbalance related predisposition to mastitis was studied in group-fed postpartum dairy cows (n = 333) kept in 4 large-scale units and producing milk of low somatic cell count (SCC). Blood samples were taken on Days 1-3 after calving for assaying some metabolites and hormones related to the negative energy balance (NEB). If mastitis was diagnosed later, aseptic milk samples were taken to identify the pathogens. Considering pathogen types [contagious pathogens: Staphylococcus (S.) aureus, Gram-positive (GP) environmental pathogens, and Gram-negative (GN) environmental pathogens + mastitis with no detectable pathogens (NDP)] separately, stepwise logistic regression was used to analyse the relation between the potential prognostic value of hormones and metabolites and mastitis outbreak. Only the elevated (> or = 1.00 mmol/l) serum beta-hydroxybutyrate (BHB) levels predisposed the cows to mastitis in the subsequent 4 weeks. This prognostic value of BHB was significant only in GN + NDP mastitis and in cases caused by GP environmental pathogens, but not in S. aureus mastitis (odds ratio: 5.333, 3.600 and 1.333, respectively).     

Binding of mycoplasmas to solid phase adsorbents

The capture of mycoplasmas (M. hominis, M. buccale, M. fermentans, M. bovis, M. synoviae, M. gallisepticum and M. arthritidis) based on lipid structures and adhesion molecules present in the mycoplasmal membrane was tested using different chromatographic resins (ActiClean Etox, ClarEtox, Heparin-Actigel, Sulfated Hiflow and SulfEtox). All of the resins efficiently reduced mycoplasma concentrations in Phosphate Buffered Saline (PBS) and in Fetal Bovine Serum (FBS) by 3-8 logs in a few minutes. This technology could be used for removing mycoplasmas from tissue culture components such as serum, and for concentrating mycoplasmas in vaccine production.     

First report of target-site resistance to ACCase-inhibiting herbicides in Bromus tectorum L.

Background

The prevalent and repeated use of acetyl-coenzyme A carboxylase (ACCase)-inhibiting herbicides for Bromus tectorum L. control in fine fescue (Festuca L. spp) grown for seed has selected ACCase-resistant B. tectorum populations. The objectives of this study were to (1) evaluate the response of nine B. tectorum populations to the ACCase inhibitors clethodim, sethoxydim, fluazifop-P-butyl, and quizalofop-P-ethyl and the acetolactate synthase (ALS) inhibitor sulfosulfuron and (2) characterize the resistance mechanisms.

Results

Bromus tectorum populations were confirmed to be resistant to the ACCase-inhibiting herbicides tested. The levels of resistance varied among the populations for clethodim (resistance ratio, RR = 5.1–14.5), sethoxydim (RR = 18.7–44.7), fluazifop-P-butyl (RR = 3.1–40.3), and quizalofop-P-ethyl (RR = 14.5–36). Molecular investigations revealed that the mutations Ile2041Thr and Gly2096Ala were the molecular basis of resistance to the ACCase-inhibiting herbicides. The Gly2096Ala mutation resulted in cross-resistance to the aryloxyphenoxypropionate (APP) herbicides fluazifop-P-butyl and quizalofop-P-ethyl, and the cyclohexanedione (CHD) herbicides clethodim, and sethoxydim, whereas Ile2041Thr mutation resulted in resistance only to the two APP herbicides. All B. tectorum populations were susceptible to sulfosulfuron (RR = 0.3–1.7).

Conclusions

This is the first report of target-site mutations conferring resistance to ACCase-inhibiting herbicides in B. tectorum. The results of this study suggest multiple evolutionary origins of resistance and contribute to understanding the patterns of cross-resistance to ACCase inhibitors associated with different mutations in B. tectorum. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Lipophilicity of aminopyridazinone regioisomers

Ten pairs of pyridazinone regioisomers were prepared, and their lipophilicity was described by the logarithm of the octanol/water partition coefficient (log P) determined experimentally and calculated with prediction methods. The 4- and 5-(substituted amino)-3(2H)-pyridazinone regioisomers were synthesized by nucleophilic substitution of one of the chloro atoms of 4,5-dichloro-2-methyl-3(2H)-pyridazinone or its 6-nitro derivative. Structures of new compounds were proven by spectroscopic methods. The experimental log P values were obtained by a shake flask method in octanol and a S?rensen buffer (pH 7.4) solvent system. A consequent difference was found in the lipophilicity of regioisomers. For each isomer pair, the log P value of the 4-isomer was significantly (average by 0.75 log unit) higher than that of the 5-isomer. Some quantum chemical calculations as well as X-ray analysis of two pairs of regioisomers were also carried out to gain insight into the structural differences of regioisomers. The log P values were calculated by the fragmental approach KOWWIN and a QSPR analysis (3DNET). The a priori KOWWIN gave poor agreement, but with the programs KOWWIN with EVA (experimental value adjusted) and 3DNET, the results were generally in agreement with experiment.     

Erarbeitung eines Testverfahrens zur Bestimmung der Protektiv- und Kurativleistung von Fungiziden bei der Bek?mpfung des Erregers der Stemphylium-Laubkrankheit Stemphylium botryosum in der Spargelkultur (Asparagus officinalis L.)

The foilage of Asparagus officinalis L. has a large contact surface for different leaf diseases, which can lead to profits cuts, especially by the causal pathogen of Stemphylium leaf spot, Stemphylium botryosum. The maintaining healthy of the foliage is an important precondition to achieve an optimal harvest in the following year. Therefore, it is necessary to describe the efficacy of currently approved plant protection products against S. botryosum in order to optimize the fungicide applications. In the context of greenhouse trials with asparagus seedlings a method should be developed to evaluate the preventive and curative efficacy of fungicides. At the beginning, stable, repeatable and homogeneous infections of S. botryosum needed to be achieved. Ad hoc the factors plant age (weeks after sowing date), inoculum (density of conidia per ml and concentration of malt), infection conditions (period of high relative humidity) and day of rating, as well as their influence on the occurrence of the symptoms, were investigated. With the exception of the factor plant age, the other factors had a significant effect on the infection. For the trials to evaluate the efficacy of fungicides, the analysed factors were defined. The age of the seedlings was six weeks after sowing, the inoculum was composed of 1,00?×?10⁵ conidia per ml combined with an 0,5% malt concentration, as infection condition the ??tunnel system?? with periodically high relative humidity was chosen. 13 days past infection was the optimal day of rating. To apply the seedlings with the fungicides, the registered application rate was reduced to 1/5. The trials to evaluate fungicides were conducted with the fungicides Signum^®, Rovral^® WG and two different testing agencies. At four consecutive application days, the plants were covered with fungicides. In the following, the infestation degree of stems in percent was evaluated. Additionally, the used S. botryosum-Isolates were biomolecularly investigated and the QoI- and SDHI resistance state determined.     

Adventitious shoot regeneration from leaf thin cell layers in apple

The present investigation was conducted to study organogenesis of a conventionally easy-to-regenerate (‘Royal Gala’) and a difficult-to-regenerate (‘Freedom’) apple cultivar using leaf transverse thin cell layers (tTCLs). The position of the leaf source, explant type and light regime applied to tTCLs were investigated as was their influence on the organogenic outcome. Light during the onset of regeneration delayed morphogenesis or, in ‘Freedom’, completely inhibited organogenesis on leaf tTCLs. Genotype and initial position of the leaf also influenced percentage regeneration and number of regenerated shoots per explant. This study demonstrates the successful application of leaf tTCLs for adventitious shoot regeneration in apple, even though further studies are needed to increase the number of shoots more than control explants. Shoot regeneration from apple leaf tTCLs serves as an alternative protocol for studying fundamental and applied aspects of regeneration.     

Interaction between the three frequently co‐occurring Fusarium mycotoxins in rats

To test the complex, acute biochemical effects of combined, naturally co‐occurring fusariotoxins, a 5‐day rat study was performed. Mycotoxin treatment was invented by intraperitoneal injection: FB₁ (F): 9 µg/animal/day (approx. 30 µg/kg bw/day), DON (D): 16.5 µg/animal/day (approx. 55 µg/kg bw/day) and ZEN (Z): 12.75 µg/animal/day (approx. 42.5 µg/kg bw/day). The binary groups (FB₁ and DON [FD], FB₁ and ZEN [FZ] and DON and ZEN [DZ]) as well as the ternary (FB₁, DON and ZEN [FDZ]) group were dosed at the same combined level as the individual mycotoxins. Body weight, feed intake and mortality were not affected by any of the treatments. FB₁ and DON in combination (FD) increased the plasma aspartate aminotransferase activity synergistically (compared to the individual FB₁ and DON). In the liver, both the total glutathione (GSH) and the glutathione peroxidase (GPx) activity were increased (p < 0.05) by the binary FB₁ and ZEN (FZ) and the DON and ZEN (DZ) groups as well as the ternary FB₁, DON and ZEA group (FDZ) compared to the control. The GSH level of the ternary group was significantly increased compared to the FB₁ group, whereas the GPx activity of the ternary group was significantly increased compared to all three the individual mycotoxin groups. The Bliss independence method revealed synergism between DON and ZEN (DZ), as well as FB₁ and DON (FD) on liver GPx activity. None of the toxins alone or in combination exerted strong genotoxicity on lymphocytes, neither on the gross histopathological characteristics. However, even at these low levels acute exposure of more than one of these mycotoxins (FB₁, DON and ZEN) affected metabolic and detoxification changes.     

Local extinctions of the wetland specialist Swertia perennis L. (Gentianaceae) in Switzerland: a revisitation study based on herbarium records

We studied population extinction of the locally abundant fen plant Swertia perennis in Switzerland and used up to 127-year old herbarium records to relocate 63 sites that had once hosted this species. We recorded current site characteristics and related them to the absence or abundance of populations. Fifty-four sites (86%) were still traditionally used (extensively mown or grazed). Fifteen populations (24%) had gone extinct. Extinction was more likely at lower altitude, in the peripheral distribution range of S. perennis (58% peripheral, 9% central populations extinct), on the smallest fens (75% extinct on fens <400 m²) and on fens with intensified land use. However, even on traditionally managed wetlands 18.5% of the populations had gone extinct. Moreover, 40% of all remaining populations were smaller than 250 flowering plants. We conclude that both intensified agricultural practice and habitat fragmentation contributed to local extinction of S. perennis. Small populations, especially, may not be able to persist in the long term.