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Prevalence of pathogens from Mollicutes class in cattle affected by respiratory diseases and molecular characteristics of Mycoplasma bovis field strains
2016
Szacawa Ewelina | Szymańska-Czerwińska Monika | Niemczuk Krzysztof | Dudek Katarzyna | Woźniakowski Grzegorz | Bednarek Dariusz
Introduction: Mycoplasma bovis is one of the main pathogens involved in cattle pneumonia. Other mycoplasmas have also been directly implicated in respiratory diseases in cattle. The prevalence of different Mycoplasma spp. in cattle affected by respiratory diseases and molecular characteristics of M. bovis field strains were evaluated. Material and Methods: In total, 713 nasal swabs from 73 cattle herds were tested. The uvrC gene fragment was amplified by PCR and PCR products were sequenced. PCR/DGGE and RAPD were performed. Results: It was found that 39 (5.5%) samples were positive for M. bovis in the PCR and six field strains had point nucleotide mutations. Additionally, the phylogenetic analysis of 20 M. bovis field strains tested with RAPD showed two distinct groups of M. bovis strains sharing only 3.8% similarity. PCR/DGGE analysis demonstrated the presence of bacteria belonging to the Mollicutes class in 79.1% of DNA isolates. The isolates were identified as: Mycoplasma bovirhinis, M. dispar, M. bovis, M. canis, M. arginini, M. canadense, M. bovoculi, M. alkalescens, and Ureaplasma diversum. Conclusion: Different Mycoplasma spp. strains play a crucial role in inducing respiratory diseases in cattle.
Show more [+] Less [-]Prevalence of pathogens from Mollicutes class in cattle affected by respiratory diseases and molecular characteristics of Mycoplasma bovis field strains
2016
Szacawa, Ewelina | Szymańska-Czerwińska, Monika | Niemczuk, Krzysztof | Dudek, Katarzyna | Woźniakowski, Grzegorz | Bednarek, Dariusz
Introduction: Mycoplasma bovis is one of the main pathogens involved in cattle pneumonia. Other mycoplasmas have also been directly implicated in respiratory diseases in cattle. The prevalence of different Mycoplasma spp. in cattle affected by respiratory diseases and molecular characteristics of M. bovis field strains were evaluated. Material and Methods: In total, 713 nasal swabs from 73 cattle herds were tested. The uvrC gene fragment was amplified by PCR and PCR products were sequenced. PCR/DGGE and RAPD were performed. Results: It was found that 39 (5.5%) samples were positive for M. bovis in the PCR and six field strains had point nucleotide mutations. Additionally, the phylogenetic analysis of 20 M. bovis field strains tested with RAPD showed two distinct groups of M. bovis strains sharing only 3.8% similarity. PCR/DGGE analysis demonstrated the presence of bacteria belonging to the Mollicutes class in 79.1% of DNA isolates. The isolates were identified as: Mycoplasma bovirhinis, M. dispar, M. bovis, M. canis, M. arginini, M. canadense, M. bovoculi, M. alkalescens, and Ureaplasma diversum. Conclusion: Different Mycoplasma spp. strains play a crucial role in inducing respiratory diseases in cattle.
Show more [+] Less [-]Chlamydia psittaci reference genes for normalisation of expression data differ depending on the culture conditions and selected time points during the chlamydial replication cycle
2016
Van Lent, Sarah | Vanrompay, Daisy
Introduction: Chlamydia psittaci is a gram-negative obligate intracellular pathogen of birds. Poultry infections lead to economic losses and can be transmitted to humans. No vaccine is available and the bacterium-host cell interaction is not completely understood. Replicating bacteria cause pneumonia, but C. psittaci can also be non-replicating and persistent inside the cytoplasm of avian cells. RT-qPCR provides insight into the molecular pathogenesis of both active replicating and persistent Chlamydia psittaci in birds, but requires identification of stably expressed reference genes to avoid biases. Material and Methods: We investigated the expression stability of 10 C. psittaci candidate reference genes for gene expression analysis during normal growth and penicillin-induced persistence. C. psittaci Cal10 was cultured in HeLa229 and RNA was extracted. The expression level of each candidate was examined by RT-qPCR and Cq values were analysed using geNorm. Results: The genes tyrS, gidA, radA, and 16S rRNA ranked among the most stably expressed. The final selected reference genes differed according to the bacterial growth status (normal growth versus persistent status), and the time points selected during the duration of the normal chlamydial developmental cycle. Conclusion: The study data show the importance of systematic validation of reference genes to confirm their stability within the strains and under the conditions selected.
Show more [+] Less [-]Pathogenicity of local isolates of Mycoplasma ovipneumoniae and Mycoplasma arginini in experimental West African Dwarf goats
2016
Chinedu Adive Akwuobu | Kennedy Foinkfu Chah | Stephen Ike Oboegbulem | Jude S Rabo
Objective: This study was carried out to assess the pathogenicity of local isolates of Mycoplasma ovipneumoniae and M. arginini in West African dwarf goats (kids) in Nigeria. Materials and methods: A total of 22 goats aged less than 1-year were purchased from markets. The goats were divided into six groups comprising of four experimental groups (EG; 4 in each) and two control groups (CG; 3 in each). The goats were fed ad libitum with standard diets and safe water. Groups EG1 and EG2 were infected with M. ovipneumoniae through trans-tracheal (TT) and intravenous (IV) routes, respectively, while those in groups EG3 and EG4 were infected with M. arginini through the same routes. Goats in groups CG1 and CG2 were inoculated with sterile Mycoplasma broth through TT and IV routes, respectively. In all cases, the amount of bacteria inoculated was 1.5x108 cells/mL. After the onset of the disease in goats, re-isolation of Mycoplasma was performed by culturing on mycoplasma agar supplemented with mycoplasma supplement G. The goats were monitored for 14 days post-infection (PI) to observe respiratory signs and mortality. Post-mortem (PM) examination was performed on each animal that died, while one surviving goat from each of the groups was sacrificed at 14 days PI for PM. After PM, histopathology was performed to observe the changes in tissues. Results: Cough and nasal discharges were observed in all the experimentally infected goats seven days PI. Mortalities were recorded in goats in EG1 (two goats), EG2 (one goats), EG3 (two goats) and EG4 (one goat). At PM, pneumonic lesions were observed in the lungs of all the experimentally infected goats. Conclusion: This study provides evidence that the local isolates of M. ovipneumoniae and M. arginini strains are pathogenic for goats in Nigeria. [J Adv Vet Anim Res 2016; 3(3.000): 242-251]
Show more [+] Less [-]Para influenza virus 3 infection in cattle and small ruminants in Sudan
2016
Intisar Kamil Saeed | Yahia Hassan Ali | Khalid Mohammed Taha | Nada ElAmin Mohammed | Yasir Mehdi Nouri | Baraa Ahmed Mohammed | Osama Ishag Mohammed | Salma Bushra Elmagboul | Fahad AlTayeb AlGhazali
Objective: This study was aimed at elucidating the association between Para influenza virus 3 (PIV3) and respiratory infections in domestic ruminants in different areas of Sudan.Materials and methods: During 2010-2013, five hundred sixty five lung samples with signs of pneumonia were collected from cattle (n=226), sheep (n=316) and goats (n=23) from slaughter houses in different areas in Sudan. The existence of PIV3 antigen was screened in the collected samples using ELISA and Fluorescent antibody technique. PIV3 genome was detected by PCR, and sequence analysis was conducted.Results: Positive results were found in 29 (12.8%) cattle, 31 (9.8%) sheep and 11 (47.8%) goat samples. All the studied areas showed positive results. Highest prevalence (66.7%) was detected in the sheep and goats in Khartoum, followed by in goats in Nyala (33.3%) at western Sudan. Sequence analyses of PIV3 of different regions of Sudan indicated that these were similar in sequence and length. The BLAST analysis indicated that the test sequences were closely related to the available annotated sequences at the GenBank. All these sequences matched with Bovine parainfluenza virus 3 except two those were matching with Swine parainfluenza virus 3. Conclusion: The results prove the existence of PIV3 infection in cattle, sheep and goats in the studied areas in Sudan and suggest its possible role in the respiratory infections. Genetic analysis indicate that the virus is mostly similar with bovine PIV3. [J Adv Vet Anim Res 2016; 3(3.000): 236-241]
Show more [+] Less [-]Clinical disease and lung lesions in calves experimentally inoculated with Histophilus somni five days after metaphylactic administration of tildipirosin or tulathromycin
2016
Confer, Anthony W. | Snider, Timothy A. | Taylor, Jared D. | Montelongo, Marie | Sorensen, Nicholas J.
OBJECTIVE To compare clinical disease and lung lesions in calves experimentally inoculated with Histophilus somni 5 days after metaphylactic administration of tildipirosin or tulathromycin. ANIMALS Twenty-four 3-month-old Holstein and Holstein-crossbreed steers. PROCEDURES Calves were randomly allocated to 3 groups of 8 calves. On day 0, calves in group 1 received tildipirosin (4 mg/kg, SC), calves in group 2 received tulathromycin (2.5 mg/kg, SC), and calves in group 3 received isotonic saline (0.9% NaCl) solution (1 mL/45 kg, SC; control). On day 5, calves were inoculated with 10 mL of a solution containing H somni strain 7735 (1.6 × 109 CFUs/mL, intrabronchially; challenge). Calves were clinically evaluated on days 5 through 8 and euthanized on day 8. The lungs were grossly evaluated for evidence of pneumonia, and bronchial secretion samples underwent bacteriologic culture. RESULTS The mean clinical score for each group was significantly increased 12 hours after challenge, compared with that immediately before challenge, and was significantly lower for tildipirosin-treated calves on days 6, 7, and 8, compared with those for tulathromycin-treated and control calves. The mean percentage of lung consolidation for tildipirosin-treated calves was significantly lower than those for tulathromycin-treated and control calves. Histophilus somni was isolated from the bronchial secretions of some tulathromycin-treated and control calves but was not isolated from tildipirosin-treated calves. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that metaphylactic administration of tildipirosin to calves 5 days prior to H somni challenge prevented subsequent culture of the pathogen from bronchial secretions and was more effective in minimizing clinical disease and lung lesions than was metaphylactic administration of tulathromycin.
Show more [+] Less [-]Genetic diversity of Streptococcus suis serotype 2 isolated from pigs in Brazil
2016
Doto, Daniela Sabatini | Moreno, Luisa Zanolli | Calderaro, Franco Ferraro | Matajira, Carlos Emilio Cabrera | Moura Gomes, Vasco Tulio De | Ferreira, Thais Sebastiana Porfida | Mesquita, Renan Elias | Timenetsky, Jorge | Gottschalk, Marcelo | Moreno, Andrea Micke
Streptococcus suis is an emerging zoonotic pathogen that causes septicemia, meningitis, arthritis, and pneumonia in swine and humans. The present study aimed to characterize the genetic diversity of S. suis serotype 2 isolated from pigs showing signs of illness in Brazil using pulsed-field gel electrophoresis (PFGE), single-enzyme amplified fragment length polymorphism (SE-AFLP), and profiling of virulence-associated markers. A total of 110 isolates were studied, 62.7% of which were isolated from the central nervous system and 19.1% from the respiratory tract. Eight genotypes were obtained from the combination of virulence genes, with 43.6% and 5.5% frequencies for the mrp (+) /epf (+) /sly (+) and mrp (-) /epf (-) /sly (-) genotypes, respectively. The presence of isolates with epf gene variation with higher molecular weight also appears to be a characteristic of Brazilian S. suis serotype 2. The PFGE and SE-AFLP were able to type all isolates and, although they presented a slight tendency to cluster according to state and year of isolation, it was also evident the grouping of different herds in the same PFGE subtype and the existence of isolates originated from the same herd classified into distinct subtypes. No further correlation between the isolation sites and mrp/epf/sly genotypes was observed.
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