Seroprevalence of Coxiella burnetii antibodies among students of the Faculty of Medicine in Kosice (Slovakia).

Titers of immunoglobulin IgG against phases I and II of Coxiella burnetii were determined in 241 students of the Faculty of Medicine by ELISA method and the respective risk factors were evaluated, e.g., rural and urban life, consumption of milk, contact with animals and gender, which may be associated with exposure to C. burnetii. Phase I antibodies (Abs) were detected in 59 serum samples (24.4 %) at antibody level of 1: 100-1: 400. Phase II Abs were found in 179 persons (74.2 %). The titers were in the range of 1: 100-1: 1600. The titer > or =1: 800 of IgG was used as a cut-off level, and was detected only in 20 students (8.2 %). No significant difference in the prevalence of Abs was detected either between the students living in rural and urban environment (78.8 and 73.2 %, respectively) or between males and females (74.0 and 74.7 %, respectively). Abs were detected more frequently in raw milk consumers (68.1 %) and in students who kept some animals (73.7 %).

Serotypes, virulence genes, intimin types and PFGE profiles of Escherichia coli isolated from piglets with diarrhoea in Slovakia.

Two hundred and fifty Escherichia coli isolates from diarrhoeic and healthy piglets were serotyped and tested for the presence of virulence genes for fimbriae, intimin, heat-labile (LT) and heat-stable (STa and STb) enterotoxins, Stx toxins, and enteroaggregative heat-stable 1 (EAST1) enterotoxin by polymerase chain reaction (PCR). Although 220 isolates from diarrhoeic piglets belonged to 43 O serogroups and 77 O:H serotypes, 60% were of one of the 10 serogroups O2, O8, O15, O54, O84, O101, O141, O147, O149 and O157, and 60% belonged to only 10 serotypes (O8:H-, O54:H-, O84:H7, O101:H-, O141:H-, O141:H4, O147:H-, O149:H10, O163:H-, and ONT:H-). PCR showed that 79% of 220 isolates carried genes for at least one of the virulence factors tested. The gene encoding for EAST1 was the most prevalent (65%) followed by those encoding for STb (49%), LT (42%), STa (13%), and Stx2e (4%). Eighty-three (38%) of the 220 E. coli isolates carried the gene for F4 (K88), whereas genes for F18, F5 (K99), F41, F6 (P987), F17, and intimin (eae) were detected in 9%, 3%, 3%, 3%, 1%, and 3%, respectively. Seropathotype O149:H10:F4:LT/STb/EAST1 (70 isolates) was the most common, representing 32% of isolates. Pulsed-field gel electrophoresis (PFGE) analysis with XbaI of 15 O149:H10 representative isolates from diarrhoeic piglets distinguished 14 types. The 15 isolates exhibited a wide variability of distinct restriction patterns though all belonged to the same serotype (O149:H10), and all but one showed identical virulence determinants (F4, LT, STb, and EAST1). Among 30 isolates from healthy piglets only two virulence genes were detected: EAST1 (26%) and eae (17%). In total, 12 isolates were positives for the eae gene: five isolates had intimin beta1, four possessed intimin theta and three showed intimin type xiB. This is believed to be the first study describing the presence of intimin type xiB in E. coli of porcine origin.

Predictions for the timing and use of culling or vaccination during a foot-and-mouth disease epidemic.

First-fortnight incidence (FFI) is a modelling parameter that can be used to predict both the prevalence and duration of a foot-and-mouth disease (FMD) epidemic at regional and national levels. With an indication of how long an epidemic may last by the end of week two, it becomes possible to estimate whether vaccination would be economically viable from the start of an epidemic. Where FFI indicates that an epidemic is unlikely to last for as long as an export ban on agricultural produce, it may be inappropriate to implement a policy of 'vaccination to live'. Alternatively where FFI indicates that an epidemic will equal or exceed the ban length, then the benefits of vaccination should be considered at an early stage, during or after the first fortnight. Since blanket vaccination of the national or regional herds and flocks would be both costly and heighten the risk of producing carrier animals, targetting vaccination through risk assessment becomes useful.

Diabetes mellitus and candidiases.

Patients in various clinical states of diabetes mellitus (according to the recommendation of the American Diabetes Association) as a primary diagnosis were examined for fungal infections by Candida species. Candida spp. were detected in urine, in the material taken from the mouth cavity, nails, skin lesions, ears and eyes, by cultivation on the Sabouraud agar, CHROMagar Candida, and by saccharide assimilation. In the group of diabetics with symptoms of oral candidiasis and denture stomatitis C. albicans was identified in 8 cases, C. tropicalis in 3, C. parapsilosis in 2; 1 strain of C. guilliermondii was also isolated. In patients with urinary tract infections the presence of C. albicans was shown in 12 cases; C. parapsilosis was detected in 6 cases and two strains of each C. tropicalis and C. krusei were also isolated. In patients with leg ulcers C. albicans (25 cases), C. parapsilosis (5), C. tropicalis (3) and one strain of each C. krusei and C. robusta were isolated. Otomycosis was associated with one strain of C. albicans, C. parapsilosis, C. tropicalis and C. guilliermondii. C. albicans was most frequently associated with onychomycosis, paronychia and endophthalmitis; C. parapsilosis was the second most rated yeast.

PCR assay for detection and differentiation of K88ab(1), K88ab(2), K88ac, and K88ad fimbrial adhesins in E. coli strains isolated from diarrheic piglets.

Primers were designed and prepared and conditions were determined for PCR detection and differentiation of enterotoxigenic E. coli bacterial strains isolated from diarrheic pigs. Primers K88/1 and K88/2 are 25 bp oligomers that correspond to a region of genes encoding one of serological variants of the K88 antigen (K88ab(1), K88ab(2), K88ac or K88ad). A positive result of PCR is an amplificate of 792 bp in size for K88ab and K88ad variant or 786 bp for K88ac variant. The individual serological variants of genes of the K88 antigen could be differentiated by cutting the obtained PCR amplificates by restriction endonucleases. The PCR analysis of 674 E. coli strains isolated from diarrheic pigs showed that 184 strains were K88 positive. By using restriction endonucleases the K88-positive strains were in 4 cases classified as K88ab variant, 180 as K88ac variant and none contained gene for the K88ad variant. Ninety-five % coincidence with serological examination using K88ab, K88ac and K88ad specific antibodies was shown.

Distribution of virulence genes in Escherichia coli strains isolated from diarrhoeic piglets in the Slovak Republic.

Ninety-two Escherichia coli isolates from 14 to 28-day-old piglets that died because of diarrhoea were examined for genes for fimbriae (F4, F5, F6, F18 and F41), enterotoxins (STa, STb and LT), verotoxin (VT2e or Stx2e) and enteroaggregative heat-stable enterotoxin 1 (EAST1) by polymerase chain reaction. Twenty-two strains (24%) carried a gene for F4, whereas genes for F18, F6 and F5 + F41 were detected in 10.8, 3.3 and 1.1% of strains respectively. Genes for STb, LT, STa and Stx2e were detected in 40.2, 26.1, 14.1 and 1.1% of strains respectively. The astA gene was detected in 49 (53.3%) isolates, 35 of which also carried genes for enterotoxins and/or fimbriae. The major genotypes reached at (in decreasing order of prevalence) were F4/STb/LT/EAST1, F18/STa/STb/EAST1, STb/EAST1, F6/STa/STb/EAST1 and F18/STb/EAST1.

Genotyping of fimbrial adhesins in Escherichia coli strains isolated from Slovak piglets suffering from diarrhea.

One-hundred sixty Escherichia coli isolates obtained from piglets with diarrhea from different parts of Slovakia were examined for the presence of genes coding for F4, F5, F6 and F41 fimbrial adhesins, and hemolytic activity. According to polymerase chain reaction tests 74 (46%) E. coli isolates were positive for primers that detected genes coding for fimbrial adhesins. Of these 74 isolates, 64 were positive for genes encoding for F4+, four for F5+, five for F6+, and one for both F41+ and F5+ adhesins.

Occurrence of Candida strains in cases of paronychia.

A total of 43 patients, 11 males and 32 females, with paronychia of the fingernails were examined for the presence of Candida spp. The yeast species isolated were identified using standard laboratory methods, including germ-tube production, morphology on rice agar with Tween 80, and mainly fermentation and assimilation of saccharides. In the male group, two Candida species were detected: C. albicans as the dominant species in 9 patients and C. parapsilosis in 2 cases. Similarly, C. albicans was the prevalent species also in females (n = 17); other Candida species detected were C. parapsilosis (n = 7), C. tropicalis (5) and C. krusei (3). In addition to the genus Candida, the following anaerobic and aerobic microorganisms were isolated from patients of both groups: Fusobacterium spp., Bacteroides spp., Staphylococcus aureus, alpha-hemolytic streptococci, group A beta-hemolytic streptococci, Klebsiella pneumoniae, Neisseria spp. and Pseudomonas aeruginosa.

Otomycoses of candidal origin in eastern Slovakia.

Mycological analysis of swabs and scraping samples from the external ear canals of 40 patients with clinically diagnosed otomycosis (10 neonates, 30 adults) revealed the presence of fungi as etiological agents. They were investigated microscopically using 20 % potassium hydroxide, and by cultivation on Sabouraud's glucose agar. The Candida species were identified using the germ-tube test, micromorphology observations of colonies on rice agar, and particularly by the commercial kit AUXAcolor. The following Candida species were identified in the aural material examined: C. albicans (n = 21; 52.5 %), C. parapsilosis (11; 27.5), C. tropicalis (3; 7.5), C. krusei (3; 7.5), C. guilliermondii (2; 5.0). The above yeasts were present in samples together with Staphylococcus epidermidis (31), S. aureus (16), alpha-hemolytic streptococci (14), Neisseria spp. (14), Proteus mirabilis (3), Pseudomonas aeruginosa (3), Escherichia coli (1) and Haemophilus influenzae (1). The most frequent predisposing factors for otomycosis were swimming in public pools and/or bath, spa and diabetes mellitus.

The detection of anti-Candida antibodies on experimental animal models.

The levels of anti-Candida antibodies were determined in experimental animals immunized with 4 different yeast doses (0.3, 0.6, 1.2, 2.4 x 10(8) CFU) at weekly intervals. After immunization (sampling intervals 1, 2, and 3 months), the intravenous blood was examined for the presence of serum anti-Candida antibodies. After one month, the titers of anti-Candida antibodies reached 1:40-1: 1280 and remained at the same level after two months in the majority of animals; only in a few of them the titers increased or were detected de novo. After three months, when the animals were no longer immunized, a decreasing trend in antibody titers was detected in the majority of animals.

Candida--agent of the diaper dermatitis?

Occurrence of Candida spp. was determined in a population of 60 infants, 1-15-month-old, with diaper dermatitis, admitted to a neonatal intensive care unit in Hospital Saca (Kosice, Slovakia). Specimens were obtained from the perianal, pubic, inguinal, or gluteal areas that showed signs of secondary infection as manifested by erythema, oozing, vesiculopustular lesions, and pus formation. The most frequently isolated species was C. albicans (41), followed by C. parapsilosis (8), C. tropicalis (4), C. pulcherrima (4), C. guilliermondii (2), and C. zeylanoides (1). Other organisms present in the mixed culture from the diaper area were Staphylococcus aureus (6), Escherichia coli (3), and 2 strains of each group B and D streptococci, and Proteus mirabilis. Infants diapered exclusively in disposable diapers showed less rash than those diapered exclusively or sometimes in cloth diapers.

Effect of vaccination therapy for acne, using a staphylococci antigenic complex in combination with clindamycin.

A group of 46 patients with acne vulgaris were prescribed clindamycin in combination with a vaccination therapy using a staphylococci antigenic complex (Polystafana; Sevapharma, Czechia). Acne papulopustulosa was diagnosed in 36 patients and acne indurata in 10 patients. The clinical effect of clindamycin and Polystafana was determined on the basis of the regression of the inflammatory manifestation of acne: regression by 0-30% was considered unsuccessful, by 30-80% satisfactory, and above 80% excellent; the immunomodulation effect of the treatment was evaluated in parallel. The clinical effect was excellent in 21 patients with acne papulopustulosa and in 6 with acne indurata; it was accompanied by pronounced immunomodulation effect as both cellular and humoral immunity was also tended to return to normal values. A satisfactory clinical effect was observed in 15 patients with acne papulopustulosa, whose cellular immunity component was also stimulated. Only 4 patients with acne indurata failed to respond successfully. These were the patients who showed a pronounced deficit of the cellular immunity component persisting even after the treatment. The administration of clindamycin in combination with Polystafana vaccination can be recommended as an effective means for the treatment of acne vulgaris.

Spreading of Salmonella enteritidis in the cecum of chickens.

Adhesion and colonization of high (2 x 10(8) CFU) and low doses (2 x 10(2) CFU) of Salmonella enteritidis (phage type 4) was determined in the ceca collected 6 h-4 weeks after inoculation (pi), of 1-d-old White Plymouth Rock orally-inoculated chickens. S. enteritidis was associated with the epithelial surface of the villi in the low-dose group 18 h-7 d pi, the penetration in the cecal lamina propria was observed on day 1 and 10 pi. In the high-dose group, adhesion and colonization was observed in all birds killed 6 h-14 d pi; penetration of the bacteria into the cecal lamina propria was seen 1-21 d pi. Large numbers of macrophage-like cells containing S. enteritidis were observed in the cecal lamina propria on days 3-21 pi. Colonization and migration by S. enteritidis in the intestinal tract of chickens was shown to be dose dependent.

Comparative study of the topographic localization of NADPH diaphorase positive cells in the rabbit and pheasant thymuses.

Nicotinamide adenine dinucleotide hydrogen phosphatediaphorase (NADPHd) histochemistry was used as a marker for nitric oxide synthase (NOS). In the rabbit thymus, NADPHd staining was observed between the capsule and corticomedullary junction in radially oriented blood vessels in the cortex. The outer surface of the thymic lobule and interlobular septa showed adipocytes clumped together. There was a high density of NADPHd positive cells in the medulla, without a sharp boundary in corticomedullary space. In addition to radially oriented blood vessels in the cortex, they were also found as solitary profiles with well stained walls in the medulla. Neuronal plexuses were localized in perivascular topography. In the pheasant thymus, NADPHd positive cells were present as clusters which were distributed in the medulla and the corticomedullar area. NADPHd positive nerve fibres were localized in perivascular topography.

Detection of anti-Candida antibodies in neonates from a Neonatal Intensive Care Unit.

Anti-Candida antibodies were determined in a group of preterm neonates from a neonatal intensive care unit with serious diseases including candidemia. Antibodies to C. albicans blastospores, i.e. antibodies to C. albicans surface mannan and to C. albicans germ tubes were detected. Higher titers of antibodies to blastospores (1:320) occurred in all patients examined while antibodies to C. albicans germ tubes (with the highest titer of 1:160) were present in 32 out of 66 neonates examined. The highest titers of both anti-C. albicans blastospore antibodies and anti-C. albicans germ tube antibodies were detected in neonates with candidemia and disorders of saccharide metabolism.

Fungal diseases of the respiratory tract.

The proportion of Candida and non-Candida species in the clinical material from patients with respiratory-tract diseases was determined. C. albicans was isolated in 102 cases. An additional 89 strains of yeasts, isolated in association with respiratory diseases, belonged to 10 non-albicans Candida spp. and Cryptococcus spp. The prevailing species, which occurred in 47 cases, was C. parapsilosis. C. tropicalis, C. glabrata, and C. guilliermondii were isolated in 12, 10, and 9 cases, respectively. Four strains of C. krusei and three strains of C. lusitaniae and one strain each of C. freyschussii, C. robusta, C. zeylanoides, and Cryptococcus neoformans were also isolated.

Candida species isolated from cerebrospinal fluid.

In the period from November 1998 to June 2001 13 cases of nosocomial meningitis were reported. Candida albicans was isolated from 54% of the patients (7); C. parapsilosis from 23% (3); C. tropicalis from 15% (2) and C. krusei from 8% (1). C. albicans was isolated from the cerebrospinal fluid (CSF) of five children with the following diagnoses: nonspecified tumor of the central nervous system, Hodgkin's disease, meningitis, suspect neuroinfection, and sepsis. Examination of CSF allowed us to detect 2 strains of C. albicans from adult patients, one after neurosurgery because of a brain tumor and one with a vascular disease of the brain. C. parapsilosis was found in CSF from two premature children and one child with epilepsy. Two isolates of C. tropicalis were obtained from both blood and the CSF of a child from the neonatal intensive care unit and from a child from pediatric oncology with multiple malignant neoplasms. Only one strain of C. krusei was found in the oral cavity and CSF of a patient after neurosurgery performed after head trauma.

Candidal urinary tract infections caused by non-albicans Candida species.

The incidence of non-albicans Candida and non-Candida species isolated from the urine of patients admitted to various departments of the Faculty Hospital of the Medical Faculty of Safárik University in Kosice was examined. From a total of 94 samples of analyzed urine 58 strains of C. albicans and 36 strains of yeasts belonging to 6 species of non-albicans Candida and non-Candida spp. were detected: C. parapsilosis (n = 23), C. tropicalis (6), C. krusei (3), C. robusta (2), C. catenulata (1) and Cryptococcus neoformans (1). In relation to the diagnosis, the yeasts were isolated from patients suffering from a kidneys disease, epididymitis, diabetes, neoplastic diseases, urogenital anomalies, obstructive uropathy, cystitis, prostatitis, hemolytic-uremic syndrome, and others.

Detection of anti-Candida antibodies by the indirect immunofluorescence assay in patients with cancer in the orofacial region.

An indirect immunofluorescence assay was performed to detect antibodies to Candida albicans blastospores and germ tubes. Serum specimens were obtained from 82 patients with neoplastic diseases in the orofacial region and thrush of the oral mucosa. C. albicans was identified in the oral cavity of 63 patients investigated but serum anti-Candida antibodies were detected in only 23 of them. Serological examination showed that titers of antibodies to C. albicans blastospores ranged from 1:20 to 1:1280. High titers from 1:640 to 1:1280 were detected in patients without antibiotic, cytostatic, or radiotherapeutic treatment. The titers of antibodies to C. albicans germ tubes ranged from 1:20 to 1:640. Our results indicate that titers of antibodies to the C. albicans germ tubes were lower and were detected in a smaller number of patients.

Yeast-like microorganisms in eye infections.

The proportion of yeast species involved in eye infections in 11 patients was examined. The presence of yeast organisms as causative agents of endophthalmitis was found in corneal smears (n = 4), conjunctival swabs (4), and vitreous fluid (3). Altogether 5 strains of Candida albicans, 2 strains of C. krusei and one strain each of C. guilliermondii, C. parapsilosis, C. tropicalis and Cryptococcus neoformans were isolated from the clinical material. The hematogenic origin of endophthalmitis was proved in 7 cases on the basis of positive blood samples and in 2 cases by the isolation of yeasts from the tip of an intravenous catheter. Endophthalmitis-supporting risk factors such as indwelling intravenous catheters, prolonged use of broad-spectrum antibiotics and chemotherapy, surgical intervention, diabetes mellitus, and malignancy were observed in the patients.