Suitability of SSCP-PCR analysis for molecular detection of quinolone resistance in Campylobacter jejuni

Foodborne infections with Campylobacter spp. are increasing, especially antibiotic resistant strains are emerging. Quinolone resistant isolates can cause failure of therapy in severe clinical infections. Molecular characterisation is needed for the detection of resistant variants of C. jejuni. Therefore 23 isolates from poultry and human medicine as well as three control strains were tested for their minimal inhibitory concentration, their Single-Strand-Conformation-Polymorphism (SSCP)-PCR pattern (a method for the detection of resistance determining point mutations), and their sequence of the quinolone resistance determining region (QRDR). Six different SSCP types could be identified: two types for quinolone resistant isolates and other types containing so called silent mutations without influence on the resistance. A genotypic monitoring of the quinolone resistance in C. jejuni can be useful for the early detection of new resistance variants. As a screening method for detection of point mutations in the QRDR the SSCP-PCR can be applied. Compared to other genotypic methods the SSCP-PCR is less time and cost consuming and needs only standard technical equipment.     

Comparison of antibody values in sera of pigs vaccinated with a subunit or an attenuated vaccine against classical swine fever

Ten pigs, aged 85 days, were vaccinated with a subunit vaccine containing 32 g of classical swine fever virus glycoprotein E2 (gp E2) (group 1), and a further 10 pigs were vaccinated with a C strain vaccine (10^4±0.15 TCID₅₀/ml), produced by amplification in minipig kidney (MPK) cell culture (group 2). Nine non-vaccinated pigs served as a control group (group 3). Serum samples were collected before (day 0) and at 4, 10, 21 and 28 days after vaccination and were analysed by two commercially available enzyme immunoassays and by a neutralizing peroxidase-linked assay (NPLA). At the same times, peripheral blood was taken for determining the total leukocyte count and the body temperature was taken daily. Antibodies were not detected in serum samples collected before vaccination (day 0), and no side-effects that could be connected with vaccination were observed during the trial. Ten days after vaccination 6/10 pigs vaccinated with the subunit vaccine were seropositive. On days 21 and 28, the ratios of serologically positive to vaccinated pigs were 9/10 and 10/10, respectively. Four of the ten pigs that were vaccinated with the C strain vaccine were positive on day 21 and 9/10 on day 28. However, the results of the NPLA showed that only 4/10 pigs had an antibody titre >1:32 at the end of the trial in both the vaccinated groups, even though the subunit vaccine initiated an earlier and higher level of neutralizing antibodies than the vaccine produced from the C strain. Challenge was performed 28 days after vaccination on four randomly selected pigs from both vaccinated groups. The pigs survived the challenge without showing any clinical signs of classical swine fever (CSF), while two nonvaccinated control pigs died on the 10th and 12th days after infection.     

Evaluation of the perioperative stress response in dogs administered medetomidine or acepromazine as part of the preanesthetic medication

OBJECTIVE: To compare the perioperative stress response in dogs administered medetomidine or acepromazine as part of the preanesthetic medication. ANIMALS: 42 client-owned dogs that underwent elective ovariohysterectomy. PROCEDURE: Each dog was randomly allocated to receive medetomidine and butorphanol tartrate (20 microgram/kg and 0.2 mg/kg, respectively, IM) or acepromazine maleate and butorphanol (0.05 and 0.2 mg/kg, respectively, IM) for preanesthetic medication. Approximately 80 minutes later, anesthesia was induced by administration of propofol and maintained by use of isoflurane in oxygen. Each dog was also given carprofen before surgery and buprenorphine after surgery. Plasma concentrations of epinephrine, norepinephrine, cortisol, and beta-endorphin were measured at various stages during the perioperative period. In addition, cardiovascular and clinical variables were monitored. RESULTS: Concentrations of epinephrine, norepinephrine, and cortisol were significantly lower for dogs administered medetomidine. Concentrations of beta-endorphin did not differ between the 2 groups. Heart rate was significantly lower and mean arterial blood pressure significantly higher in dogs administered medetomidine, compared with values for dogs administered acepromazine. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that for preanesthetic medications, medetomidine may offer some advantages over acepromazine with respect to the ability to decrease perioperative concentrations of stress-related hormones. In particular, the ability to provide stable plasma catecholamine concentrations may help to attenuate perioperative activation of the sympathetic nervous system.     

In vitro antimicrobial susceptibility of 183 Pseudomonas aeruginosa strains isolated from dogs to selected antipseudomonal agents

During the years from 1993 to 2000, 183 strains of Pseudomonas aeruginosa were isolated from different pathological specimens originating from dogs. Antimicrobial susceptibility patterns against 10 antipseudomonal agents were obtained on 183 P. aeruginosa strains. In vitro antimicrobial susceptibility testing was performed using the disk diffusion method (Kirby-Bauer). Antimicrobial susceptibility profiles showed that among beta-lactam antibiotics, imipenem was the most active compound. Out of the 183 strains tested, 96.7% were sensitive to imipenem. Cefoperazone showed good in vitro activity against 86.9% of the tested strains. Against ceftazidime, 77.0% of strains showed sensitivity. An old penicillin, carbenicillin, gave only 71.6% sensitive strains. Sensitivity to amikacin was 87.4% and it was 83.1% to gentamicin. Pipimedic acid, a first-generation quinolone, was the least active compound of all those tested, 47.0% were resistant. The in vitro sensitivity against enrofloxacin showed that 71.0% strains were sensitive and 26.2% showed resistance. Almost all strains tested, 93.4%, were susceptible to ciprofloxacin and marbofloxacin. Besides imipenem, the quinolone antibiotics, marbofloxacin and ciprofloxacin were the most effective against P. aeruginosa strains isolated from dogs.     

Occurrence of enteric redmouth disease in rainbow trout (Oncorhynchus mykiss) on farms in Croatia

During the spring of 1996 and autumn of 1997 unusual mortality outbreaks among rainbow trout fry and yearlings occurred at two different trout farms, resulting in mortality of 20 and 10 per cent, respectively. Generally, the affected fish, swimming at the water surface, were reluctant to eat and were dark pigmented with visible haemorrhages around and within the oral cavity. Bacterial isolates from moribund fish from both cases were identified as Yersinia ruckeri by standard biochemical tests and API 20E. The isolated strains were found to be sensitive to tetracycline, chloramphenicol, co-trimoxazole, nalidixic acid, flumequine, enrofloxacin, carbenicillin and gentamicin. Microplate agglutination assay confirmed that both isolates belonged to serotype O1. The pathogenicity of the isolated bacteria was confirmed by challenge experiment. Titres of specific antibodies were determined in the sera of survivors. The titre was highest on the 21st day postchallenge and was detectable until the 81st day.     

A survey of chickens for viable toxoplasms in Croatia

Brain tissues of 716 slaughtered domestic chickens (524 broilers and 192 hens) were bioassayed for viable toxoplasms. Each tissue was homogenized and subcutaneously injected into 4 SPF mice. Six weeks later the mice were euthanatized and their brains microscopically examined for Toxoplasma gondii tissue cysts. Three (0.4%) out of a total of 716 birds were positive. All positive cases were hens. This is the first isolation of T. gondii from chickens in Croatia.     

Parasitoid complex (Hymenoptera, Braconidae, Aphidiinae) of Aphis craccivora Koch (Hemiptera: Aphidoidea) in Iran

The parasitoids (Hymenoptera, Braconidae, Aphidiinae) of the cowpea aphid, Aphis craccivora Koch, were investigated throughout two far distant provinces, Tehran and Sistan-Baluchestan. Colonies of the cowpea aphid were collected from different host plants and reared under laboratory conditions until the parasitoids emerged. The aphids were parasitized by an expected range of parasitoids. The species were Aphidius colemani Viereck, Lysiphlebus fabarum (Marshall), Lysiphlebus confusus Tremblay and Eady, Lysiphlebus testaceipes (Cresson), Binodoxys acalephae (Marshall), Binodoxys angelicae (Haliday), Praon volucre (Haliday) and Ephedrus persicae Froggatt. L. testaceipes is reported here for the first time in Iran. The most common species was L. fabarum followed by B. acalephae and L. confusus. A key is provided for identification and host plant associations are considered.     

Bemerkungen über die Populationsbewegungen der Mittelmeerfruchtfliege (Ceratitis capitata Vied.) an der jugoslawischen Adriaküste

Zusammenfassung Untersuchungen über den Massenwechsel und die Biologie der MittelmeerfruchtfliegeCeratitis capitata Vied. während der letzen 5 Jahre führten zu folgenden Ergebnissen:1. Die Mittelmeerfruchtfliege, die bis 1947 in ganz Jugoslawien unbekannt war, kommt seit mindestens zehn Jahren auch an der Adriaküste vor.2. Erst im Jahre 1958 erwies sich die Fruchtfliege als Schädling verschiedener Obstarten.3. Wie aus Beobachtungen in Obstanlagen an der Adriaküste hervorgeht, befällt die Fliege in erster Linie Pfirsich, ferner auch Birne, Kaki, Feige, Apfel, Pflaume, Orange und grünen Paprika. Im Laboratorium gelang die Zucht außerdem noch an Banane, nicht aber an Tomate und Trauben.4. ObwohlC. capitata ein polyphages Insekt ist, befällt sie an einigen Stellen, wo sie regelmäßig große Häufigkeit aufweist, nur Pfirsich (Split, Debeli Rti) bzw. nur Kaki (Ankaran). In manchen Obstgärten lebt sie aber an verschiedenen Früchten (Podstrana, Stroanac). Es gibt bei der Fruchtfliege also auf Obstarten spezialisierte Populationen und solche, die an verschiedenen Obstarten vorkommen.5. Unter den in unserem Küstenland herrschenden Klimabedingungen kann die Fruchtfliege in einem Jahr 4–5 Generationen entwickeln. Die Herbstgeneration verpuppt sich Ende Oktober/Anfang November in der Erde, die Imagines verlassen bis Januar die Puppen und warten auf das Reifen der ersten, frühen Pfirsichsorten. Die gleiche Überwinterungsweise wurde auch an der Olivenfliege beobachtet.6. Mit Angelica-Öl versehene Fanggläser brachten nur zum Teil befriedigende Ergebnisse, da dieser Lockstoff nur die Männchen anzieht, aber auch nur in der Zeit, in der die Fliegen überhaupt selten sind. Im Spätsommer und Herbst wirkt ein aus Kleie und Biammonphosphat bereiteter Köder auf beide Geschlechter stark anlockend.     

Laser ablation-combustion-GC-IRMS--a new method for online analysis of intra-annual variation of delta13C in tree rings

We present a new, rapid method for high-resolution online determination of delta13C in tree rings, combining laser ablation (LA), combustion (C), gas chromatography (GC) and isotope ratio mass spectrometry (IRMS) (LA-C-GC-IRMS). Sample material was extracted every 6 min with a UV-laser from a tree core, leaving 40-microm-wide holes. Ablated wood dust was combusted to CO2 at 700 degrees C, separated from other gases on a GC column and injected into an isotope ratio mass spectrometer after removal of water vapor. The measurements were calibrated against an internal and an external standard. The tree core remained intact and could be used for subsequent dendrochronological and dendrochemical analyses. Cores from two Scots pine trees (Pinus sylvestris spp. sibirica Lebed.) from central Siberia were sampled. Inter- and intra-annual patterns of delta13C in whole-wood and lignin-extracted cores were indistinguishable apart from a constant offset, suggesting that lignin extraction is unnecessary for our method. Comparison with the conventional method (microtome slicing, elemental analysis and IRMS) indicated high accuracy of the LA-C-GC-IRMS measurements. Patterns of delta13C along three parallel ablation lines on the same core showed high congruence. A conservative estimate of the precision was +/- 0.24 per thousand. Isotopic patterns of the two Scots pine trees were broadly similar, indicating a signal related to the forest stand's climate history. The maximum variation in delta13C over 22 years was about 5 per thousand, ranging from -27 to -22.3 per thousand. The most obvious pattern was a sharp decline in delta13C during latewood formation and a rapid increase with spring early growth. We conclude that the LA-C-GC-IRMS method will be useful in elucidating short-term climate effects on the delta13C signal in tree rings.