Molecular cloning of the OMP19 gene from Brucella melitensis strain H38 and its antigenicity compared to that of commercial OMP19.

Brucellosis is a worldwide zoonosis, that can still be classified as endemic despite its ancient origins which causes economic losses and public health problems. Although effectively controlled by vaccination in animals, there is currently no vaccine for use in humans. Outer Membrane Proteins (OMP) that play an active immunogenic and protective role in the Brucellae family. OMP19 is present in all Brucella species as a surface antigen and is a potent immunogen responsible for Brucellosis intracellular infection. For this reason, the study was aimed to be used safely as a potential recombinant vaccine candidate against all Brucella infections, especially in humans and pregnant animals. This study evaluated a Brucella lipoprotein antigen, i.e. 19 kilodalton (kDa) outer membrane protein (OMP19), which was amplified and cloned into the pETSUMO vector system. The immunogenic power of the purified recombinant OMP19 antigen against brucellosis was compared with that of OMP19 (Raybiotech Inc, USA) in a mouse model and the obtained rOMP19 antigen was found to be similar to the commercially available recombinant protein.

Bactericidal efficacy of Er,Cr:YSGG laser irradiation against Enterococcus faecalis compared with NaOCl irrigation: an ex vivo pilot study.

AIM: To compare the efficacy of a standard NaOCl irrigation procedure with that of Er,Cr:YSGG laser irradiation in contaminated root canals having small and large apical foramina. METHODOLOGY: Forty root canals of extracted central incisor teeth with straight roots were chosen so that their apical foramina just permitted the tip of a size 20-K file to pass through. The canals were then enlarged with files to size 60 and randomly divided into four groups of 10 teeth each. The apical foramina of one group were widened further so that the tip of a size 45-K file could just pass through. After sterilization, all roots were inoculated with Enterococcus faecalis for 48 h at 37 degrees C. The first group was used as a control, the second group was irrigated with 3% NaOCl solution for 15 min, and the last two groups having different sizes of apical foramina were irradiated with the Er,Cr:YSGG laser at output power from 0.5 W, with 20% air and water levels. The disinfecting efficacy of the groups was tested by collecting dentine chips from the inner canal walls of the specimens and counting viable E. faecalis on Mueller-Hinton agar plates. RESULTS: The differences in the mean number of viable colonies between the control and laser groups were statistically significant (P < 0.05). The control specimens had the highest number of microorganisms (153 x 10(3) +/- 39 x 10(3)). Complete sterilization was achieved in the 3% NaOCl group. The mean colony forming units (CFU) values obtained after Er,Cr:YSGG laser irradiation were 6.6 x 10(3) CFU and 6.5 x 10(3) CFU in root canals having large and small apical foramina respectively. CONCLUSION: In teeth with straight roots the Er,Cr:YSGG laser reduced the viable microbial population in root canals with small and large apical foramina but did not eradicate all bacteria. Three percent NaOCl inhibited the growth of E. faecalis and effectively sterilized all root canals.

Field evaluation of a PCR for the diagnosis of chlamydial abortion in sheep.

A total of 94 vaginal swab samples and 195 serum samples collected from aborted ewes in 15 flocks were examined by pcr and a complement fixation test, respectively. In addition, 172 samples of stomach contents from fetuses from different flocks submitted for the diagnosis of abortion during the four lambing periods between 2000 and 2004 were tested by pcr. Chlamydial dna was detected in seven vaginal swabs obtained from five of the 15 flocks and in six samples of fetal stomach contents. The results of pcr and flock serology for Chlamydia were positive in five of the 15 flocks and negative in eight.

Efficacies of liposome-encapsulated enrofloxacin against Staphylococcus aureus infection in Anatolian shepherd dog monocytes in vitro.

The aim of this study was to evaluate the intracellular activity of two types of liposome-encapsulated enrofloxacin (LE) compared with free enrofloxacin and non-treated control against Staphylococcus aureus, phagocytosed by monocytes in healthy Anatolian Shepherd dogs. Enrofloxacin was encapsulated with two different types of liposome in multilamellar large vesicles (MLV). Type A MLV were composed of 15 mg phosphatidylcholine and 35 mg cholesterol, Type B MLV were composed of phosphatidylcholine, cholesterol and enrofloxacin in a molar ratio of 1:1:1. Intracellular activity was estimated by comparing the numbers of bacteria surviving intracellularly in monocytes exposed to free enrofloxacin and LE for 4 h at the doses of 0.25, 0.5 and 1 microg/ml, with those surviving intracellularly in untreated control monocytes. All three forms of enrofloxacin (free, Type A and B liposomes) increased the intracellular killing of S. aureus in a concentration dependent manner. Comparison of 1 microg/ml Type B LE revealed that killing activity was significantly higher than those of other concentrations. The results showed that LE was superior in reducing the number of intracellularly located bacteria compared to the free drug and control. The beneficial effect of liposomal encapsulation is presumably due to the fact that both liposomes and bacteria are localized at the same spot in phagocytic cells.

Comparison of antibacterial activity of two dentin bonding systems using agar well technique and tooth cavity model.

OBJECTIVE: This study compared the antibacterial activities of two dentin bonding systems (ABF, Kuraray and Reactmer Bond, Shofu) by a conventional agar well technique and a newly designed in vitro test using tooth model. METHODS: In the agar well technique, the test materials were filled in the wells of Muller Hinton agar plates inoculated with Streptococcus mutans NCTC10449, and the diameters of inhibition zones produced around the materials were measured after 24h of incubation. For the tooth model test, three cavities (diameter 1mm, depth 2mm) were prepared in the flat occlusal dentin of human extracted molar. After sterilization, the teeth were left in broth culture of 1.56 x 10(8)CFU/ml of S. mutans at 37 degrees C for 72h for allowing bacteria to invade the cavity. The dentin bonding systems were applied separately to each of the two infected cavities, and the third cavity was left unapplied for control. After sealing the occlusal surfaces, the teeth were kept in physiologic saline solution at 37 degrees C for 72h. The standardized amounts of dentin chips (120+/-5mg) were obtained from the cavity walls and the number of bacteria recovered was determined. The results were analyzed by One Way ANOVA, Kruskal Wallis and Mann-Whitney's U tests. RESULTS: The primer of ABF and Reactmer Bond produced inhibition zones with similar sizes (p>0.05), but the bonding resin of ABF did not produce any inhibition. When tested by the model cavity method, the application of ABF resulted in significantly less bacterial recovery than Reactmer Bond (p<0.05), demonstrating substantial antibacterial effects. CONCLUSIONS: The tooth model method used in this study was effective for evaluating the substantial antibacterial effects of dentin bonding agents, and the experimental dentin bonding system ABF was demonstrated to be able to inactivate the bacteria in the cavity effectively in comparison with little antibacterial activity shown by Reactmer Bond.

A comparative study on detection of Ornithobacterium rhinotracheale antibodies in meat-type turkeys by dot immunobinding assay, rapid agglutination test and serum agglutination test.

The objectives of the present study were to develop a dot-immunobinding assay (DIA) and a serum agglutination test (SAT) for detection of Ornithobacterium rhinotracheale, to compare the rapid agglutination test (RAT) and the SAT, and to make a serosurvey of O. rhinotracheale exposure on turkey farms in Turkey. Antiserum against O. rhinotracheale bacterin was prepared in rabbits and 72 serum samples were collected from turkeys with respiratory signs on four farms. Comparison of the tests showed that 55.5, 48.6 and 40.3% of serum samples were positive by RAT, SAT and DIA, respectively. The sensitivity of the DIA appeared to be lower than that of the agglutination tests but the specificity is not known.

Clinical efficacy of florfenicol in the treatment of calf respiratory tract infections.

This paper reports on a study of the aetiology of calf pneumonia and the clinical efficacy of florfenicol, a new antibiotic in Turkey. Twenty-seven weaned and unweaned calves (13 males and 14 females) between 1 and 16 months of age brought to the clinics of Selçuk University, Faculty of Veterinary Science. Broncho-alveolar lavage (BAL) fluid samples were taken from the animals diagnosed to have upper respiratory tract infection associated with bronchitis (N=2), bronchitis (N=5), bronchopneumonia (N=4), pneumonia (N=3), pleuropneumonia (N=11), bronchopneumonia plus pulmonary oedema (N=2) based on the results of the clinical and laboratory examinations. Then microbiological isolation and antibiotic culturing were performed. The animals were treated with 1 ml/15 kg (20 mg/kg) florfenicol (Nuflor, DIF) twice within 48 hours via intramuscular injection. At the end of the treatment, 23 of the weaned and unweaned calves were completely healed, 1 calf had died and 3 calves showed no healing. The results of BAL samples and microbiological examinations of the 3 calves that did not respond to the treatment indicated that these cases were affected by mixed infections of yeasts, fungi, and bacteria. Widespread pleuropneumonia was observed. According to the results of the microbiological examination of the BAL samples, Mannheimia (Pasteurella) haemolytica had the highest isolation rate (25%) compared with the other isolated bacteria, namely, Klebsiella pneumonia (20%), Actinomyces pyogenes (15%), beta-hemolytic streptococci. (10%), Staphylococcus spp. (5%), and E. coli (5%). The study also revealed fungi [Penicillum spp. (5%) and Aspergillus spp. (5%)] and two calves (10%) had a yeast infection.. We conclude that florfenicol has a high bacteriological and clinical efficacy (100% and 96% respectively) in the treatment of calf respiratory tract diseases.

Effects of dietary levels of vitamin A on the egg yield and immune responses of laying hens.

This research, which was designed and carried out as two consecutive experiments, investigated the effects of four different levels (0, 4,000, 12,000, and 24,000 IU/kg) of vitamin A supplementation on egg yield, plasma vitamin A levels, and immune responses of laying hens. Transmission of maternal immunity to their descendants was also studied. In the first experiment, egg yield, blood vitamin A levels, and various parameters of the immune system such as T lymphocyte levels in the peripheral blood, plasma cell counts in the spleen, and antibody titers against Newcastle Disease Virus (NDV) in the sera were investigated for a 1-yr period. A total of 864 Hisex-brown laying hens were used in this experiment. The chicks were reared as commercial flocks until the 18th wk of age. No significant differences occurred among the parameters of the different diet groups. In the second experiment, maternal immunity was assessed in the chickens, supplied by hatching the eggs from hens in the first experiment. Maternal immunity was assayed by using the parameters as in Experiment 1. For this purpose, both blood and tissue samples were taken on the 2nd, 7th, and 10th d posthatch. Vitamin A supplementation had no significant effects on maternally, derived antibody titers or histologic structure of the lymphoid organs.

An in vitro test model for investigation of disinfection of dentinal tubules infected with Enterococcus faecalis.

The aim of the present study was to develop an in vitro test model from human teeth to comparatively examine antibacterial effectiveness of calcium hydroxide, parachlorophenol (PCP) and camphorated parachlorophenol (CPCP) against Enterococcus faecalis in infected root canals. Cylindrical dentin specimens were prepared from freshly extracted human maxillary anterior teeth. The specimens were inoculated with E. faecalis and then medicated with either CPCP, PCP or Ca(OH)2. The disinfecting efficacy of these agents was tested by collecting dentin chips from the inner ("canal") walls of the specimens and counting viable E. faecalis. The dentin chips were diluted and a classical bacterial count technique was used for recovery of E. faecalis strains of 5% sheep blood agar. The effectiveness of CPCP and PCP at one day was superior to the effectiveness of Ca(OH)2. In the three-day group, CPCP was the most effective, followed by Ca(OH)2. The experimental model used in this study may be useful for investigation of the effect of intracanal medicaments on microorganisms lodged in the root dentinal tubules.

Suitability of lactate dehydrogenase activity and somatic cell counts of milk for detection of subclinical mastitis in Merino ewes (short communication).

The relationship between somatic cell counts (SCC) and LDH activity in milk was examined in Turkey to find out the suitability of these variables for early detection of subclinical mastitis in Merino ewes. A significant positive correlation was found between LDH activity and SCC in ewes' milk. LDH activity in milk samples appeared to be a sensitive and specific indicator of subclinical mastitis in ewes: it was significantly higher in milk from inflamed (mastitic) udders than in normal milk.

Hemagglutination, hydrophobicity, enterotoxigenicity, and drug-resistance characteristics of avian Escherichia coli.

A total of 35 Escherichia coli isolates obtained from necropsy materials of hens with septicemia in the Konya region of Turkey were examined for hemagglutination (HA), cell-surface hydrophobicity, enterotoxigenicity, and drug resistance. HA tests were performed on live cultures with human (group A), bovine, avian (chicken), and guinea pig erythrocytes with and without mannose. Nine HA patterns were observed. Of the 35 isolates, 62.8% exhibited mannose sensitive hemagglutination (MSHA), 8.6% exhibited mannose resistant hemagglutination (MRHA), and 28.6% did not hemagglutinate. Of the isolates, 85.7% were hydrophobic by a salt aggregation test (SAT). Only three isolates were enterotoxigenic by a suckling mouse assay. The majority of the isolates were resistant to chloramphenicol, tetracycline, streptomycin, ampicillin, erythromycin, and trimethoprim + sulfamethoxazole but were highly sensitive to gentamicin and nalidixic acid.

Isolation of Acinetobacter lwoffi from hens with septicemia.

Acinetobacter lwoffi was isolated from the lungs of two hens. The isolate was sensitive to chloramphenicol, ampicillin, erythromycin, oxytetracycline but resistant to penicillin and nitrofurantoin.