RESUMEN
Vaccination is the most effective, reliable, and economical way of preventing or reducing the effect of infectious diseases. When preparing inactive vaccines, a range of additives called adjuvants are necessary to enhance the magnitude of the immune response. Boron has a wide range of industrial and medical applications, and its positive effects on distinct functions have been described in plants, humans, and animals. However, no studies exist about the possible adjuvant activities of boron compounds in vaccines. Hence, in this study, the potential adjuvant effect of boric acid was explored and compared with common veterinary adjuvants in a mice model. Staphylococcus aureus (S. aureus) used as vaccine antigen was isolated from dairy cows with bovine mastitis. Vaccines adjuvanted with boric acid, aluminum hydroxide, Montanide ISA 50 and ISA 206, and Montanide + boric acid combinations were prepared. The efficacy of vaccines was evaluated according to local reactions at the injection site, C-reactive protein, total Ig G, total Ig M, and anti-S. aureus antibody levels in mice. Boric acid reduced local inflammatory reactions induced by the Montanide adjuvants. Moreover, mice vaccinated with boric acid-adjuvanted vaccine had higher levels of anti-S. aureus antibody than those in the controls (P < 0.05) and were similar to the levels found in mice sensitized with aluminum hydroxide. Total Ig G and Ig M results were, however, unsuitable for the assessment of adjuvant activity for this study. In conclusion, this study revealed that boric acid has an adjuvant potential in inactive bacterin vaccines, but further target animal studies are needed.
Asunto(s)
Vacunas Bacterianas , Mastitis Bovina , Adyuvantes Inmunológicos/farmacología , Animales , Boro/farmacología , Bovinos , Femenino , Ratones , Staphylococcus aureusRESUMEN
Mycoplasma spp. can cause diseases of the respiratory system as well as urogenital infections, infertility, and anemia. The members of this genus have a low Gâ¯+â¯C content compared to other bacteria. Because primers used in the random amplified polymorphic DNA (RAPD) technique are only 10â¯bp long and have high GC content, this method can be inadequate for genotyping Mycoplasma spp. isolates. The aim of this study was to develop and evaluate multiple-locus variable number tandem repeat analysis (MLVA) and two-primer RAPD (TP-RAPD) procedures for subtyping Mycoplasma cynos isolates. A total of 55â¯M. cynos isolates obtained from 162 bronchoalveolar lavage fluid samples from shelter and pet dogs were used in this study. Seventy-four tandem repeat regions were detected in the M. cynos genome, and two of these loci were determined to be suitable and used for development of the MLVA scheme. The results of variable number tandem repeat (VNTR) analysis and TP-RAPD-PCR were compared with RAPD-PCR. The discriminatory power of TP-RAPD-PCR (Hunter-Gaston diversity index [HGDI]â¯=â¯0.84) was higher than those of RAPD-PCR (HGDIâ¯=â¯0.727), VNTR1 (HGDIâ¯=â¯0.8), and VNTR3 (HGDIâ¯=â¯0.757). We observed that the TP-RAPD-PCR and MLVA methods provide clearer data and are more successful in determining genetic diversity, in contrast to the RAPD-PCR method for this species.
Asunto(s)
Repeticiones de Minisatélite , Mycoplasma/genética , Técnica del ADN Polimorfo Amplificado Aleatorio/métodos , Animales , Composición de Base , Líquido del Lavado Bronquioalveolar/microbiología , Cartilla de ADN , ADN Bacteriano/genética , Perros , Variación Genética , Genotipo , Técnicas de Genotipaje , Mycoplasma/aislamiento & purificación , Infecciones por Mycoplasma/microbiología , Infecciones por Mycoplasma/veterinaria , Reacción en Cadena de la Polimerasa , Secuencias Repetidas en TándemRESUMEN
Extraction of DNA from Mycoplasma cultured on agar medium is difficult because the plasticity of these microorganisms enables agar penetration. This eventually causes cell loss during harvesting of colonies from the agar surface. Here, we used the GenElute™ gel extraction kit, which is usually used to purify polymerase chain reaction products, for extracting DNA from Mycoplasma. We compared the DNA extraction efficiency of the GenElute™ gel extraction kit from Mycoplasma cynos cultured in agar medium with four other DNA extraction methods. The results were evaluated based on the purity and amount of DNA obtained from one Mycoplasma colony. Eight strains of Mycoplasma cynos isolated from the broncho-alveolar lavage fluid of dogs were used. The GenElute™ gel extraction protocol was the most efficient among all the methods tested in this study as it yielded the highest amount and the purest quality of DNA (199.3±0.744ng/µl) from a single colony. Among the methods tested, the GenElute™ gel extraction method is the most rapid, sensitive, and simple method for DNA extraction from Mycoplasma. This procedure may also prove useful for extracting DNA from other Mycoplasma species.
Asunto(s)
Técnicas Bacteriológicas/métodos , ADN Bacteriano/aislamiento & purificación , Mycoplasma/genética , Agar , Animales , Técnicas Bacteriológicas/instrumentación , Medios de Cultivo , ADN Bacteriano/genética , Enfermedades de los Perros/diagnóstico , Perros , Infecciones por Mycoplasma/diagnóstico , Infecciones por Mycoplasma/veterinaria , Reacción en Cadena de la Polimerasa/métodos , Sensibilidad y EspecificidadRESUMEN
Boron (B) compounds are used in many fields ranging from medicine to industry. In this study, boric acid (BA) and disodium octaborate tetrahydrate (DOT) were evaluated for their antibacterial effects and antibiofilm capacities on selected strains of clinical and type cultures that are of veterinary concern (Staphylococcus aureus ATCC 25923, Aeromonas hydrophila ATCC 19570, Pseudomonas aeruginosa ATCC 27853, Brucella melitensis Rev1 and field isolates of Vibrio anguillarum, Aeromonas hydrophila, Yersinia ruckeri, Pseudomonas aeruginosa, Lactococcus garvieae, and Brucella abortus). Also, the inhibition of biofilm was monitored by scanning electron microscopy. The lowest MIC values of BA and DOT were measured, by broth method using microdilution, from Pseudomonas aeruginosa ATCC 27853, and were 0.385 and 0.644 mg/ml, respectively. Staphylococcus aureus was the most resistant to both BA and DOT. Using the microplate method, we observed that the strongest positivities for biofilm production were presented by Pseudomonas aeruginosa ATCC 27853 and also a clinical isolate of Lactococcus garviea. Lower values in the MIC scores for both B compounds were tested by measuring the inhibitory effect on biofilm production. We found that all the bacterial strains inhibited biofilm formation with the exception of the Pseudomonas aeruginosa strains for BA only and an isolate of Lactococcus garviea for DOT only. Such effects by BA and DOT are worth discussing in order to find novel approaches for different functions in medicine and industry using the bacteria tested.
Asunto(s)
Antibacterianos/farmacología , Bacterias/crecimiento & desarrollo , Fenómenos Fisiológicos Bacterianos/efectos de los fármacos , Biopelículas/crecimiento & desarrollo , Boro/farmacologíaRESUMEN
Brucella canis which is the main etiologic agent of brucellosis in dogs, can be transmitted to man. It causes mild or asymptomatic infection in human compared with other Brucella species. B.canis can be transmitted to man either by laboratory accidents or contact with infected dogs. Since B.canis infections in humans are not routinely investigated in hospitals in Turkey, the data are limited to reveal the current status of B.canis infections in people in our country. The purpose of this study was to determine the seroprevalence of B.canis infection in brucellosis-suspected cases. The study was conducted at Konya Education and Research Hospital, (located at Central Anatolia of Turkey) during March-August 2010 period. Serum samples were obtained from 1000 patients (age range: 15-65 years; 652 of them were women) presented with brucellosis-like symptoms, including fever, headache, night sweats, appetite loss, weakness, arthralgia and myalgia. Rose Bengal Plate Tests (Seromed, Turkey) for smooth Brucella species were negative in all serum samples. Rough type B.canis antigen was prepared with B.canis NCTC 10854 strain for serodiagnosis. Antibody responses to B.canis in the serum samples were investigated by rapid slide agglutination test (SAT) and modified plate agglutination test (MPAT). Of the 1000 sera tested, 34 (0.34%) were found to be positive with SAT while the remaining were found negative. MPAT was used for the detection of antibody titer and 22 (0.22%) out of 1000 sera were found positive with MPAT (one had 1/48, five had 1/96, six had 1/192, six had 1/384, four had 1/768 titers). Among 22 positive patients, 17 were female and five were male, and the difference between the genders was found statistically significant (p< 0.05). It was concluded the use of both S and R antigens in the serological tests applied for the diagnosis of brucellosis in our country will supplement both diagnosis and seroepidemiological data related to brucellosis.