Detection of Known and Novel Adenoviruses in Cattle Wastes via Broad-Spectrum Primers

Abstract

The critical assessment of bovine adenoviruses (BAdV) as indicators of environmental fecal contamination requires improved knowledge of their prevalence, shedding dynamics, and genetic diversity. We examined DNA extracted from bovine and other animal waste samples collected in Wisconsin for atadenoviruses and mastadenoviruses using novel, broad-spectrum PCR primer sets. BAdV were detected in 13% of cattle fecal samples, 90% of cattle urine samples, and 100% of cattle manure samples; 44 percent of BAdV-positive samples contained both Atadenovirus and Mastadenovirus DNA. Additionally, BAdV were detected in soil, runoff water from a cattle feedlot, and residential well water. Overall, we detected 8 of 11 prototype BAdV, plus bovine, rabbit, and porcine mastadenoviruses that diverged significantly from previously reported genotypes. The prevalence of BAdV shedding by cattle supports targeting AdV broadly as indicators of the presence of fecal contamination in aqueous environments. Conversely, several factors complicate the use of AdV for fecal source attribution. Animal AdV infecting a given livestock host were not monophyletic, recombination among livestock mastadenoviruses was detected, and the genetic diversity of animal AdV is still underreported. These caveats highlight the need for continuing genetic surveillance for animal AdV and for supporting data when BAdV detection is invoked for fecal source attribution in environmental samples. To our knowledge, this is the first study to report natural BAdV excretion in urine, BAdV detection in groundwater, and recombination in AdV of livestock origin.

Detection of human adenovirus, rotavirus and enterovirus in water samples collected on dairy farms from Tenente Portela, Northwest of Rio Grande do Sul, Brazil

“…Enteric viruses have a number of characteristics that make them excellent markers for fecal contamination of water: i) they are extremely resistant in the environment due to its non-enveloped structure, ii) they are eliminated in large quantities in the feces of humans and animals sick or subclinical infections in iii) in most cases these viruses are host-specific and thus allow screening of the species which is the source of fecal contamination (Fong and Lipp, 2005; Silva et al , 2011; Wolf et al , 2010; Wu et al , 2011). Among the enteric viruses three of the most studied as environmental contaminants are the adenoviruses (AdV, Adenoviridae family, Mastadenovirus genus, double-stranded DNA), enteroviruses (EV, Picornavirales order, Picornaviridae family, Enterovirus genus, single-stranded RNA, positive sense) and rotaviruses (RV, Reoviridae family, Sedoreovirinae subfamily, genus Rotavirus ) (Comerlato et al , 2011; Fong and Lipp, 2005; Matthijnssens et al , 2008; Sibley et al , 2011). These agents are transmitted by the fecal-oral route, being associated with a number of diseases, especially gastroenteritis, either in human beings or animals (Ahmad et al , 2009; Hamza et al , 2011).…”

A Novel Strain of Porcine Adenovirus Detected in Urinary Bladder Urothelial Cell Culture

“…For this purpose a broad range of adenovirus primers was used, enabling successful amplification of the hexon and polymerase genes of various types of mammal adenoviruses. The sequencing and sequence analysis of the amplified hexon gene fragment of adenovirus isolate showed the highest similarity to recently described novel porcine adenovirus strain PAdV-WI, which was so far detected only in pen wash water of newborn to finisher pigs and has been proposed as a prototype of a new species in the Mastadenovirus genus [14]. According to the short nucleotide sequence of the hexon gene analyzed in this study and the comparison to the initially recognized PAdV-WI, both strains showed a clear separation from other, even the most similar representatives in the Mastadenovirus genus obtained from the GenBank (Figure 4B, Table 1).…”

Viruses Associated With Foodborne Infections

“…A broad-spectrum PCR assay was developed by Sibley et al (2011) for the detection of Mastadenovirus (MaAdV) and Atadenovirus (AtAdV), based on the adenovirus hexon gene. MaAdV, which comprises human and bovine adenoviruses and a large variety of mammalian adenoviruses.…”