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Potential role of different fish species as vectors of koi herpesvirus (CyHV-3) infection
2019
Matras, Marek | Stachnik, Magdalena | Borzym, Ewa | Maj-Paluch, Joanna | Reichert, Michał
Koi herpesvirus (KHV) has infected farmed common carp in Poland clinically and asymptomatically since 2004. The role of non-carp species as vectors of virus transmission is well known except for in the case of KHV. The aim was to better understand this virus’ infection and transmission pathways in common carp, looking at the potential vector role of fishes kept with them. Eight species were experimentally infected with KHV by immersion in a suspension at 20°C ±1 and transferred to a tank after 45 minutes. Specimens were euthanised at intervals up to 56 days post infection (dpi) and tissue was examined for KHV DNA. Surviving infected fishes were introduced at intervals, each time into a separate tank, to naïve common carp for experimental infection. These were observed daily for symptoms, sacrificed along with controls after three months, and dissected to provide tissue samples. Also fish from 14 species collected from a farm with a history of KHV were sampled from 3 to 22 months after disease was confirmed. Organ sections from single fish were collected in a single tube. Viral DNA was detected in tench and roach samples up to 49 dpi, but in three-spined stickleback and stone maroko samples only up to 14 dpi. Transmission of KHV to naïve carp occurred after cohabitation. KHV DNA was detected in three fish species three months after the farm outbreak. We confirmed that grass and Prussian carp, tench, roach, and brown bullhead can transfer the virus to naïve common carp.
Mostrar más [+] Menos [-]Evaluation of the possibility of C. burnetii transmission by the alimentary route in a guinea pig model
2019
Jodełko, Agnieszka | Szymańska-Czerwińska, Monika | Kycko, Anna | Niemczuk, Krzysztof
Q fever (coxiellosis) is an infectious disease of animals and humans, caused by.C. burnetii and widely distributed throughout the world. It is known that people and animals acquire the disease predominantly.via inhalation of infectious aerosols. The possibility of transmission of the pathogen by the alimentary route is still a matter of debate and remains controversial. Therefore the aim of this study was to fill the gaps in knowledge of oral transmission of.C. burnetii by conducting biological tests on the guinea pig model. Guinea pigs, divided into five groups comprising a negative control and four experimental groups, received specified concentrations of.C. burnetii per os. To determine the presence of specific antibodies, blood samples were tested using CFT. Also, internal organs collected during necropsy were screened by a real-time PCR targeting I.1111. Additionally, histopathological evaluation of the tissues was performed. The presence of antibodies and pathogen DNA in caecum was confirmed in one guinea pig from experimental group IV..C. burnetii was also detected in testicular tissue collected from one animal of experimental group II. The presence of pathogen DNA in the testicular tissue indicates that infection spreads haematogenously. In the majority of experimental animals specific antibodies and genetic material of.C. burnetii were not detected. This fact suggests that development of infection depends on many factors, such as animal immune status.
Mostrar más [+] Menos [-]Amorphus globosus foetuses in Polish Holstein cattle: anatomical, histological, and genetic studies
2019
Gehrke, Marek | Blaszak, Beata | Stachowiak, Monika | Szczerbal, Izabela | Stefanska, Barbara | Jaśkowski, Jędrzej M. | Nowak, Włodzimierz | Świtoński, Marek
A comprehensive description is presented of four novel cases ofamorphus globosus (ag) foetuses originating from multiple pregnancies of Polish Holstein cows. Four amorphic foetuses were delivered by three cows. Tissue samples were collected during autopsy, embedded in paraffin, sectioned, and stained with haematoxylin and eosin. Genomic DNA was isolated from tissue samples of abnormal foetuses and from blood leukocytes of their healthy siblings. PCR reactions were used to reveal the presence of Y-linked genes (SRY and AMELY) and an X-linked gene (AMELX). All foetuses were classified to the groupholoacardius amorphous (anideus). Molecular analysis clearly showed that at 17 microsatellite loci, the studied amorphous foetuses had identical genotypes to the viable co-twins. Foetuses had monozygotic origin. Histological analysis showed a low level of development of tissues of meso- and ectodermal origin, as well as features of degrading patterns.
Mostrar más [+] Menos [-]Elaboration of triplex PCR for detection of selected viral infections in waterfowl
2019
Kozdruń, Wojciech | Czekaj, Hanna | Styś-Fijoł, Natalia | Piekarska, Karolina | Samanta Niczyporuk, Jowita
Viral infections are the greatest threat to waterfowl and cause significant economic losses. Diagnosis and differentiation of three goose viruses is difficult in the field and often requires laboratory confirmation. Therefore, the aim of the study was to develop a triplex PCR and optimise its parameters for simultaneous detection of DNA of goose parvovirus (GPV), goose polyomavirus (GHPV), and goose circovirus (GoCV). The DNA of viruses isolated from field cases from the National Veterinary Research Institute’s own collection was used for the study. The primer attachment temperature, the number of reaction cycles, and the Taq DNA polymerase and Mg²⁺ concentrations were optimised. The sensitivity and specificity of this triplex PCR was also determined. Based on the obtained results, triplex PCR parameters were optimised for simultaneous detection of DNA of GPV, GHPV, and GoCV in one sample. The following PCR products of the expected size were obtained: GPV DNA of 806 bp, GoCV DNA of 571 bp, and GHPV DNA of 180 bp. The developed triplex PCR method proved to be useful for simultaneous detection of infections with three waterfowl viruses and will be used in relevant laboratory diagnostics.
Mostrar más [+] Menos [-]Investigation of effects of omeprazole on the fecal and gastric microbiota of healthy adult horses
2019
Tyma, Jesse F. | Epstein, Kira L. | Whitefield-Cargile, Canaan M. | Cohen, Noah D. | Giguère, S. (Steeve)
OBJECTIVE To determine the effects of oral omeprazole administration on the fecal and gastric microbiota of healthy adult horses. ANIMALS 12 healthy adult research horses. PROCEDURES Horses were randomly assigned to receive omeprazole paste (4 mg/kg, PO, q 24 h) or a sham (control) treatment (tap water [20 mL, PO, q 24 h]) for 28 days. Fecal and gastric fluid samples were collected prior to the first treatment (day 0), and on days 7, 28, 35, and 56. Sample DNA was extracted, and bacterial 16S rRNA gene sequences were amplified and sequenced to characterize α and β diversity and differential expression of the fecal and gastric microbiota. Data were analyzed by visual examination and by statistical methods. RESULTS Composition and diversity of the fecal microbiota did not differ significantly between treatment groups or over time. Substantial variation in gastric fluid results within groups and over time precluded meaningful interpretation of the microbiota in those samples. CONCLUSIONS AND CLINICAL RELEVANCE Results supported that omeprazole administration had no effect on fecal microbiota composition and diversity in this group of healthy adult horses. Small sample size limited power to detect a difference if one existed; however, qualitative graphic examination supported that any difference would likely have been small and of limited clinical importance. Adequate data to evaluate potential effects on the gastric microbiota were not obtained. Investigations are needed to determine the effects of omeprazole in horses with systemic disease or horses receiving other medical treatments.
Mostrar más [+] Menos [-]Use of standard diagnostic techniques to determine eradication of infection in experimental equine septic arthritis
2019
Koziv, R. V. | Yoshimura, S. | Dickinson, R. | Rybicka, J. M. | Moshynskyy, I. | Ngeleka, M. | Bracamonte, J. L. | Simko, E.
Septic arthritis is an important disease in horses, necessitating aggressive and prolonged therapy. In order to guide therapy, reliable methods of detecting the eradication of infection are needed. Therefore, the objective of this study was to investigate detection of eradication of infection in an experimental model of equine septic arthritis using standard diagnostic techniques. For this purpose, 17 adult horses were assigned to 3 experimental groups. The middle carpal joint of each horse was injected with Escherichia coli (Septic group, n = 8), lipopolysaccharide (LPS) (LPS group, n = 6), or sterile saline (Control group, n = 3) at day 0. Contralateral joints were not injected. Standard therapy was applied to all joints except non-injected joints in the Control group at day 1. Sequential samples of synovial fluid (SF) were collected for bacterial culture using 3 culture media [Columbia blood agar (CBA), brain heart infusion broth (BHI), and Signal blood culture medium] and for cytological evaluation [percentage neutrophils (PN), total nucleated cell count (TNCC), and total protein (TP)]. Escherichia coli-specific polymerase chain reaction (PCR) was carried out to detect E. coli DNA in synovial fluid. Culture and PCR were positive for E. coli in all joints injected with E. coli at day 1 and 1 joint was positive on BHI at day 4. Based on the results of bacterial culture, PCR, and TNCC, the elimination of infection in our experimental model occurred by day 4 post-infection in 6 out of 7 cases. Total protein (TP) and PN remained elevated at clinical threshold used for diagnosis of septic arthritis until day 14. In our experimental model of E. coli-induced arthritis, we conclude that TP and PN may not be good indicators for detecting the eradication of bacterial infection caused by E. coli from infected and subsequently treated joints.
Mostrar más [+] Menos [-]Comparative molecular analysis of fecal microbiota of bobcats (Lynx rufus) and domestic cats (Felis catus)
2019
Eshar, D. | Lee, C. | Weese, J. S.
The goal of this study was to explore and describe fecal microbiota of captive and wild bobcats (Lynx rufus) and compare the results to those of domestic cats (Felis catus). Fecal samples from 27 bobcats (8 wild, 19 zoo-kept) were used for novel bacterial deoxyribonucleic acid (DNA) identification using next-generation sequencing of the V4 region of the bacterial 16S ribosomal ribonucleic acid (rRNA) gene, analyzed by Illumina sequencing, and then compared to data obtained from a colony of 10 domestic cats. In this study, the microbiota of both species was dominated by Firmicutes, followed by Proteobacteria, Actinobacteria, Bacteroidetes, and Verrucomicrobia. When compared, fecal samples from bobcats harbored more Proteobacteria and Actinobacteria than fecal samples from domestic cats. There was a remarkable inter-bobcat variation in the relative abundances of the main bacterial genera. There were no significant differences, however, between the main phyla of the microbiota of the wild and domestic bobcats. Proteobacteria in wild bobcats (P = 0.079) and Firmicutes in zoo-kept bobcats (P = 0.079) approached significance. There were no differences in predominant genera between wild and captive bobcats. The results of this study showed that there are notable differences in fecal bacterial communities between domestic cats and both captive and wild bobcats. The lack of significant differences in bacterial communities between wild and zoo-kept bobcats suggests that the varied diet provided for these felids can result in a fecal microbiota resembling that generated by a wild diet.
Mostrar más [+] Menos [-]Survival of Mycobacterium bovis during forage ensiling
2019
Grooms, Dan | Bolin, Steven R. | Plastow, Jessica L. | Lim, Ailam | Hattey, Joseph | Durst, Phillip T. | Rust, Steven R. | Allen, Michael S. | Buskirk, Daniel D. | Smith, Richard W.
OBJECTIVE To determine whether Mycobacterium bovis remains viable in ensiled forages. SAMPLE Alfalfa, mixed mostly grass, and corn silages. PROCEDURES For each of 10 sampling days, six 250-g replicate samples of each feedstuff were created and placed in a film pouch that could be vacuum sealed to simulate the ensiling process. Within each set of replicate samples, 4 were inoculated with 10 mL of mycobacterial liquid culture medium containing viable M bovis and 2 were inoculated with 10 mL of sterile mycobacterial liquid culture medium (controls) on day 0. Pouches were vacuum sealed and stored in the dark at room temperature. On the designated sampling day, 1 control pouch was submitted for forage analysis, and the other pouches were opened, and forage samples were obtained for M bovis culture and analysis with a PCR assay immediately and 24 hours later. RESULTS None of the control samples had positive M bovis culture or PCR assay results. Among M bovis-inoculated samples, the organism was not cultured from alfalfa and corn silage for > 2 days but was cultured from mixed mostly grass silage for 28 days after inoculation and ensiling initiation. Mycobacterium bovis DNA was detected by PCR assay in samples of all 3 feedstuffs throughout the 112-day observation period. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that properly ensiled forages would be an unlikely source for M bovis transmission to cattle. Further research is necessary to determine whether ensiling kills M bovis or forces it to become dormant and, if the latter, elucidate the conditions that cause it to revert to an infectious state.
Mostrar más [+] Menos [-]A missense mutation in MYO7A is associated with bilateral deafness and vestibular dysfunction in the Doberman pinscher breed
2019
Webb, A. A. | Ruhe, A. L. | Neff, M. W.
Bilateral deafness with concurrent vestibular dysfunction was first reported in the Doberman pinscher in 1980. Here, we identify a coding mutation in the MYO7A gene that is perfectly associated with the disorder. The lack of visual deficits in affected dogs suggests that, like rodents but unlike humans, MYO7A is not required for retinal function. DNA testing of the mutation will enable dog breeders to manage the incidence of this genetic defect.
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