RESUMO
Woodchip filters have received attention in recent years for their ability to sustain denitrification activity across multiyear time frames. However, in some freshwater aquatic ecosystems, P rather than N is the nutrient considered most responsible for eutrophication. Previous studies have indicated that woodchip filters have limited ability to remove dissolved P, but in agricultural terrain, P export in watercourses is often dominated by particulate P (PP). Woodchip media, because of their high porosity and permeability and the surface roughness of the particles, could be effective for PP removal. In this study, we tested a woodchip filter for its ability to remove suspended sediment and associated PP at a farm in southern Ontario, Canada, where vegetable wash water with extremely high total suspended solids (TSS) was generated. The treatment system consisted of a 12.3-m concrete sedimentation tank and a slightly larger woodchip filter (16.1 m) installed in a subsurface trench. During 7 mo of full-scale operation, treating 10.8 m d, the filter system removed 71% of influent total P (TP) averaging 8.8 mg L and 99% of TSS averaging 5800 mg L, with most of the removal occurring in the tank and a lesser amount (6-16%) occurring in the woodchip filter. Almost all of the TP removal was associated with PP (91% removal) because dissolved P, averaging 1.5 mg L in the wash water, was little changed. Woodchip filters, when coupled with a solids settling tank, have the potential to provide high-capacity, low-maintenance treatment of suspended solids and associated particulate P in turbid waters.
Assuntos
Agricultura , Fósforo/análise , Purificação da Água , Canadá , Desnitrificação , Filtração , MadeiraRESUMO
A real-time quantitative polymerase chain reaction (Q-PCR) assay was developed for detecting and quantifying Escherichia coli in water samples from agricultural watersheds. The assay included optimization of DNA extraction and purification from water samples, and Q-PCR amplification conditions using newly designed species-specific oligonucleotide primers derived from conserved flanking regions of the 16S rRNA gene, the internal transcribed spacer region (ITS) and the 23S rRNA gene. The assay was optimized using a pure culture of E. coli with known quantities spiked into autoclaved agricultural water samples. The optimized assay was capable of a minimum quantification limit of 10 cells/ml of E. coli in the spiked agricultural water samples. A total of 121 surface water samples from three agricultural watersheds across Canada were analyzed, and results were compared with conventional culture-based enumerations of E. coli. The Q-PCR assay revealed significantly higher numbers of E. coli in water samples than the culture-based assay in each agricultural watershed. The new Q-PCR assay can facilitate the quantification of E. coli in a single water sample in < 3 h, including melt curve analysis, across a range of agricultural water quality conditions.
Assuntos
Agricultura , Escherichia coli/isolamento & purificação , Água Doce/microbiologia , Reação em Cadeia da Polimerase/métodos , Animais , Gatos , Bovinos , Criança , Contagem de Colônia Microbiana , Meios de Cultura , DNA Bacteriano/análise , DNA Bacteriano/isolamento & purificação , DNA Espaçador Ribossômico/análise , Cães , Escherichia coli/genética , Escherichia coli/crescimento & desenvolvimento , Cobaias , Humanos , Dados de Sequência Molecular , RNA Ribossômico 16S/genética , RNA Ribossômico 23S/genética , Análise de Sequência de DNA , Fatores de TempoRESUMO
Surface pasteurization for inactivation of Listeria monocytogenes was evaluated for radiant heat prepackage pasteurization, submersed water postpackage pasteurization, and combinations of the two techniques on various types of ready-to-eat deli turkey products obtained from at least four different manufacturers. Products were inoculated either by in-package liquid inoculum or surface sponge-contact with approximately 10(9) CFU of L. monocytogenes. Additional testing of radiant heat pasteurization was performed with low-level inoculation of product undersides with approximately 100 CFU of L. monocytogenes followed by enrichment recovery after pasteurization. Prepackage pasteurization provided 2.0 to 2.8 log reductions when processed for 60 s and 2.8 to 3.8 log reductions when processed for 75 s. An improved radiant oven provided 3.53 (60 s) and 4.76 (75 s) log reductions of L. monocytogenes. No positive samples were detected after enrichment when 40 samples of deli turkey (4 to 4.5 kg) undersides were inoculated at low levels and processed for 75 s. Submersed water postpackage pasteurization provided 1.95 to 3.0 log reductions when processed for 2, 3, 4, or 5 min, and combinations of the two processes gave 3.0 to 4.0 log inactivation of L. monocytogenes using either 60 + 60 s or 60 + 90 s for the prepackage and postpackage pasteurization processes, respectively. These processes, either individually or in combination, can provide postprocess elimination of bacteria for the manufacture of safe ready-to-eat deli meats.
Assuntos
Embalagem de Alimentos/métodos , Conservação de Alimentos/métodos , Temperatura Alta , Listeria monocytogenes/crescimento & desenvolvimento , Produtos da Carne/microbiologia , Animais , Contagem de Colônia Microbiana , Qualidade de Produtos para o Consumidor , Manipulação de Alimentos/métodos , Microbiologia de Alimentos , Fatores de Tempo , PerusRESUMO
Campylobacter species contribute to an enormous burden of enteric illnesses around the world. This study compared two different methods for detecting Campylobacter species in surface water samples from agricultural watersheds across Canada. One method was based on membrane filtration (MF) of 500 ml water samples followed by selective microaerophilic enrichment at 42 degrees C in Bolton broth, isolation of Campylobacter on CCDA, and subsequent identification confirmation by a PCR assay. The second method was based on centrifugation (CF) of 1000 ml water samples, followed by selective microaerophilic enrichment at 42 degrees C in Bolton broth, isolation of Campylobacter on Modified Karmali Agar, and subsequent identification confirmation by a different PCR assay. Overall comparison of the CF and MF methods indicated that both methods found Camylobacterjejuni to be the most commonly detected Campylobacter species in 699 water samples from four agricultural watersheds across Canada, and that C. jejuni frequency of occurrence was similar by both methods. However, the CF method detected significantly higher frequencies of Campylobactercoli (17%) and other Campylobacter species (13%) than the MF method (11% and 3%, respectively). It was frequently found that one method would detect Campylobacter in a water sample when the other method would not for a simultaneously collected, duplicate water sample. This study indicates that methods can have significantly different recovery efficiencies for Campylobacter species, and that caution is needed when comparing studies that report on the frequency of occurrence of waterborne Campylobacter at the genus level when different detection methods are used.
Assuntos
Técnicas de Tipagem Bacteriana/métodos , Campylobacter/isolamento & purificação , Água Doce/microbiologia , Microbiologia da Água , Campylobacter/genética , Centrifugação/métodos , Meios de Cultura , Eletroforese em Gel de Ágar , Filtração/métodos , Genes Bacterianos , Reação em Cadeia da Polimerase/métodos , RNA Ribossômico 16S/genéticaRESUMO
Recent molecular evidence suggests that different species and/or genotypes of Cryptosporidium display strong host specificity, altering our perceptions regarding the zoonotic potential of this parasite. Molecular forensic profiling of the small-subunit rRNA gene from oocysts enumerated on microscope slides by U.S. Environmental Protection Agency method 1623 was used to identify the range and prevalence of Cryptosporidium species and genotypes in the South Nation watershed in Ontario, Canada. Fourteen sites within the watershed were monitored weekly for 10 weeks to assess the occurrence, molecular composition, and host sources of Cryptosporidium parasites impacting water within the region. Cryptosporidium andersoni, Cryptosporidium muskrat genotype II, Cryptosporidium cervine genotype, C. baileyi, C. parvum, Cryptosporidium muskrat genotype I, the Cryptosporidium fox genotype, genotype W1, and genotype W12 were detected in the watershed. The molecular composition of the Cryptosporidium parasites, supported by general land use analysis, indicated that mature cattle were likely the main source of contamination of the watershed. Deer, muskrats, voles, birds, and other wildlife species, in addition to sewage (human or agricultural) may also potentially impact water quality within the study area. Source water protection studies that use land use analysis with molecular genotyping of Cryptosporidium parasites may provide a more robust source-tracking tool to characterize fecal impacts in a watershed. Moreover, the information is vital for assessing environmental and human health risks posed by water contaminated with zoonotic and/or anthroponotic forms of Cryptosporidium.