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.     

Distribution and characterization of class 1 integrons in Salmonella enterica serotype Gallinarum biotype Gallinarum

Fowl typhoid caused by Salmonella enterica subsp. enterica serotype Gallinarum biotype Gallinarum is the most important chicken disease in Korea. Due to appearance of new or multiple antibiotics resistances in the recently isolated strains, it was difficult to control the disease using antibiotics in our country. Therefore, the prevalence and genetic contents of class 1 integrons in biotype Gallinarum isolated between 1992 and 2001 were investigated by PCR and direct sequencing, respectively. Out of 90 strains, 35 (39%) carried class 1 integrons. The 1.0, 1.6 and 2.0kbp amplicons were amplified in 32 strains (36%), 2 strains (2%) and 1 strain (1%), respectively. The 1.0, 1.6 and 2.0 kbp amplicons contained one (aadA1a), two (aadB-aadA1b) and three cassettes (dhfrXII-orfF-aadA2), respectively, providing resistances against aminoglycosides (aadA1a, aadA1b, aadB, and aadA2) and trimethoprim (dhfrXII). The integron-carrying strains of biotype Gallinarum appeared in 1996 and acquired additional cassettes in 2000. Although the resistances to ampicillin, tetracycline and chloramphenicol are unrelated to class 1 integrons, relatively high prevalence of integron in biotype Gallinarum may be a dormant threat to the chemotherapy of the disease in the near future because of potency to acquire additional antibiotics resistances.     

Mitochondrial transmembrane potential and reactive oxygen species generation regulate the enhanced effect of CCCP on TRAIL-induced SNU-638 cell apoptosis

TRAIL is a member of the tumor necrosis factor family and engages apoptosis via recruitment and rapid activation of caspase-8. This study investigated the effect of carbonyl cyanide m-chlorophenylhydrazone (CCCP), a classic uncoupler of oxidative phosphorylation, on TRAIL-induced apoptosis in SNU-638 cells derived from human gastric cancer cells. It was found that treatment with CCCP followed by incubation with TRAIL markedly enhanced apoptosis by 2 fold compared with treatment with TRAIL alone. This effect was accompanied by reduction in mitochondrial transmembrane potential and generation of reactive oxygen species. This sensitization was inhibited by N-acetyl-l-cysteine, which restored the mitochondrial transmembrane potential and reduced reactive oxygen species generation. Treatment with N-acetyl-L-cysteine also inhibited expression of apoptotic proteins such as Bax and Smac and abrogated caspase-8 activation. Moreover, treatment with N-acetyl-L-cysteine prior to induction with TRAIL increased expression of the anti-apoptotic Bcl-2 protein. These data indicate that CCCP enhanced TRAIL-induced apoptosis by dissipation of mitochondrial transmembrane potential and reactive oxygen species, suggesting that treatment with CCCP combined with that with TRAIL can be an efficient method to induce death of tumor cells, particularly cells that are resistant to TRAIL-induced apoptosis.     

Echinochrome A Treatment Alleviates Atopic Dermatitis-like Skin Lesions in NC/Nga Mice via IL-4 and IL-13 Suppression

Atopic dermatitis (AD) is a chronic inflammatory skin disease in which skin barrier dysfunction leads to dryness, pruritus, and erythematous lesions. AD is triggered by immune imbalance and oxidative stress. Echinochrome A (Ech A), a natural pigment isolated from sea urchins, exerts antioxidant and beneficial effects in various inflammatory disease models. In the present study, we tested whether Ech A treatment alleviated AD-like skin lesions. We examined the anti-inflammatory effect of Ech A on 2,4-dinitrochlorobenzene (DNCB)-induced AD-like lesions in an NC/Nga mouse model. AD-like skin symptoms were induced by treatment with 1% DNCB for 1 week and 0.4% DNCB for 5 weeks in NC/Nga mice. The results showed that Ech A alleviated AD clinical symptoms, such as edema, erythema, and dryness. Treatment with Ech A induced the recovery of epidermis skin lesions as observed histologically. Tewameter^® and Corneometer^® measurements indicated that Ech A treatment reduced transepidermal water loss and improved stratum corneum hydration, respectively. Ech A treatment also inhibited inflammatory-response-induced mast cell infiltration in AD-like skin lesions and suppressed the expression of proinflammatory cytokines, such as interferon-γ, interleukin-4, and interleukin-13. Collectively, these results suggest that Ech A may be beneficial for treating AD owing to its anti-inflammatory effects.     

Anaesthetic efficacy and physiological responses to clove oil-anaesthetized kelp grouper Epinephelus bruneus

The efficacy of clove oil as an anaesthetic and at producing a physiological response (plasma cortisol and glucose) was evaluated in the kelp grouper, Epinephelus bruneus . To acquire complete anaesthesia in less than 3 min and recovery in <10 min, three doses of clove oil were tested at 18, 22 and 26 °C. Although higher anaesthetic doses resulted in shorter induction times and longer recovery times, and a lower temperature resulted in longer anaesthesia induction and slower recovery, we found the optimal dose and administering temperature of clove oil to be 250–300 mg L⁻¹ at water temperature of 18 °C, 150–200 mg L⁻¹ at water temperature of 22 °C and 50–100 mg L⁻¹ at water temperature of 26 °C respectively. Following the administration of 150 mg L⁻¹ of clove oil at 22 °C, the plasma cortisol level was highest (4.24 ± 1.571 μg dL⁻¹) after 12 h and the plasma glucose was highest (92.7 ± 9.61 mg dL⁻¹) after 2 h. These results should be useful to the aquaculture industry, where anaesthesia is necessary for a range of activities.     

Epidemiological and microbiological study of an outbreak of infectious keratoconjunctivitis in sheep

After several thousand sheep had been imported from Australia and New Zealand to Croatia during 1995, many native sheep that had been in contact with the imported animals acquired a severe ocular disease closely resembling infectious keratoconjunctivitis. In affected flocks glucose-fermenting mycoplasma were isolated from 48 per cent of conjunctival swabs and Branhamella ovis from 58 per cent. Twelve of 42 culturally and biochemically identical isolates were identified as Mycoplasma conjunctivae by polymerase chain reaction. From the conjunctivae of two animals M conjunctivae and M arginini were isolated in mixed culture. For many reasons most farmers removed the imported animals from their flocks and only sporadic cases of the disease were recognised in 1996. At the end of 1997, six flocks which were clinically free of the disease but had been affected during 1995, and five flocks with no history of the severe ocular disease were examined clinically and microbiologically, and were found to be free of M conjunctivae infection. At the time, B ovis was cultured almost exclusively from sheep originating from flocks which had been affected during 1995 and/or 1996. It was usually isolated in pure culture or as the predominant bacterial species, and was often accompanied by mild conjunctivitis. There were no microbiologically confirmed new cases of infectious keratoconjunctivitis during 1998 and 1999.     

Back-fat thickness as a primary index reflecting the yield and overall acceptance of pork meat

Predicting aspects of lean meat yield and eating quality of pork based on some particular carcass traits become increasingly important from an economic profitability point of view. The present study aimed to determine the relationship between the back-fat (BF) thickness with lean meat yield and meat quality traits of crossbred pigs. A total of 220 crossbred pigs [(Landrace × Yorkshire) ♀ × Duroc ♂] reared under identical conditions and harvested at 180 days old were slaughtered and screened for BF thickness. Four BF thickness groups: 12–15, 16–20, 21–25, and 26–30 mm were classified, and longissimus dorsi (LD) muscles from these groups were used to analyze meat quality characteristics. Results showed that increasing BF thickness decreased the lean meat yield and loin lean area. Increasing BF thickness up to 26–30 mm increased intramuscular fat content and pH of LD muscles. The LD muscles of the thicker BF (21–30 mm) groups received significantly higher flavor, juiciness, and overall acceptability scores than those of the thinner back-fat (12–20 mm) groups. Overall, when all the pre-harvest factors are kept the same, BF thickness could be considered as a primary index for predicting lean meat yield, and the minimal back-fat thickness of 21–30 mm is required to improve the overall acceptability of pork meat.     

Clove oil as an anaesthetic for common octopus (Octopus minor, Sasaki)

We evaluated the anaesthetic effect of clove oil [2‐methoxy‐4‐2‐(2‐propenyl)‐phenol] on the common octopus (Octopus minor), in terms of the time required to become anaesthetized (‘anaesthetic time’) and recovery time. We used a factorial experimental design and administered clove oil at different temperatures (15, 20 and 25°C) and concentrations (50, 100, 150, 200, 250 and 300 mg L⁻¹). We observed a significant (P<0.05) relationship between concentration and temperature, and each variable was effective (P<0.05). Anaesthetic time linearly decreased as the concentration and temperature increased. However, recovery time increased as the concentration increased and temperature decreased. There was no mortality. A concentration of 200 mg L⁻¹ clove oil showed rapid anaesthetic and recovery times in the common octopus, indicating its suitability for this species.     

Yersiniosis in zoo marmosets (Callitrix jacchuss) caused by Yersinia enterocolitica 4/O:3

The aim of this research was to describe two fatal cases of Yersinia enterocolitica bioserotype 4/O:3 infection in non-human primates and to characterise the isolates by PCR and PFGE. In July 2004, two marmosets (Callitrix jacchuss) born in captivity in Zagreb Zoo, died following a few days of intermittent diarrhoea in intervals of 2 weeks. The pathomorphological diagnosis of the female (born in 1997) and the male (born in 1995) marmoset, was disseminated miliary necrosis of the liver. Y. enterocolitica 4/O:3 was isolated from both livers showing that monkeys are susceptible to this bioserotype. The ail gene, which is an essential chromosomal virulence factor in pathogenic Y. enterocolitica isolates, was present in the marmoset isolates. Two different PFGE patterns were obtained from the isolates of the male liver with NotI enzyme. One genotype of the male marmoset isolate was indistinguishable from the genotype of the female marmoset isolate when NotI, ApaI and XhoI enzymes were used indicating a common infection source for the marmosets. The genotypes of the marmoset isolates differed only slightly from one human (of seven Croatian isolates) and from one pig isolate (representing a common genotype found among human and porcine isolates in Germany) suggesting that raw pork fed to the marmoset could have been the infection source.