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.

Enteric Viruses of Humans and Animals in Aquatic Environments: Health Risks, Detection, and Potential Water Quality Assessment Tools

“…In addition, some enteric viruses have been linked to chronic diseases such as myocarditis and insulindependent diabetes (60,90). Enteric virus infections in animals such as cattle and swine are normally asymptomatic but can lead to abortion, neurological disorders, and mortality (80,83,97,105).…”

A Survey of Porcine Picornaviruses and Adenoviruses in Fecal Samples in Spain

“…Nevertheless, when comparing both studies, one should bear in mind that they differ in 2 important aspects: 1) the number of isolates analyzed (206 vs. 33) and 2) the sampling period, comprising 10 years in the Italian study (1992)(1993)(1994)(1995)(1996)(1997)(1998)(1999)(2000)(2001)(2002) and only 2 years in the present study (2004 and 2005). The current study was focused on members of the Picornaviridae and Adenoviridae families, because viruses belonging to these groups have been detected previously in fecal and environmental samples from pig populations in the absence of disease, 2,4,5,8 However, a still substantial percentage (40%) of the isolates could not be identified. These isolates are thus cytopathic viruses not belonging to the aforementioned viral groups, and their identification would merit further investigation.…”

“…Porcine adenoviruses (family Adenoviridae) have also been frequently found in pig fecal and environmental samples. 8 Among the Picornaviridae family, Swine vesicular disease virus (SVDV), a subtype of Human enterovirus B and belonging to the coxsackievirus B group, causes a disease of high relevance for trade and is included in the World Organization for Animal Health (OIE) list of notifiable animal diseases. 13 In Spain, a specific program for surveillance of swine diseases is being implemented.…”

“…This conventional RT-PCR is able to distinguish PTV and PSV by the size of the band amplified. To control the specificity of the technique, samples corresponding to PSV and PTV reference strains (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11) c,d were assessed, giving bands of the expected sizes (Table 1). When the assay was applied to all 206 virus isolates, 18 PTV-negative isolates were identified as PSV, whereas only 48 of 97 PTV-positive isolates were found positive for PTV in this second test; this latter result possibly reflects the higher sensitivity of the real-time RT-PCR protocol applied in the first analysis for the detection of PTV.…”

“…Next, 2 more tests were applied for 2 groups of cytopathic porcine enteric viruses likely to be found in swine fecal samples: an RT-PCR assay for enteroviruses that detects Porcine enterovirus B (PEV-B) 5,9 and a specific assay for the detection of Porcine adenovirus A-C (PAdV-A-C; protease gene). 8 Both tests were assessed using specific viral material as a performance control: in the first method, 2 serotypes of PEV-B (1 and 2 reference strains) c were used, whereas in the second method, a PAdV isolate e was used. Each technique was able to detect its homologous virus and did not cross-react with other viruses analyzed in the present study ( Table 1).…”