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Evaluation of direct metagenomics and target enriched approaches for high-throughput sequencing of field rabies viruses Full text
2019
Orłowska Anna | Iwan Ewelina | Smreczak Marcin | Rola Jerzy
Evaluation of direct metagenomics and target enriched approaches for high-throughput sequencing of field rabies viruses Full text
2019
Orłowska Anna | Iwan Ewelina | Smreczak Marcin | Rola Jerzy
High-throughput sequencing (HTS) identifies random viral fragments in environmental samples metagenomically. High reliability gains it broad application in virus evolution, host-virus interaction, and pathogenicity studies. Deep sequencing of field samples with content of host genetic material and bacteria often produces insufficient data for metagenomics and must be preceded by target enrichment. The main goal of the study was the evaluation of HTS for complete genome sequencing of field-case rabies viruses (RABVs).
Show more [+] Less [-]Evaluation of direct metagenomics and target enriched approaches for high-throughput sequencing of field rabies viruses Full text
2019
Orłowska, Anna | Iwan, Ewelina | Smreczak, Marcin | Rola, Jerzy
High-throughput sequencing (HTS) identifies random viral fragments in environmental samples metagenomically. High reliability gains it broad application in virus evolution, host-virus interaction, and pathogenicity studies. Deep sequencing of field samples with content of host genetic material and bacteria often produces insufficient data for metagenomics and must be preceded by target enrichment. The main goal of the study was the evaluation of HTS for complete genome sequencing of field-case rabies viruses (RABVs). The material was 23 RABVs isolated mainly from red foxes and one European bat lyssavirus-1 isolate propagated in neuroblastoma cells. Three methods of RNA isolation were tested for the direct metagenomics and RABV-enriched approaches. Deep sequencing was performed with a MiSeq sequencer (Illumina) and reagent v3 kit. Bioinformatics data were evaluated by Kraken and Centrifuge software and de novo assembly was done with metaSPAdes. Testing RNA extraction procedures revealed the deep sequencing scope superiority of the combined TRIzol/column method. This HTS methodology made it possible to obtain complete genomes of all the RABV isolates collected in the field. Significantly greater rates of RABV genome coverages (over 5,900) were obtained with RABV enrichment. Direct metagenomic studies sequenced the full length of 6 out of 16 RABV isolates with a medium coverage between 1 and 71. Direct metagenomics gives the most realistic illustration of the field sample microbiome, but with low coverage. For deep characterisation of viruses, e.g. for spatial and temporal phylogeography during outbreaks, target enrichment is recommended as it covers sequences much more completely.
Show more [+] Less [-]Plasma metabolic characterisation of dairy cows with inactive ovaries and oestrus during the peak of lactation Full text
2019
Song Yu-Xi | Hu Pan | Bai Yun-Long | Zhao Chang | Xia Cheng | Xu Chuang
Differential metabolites (DMs) between cows with inactive ovaries (IO) and oestrous (E) cows were screened and metabolic pathways of DMs associated with IO were determined.
Show more [+] Less [-]Effects of tiletamine-xylazine-tramadol combination and its specific antagonist on AMPK in the brain of rats Full text
2019
Ma Ning | Li Xin | Wang Hong-bin | Gao Li | Xiao Jian-hua
Effects of tiletamine-xylazine-tramadol combination and its specific antagonist on AMPK in the brain of rats Full text
2019
Ma Ning | Li Xin | Wang Hong-bin | Gao Li | Xiao Jian-hua
Introduction: Tiletamine-xylazine-tramadol (XFM) has few side effects and can provide good sedation and analgesia. Adenosine 5’-monophosphate-activated protein kinase (AMPK) can attenuate trigeminal neuralgia. The study aimed to investigate the effects of XFM and its specific antagonist on AMPK in different regions of the brain. Material and Methods: A model of XFM in the rat was established. A total of 72 Sprague Dawley (SD) rats were randomly divided into three equally sized groups: XFM anaesthesia (M group), antagonist (W group), and XFM with antagonist interactive groups (MW group). Eighteen SD rats were in the control group and were injected intraperitoneally with saline (C group). The rats were sacrificed and the cerebral cortex, cerebellum, hippocampus, thalamus, and brain stem were immediately separated, in order to detect AMPKα mRNA expression by quantitative PCR. Results: XFM was able to increase the mRNA expression of AMPKα1 and AMPKα2 in all brain regions, and the antagonist caused the opposite effect, although the effects of XFM could not be completely reversed in some areas. Conclusion: XFM can influence the expression of AMPK in the central nervous system of the rat, which can provide a reference for the future development of anaesthetics for animals.
Show more [+] Less [-]Effects of tiletamine-xylazine-tramadol combination and its specific antagonist on AMPK in the brain of rats Full text
2019
Ma, Ning | Li, Xin | Wang, Hong-bin | Gao, Li | Xiao, Jian-hua
Introduction: Tiletamine-xylazine-tramadol (XFM) has few side effects and can provide good sedation and analgesia. Adenosine 5’-monophosphate-activated protein kinase (AMPK) can attenuate trigeminal neuralgia. The study aimed to investigate the effects of XFM and its specific antagonist on AMPK in different regions of the brain. Material and Methods: A model of XFM in the rat was established. A total of 72 Sprague Dawley (SD) rats were randomly divided into three equally sized groups: XFM anaesthesia (M group), antagonist (W group), and XFM with antagonist interactive groups (MW group). Eighteen SD rats were in the control group and were injected intraperitoneally with saline (C group). The rats were sacrificed and the cerebral cortex, cerebellum, hippocampus, thalamus, and brain stem were immediately separated, in order to detect AMPKα mRNA expression by quantitative PCR. Results: XFM was able to increase the mRNA expression of AMPKα1 and AMPKα2 in all brain regions, and the antagonist caused the opposite effect, although the effects of XFM could not be completely reversed in some areas. Conclusion: XFM can influence the expression of AMPK in the central nervous system of the rat, which can provide a reference for the future development of anaesthetics for animals.
Show more [+] Less [-]Carbapenem-resistant Pseudomonas aeruginosa originating from farm animals and people in Egypt Full text
2019
Elshafiee Esraa A. | Nader Sara M. | Dorgham Sohad M. | Hamza Dalia A.
Carbapenem-resistant Pseudomonas aeruginosa originating from farm animals and people in Egypt Full text
2019
Elshafiee Esraa A. | Nader Sara M. | Dorgham Sohad M. | Hamza Dalia A.
Carbapenem-resistant Pseudomonas aeruginosa (CRPA) has become the leading cause of health care-associated infections. Treatment is difficult due to the lack of an effective antimicrobial therapy, and mortality is high. This study investigated the occurrence of CRPA in farm animals (buffaloes and cattle), livestock drinking water, and humans in Egypt.
Show more [+] Less [-]Carbapenem-resistant Pseudomonas aeruginosa originating from farm animals and people in Egypt Full text
2019
Elshafiee, Esraa A. | Nader, Sara M. | Dorgham, Sohad M. | Hamza, Dalia A.
Carbapenem-resistant Pseudomonas aeruginosa (CRPA) has become the leading cause of health care-associated infections. Treatment is difficult due to the lack of an effective antimicrobial therapy, and mortality is high. This study investigated the occurrence of CRPA in farm animals (buffaloes and cattle), livestock drinking water, and humans in Egypt. A total of 180 samples were examined: 50 faecal each from buffaloes and cattle, 30 of livestock drinking water, and 50 stool from humans. The samples were cultured on cetrimide agar and the plates were incubated aerobically at 37°C for 24 h. The isolates were examined for the presence of the blaKPC, blaOXA₋₄₈, and blaNDM carbapenemase-encoding genes using PCR and investigated for the exotoxin A (toxA) gene. The toxA gene from carbapenem- group resistant isolates was phylogenetically analysed. P. aeruginosa was isolated from buffaloes, cattle, drinking water, and humans, with occurrences of 40%, 34%, 10%, and 20%, respectively. Carbapenem resistance genes were found in 60%, 59%, 67%, and 70% in buffalo, cattle, water and human samples, respectively. The toxA gene was detected in 80% of samples. The phylogenetic analysis showed that cattle and water sequences were in one cluster and more related to each other than to human isolates. Occurrence of CRPA among farm animals, drinking water, and humans was high, reflecting the environmental origin of P. aeruginosa and highlighting contaminated water as a potential transmitter of CRPA to livestock and next to humans.
Show more [+] Less [-]Bivagina pagrosomi Murray (1931) (Monogenea: Polyopisthocotylea), a microcotylid infecting the gills of the gilt-head sea bream Sparus aurata (Sparidae) from the Red Sea: morphology and phylogeny Full text
2019
Dajem Saad Bin | Morsy Kareem | Shati Ali | Ezzat Ahmed | El-Said Fahmy | Abdel-Gaber Rewaida
Monogenea is a class of ectoparasitic flatworms on the skin, gills, or fins of fish. Microcotylidae is a family of polyopisthocotylean monogeneans parasitising only marine fishes. This work describes and taxonomically determines a microcotylid polyopisthocotylean monogenean in an important fish in Saudi aquaculture.
Show more [+] Less [-]Accessory genital glands in the New Zealand White rabbit: a morphometrical and histological study Full text
2019
Skonieczna Joanna | Madej Jan P. | Będziński Romuald
Accessory genital glands in the New Zealand White rabbit: a morphometrical and histological study Full text
2019
Skonieczna Joanna | Madej Jan P. | Będziński Romuald
Introduction: The aim of this research was to provide a detailed description of the morphology, topography, and histometry of rabbit accessory genital glands. Material and Methods: Seven male New Zealand White rabbits, 3–4 months of age and weighing 2.1–3 kg were used for the study. The whole urethra from the urinary bladder to the external urethral orifice accompanied by accessory genital glands was sliced at intervals of 1 mm. The serial sections were prepared with haematoxylin-eosin (H&E) and Movat–Russell modified pentachrome stain. Results: A detailed description of the morphology and morphometry was provided. The topography of the organs was explained on the basis of characteristic cross-sections on histological slides. The inconsistent nomenclature and descriptions of these glands by different authors were also discussed. Conclusion: The morphometric analysis indicated that some of the glands described have similar dimensions in different individuals, while others like paraprostates revealed high diversity in the number of lobes, their size, and their structure. The accessory glands are also good topographic markers which precisely define the segment of the urethra. The terms “proprostate”, “prostate”, and “paraprostates” as the nomenclature of the prostate complex reflect the location of these glands well and indicate their common origin and function.
Show more [+] Less [-]Accessory genital glands in the New Zealand White rabbit: a morphometrical and histological study Full text
2019
Skonieczna, Joanna | Madej, Jan P. | Będziński, Romuald
Introduction: The aim of this research was to provide a detailed description of the morphology, topography, and histometry of rabbit accessory genital glands. Material and Methods: Seven male New Zealand White rabbits, 3–4 months of age and weighing 2.1–3 kg were used for the study. The whole urethra from the urinary bladder to the external urethral orifice accompanied by accessory genital glands was sliced at intervals of 1 mm. The serial sections were prepared with haematoxylin-eosin (H&E) and Movat–Russell modified pentachrome stain. Results: A detailed description of the morphology and morphometry was provided. The topography of the organs was explained on the basis of characteristic cross-sections on histological slides. The inconsistent nomenclature and descriptions of these glands by different authors were also discussed. Conclusion: The morphometric analysis indicated that some of the glands described have similar dimensions in different individuals, while others like paraprostates revealed high diversity in the number of lobes, their size, and their structure. The accessory glands are also good topographic markers which precisely define the segment of the urethra. The terms “proprostate”, “prostate”, and “paraprostates” as the nomenclature of the prostate complex reflect the location of these glands well and indicate their common origin and function.
Show more [+] Less [-]Occurrence of enterococci in mastitic cow’s milk and their antimicrobial resistance Full text
2019
Różańska Hanna | Lewtak-Piłat Aleksandra | Kubajka Maria | Weiner Marcin
Occurrence of enterococci in mastitic cow’s milk and their antimicrobial resistance Full text
2019
Różańska Hanna | Lewtak-Piłat Aleksandra | Kubajka Maria | Weiner Marcin
Introduction: The aim of the study was to evaluate the occurrence of enterococci in inflammatory secretions from mastitic bovine udders and to assess their antimicrobial resistance.
Show more [+] Less [-]Occurrence of enterococci in mastitic cow’s milk and their antimicrobial resistance Full text
2019
Różańska, Hanna | Lewtak-Piłat, Aleksandra | Kubajka, Maria | Weiner, Marcin
Introduction: The aim of the study was to evaluate the occurrence of enterococci in inflammatory secretions from mastitic bovine udders and to assess their antimicrobial resistance. Material and Methods: A total of 2,000 mastitic milk samples from cows were tested in 2014–2017. The isolation of enterococci was performed by precultivation in buffered peptone water, selective multiplication in a broth with sodium azide and cristal violet, and cultivation on Slanetz and Bartley agar. The identification of enterococci was carried out using Api rapid ID 32 strep kits. The antimicrobial susceptibility was evaluated using the MIC technique. Results: Enterococci were isolated from 426 samples (21.3%). Enterococcus faecalis was the predominant species (360 strains), followed by E. faecium (35 isolates), and small numbers of others. The highest level of resistance was observed to lincomycin, tetracycline, quinupristin/dalfopristin (Synercid), erythromycin, kanamycin, streptomycin, chloramphenicol, and tylosin. Single strains were resistant to vancomycin and ciprofloxacin. All isolates were sensitive to daptomycin. E. faecalis presented a higher level of resistance in comparison to E. faecium, except to nitrofurantoin. Conclusion: The results showed frequent occurrence of enterococci in mastitic cow’s milk and confirmed the high rate of their antimicrobial resistance.
Show more [+] Less [-]Epidemiology and antibiogram of Riemerella anatipestifer isolated from waterfowl slaughterhouses in Taiwan Full text
2019
Chang Fei-Fei | Chen Chang-Chieh | Wang Shao-Hung | Chen Chiou-Lin
Epidemiology and antibiogram of Riemerella anatipestifer isolated from waterfowl slaughterhouses in Taiwan Full text
2019
Chang Fei-Fei | Chen Chang-Chieh | Wang Shao-Hung | Chen Chiou-Lin
Introduction: Laryngeal swab samples collected from three waterfowl slaughterhouses in central Taiwan were cultured and suspected isolates of Riemerella anatipestifer were identified by API 20NE and 16S rDNA PCR.
Show more [+] Less [-]Epidemiology and antibiogram of Riemerella anatipestifer isolated from waterfowl slaughterhouses in Taiwan Full text
2019
Chang, Fei-Fei | Chen, Chang-Chieh | Wang, Shao-Hung | Chen, Chiou-Lin
Introduction: Laryngeal swab samples collected from three waterfowl slaughterhouses in central Taiwan were cultured and suspected isolates of Riemerella anatipestifer were identified by API 20NE and 16S rDNA PCR. Material and Methods: Serum agglutination was used for serotyping, and antimicrobial susceptibility was tested. Results: Seventy-six R. anatipestifer isolates were detected, and the prevalences in the ducks and geese were 12.3% (46/375) and 8.0% (30/375), respectively. The positive isolation rates were 65.6% for all arriving waterfowl, 76.0% for birds in the holding area, 1.6% for defeathered carcasses, but zero for degummed carcasses. A PCR examination detected R. anatipestifer in the slaughtering area frequently. Serotype B was dominant in both duck (34.8%) and goose (46.7%) isolates, but the wide serotype distribution may very well impede vaccination development. All isolates were resistant to colistin, and 79.7% were resistant to more than three common antibiotics. Conclusion: The results proved that most ducks had encountered antibiotic-resistant R. anatipestifer in rearing, which suggests that the bacterium circulates in asymptomatic waterfowl. It is worth noting that most waterfowl farms were found to harbour R. anatipestifer, and contaminated slaughterhouses are a major risk factor in its spread. Effective prevention and containment measures should be established there to interrupt the transmission chain of R. anatipestifer.
Show more [+] Less [-]Evaluation of a canine transmissible venereal tumour cell line with tumour immunity capacity but without tumorigenic property Full text
2019
Zayas Yareellys Ramos | Molina Moisés Armides Franco | Guerra Reyes Tamez | Padilla Cristina Rodríguez
Evaluation of a canine transmissible venereal tumour cell line with tumour immunity capacity but without tumorigenic property Full text
2019
Zayas Yareellys Ramos | Molina Moisés Armides Franco | Guerra Reyes Tamez | Padilla Cristina Rodríguez
Introduction: Canine transmissible venereal tumour (CTVT) is a sexually transmitted tumour affecting dogs worldwide, imposing a financial burden on dog owners. A stable culture cell line in continuous passages for >18 months has only been achieved once. The present study investigated a stable CTVT cell line isolated from a bitch and its potential as a vaccine. Material and Methods: A biopsy from a 2-year-old mongrel bitch with CTVT was obtained for histopathological confirmation and isolation of tumour cells. The isolated cells were cultured to passage 55 and characterised by flow cytometry, with karyotyping by GTG-banding and by PCR detection of myc S-2 and LINE AS1. The isolated CTVT cell line was also used as a preventive vaccine in a canine model. Results: Histopathological analysis of the isolated tumour cells revealed typical CTVT characteristics. Constant proliferation and stable morphological characteristics were observed during culture. Phenotypic analysis determined the expression of HLA-DR+, CD5.1+, CD14+, CD45+, CD83+, CD163+, and Ly-6G-Ly-6C+. GTG-banding revealed a mean of 57 chromosomes in the karyotype with several complex chromosomal rearrangements. LINE-c-myc insertion in the isolated CTVT cell line at 550 bp was not detected. However, a 340-bp band was amplified. Isolated CTVT cell line inoculation at a concentration of 1×108 did not induce tumour growth in bitches, nor did a challenge with primary CTVT cells. Conclusion: The present study successfully identified and isolated a stable CTVT cell line that may be useful in CTVT prevention.
Show more [+] Less [-]Evaluation of a canine transmissible venereal tumour cell line with tumour immunity capacity but without tumorigenic property Full text
2019
Zayas, Yareellys Ramos | Molina, Moisés Armides Franco | Guerra, Reyes Tamez | Padilla, Cristina Rodríguez
Introduction: Canine transmissible venereal tumour (CTVT) is a sexually transmitted tumour affecting dogs worldwide, imposing a financial burden on dog owners. A stable culture cell line in continuous passages for >18 months has only been achieved once. The present study investigated a stable CTVT cell line isolated from a bitch and its potential as a vaccine. Material and Methods: A biopsy from a 2-year-old mongrel bitch with CTVT was obtained for histopathological confirmation and isolation of tumour cells. The isolated cells were cultured to passage 55 and characterised by flow cytometry, with karyotyping by GTG-banding and by PCR detection of myc S-2 and LINE AS1. The isolated CTVT cell line was also used as a preventive vaccine in a canine model. Results: Histopathological analysis of the isolated tumour cells revealed typical CTVT characteristics. Constant proliferation and stable morphological characteristics were observed during culture. Phenotypic analysis determined the expression of HLA-DR⁺, CD5.1⁺, CD14⁺, CD45⁺, CD83⁺, CD163⁺, and Ly-6G-Ly-6C⁺. GTG-banding revealed a mean of 57 chromosomes in the karyotype with several complex chromosomal rearrangements. LINE-c-myc insertion in the isolated CTVT cell line at 550 bp was not detected. However, a 340-bp band was amplified. Isolated CTVT cell line inoculation at a concentration of 1×10⁸ did not induce tumour growth in bitches, nor did a challenge with primary CTVT cells. Conclusion: The present study successfully identified and isolated a stable CTVT cell line that may be useful in CTVT prevention.
Show more [+] Less [-]Retrospective liver histomorphological analysis in dogs in instances of clinical suspicion of congenital portosystemic shunt Full text
2019
Sobczak-Filipiak Małgorzata | Szarek Józef | Badurek Iwona | Padmanabhan Jessica | Trębacz Piotr | Januchta-Kurmin Monika | Galanty Marek
Retrospective liver histomorphological analysis in dogs in instances of clinical suspicion of congenital portosystemic shunt Full text
2019
Sobczak-Filipiak Małgorzata | Szarek Józef | Badurek Iwona | Padmanabhan Jessica | Trębacz Piotr | Januchta-Kurmin Monika | Galanty Marek
Introduction: The clinical symptoms of portosystemic shunts (PSSs) and hepatic microvascular dysplasia (HMD) – portal vein hypoplasia (PVH) in dogs are similar. PSSs are abnormal vascular connections between the portal vein system and systemic veins. HMD is a very rare developmental vascular anomaly, recognisable during histopathological examination. The study aim was to assess the prevalence of HMD–PVH and hepatocellular and vascular pathologies in the liver. Material and Methods: Liver biopsies from 140 dogs (of different breeds and both sexes) arousing clinical suspicion of PSS were examined histopathologically. Results: An initial PSS diagnosis was confirmed in 125 dogs (89.29%). HMD–PVH was found in 12.32% of dogs, as an isolated disease in 9.29%, especially in Yorkshire terriers, and with extrahepatic PSS in 6.67%. Histopathological analysis of muscles around sublobular veins showed that HMD cases presented hypertrophy or hypertrophy with fibrosis. In 2.17% of all dogs with liver vascular developmental disorders calcification was visible around vessels (without correlation by degenerative changes in those vessels), suggesting prior onset of deep metabolic disorders. Clinical suspicion of PSS was also formed upon quite different pathological processes in young dogs. Conclusion: Histopathological findings diagnosed the type of vascular anomalies (PSS or HMD–PVH) or other pathological changes conclusively, therefore detailed hepatic histopathology is an indispensable component of the clinical diagnostic process.
Show more [+] Less [-]Retrospective liver histomorphological analysis in dogs in instances of clinical suspicion of congenital portosystemic shunt Full text
2019
Sobczak-Filipiak, Małgorzata | Szarek, Józef | Badurek, Iwona | Padmanabhan, Jessica | Trębacz, Piotr | Januchta-Kurmin, Monika | Galanty, Marek
Introduction: The clinical symptoms of portosystemic shunts (PSSs) and hepatic microvascular dysplasia (HMD) – portal vein hypoplasia (PVH) in dogs are similar. PSSs are abnormal vascular connections between the portal vein system and systemic veins. HMD is a very rare developmental vascular anomaly, recognisable during histopathological examination. The study aim was to assess the prevalence of HMD–PVH and hepatocellular and vascular pathologies in the liver. Material and Methods: Liver biopsies from 140 dogs (of different breeds and both sexes) arousing clinical suspicion of PSS were examined histopathologically. Results: An initial PSS diagnosis was confirmed in 125 dogs (89.29%). HMD–PVH was found in 12.32% of dogs, as an isolated disease in 9.29%, especially in Yorkshire terriers, and with extrahepatic PSS in 6.67%. Histopathological analysis of muscles around sublobular veins showed that HMD cases presented hypertrophy or hypertrophy with fibrosis. In 2.17% of all dogs with liver vascular developmental disorders calcification was visible around vessels (without correlation by degenerative changes in those vessels), suggesting prior onset of deep metabolic disorders. Clinical suspicion of PSS was also formed upon quite different pathological processes in young dogs. Conclusion: Histopathological findings diagnosed the type of vascular anomalies (PSS or HMD–PVH) or other pathological changes conclusively, therefore detailed hepatic histopathology is an indispensable component of the clinical diagnostic process.
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