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.

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.

Detection and identification of Mycoplasma conjunctivae in infectious keratoconjunctivitis by PCR based on the 16S rRNA gene.

A specific PCR assay based on unique sequences of the rrs genes (16S rRNA) of Mycoplasma conjunctivae was developed for direct detection and identification of this pathogen from clinical material. DNA from eye swabs was amplified after a simple lysis step by either a single PCR with the M. conjunctivae specific primer pair McoR1 and McoF1, or by a nested PCR with the Mycoplasma genus specific primer pair MOLIGEN1-L and 16UNI-R in the first step and McoR1 and McoF1 in the second step. The specificity of the primer pair McoR1 and McoF1 was verified with purified DNA from the type strain, from 17 field isolates of M. conjunctivae and from several Mollicutes which are phylogenetically related to M. conjunctivae or which can be isolated from the same host animals. This method identified mycoplasma isolates from goat, sheep, ibex and chamois originating from different countries as M. conjunctivae. No cross amplifications with other mycoplasmas which are related to M. conjunctivae were observed. Eye swab samples containing known numbers of M. conjunctivae cells were analysed after direct lysis of the material. The detection level was estimated to be 20 cells per swab when the nested PCR procedure was used and 2 x 10(5) by the single PCR method. In an experimental infection model of sheep, the nested PCR method for detection of M. conjunctivae gave results which were comparable to mycoplasmal culture. These are the implications for diagnostic purposes: M. conjunctivae isolates can be identified by the one-step PCR method, whereas for detection and identification of M. conjunctivae in clinical material the two-step method should be used (higher sensitivity).

An outbreak of abortion in mares associated with Salmonella abortusequi infection.

An abortion outbreak occurred in a herd of 38 horses, 26 of which were pregnant mares. Twenty-one mares aborted between 5-10 months of gestation. In no case were there indications of impending abortion. Pathoanatomical, histopathological, virological and bacteriological examinations were carried out on 4 aborted fetuses. Histopathology identified Gram-negative bacteria compatible with salmonella in all 4 placentae. By subsequent bacteriological examination Salmonella abortusequi was isolated as the single causative agent in each case. Nonmotile Salmonella abortusequi with antigenic formula 4,12:-:- was isolated from one of the 4 fetuses. The described episode of equine abortion clearly indicates that Salmonella abortusequi has not been eradicated from Europe as previously thought.

Prevention of necrotic enteritis in piglets by vaccination of pregnant gilts with a Clostridium perfringens type C and D bacterin-toxoid.

On a large pig farm with a known history of necrotic enteritis 12 pregnant gilts were vaccinated s. c. 7 and 2 weeks before expected farrowing with a commercial bacterin-toxoid preparation of toxigenic strains C. perfringens type C and D (DizevakR-Pliva, Zagreb). At the farrowing the titers of beta-antitoxins in serum samples from vaccinated gilts ranged from 9.0 to 26.0 IU/ml with a mean value of 14.16 IU/ml. Colostral titers varied from 12.0 IU/ml with a mean of 16.12 IU/ml. On the second day of life the mean serum titers between litters differed greatly from 4.75 to 24.0 IU/ml. By the age of 7 days the average serum titers were commonly lower and varied between the litters from 2.25 to 15.0 IU/ml, with a low of 1.5 to a high of 16.0 IU/ml in single animals. Ten (8.47%) out of a total of 118 piglets from vaccinated gilts died during the first 7 days of life but the losses were not caused by C. perfringens infection. In unvaccinated control animals 18 (15.9%) of 113 piglets died, eleven of them with clinical and pathoanatomical signs of necrotic enteritis. The affected piglets predominantly succumbed in the first 4 days of life. These data indicate that the investigated bacterin-toxoid can be successfully used in immunoprophylaxis of necrotic enteritis in piglets.

Susceptibility to antimicrobial agents of Streptococcus suis capsular type 2 strains isolated from pigs.

The minimal inhibitory concentrations (MICs) for thirty-three epidemiologicaly unrelated clinical isolates of Streptococcus suis capsular type 2 were determined in relation to ampicillin, ampicillin-sulbactam, amoxicillin, clavulanate-amoxicillin, penicillin G, cephalexin, gentamicin, streptomycin, erythromycin, tylosin and doxycycline, using the microtitre broth dilution procedure described by the U.S. National Committee for Clinical Laboratory Standards (NCCLS). Gentamicin was the most active compound tested, with an MIC for 90% of the strains tested (MIC(90)) of 0.4 mg/L. Overall, 70% of strains were resistant to doxycycline (MIC(90) > or = 100.0 mg/L), followed by penicillin G (51% of strains) (MIC(90) + or = 100.0 mg/L). Resistance to amoxicillin and ampicillin was 36.4% (MIC(90) 12.5 mg/L) and 33.3% (MIC(90) 50.0 mg/L), respectively. 15.2% of S. suis strains were resistant to streptomycin, tylosin and cephalexin with MIC90 values of 25.0 mg/L, 12.5 mg/L and 25.0 mg/L, respectively. A combination of ampicillin and sulbactam (MIC(90) 6.3 mg/L) and a combination of amoxicillin and clavulanate (MIC(90) 3.1 mg/L) as well as erythromycin (1.6 mg/L) were of the same efficacy, with a total of 9.1% resistant S. suis strains. This high percentage of resistance to doxycycline and penicillin G precludes the use of these antibiotics as empiric therapy of swine diseases.

Vibriosis in rainbow trout cultured in the Krka estuary, Croatia: occurrence and comments.

The frequency of vibriosis in cultured rainbow trout, maintained under different rearing conditions in the Krka estuary, was examined over a 6 yr period. Annual studies commenced regularly in October and ended in June of the following year. Every month during study periods, 37-75 trouts were randomly taken from each of 4 farm sites for routine examination. Twenty fish from these samples were subsequently employed for bacteriological analyses. Based on morphological and biochemical properties, the bacterial isolates were identified as Vibrio anguillarum, (biotype I). The findings demonstrate high occurrence of vibriosis in trout cultivated in the Krka estuary. Furthermore, there was a direct relationship between water quality parameters and the severity of vibriosis epizootics. Moreover, the causative agent has been isolated from free-living species, fish, notably eel and mullet, which are abundant to the Krka aquatorium. The findings from these long-term studies will be considered with reference to the developing salmon farming industry of Croatia.

Mycoplasmas associated with bovine conjunctivitis and keratoconjunctivitis.

In two separate herds of fattening calves a sudden-onset outbreak of ocular disease with profuse lacrimation occurred. The disease resembled the early stage of infectious bovine keratoconjunctivitis but after a few days the clinical signs of bronchopneumonia appeared. From conjunctival swabs Mycoplasma (M.) bovigenitalium, M. bovirhinis and infectious bovine rhinotracheitis (IBR) virus were isolated. Moraxella bovis infection was not established. In one of the herds M. bovigenitalium was also found in the pneumonic lungs of dead calves. In one herd M. bovoculi was isolated from a cow with chronic keratoconjunctivitis, housed together with affected calves. Mycoplasmas were not isolated from ocular swabs of six bulls originating from a Reproductive Centre with temporary occurrence of unilateral serous conjunctivitis resistant to antibiotic therapy.

Natural uterine Mycoplasma pulmonis infection in female rats.

Uterine washings from 124 apparently healthy and non-pregnant female Wistar rats of different ages were cultured for mycoplasmas and bacteria. The animals originated from four conventional breeding colonies which were known to be chronically infected with M. pulmonis from the previous microbiological examination. Mycoplasmas were isolated from the uterus in 30.6% of examined females. All the isolates were biochemically and seriologically identified as M. pulmonis. Uterine colonization with this organism was first evidenced in non-mated female rats at the age of three months. After mating the number of infected females rapidly increased. This observation points out the microbiologically uncontrolled mating as an important factor in the distribution of genital infection within the colony. Bacterial examination of uterine washings revealed only ubiquitous organisms in some animals. Gross lesions in the form of purulent salpingitis and mild endometritis were observed only in two animals.

[Aspergillus oryzae as a cause of keratomycosis in the horse]. / Aspergillus oryzae als Ursache einer Keratomykose beim Pferd.

A case of a spontaneous mycokeratitis of a previously injured cornea in a horse is described. The infection was caused by Aspergillus oryzae. After application of Chloramphenicol ophthalmic ointment a corneal clouding was found in the centre which was circularly sharply defined and which - after dispensing Dexamethason-Neomycin eye drops - expanded all over to a purulent keratitis. The demarcated and initially non purulent mycotic lesions largely improved after the application of tincture of iodine, whereas the purulent keratitis could be completely cured only by a lasting treatment with Lugol's solution.