scholarly journals Characterization of Shedding Patterns of Porcine Circovirus Types 2a and 2b in Experimentally Inoculated Mature Boars

2008 ◽  
Vol 20 (6) ◽  
pp. 725-734 ◽  
Author(s):  
Darin M. Madson ◽  
Sheela Ramamoorthy ◽  
Chris Kuster ◽  
Narinder Pal ◽  
Xiang-Jin Meng ◽  
...  
Keyword(s):  
Sperm Morphology ◽  
Porcine Circovirus ◽  
Serum Samples ◽  
Treatment Groups ◽  
Swine Pathogen ◽  
Polymerase Chain ◽  
Swine Herds ◽  
Negative Controls ◽  
Group 1

Porcine circovirus-2 (PCV-2) is an economically important swine pathogen and causes PCV-associated disease (PCVAD) in pigs worldwide. Currently, 2 genotypes of PCV-2, PCV-2a and −2b, are circulating in U.S. swine herds. The objectives of the current study were to evaluate the amount of PCV-2 DNA present in semen over time, compare and correlate incidence and amount of PCV-2 present in semen samples to that present in serum samples and blood swabs, and determine if there are differences in shedding patterns between PCV-2a and −2b. Fifteen 7-month-old PCV-2-naïve Landrace boars ( Sus scrofa) were randomly allocated to 3 treatment groups. The boars in group 1 ( n = 3) served as negative controls, and those in groups 2 ( n = 6) and 3 ( n = 6) were intranasally and intramuscularly inoculated with PCV-2a and −2b, respectively. Semen, serum, and blood swab samples were collected up to 90 days postinoculation (DPI), and necropsies were performed on DPI 23,48, and 90. Larger quantities of both PCV-2a and − 2b DNA were detected earlier in serum and blood swab samples than in raw semen of experimentally inoculated boars. The incidence and duration of presence of PCV-2 DNA in semen varied among boars; however, intermittent shedding was not observed. In all sex glands, PCV-2 DNA was detected by polymerase chain reaction; however, PCV-2 antigen was not detected by immunohistochemistry, and PCV-2 had no effect on sperm morphology. Differences in shedding patterns between PCV-2a and −2b were not observed under the study conditions.

scholarly journals Prevalence and genetic characterization of Toxoplasma gondii in naturally infected backyard pigs intended for familial consumption in Romania

2019 ◽  
Vol 12 (1) ◽  
Author(s):  
Anamaria Ioana Paştiu ◽  
Anamaria Cozma-Petruț ◽  
Aurélien Mercier ◽  
Anamaria Balea ◽  
Lokman Galal ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Rural Areas ◽  
Antibody Test ◽  
Antibody Detection ◽  
Serum Samples ◽  
Potential Threat ◽  
Polymerase Chain ◽  
Pcr Targeting ◽  
Backyard Pigs

Abstract Background Foodborne toxoplasmosis in humans can be due to the exposure to tissue cysts of Toxoplasma gondii through the consumption of meat, including pork, of infected animals. Traditional Romanian food habits include pork as the preferred meat, while backyard pig rearing remains a common practice in many rural areas of Romania. The aims of the present study were to estimate the prevalence of T. gondii infection in naturally infected backyard pigs slaughtered for familial consumption and to genetically characterize the T. gondii strains obtained. Methods Paired blood and heart samples were collected from 94 backyard pigs, home slaughtered for private consumption. Serum samples were analyzed using the immunofluorescence antibody test (IFAT) for anti-T. gondii antibody detection. Heart samples were screened by polymerase chain reaction (PCR) targeting the 529-bp repeat region (REP529) for T. gondii detection. In addition, heart samples from IFAT positive animals were bioassayed in mice. The T. gondii isolates were genotyped by the analysis of 15 microsatellite markers. Results The results showed that almost half of the pigs investigated were T. gondii seropositive (46.8%, 95% confidence interval (CI): 36.4–57.4%) and in more than a quarter of the pigs (26.6%, 95% CI: 18.0–36.7%), the parasite was detected by PCR. Three (3/44) T. gondii strains were isolated from hearts of seropositive pigs and they all belonged to genotype II. Conclusions The present study showed the presence of T. gondii infection in backyard pigs in Romania, which suggests that consumption of pork from animals reared and slaughtered at home may pose a potential threat to human health and should be given attention. In addition, to our knowledge, this is the first study to provide data concerning T. gondii strains circulating in pigs from Romania.

scholarly journals Genotype 1 of human parvovirus B19 in clinical cases

2017 ◽  
Vol 63 (3) ◽  
pp. 224-228 ◽  
Author(s):  
Maria Isabel de Oliveira ◽  
Ana Maria Sardinha Afonso ◽  
Suely Pires Curti ◽  
Patrícia Evelin Silva ◽  
Tamyris Fernanda Barbosa ◽  
...  
Keyword(s):  
Parvovirus B19 ◽  
Infectious Agent ◽  
Immunoglobulin M ◽  
Human Parvovirus B19 ◽  
Genotype 1 ◽  
Serum Samples ◽  
Chronic Anemia ◽  
Virus Surveillance ◽  
Polymerase Chain

Summary Introduction: Virus surveillance strategies and genetic characterization of human parvovirus B19 (B19V) are important tools for regional and global control of viral outbreak. In São Paulo, Brazil, we performed a study of B19V by monitoring the spread of this virus, which is an infectious agent and could be mistakenly reported as a rash and other types of infection. Method: Serum samples were subjected to enzyme immunoassay, real time polymerase chain reaction, and sequencing. Results: From the 462 patients with suspected cases of exanthematic infections, the results of the 164 serum samples were positive for B19V immunoglobulin M. Among these cases, there were 38 patients with erythema infections and B19-associated with other infections such as encephalitis, hydrops fetalis, chronic anemia, hematological malignancies. These samples were sequenced and identified as genotype 1. Conclusion: This study showed patients with infections caused by B19V and sequencing genotype 1. Continuous monitoring is necessary to detect all known genotypes, and the emergence of new genotypes of these viruses for case management in public health control activities.

scholarly journals Detection and in vitro and in vivo characterization of porcine circovirus DNA from a porcine-derived commercial pepsin product

2004 ◽  
Vol 85 (11) ◽  
pp. 3377-3382 ◽  
Author(s):  
M. Fenaux ◽  
T. Opriessnig ◽  
P. G. Halbur ◽  
Y. Xu ◽  
B. Potts ◽  
...  
Keyword(s):  
Porcine Circovirus Type ◽  
Human Infection ◽  
Porcine Circovirus ◽  
Specific Pathogen ◽  
Group 3 ◽  
Group 2 ◽  
Group 1

Non-pathogenic porcine circovirus type 1 (PCV1) and pathogenic PCV2 are widespread in swine herds. In this study, the detection and characterization of PCV1 and PCV2 DNA from porcine-derived commercial pepsin are reported. The complete genomic sequences of the pepsin-derived PCV1 and PCV2 share 76 % nucleotide sequence identity with each other and 95–99 % identity with respective North American PCV1 and PCV2 isolates. However, the PCV-contaminated pepsin lacks infectivity in PK-15 cells. To further assess the infectivity of the contaminating pepsin in vivo, 16 5-week-old, specific-pathogen-free pigs were divided randomly into three groups: pigs in group 1 (n=5) were each inoculated intramuscularly and intranasally with 4 ml PBS buffer as negative controls, those in group 2 (n=6) were each inoculated with 400 mg contaminated pepsin dissolved in 4 ml PBS and those in group 3 (n=5) were each inoculated with 4×104·3 TCID50 PCV2 as positive controls. PCV2 viraemia, seroconversion and pathological lesions were detected in group 3 pigs, but not in group 1 or 2 pigs, confirming that the contaminating PCVs were non-infectious. Nevertheless, the detection of PCV DNA in a porcine-derived commercial product raises concern for potential human infection through xenotransplantation.

scholarly journals Serological diagnosis and molecular characterization of Leptospira spp. in the blood and urine of bovine females from refrigerated slaughterhouses

2018 ◽  
Vol 39 (3) ◽  
pp. 1125
Author(s):  
Aliny Fernanda de Oliveira ◽  
Roberta Torres Chiderolli ◽  
Luciano Seraphim Gasques ◽  
Arianne Peruzo Pires Gonçalves ◽  
Érica Dourado Neves ◽  
...  
Keyword(s):  
One Health ◽  
Preventive Measures ◽  
Agglutination Test ◽  
Economic Losses ◽  
Chain Reaction ◽  
Serum Samples ◽  
Leptospira Spp ◽  
Blood And Urine Samples ◽  
Polymerase Chain

Leptospirosis is an important socioeconomic disease in humans, as well as in domestic and wild animals, being caused by Leptospira spp. Bovine animals are considered reservoirs of this disease, because they intermittently disseminate the bacteria into the environment through their urine. In this way, the cattle an important source of Leptospira infection. The objective of this study was to detect Leptospira spp. antibodies and DNA in bovine females from two refrigerated slaughterhouses in the microregion of Umuarama, Paraná, Brazil. In particular, blood and urine samples from 52 crossbred bovine females older than 36 months from the two slaughterhouses were used. The microscopic agglutination test (MAT) was used to detect leptospiral antibodies, and the polymerase chain reaction (PCR) and subsequent sequencing were used to detect Leptospira DNA. The MAT yielded 22 (42.3%) serum samples considered reagent, while the nested PCR test resulted in one amplified sample (1.9%) of 289 bp. This single sample was then amplified again using primers for the SecY gene (549 bp). Sequencing of this gene characterized the bacteria as L. borgpetersenii that were similar to the serovar Hardjo of the genotype Hardjobovis. This is the first molecular confirmation of Hardjobovis-like L. borgpetersenii in the urine of crossbred bovine females older than 36 months from slaughterhouses in the microregion of Umuarama. This study’s results show that it is important to combine serological and molecular diagnosis in the detection of Leptospira spp. Therefore, both methods were used to improve our understanding of the epidemiology of this disease in bovine animals from the microregion of Umuarama. In addition, the analysis informed the subsequent adoption of preventive measures and educational One Health actions to prevent economic losses related to the herd, as well as social losses related to workers and the environment.

scholarly journals Comparative Clinical Evaluation of the Roche Elecsys and Abbott Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Serology Assays for Coronavirus Disease 2019 (COVID-19)

2020 ◽  
Vol 145 (1) ◽  
pp. 32-38 ◽  
Author(s):  
Shaun S. Tan ◽  
Sharon Saw ◽  
Ka Lip Chew ◽  
Cindy Wang ◽  
Anastacia Pajarillaga ◽  
...  
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Clinical Symptoms ◽  
Test Results ◽  
Serum Samples ◽  
False Negatives ◽  
Antiretroviral Medication ◽  
Polymerase Chain ◽  
Negative Controls ◽  
Level Of Agreement ◽  
Abbott Architect

Context.— The use of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) serologic tests detects antibodies in the host, contributing to the identification of individuals who have been exposed to coronavirus disease 2019 (COVID-19). Objective.— To critically evaluate 2 commercially available SARS-CoV-2 serology tests. Design.— A total of 333 unique, nonduplicated serum samples obtained from COVID-19 patients (n = 170) and negative controls (n = 163) obtained before December 2019 were used in the study. Samples were tested on the Roche E411 and Abbott Architect i4000SR platforms, and results were correlated to reverse transcription polymerase chain reaction (PCR) results and clinical symptoms. Results.— There was a strong level of agreement in the qualitative results between both assays, with a Cohen κ value of .840, P < .001. The specificity for both Roche and Abbott were excellent at 100%. Roche exhibited marginally better performance in the 21 days or more group with a sensitivity of 90.6% (95% CI, 75.8%–96.8%) versus an Abbott sensitivity of 84.4% (95% CI, 68.3%–93.1%), as well as in the 14- to 20-day group with a sensitivity of 85.7% (95% CI, 65.4%–95.0%) versus an Abbott sensitivity of 81.0% (95% CI, 60.0%–92.3%). Less than 14 days of symptoms groups exhibited poor sensitivity at less than 50% for both assays. The areas under curve (± standard error) for Roche (0.894 ± 0.025, P < .001) and Abbott (0.884 ± 0.026, P < .001) were very similar. Potential confounders for negative serologic results include antiretroviral medication use and pauci-symptomatic patients. Conclusions.— Specificities for high-throughput Roche and Abbott immunoassays are excellent, but users need to be cautious to interpret serologic test results after 14 days of symptoms to avoid false negatives.

scholarly journals High Co-infection Status of Novel Porcine Parvovirus 7 With Porcine Circovirus 3 in Sows That Experienced Reproductive Failure

2021 ◽  
Vol 8 ◽  
Author(s):  
Jinhui Mai ◽  
Dongliang Wang ◽  
Yawen Zou ◽  
Sujiao Zhang ◽  
Chenguang Meng ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Reproductive Failure ◽  
Porcine Circovirus ◽  
Reproductive Disorders ◽  
Serum Samples ◽  
Positive Rate ◽  
Porcine Circoviruses ◽  
Swine Herds ◽  
Pcv2 Replication

Porcine parvoviruses (PPVs) and porcine circoviruses (PCVs) infect pigs worldwide, with PPV1–7 and PCV2 infections common in pigs. Although PPV7 was only identified in 2016, co-infection of PPV7 and PCV2 is already common, and PPV7 may stimulate PCV2 replication. PCV3, a novel type of circovirus, is prevalent in pig populations worldwide and considered to cause reproductive disorders and dermatitis nephrotic syndrome. In recent studies, pigs were commonly infected with both PCV3 and PPV7. Our objective was to investigate the co-infections between PPV7 and PCV3 in samples from swine on farms in Hunan, China, and assess the potential impacts of PPV7 on PCV3 viremia. A total of 209 samples, known to be positive (105) or negative (104) for PCV3, were randomly selected from serum samples that were collected from commercial swine herds in seven regions from 2016 to 2018 in our previous studies; these samples were subjected to real-time PCR to detect PPV7. Of these samples, 23% (48/209) were positive for PPV7. Furthermore, the PPV7 positive rate was significantly higher in PCV3 positive serum (31.4%, 33/105) than in PCV3 negative serum (14.4%, 15/104). Another 62 PCV3 positive sow serum samples and 20 PCV3 positive aborted fetuses were selected from 2015 to 2016 in our other previous study. These samples were designated as being from farms with or without long-standing histories of reproductive failure (RF or non-RF), respectively, and they were also subjected to real-time PCR to detect PPV7 and to determine whether PPV7 affected PCV3 viremia. Among the 62 serum samples (39 PCV3 positive RF-serum and 23 PCV3 positive non-RF-serum), 45.1% (28/62) were positive for PPV7 and PCV3, and the PPV7 positive rate was significantly higher in PCV3 positive RF-serum (51.2%, 20/39) than in PCV3 positive non-RF-serum (34.8%, 8/23). In addition, there was a higher positive rate of PPV7 (55%, 11/20) in PCV3 positive aborted fetus samples. In addition, the copy number of PCV3 in PPV7 positive samples was significantly higher than that in PPV7 negative serum samples. Based on these findings, we concluded that PPV7 may stimulate PCV3 replication.

The presence and genetic characteristics of porcine circovirus 3 from pigs in Southern and Central provinces of Vietnam

2020 ◽  
pp. 26-31
Author(s):  
Giao N. P. Trinh
Keyword(s):  
Gene Segment ◽  
Porcine Circovirus Type ◽  
Porcine Circovirus ◽  
Nucleotide Sequencing ◽  
Duplex Pcr ◽  
Serum Samples ◽  
Swine Industry ◽  
Genetic Characteristics ◽  
Swine Herds ◽  
Type 3

Porcine circovirus type 3 (PCV3) is an emerging circovirus species that has recently been reported in different countries around the world, suggesting a widespread circulation. This study was carried out in order to investigate the presence and further genetic characteristics of PCV3 from swine herds in Southern and Central provinces of Vietnam. A duplex PCR assay for rapid detection of PCV3 in pigs was established with a pair of specific primers designed between rep and cap gene segment to amplify full-length ORF2 and another set of primers binding to COX1gene serving as an internal amplification control (IAC). The resulting duplex PCR was used to examine PCV3 presence in 94tissue and serum samples. Subsequently, PCV3 was detected in 10 out of 94 cases (10.6%). The infection rate in sows (14.3%) was higher than that in grower pigs (7.7%). Regarding nucleotide sequence comparison, 10 ORF2 genes were selected for nucleotide sequencing and their alignment showed 97.2% - 99.5% homology. According to the phylogenetic analysis and sequence alignment of cap gene, all the sequences were clustered into group PCV3a,including 9 strains of sub-group PCV3a1 and only one strain of subgroup PCV3a2. These findings indicated that the PCV3a group is circulating in swine farms in Vietnam. This study provides better insights into epidemiology of this pathogen in the national swine industry.

scholarly journals Characterization of a hospital-based gastroenteritis outbreak caused by GII.6 norovirus in Jinshan, China

2020 ◽  
Vol 148 ◽  
Author(s):  
X. F. Zhang ◽  
J. R. Chen ◽  
C. L. Song ◽  
D. J. Xie ◽  
M. Tan ◽  
...  
Keyword(s):  
Host Susceptibility ◽  
Enzyme Linked Immunosorbent Assay ◽  
Blood Group Antigens ◽  
Chain Reaction ◽  
Serum Samples ◽  
Polymerase Chain ◽  
Paired Serum ◽  
Asymptomatic Individuals ◽  
Binding Interfaces

Abstract An acute gastroenteritis (AGE) outbreak caused by a norovirus occurred at a hospital in Shanghai, China, was studied for molecular epidemiology, host susceptibility and serological roles. Rectal and environmental swabs, paired serum samples and saliva specimens were collected. Pathogens were detected by real-time polymerase chain reaction and DNA sequencing. Histo-blood group antigens (HBGA) phenotypes of saliva samples and their binding to norovirus protruding proteins were determined by enzyme-linked immunosorbent assay. The HBGA-binding interfaces and the surrounding region were analysed by the MegAlign program of DNAstar 7.1. Twenty-seven individuals in two care units were attacked with AGE at attack rates of 9.02 and 11.68%. Eighteen (78.2%) symptomatic and five (38.4%) asymptomatic individuals were GII.6/b norovirus positive. Saliva-based HBGA phenotyping showed that all symptomatic and asymptomatic cases belonged to A, B, AB or O secretors. Only four (16.7%) out of the 24 tested serum samples showed low blockade activity against HBGA-norovirus binding at the acute phase, whereas 11 (45.8%) samples at the convalescence stage showed seroconversion of such blockade. Specific blockade antibody in the population played an essential role in this norovirus epidemic. A wide HBGA-binding spectrum of GII.6 supports a need for continuous health attention and surveillance in different settings.

scholarly journals Limited replication of yellow fever 17DD and 17D-Dengue recombinant viruses in rhesus monkeys

2008 ◽  
Vol 80 (2) ◽  
pp. 311-321 ◽  
Author(s):  
Gisela F. Trindade ◽  
Renato S. Marchevsky ◽  
Ana M.B. de Fillipis ◽  
Rita M.R. Nogueira ◽  
Myrna C. Bonaldo ◽  
...  
Keyword(s):  
Real Time ◽  
Yellow Fever ◽  
Rhesus Monkeys ◽  
Plaque Assay ◽  
Serum Samples ◽  
Recombinant Viruses ◽  
Peak Titer ◽  
Dengue Virus Serotypes ◽  
Polymerase Chain

For the development of safe live attenuated flavivirus vaccines one of the main properties to be established is viral replication. We have used real-time reverse transcriptase-polymerase chain reaction and virus titration by plaque assay to determine the replication of yellow fever 17DD virus (YFV 17DD) and recombinant yellow fever 17D viruses expressing envelope proteins of dengue virus serotypes 2 and 4 (17D-DENV-2 and 17D-DENV-4). Serum samples from rhesus monkeys inoculated with YFV 17DD and 17D-DENV chimeras by intracerebral or subcutaneous route were used to determine and compare the viremia induced by these viruses. Viral load quantification in samples from monkeys inoculated by either route with YFV 17DD virus suggested a restricted capability of the virus to replicate reaching not more than 2.0 log10 PFU mL-1 or 3.29 log10 copies mL-1. Recombinant 17D-dengue viruses were shown by plaquing and real-time PCR to be as attenuated as YF 17DD virus with the highest mean peak titer of 1.97 log10 PFU mL-1 or 3.53 log10 copies mL-1. These data serve as a comparative basis for the characterization of other 17D-based live attenuated candidate vaccines against other diseases.

Characterization of a cDNA for Rhesus Monkey Protein C Inhibitor – Evidence for N-Terminal Involvement in Heparin Stimulation

1995 ◽  
Vol 74 (04) ◽  
pp. 1079-1087 ◽  
Author(s):  
Klaus-P Radtke ◽  
José A Fernández ◽  
Bruno O Villoutreix ◽  
Judith S Greengard ◽  
John H Griffin
Keyword(s):  
Rhesus Monkey ◽  
Protein C ◽  
Activated Protein C ◽  
Pcr Amplification ◽  
Amino Acid Sequences ◽  
Heparin Binding ◽  
Reactive Center ◽  
Protein C Inhibitor ◽  
Polymerase Chain

SummarycDNAs for protein C inhibitor (PCI) were cloned from human and rhesus monkey 1 liver RNAs by reverse transcription and polymerase chain reaction (PCR) amplification. Sequencing showed that rhesus monkey and human PCI cDNAs were 93% identical. Predicted amino acid sequences differed at 26 of 387 residues. Pour of these differences (T352M, N359S, R362K, L3631) were in the reactive center loop that is important for inhibitory specificity, and two were in the N-terminal helix (M8T, E13K) that is implicated in glycosaminoglycan binding. PCI in human or rhesus monkey plasma showed comparable inhibitory activity towards human activated protein C in the presence of 10 U/ml heparin. However, maximal acceleration of the inhibition of activated protein C required 5-fold lower heparin concentration for rhesus monkey than for human plasma, consistent with the interpretation that the additional positive charge (E13K) in a putative-heparin binding region increased the affinity for heparin.

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