Detection of Bovine and Porcine Adenoviruses for Tracing the Source of Fecal Contamination

ABSTRACT In this study, a molecular procedure for the detection of adenoviruses of animal origin was developed to evaluate the level of excretion of these viruses by swine and cattle and to design a test to facilitate the tracing of specific sources of environmental viral contamination. Two sets of oligonucleotides were designed, one to detect porcine adenoviruses and the other to detect bovine and ovine adenoviruses. The specificity of the assays was assessed in 31 fecal samples and 12 sewage samples that were collected monthly during a 1-year period. The data also provided information on the environmental prevalence of animal adenoviruses. Porcine adenoviruses were detected in 17 of 24 (70%) pools of swine samples studied, with most isolates being closely related to serotype 3. Bovine adenoviruses were present in 6 of 8 (75%) pools studied, with strains belonging to the genera Mastadenovirus and Atadenovirus and being similar to bovine adenoviruses of types 2, 4, and 7. These sets of primers produced negative results in nested PCR assays when human adenovirus controls and urban-sewage samples were tested. Likewise, the sets of primers previously designed for detection of human adenovirus also produced negative results with animal adenoviruses. These results indicate the importance of further studies to evaluate the usefulness of these tests to trace the source of fecal contamination in water and food and for environmental studies.

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Identification of Human and Animal Adenoviruses and Polyomaviruses for Determination of Sources of Fecal Contamination in the Environment

Applied and Environmental Microbiology ◽

10.1128/aem.01090-06 ◽

2006 ◽

Vol 72 (12) ◽

pp. 7886-7893 ◽

Cited By ~ 114

Author(s):

Ayalkibet Hundesa ◽

Carlos Maluquer de Motes ◽

Silvia Bofill-Mas ◽

Nestor Albinana-Gimenez ◽

Rosina Girones

Keyword(s):

River Water ◽

Water Samples ◽

Environmental Samples ◽

Fecal Contamination ◽

Source Tracking ◽

Animal Origin ◽

Urban Sewage ◽

River Water Samples ◽

Negative Results ◽

Wastewater Samples

ABSTRACT The Adenoviridae and Polyomaviridae families comprise a wide diversity of viruses which may be excreted for long periods in feces or urine. In this study, a preliminary analysis of the prevalence in the environment and the potential usefulness as source-tracking tools of human and animal adenoviruses and polyomaviruses has been developed. Molecular assays based on PCR specifically targeting human adenoviruses (HAdV), porcine adenoviruses (PAdV), bovine adenoviruses (BAdV), and bovine polyomaviruses (BPyV) were applied to environmental samples including urban sewage, slaughterhouse, and river water samples. PAdV and BPyV were detected in a very high percentage of samples potentially affected by either porcine or bovine fecal contamination, respectively. However, BAdV were detected in only one sample, showing a lower prevalence than BPyV in the wastewater samples analyzed. The 22 slaughterhouse samples with fecal contamination of animal origin showed negative results for the presence of HAdV. The river water samples analyzed were positive for the presence of both human and animal adenoviruses and polyomaviruses, indicating the existence of diverse sources of contamination. The identities of the viruses detected were confirmed by analyses of the amplified sequences. All BPyV isolates showed a 97% similarity in nucleotide sequences. This is the first time that PAdV5, BAdV6, and BPyV have been reported to occur in environmental samples. Human and porcine adenoviruses and human and bovine polyomaviruses are proposed as tools for evaluating the presence of viral contamination and for tracking the origin of fecal/urine contamination in environmental samples.

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Development of Nested PCR Assays for Detection of Bovine Respiratory Syncytial Virus in Clinical Samples

Journal of Clinical Microbiology ◽

10.1128/jcm.32.11.2887-2887.1994 ◽

1994 ◽

Vol 32 (11) ◽

pp. 2887-2887

Keyword(s):

Respiratory Syncytial Virus ◽

Nested Pcr ◽

Bovine Respiratory Syncytial Virus ◽

Clinical Samples ◽

Pcr Assays ◽

Syncytial Virus ◽

Nested Pcr Assays

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Nested PCR assays with novel primers yield greater sensitivity to Tanzanian HIV-1 samples than a commercial PCR detection kit

Journal of Virological Methods ◽

10.1016/s0166-0934(96)02094-0 ◽

1996 ◽

Vol 62 (2) ◽

pp. 131-141 ◽

Cited By ~ 4

Author(s):

Olov Grankvist ◽

Lilian Walther ◽

Ulla Bredberg-Rådén ◽

Eligius Lyamuya ◽

Fred Mhalu ◽

...

Keyword(s):

Nested Pcr ◽

Pcr Detection ◽

Pcr Assays ◽

Nested Pcr Assays ◽

Hiv 1

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Malaria Parasites in Macaques in Thailand: Stump-Tailed Macaques (Macaca Arctoides) are New Natural Hosts for Plasmodium Knowlesi, P. Inui, P. Coatneyi and P. Fieldi.

10.21203/rs.3.rs-26507/v1 ◽

2020 ◽

Author(s):

Wirasak Fungfuang ◽

Chanya Udom ◽

Daraka Tongthainan ◽

Khamisah Abdul Kadir ◽

Balbir Singh

Keyword(s):

Nested Pcr ◽

Plasmodium Knowlesi ◽

Sampling Site ◽

Macaca Arctoides ◽

Malaria Parasites ◽

Zoonotic Malaria ◽

Natural Hosts ◽

New Locations ◽

Pcr Assays ◽

Nested Pcr Assays

Abstract Background:Certain species of macaques are natural hosts ofPlasmodium knowlesi and P. cynomolgi, which can both cause malaria in humans, and P. inui, which can be experimentally transmitted to humans. A significant number of zoonotic malaria cases have been reported in humans throughout Southeast Asia, including Thailand. There have been only two studies undertaken in Thailand to identify malaria parasites in non-human primates in 6 provinces. The objective of this study was to determine the prevalence of P. knowlesi, P. coatneyi, P. cynomolgi, P. inui and P. fieldiin non-human primates from 4 new locations in Thailand. Methods:A total of 93 blood samples from Macaca fascicularis, M. leonina and M. arctoides were collected from four locations in Thailand: 32 were captive M. fascicularisfrom Chachoengsao Province (CHA), 4 were wild M. fascicularis from Ranong Province (RAN), 32 were wildM. arctoidesfromPrachuap Kiri Khan Province (PRA), and 25 were wild M. leoninafrom Nakornratchasima Province (NAK). DNA was extracted from these samples and analysed by nested PCR assays to detect Plasmodium, and subsequently to detectP. knowlesi, P. coatneyi, P. cynomolgi, P. inui and P. fieldi.Results:Twenty-seven of the 93 (29%) samples were Plasmodium-positive by nested PCR assays. Among wild macaques, all 4 M. fascicularis at RAN were infected with malaria parasites followed by 50% of 32 M. arctoides at PRA and 20% of 25 M. leonina at NAK. Only 2 (6.3%) of the 32 captive M. fascicularisat CHA were malaria-positive. All 5 species of Plasmodium were detected and 16 (59.3%) of the 27 macaques had single infections, 9had double and 2 had triple infections.The composition of Plasmodium species in macaques at each sampling site was different. Macaca arctoides from PRA were infected with P. knowlesi, P. coatneyi, P. cynomolgi, P. inui and P. fieldi. Conclusions:The prevalence and species of Plasmodiumvaried among the wild and captive macaques, and betweenmacaques at 4 sampling sites in Thailand. Macaca arctoidesis a new natural host for P. knowlesi, P. inui,P. coatneyi and P. fieldi.

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Detection and identification of Toscana and other phleboviruses by RT-nested-PCR assays with degenerated primers

Journal of Medical Virology ◽

10.1002/jmv.10465 ◽

2003 ◽

Vol 71 (1) ◽

pp. 140-149 ◽

Cited By ~ 113

Author(s):

María-Paz Sánchez-Seco ◽

José-Manuel Echevarría ◽

Lourdes Hernández ◽

Domingo Estévez ◽

José-María Navarro-Marí ◽

...

Keyword(s):

Nested Pcr ◽

Detection And Identification ◽

Pcr Assays ◽

Nested Pcr Assays

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Simultaneous detection of fourteen respiratory viruses in clinical specimens by two multiplex reverse transcription nested-PCR assays

Journal of Medical Virology ◽

10.1002/jmv.20008 ◽

2004 ◽

Vol 72 (3) ◽

pp. 484-495 ◽

Cited By ~ 199

Author(s):

M.T. Coiras ◽

J.C. Aguilar ◽

M.L. García ◽

I. Casas ◽

P. Pérez-Breña

Keyword(s):

Reverse Transcription ◽

Nested Pcr ◽

Simultaneous Detection ◽

Respiratory Viruses ◽

Clinical Specimens ◽

Pcr Assays ◽

Nested Pcr Assays

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Development of conventional and nested PCR assays for the detection ofOphiocordyceps sinensis

Journal of Basic Microbiology ◽

10.1002/jobm.201200029 ◽

2012 ◽

Vol 53 (4) ◽

pp. 340-347 ◽

Cited By ~ 8

Author(s):

Guo-Sheng Jin ◽

Xiao-Liang Wang ◽

Yi Li ◽

Wen-Jing Wang ◽

Rui-Heng Yang ◽

...

Keyword(s):

Nested Pcr ◽

Pcr Assays ◽

Nested Pcr Assays

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Use of viral pathogens and indicators to differentiate between human and non-human fecal contamination in a microbial source tracking comparison study

Journal of Water and Health ◽

10.2166/wh.2003.0021 ◽

2003 ◽

Vol 1 (4) ◽

pp. 195-207 ◽

Cited By ~ 78

Author(s):

Rachel T. Noble ◽

Steven M. Allen ◽

Angelia D. Blackwood ◽

Weiping Chu ◽

Sunny C. Jiang ◽

...

Keyword(s):

Fecal Contamination ◽

Human Adenovirus ◽

Microbial Source Tracking ◽

Human Enterovirus ◽

Source Tracking ◽

Comparison Study ◽

Viral Pathogens ◽

Methods Comparison ◽

General Ability ◽

Negative Results

Assays for the detection and typing of adenoviruses, enteroviruses and F+ specific coliphages were performed on samples created as part of a national microbial source tracking methods comparison study. The samples were created blind to the researchers, and were inoculated with a variety of types of fecal contamination source (human, sewage, dog, seagull and cow) and mixtures of sources. Viral tracer and pathogen assays demonstrated a general ability to discriminate human from non-human fecal contamination. For example, samples inoculated with sewage were correctly identified as containing human fecal contamination because they contained human adenovirus or human enterovirus. In samples containing fecal material from individual humans, human pathogen analysis yielded negative results probably because the stool samples were taken from healthy individuals. False positive rates for the virus-based methods (0–8%) were among the lowest observed during the methods comparison study. It is suggested that virus-based source tracking methods are useful for identification of sewage contamination, and that these methods may also be useful as an indication of the public health risk associated with viral pathogens. Overall, virus-based source tracking methods are an important approach to include in the microbial source tracking ‘toolbox’.

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