Assessment of F-RNA Coliphage as a Potential Indicator of Enteric Virus Contamination of Hog Carcasses

Hepatitis E virus (HEV) is common in pigs, and some swine HEV strains are closely related to human strains. The zoonotic transmission of HEV is now well established. HEV can be detected by molecular techniques, but the significance of the presence of viral nucleic acid is questionable when foods are subjected to virus inactivation treatments. F-RNA coliphages are attractive candidates as indicators for enteric viruses because they are similar in size and survival characteristics and can be rapidly cultured. Information on the contamination of hog carcasses with enteric or hepatic viruses during slaughter is lacking. The objective of this study was to compare the incidence and levels of contamination of hog carcasses with F-RNA coliphages, HEV, total aerobic bacteria, coliforms, and Escherichia coli at different stages of the dressing process. Hog carcasses entering the commercial slaughter facility are heavily contaminated with F-RNA coliphages and HEV. Subsequent processes such as scalding, singing, and pasteurization can reduce the incidence and levels of F-RNA coliphages and HEV substantially to almost undetectable levels. Large discrepancies between the amount of viral nucleic acid and infectious F-RNA coliphage particles, both at high levels and low levels of contamination, were observed. The prevalence and levels of viable F-RNA coliphages were lower than those of total aerobic bacteria, coliforms, and E. coli in the anal area and on random sites before pasteurization. At a research abattoir, there was no overall mean reduction of viable F-RNA coliphages recovered from random sites before pasteurization and after washing, whereas overall mean reductions of 1.2, 2.6, and 2.9 log CFU for total aerobic bacteria, coliforms, and E. coli, respectively, were obtained. These findings suggest that bacteria such as coliforms and E. coli may not be suitable as indicators for enteric viruses in a meat processing environment.     

Relative value of surrogate indicators for detecting pathogens in lakes and reservoirs

This study investigated the relative behavior of pathogens, fecal indicator organisms, and particles of varying size during transport through a reservoir following a storm event inflow in Myponga Reservoir, South Australia. During the inflow, samples were collected from the river and at various locations within the reservoir to determine the fate and transport of microroganisms as they progressed through the water body. Microbiological analysis included the indicator organisms Escherichia coli, enterococci, Clostridium perfringens, aerobic spores, and somatic coliphages, the protozoan pathogens Cryptosporidium spp. and Giardia spp., and the potential physical surrogates of pathogen contamination including particle size and turbidity. Of the microbial indicator groups, C. perfringens spores were the most highly correlated with Cryptosporidium spp. concentrations (Spearman Rho = 0.58), closely followed by enterococci (Spearman Rho = 0.57). Cryptosporidium spp. oocysts were predominantly associated with small sized particles (range of 14.3-27.7 microm). All of the microbial indicator groups tested were associated with larger sized particle ranges (> 63.3 microm) except C. perfringens spores which were associated with particles in the size range of 45.5-63.3 microm. Although indicators may rank correlate with Cryptosporidium spp., the variation in settling rates of different microorganisms has significant implications for the use of surrogates to estimate pathogen attenuation within reservoirs. For example, concentrations of Cryptosporidium spp. oocysts were reduced by a factor of 3 on reaching the dam wall, whereas enterococci were reduced by a factor of 10.     

Microbiological analysis of seed sprouts in Norway

As part of larger survey of microbial contamination of fruits and vegetables in Norway, four different sprouted seed products were analysed for bacterial and parasitic contaminants (n = 300 for bacterial analyses and n = from 17 to 171 for parasite analyses, depending on parasite). Escherichia coli O157, Salmonella, Listeria monocytogenes, Cyclospora oocysts, Ascaris eggs and other helminth parasites were not detected in any of the sprout samples. Thermotolerant coliform bacteria (TCB) were isolated from approximately 25% of the sprout samples, with the highest percentage of TCB positive samples in alfalfa sprouts. Most TCB were Enterobacter spp. and Klebsiella. E. coli was isolated from 8 of 62 TCB positive mung bean sprout samples. Cryptosporidium oocysts were detected in 8% of the sprout samples and Giardia cysts were detected in 2% of the samples. All the Cryptosporidium positive samples, and most of the Giardia positive samples, were mung bean sprouts. Parasite concentrations in positive samples were low (between 1 and 3 oocysts/cysts per 50 g sprouts). Sprout irrigation water was also analysed for microbial contaminants. E. coli O157 and L. monocytogenes were not detected. TCB were isolated from approximately 40% of the water samples. Salmonella reading was isolated from three samples of spent irrigation water on 3 consecutive days. Cryptosporidium and Giardia were also isolated from spent irrigation water. Additionally, eight samples of unsprouted mung bean seed were analysed for Cryptosporidium oocysts and Giardia cysts. One or both of these parasites were detected in six of the unsprouted seed samples at concentrations of between 1 and 5 oocysts/cysts per 100 g unsprouted seed. Whilst our results support spent irrigation water as the most suitable matrix for testing for bacteria, unsprouted seed is considered the more useful matrix for analysing for parasite contaminants.     

Potential sources of microbial contamination in unpasteurized apple cider

A study was conducted to identify possible sources of microbial contamination and to assess the effect of good cleaning and sanitation practices on the microbial quality and safety of unpasteurized apple cider. Raw unwashed apples, washed apples, cleaning water, fresh cider, and finished cider samples were collected from five Ontario producers over 4 months and microbiologically tested. Total coliforms were found in 31, 71 and 38% of the unwashed apple, water, and washed apple samples, respectively. Escherichia coli was found in 40% of the water samples from one producer alone. The washing step was identified as a potential source of contamination, possibly due to water in the dump tanks seldom being refreshed, and because scrubbers, spray nozzles, and conveyors were not properly cleaned and sanitized. Higher total coliform counts (P < 0.0001) and prevalence (P < 0.0001) in fresh cider compared with those in unwashed apples and washed apples indicated considerable microbial buildup along the process, possibly explained by the lack of appropriate equipment sanitation procedures. Results showed that producers who had better sanitary practices in place had lower (P < 0.001) total coliform prevalence than the rest of the producers. Overall results show that good sanitation procedures are associated with improved microbial quality of fresh cider in terms of total coliforms and that operators who pasteurize and/or UV treat their product should still be required to have a sound good manufacturing practices program in place to prevent recontamination. Cryptosporidium parvum, an important pathogen for this industry, was found in different sample types, including washed apples, water, and fresh and finished cider.     

Comparative analysis of viral pathogens and potential indicators in shellfish

Shellfish can be responsible of outbreaks of infectious diseases and current health measures do not guarantee the absence of viral pathogens in this product. Here we examine the presence of pathogenic viruses and potential indicators in shellfish in a comparative analysis.Sixty shellfish samples collected in three areas with different levels of faecal contamination were analysed for Escherichia coli, total coliforms, Clostridium perfringens, somatic coliphages, F-specific phages of RNA (F-RNA), bacteriophages infecting Bacteroides fragilis RYC2056, human adenovirus, enterovirus and hepatitis A virus (HAV). Viruses were eluted in a glycine buffer at pH 10.The overall percentage of viral pathogens detected was 47% for human adenoviruses, 19% for enteroviruses and 24% for HAV. Since all the samples positive for enterovirus and HAV were also positives for human adenovirus, the latter may be considered useful as a molecular index of viral contamination in shellfish. No significant differences in the bioaccumulation of bacteria and bacteriophages for oysters or mussels were observed. It was found that the probability of detection of any of the pathogenic virus decreases as the temperature of shellfish growing waters increases. However, the probability of detecting viruses increases when phages of B. fragilis are found. Although more data are needed in order to fulfil the need of viral indicators for controlling the presence of human viruses in shellfish, the obtained results indicate that phages infecting B. fragilis RYC2056 could be a suitable group of bacteriophages to be used as an indicator of the presence of viruses in shellfish.     

Depuration dynamics of viruses in shellfish

The consumption of shellfish has been associated with viral infections even in cases where shellfish complied with the current regulation, which is based on bacterial analysis. In this study, depuration rates of potential indicators and human viruses have been analysed in order to study the use of complementary parameters for evaluating the microbiological quality of depurated shellfish. Depuration of naturally highly polluted mussels has been evaluated and analyses for Escherichia coli, Clostridium perfringens, somatic coliphages, F-RNA phages and bacteriophages infecting Bacteroides fragilis RYC2056 and HSP40, human adenovirus, enterovirus have been done. Seawater of the depuration tank was disinfected by UV irradiation, ozone and passed through a skimmer and a biological filter. The correct functioning of the depuration tank was monitored by the quantification of total organic carbon (TOC), NH4+ and total aerobic bacteria in the seawater. To study the relation between the bacteriophages and the human viruses analysed, a logistic regression model was applied. F-RNA phages are significantly related to human adenoviruses and enteroviruses. Thus, they can be used as a complementary parameter for evaluating the efficiency of the depuration treatment. Somatic coliphages are also significantly associated with enteroviruses. Bacteriophages infecting B. fragilis HSP40 were analysed by the double-agar-layer (DAL) method, which quantifies infectious viruses, and by nested PCR, which detects the presence of the genome of these phages. The highest sensitivity of the molecular techniques was demonstrated and the results obtained are an indicator of a close relation between positive results by PCR and the presence of infectious viral particles in shellfish. All shellfish samples were negative for human viruses by PCR after 5 days of depuration treatment and the results obtained applying a regression model also showed negative results or nearly for F-RNA phages and bacteriophages infecting B. fragilis RYC2056. Thus, in this specific depuration treatment, 5 days may be necessary to assess the sanitary quality of shellfish.     

Contamination of bivalve molluscs by Cryptosporidium oocysts: the need for new quality control standards

A yearlong study was carried out to investigate the presence and viability of Cryptosporidium oocysts in 203 samples of cultured shellfish from Galicia (NW Spain) and 38 samples imported from other European Union (EU) countries. Shellfish samples included mussels, oysters, clams and cockles. Cryptosporidium oocysts were detected, using a direct immunofluorescence antibody test (IFAT), in 34.4% of the samples analyzed; use of the fluorogenic dye propidium iodide (PI) revealed viable potentially infective oocysts in 53.0% of these samples. There was no relation between the presence of Cryptosporidium oocysts and the microbiological contamination detected in the samples expressed as Most-Probable-Number (MPN) of fecal coliforms, the different species of mollusc, or the month of sampling. One important finding was that the depuration process was ineffective in totally removing oocyst contamination. Furthermore, the existence of viable oocysts in samples with microbiological contamination levels lower than 300 fecal coliforms/100 g, which in accordance with current legislation are considered suitable for human consumption, suggests the need to include parasitological analyses in the quality control for these molluscs.     

Demonstration of Indicator Microorganisms on Surface of Vegetables on the Market in the United States and Mexico

ABSTRACT: Presence of indicator microorganisms of fecal pollution on the surface of vegetables sold in retail markets is demonstrated. Vegetables were obtained from local markets in the U.S.A. and Mexico. At least 1 of the indicators, among coliphages, fecal streptococci, total coliforms, and fecal coliforms, was detected in every sample tested. Relatively small amounts of indicators were recovered from the U.S.A. samples. Fecal streptococci showed significant correlations with all other indicators. However, some U.S.A. samples low in fecal streptococci were found to harbor certain levels of other indicators. Upon introduction of an indicator system to detect the fecal contamination of agriculture crops, it is recommended that multiple indicator microorganisms be measured.     

北京市农田回用再生水中无机阴离子的 分布规律研究

目的建立离子色谱法测定北京市农田回用再生水水体中F~-、NO_3~-、NO_2~-、Cl~-、SO_4~(2-)和PO_4~(3-)6种无机阴离子的分析方法。方法分别采集北京某再生水厂出水口、再生水灌溉渠、农田灌溉点的36份水样,经Dionex OnGuard Ⅱ Ag/H银离子交换柱处理后,采用离子色谱法测定,色谱柱为Dionex IonPac~(TM)AS11-HC,检测器为电导检测器。结果该方法能有效分离6种无机阴离子,且在0.016~10 mg/L浓度范围内线性关系良好(r~20.99),相对标准偏差在0.54%~3.65%之间。北京市农田回用再生水中检出了不同浓度水平的6种无机阴离子,其中Cl~-检出浓度最高,浓度范围为121~211 mg/L,不同位点的再生水中的浓度存在空间分布差异,NO_3~-和PO_4~(3-)在再生水出水口的浓度要高于再生水灌溉渠和农田灌溉点。结论本方法稳定性和分离效果好,准确率高,可适用于再生水中6种阴离子的监测。建议加强农田回用再生水的长期监测,并根据作物种类和土壤类型等制定更加严格的农田回用再生水水质标准。     

Bacterial growth in distribution systems: effect of assimilable organic carbon and biodegradable dissolved organic carbon

Two distribution systems, one treating water by ozonation and another treating water by nanofiltration in parallel with lime softening, were monitored for bacterial growth. Both systems kept disinfectant residuals such as chlorine and chloramine in their respective distribution systems. Bacterial growth was assessed by heterotrophic plate counts (HPC) on R2A agar. In the distribution systems fed by ozonated water, HPCs were correlated (R2 = 0.96) using an exponential model with the assimilable organic carbon (AOC) at each sampling site. Also, it was observed that ozonation caused a significant increase in the AOC concentration of the distribution system (over 100% increase) as well as a significant increase in the bacterial counts of the distribution system (average increase over 100%). The HPCs from the distribution systems fed by nanofiltration in parallel with lime-softening water also displayed an exponential correlation (R2 = 0.73) with an exponential model based on AOC. No significant correlation was found between bacteria growth on R2A agar and BDOC concentrations. Therefore, in agreement with previous work, bacterial growth in the distribution systems was found to correlate with AOC concentrations.     

Microbial quality of domestic and imported brands of bottled water in Trinidad

A cross-sectional study was conducted to determine the microbial quality of domestic and imported brands of bottled water available in Trinidad, purchased from six geographical regions in Trinidad, and representing the whole island. A sample size of 344 bottles of water was determined by using a precision rate of 2% and a Type 1 error of 5%. The membrane filter technique was used with cultures grown on m-Endo agar and m-FC agar for total coliforms and thermotolerant coliforms, respectively. Aerobic plate count (APC) was determined on nutrient agar; Pseudomonas aeruginosa was detected on MacConkey agar, Escherichia coli was isolated on eosin methylene blue (EMB) and Salmonella spp. was assayed by using standard methods.Of the 344 water samples tested, 262 (76.2%) and 82 (23.8%) were domestic and imported brands, respectively. Eighteen (5.2%) of the 344 samples contained coliforms with a mean count of 0.88+/-6.38 coliforms per 100 ml, while 5 (1.5%) samples contained E. coli. The prevalence of total coliforms in domestic brands of bottled water was 6.9% (18 of 262) as compared with 0.0% (0 of 82) detected in imported brands. The difference was statistically significant (p=0.004). Similarly, the prevalence of aerobic bacteria in domestic brands of bottled water (33.6%) was significantly higher (p=0.001) than was found in imported brands (14.8%). Twenty-six (7.6%) of the total samples of water contained Pseudomonas species, but all were negative for thermotolerant coliforms and Salmonella spp.It was concluded that based on the recommended zero tolerance for coliforms in potable water, 5% of bottled water sold in Trinidad could be considered unfit for human consumption.     

Enteric bacteriophages as potential fecal indicators in ground beef and poultry meat.

Recovery efficiencies of enteric bacteriophages (F+ RNA coliphages, somatic coliphages, and Salmonella phages) as alternative fecal indicators were determined from ground beef and chicken breast meat using amino acid eluants (glycine and threonine) and a complex eluant (3% beef extract). Levels of F+ RNA coliphages (MS2, GA, Qbeta, FI, and SP), the somatic coliphage phiX174, and three environmental isolates of Salmonella phages (isolated from raw sewage) were assayed using three respective hosts: Escherichia coli Famp, E. coli C, and Salmonella Typhimurium. When 8% polyethylene glycol and 0.1 M NaCl were used to precipitate bacteriophages eluted with five different eluants, the highest recoveries of the three phage groups were with 0.5 M threonine and 0.25 M glycine-threonine. The average recoveries of F+ RNA coliphages, somatic coliphages, and the Salmonella phages from ground beef and chicken meat were 100, 69, and 65%, respectively, with threonine (0.5 M, pH 9.0) as the eluate. Of eight market food samples tested, F+ RNA coliphages were detected in five (63%) and somatic coliphages were detected in seven (88%). The overall detection sensitivity of the method was 3 PFU/100 g of ground beef or chicken meat. Levels of bacteriophages and bacterial indicators on chicken carcass surfaces were determined at identified critical control points at a poultry plant. Through the processing steps of evisceration, washing, and chilling, the levels of F+ RNA coliphages and fecal coliforms were reduced by 1.6 and 1.9 log10 PFU or CFU/100 g, respectively. F+ RNA coliphages and perhaps other enteric bacteriophages may be effective candidate indicators for monitoring the microbiological quality of meat, poultry, and perhaps other foods during processing. The bacteriophage concentration method developed provides a simple, rapid, and practical tool for the evaluation of fecal contamination levels in ground beef and processed chicken meat.     

Surveillance of Enteric Viruses and Microbial Indicators in the Eastern Oysters (Crassostrea virginica) and Harvest Waters along Louisiana Gulf Coast

Noroviruses are the most common causative agent of viral gastroenteritis in humans, and are responsible for major foodborne illnesses in the United States. Filter‐feeding molluscan shellfish exposed to sewage‐contaminated waters bioaccumulate viruses, and if consumed raw, transmit the viruses to humans and cause illness. We investigated the occurrence of norovirus GI and GII and microbial indicators of fecal contamination in the eastern oysters (Crassostrea virginica) and water from commercial harvesting areas along the Louisiana Gulf Coast (January to November of 2013). Microbial indicators (aerobic plate count, enterococci, fecal coliforms, Escherichia coli, male‐specific coliphages, and somatic coliphages) were detected at the densities lower than public health concerns. Only one oyster sample was positive for norovirus GII at 3.5 ± 0.2 log₁₀ genomic equivalent copies/g digestive tissues. A stool specimen obtained from an infected individual associated with a norovirus outbreak and the suspected oysters (Cameron Parish, La., area 30, January 2013) were also analyzed. The norovirus strain in the stool belonged to GII.4 Sydney; however, the oysters were negative and could not be linked. In general, no temporal trend was observed in the microbial indicators. Low correlation among bacterial indicators was observed in oysters. Strongest correlations among microbial indicators were observed between enterococci and fecal coliforms (r = 0.63) and between enterococci and E. coli (r = 0.64) in water (P < 0.05); however, weak correlations were found in oysters (r < 0.45) and between oysters and harvest water (r ≤ 0.36, P > 0.05). Our results emphasize the need for regular monitoring of pathogenic viruses in commercial oyster harvesting areas to reduce the risks of viral gastroenteritis incidences.     

Monitoring the microbial populations and temperatures of fresh broccoli from harvest to retail display

Microbial populations and the temperature of fresh broccoli were monitored at several steps of a supply chain by sampling 33 distinct lots of locally grown produce over two seasons during harvest, storage, wholesale handling, and retail display. Imported broccoli was also sampled, but only at retail display. Microbiological analyses were conducted on the florets of 201 local and 60 imported broccoli samples to determine populations of total aerobic bacteria (aerobic colony count), fecal coliforms, Escherichia coli, and Listeria monocytogenes. All the samples had mean aerobic colony counts ranging between 4 and 6 log CFU/g, but L. monocytogenes was not detected (limit of detection =100 CFU/g). Fecal coliforms and E. coli (limit of detection =20 most probable number per 100 g) were found in 22 of 126 samples of local broccoli collected at various steps of the production and distribution system during the first season. None was found in 75 samples collected in the second season. Fecal coliforms and E. coli were found in 2 of 60 imported broccoli samples. Broccoli temperatures were relatively well controlled throughout the production and distribution system. No clear change in produce microbial populations was evident between harvest and retail display, during both sampling seasons. However, a large experimental variability was found, possibly associated with the high variability of the initial levels of microbial populations on broccoli at harvest.     

Bioaerosol emission rate and plume characteristics during land application of liquid class B biosolids

This study investigated bioaerosol emission rates and plume characteristics of bioaerosols generated during land application of liquid Class B biosolids. In addition, it compared the rate of aerosolization of coliphages and total coliform bacteria during land application of liquid Class B biosolids to the rate of aerosolization during land application of groundwater inoculated with similar concentrations of Escherichia coli and coliphage MS2. Air samples were taken immediately downwind of a spray applicator as it applied liquid (approximately 8% solids) biosolids to farmland near Tucson, Arizona. Air samples were also collected immediately downwind of groundwater seeded with MS2 and E. coli applied to land in an identical manner. Air samples, collected with liquid impingers, were taken in horizontal and vertical alignment with respect to the passing spray applicator. Vertical and horizontal sample arrays made it possible to calculate the flux of microorganisms through a virtual plane of air samplers, located 2 m downwind of the passing spray applicator. Neither coliphages nor coliform bacteria were detected in air downwind of spray application of liquid Class B biosolids. Based on limits of detection for the methodology, the rate of aerosolization during land application of liquid biosolids was calculated to be less than 33 plaque forming units (PFU) of coliphage and 10 colony forming units (CFU) of coliform bacteria per meter traveled by the spray applicator. The rate of aerosolization during land application of seeded groundwater was found to be, on average, 2.02 x 10(3) CFU E. coli and 3.86 x 10(3) PFU MS2 aerosolized per meter traveled by the spray applicator. This is greater aerosolization than was observed during land application of biosolids. Because concentrations of coliphages and coliforms were similar in the liquid biosolids and the seeded water, itwas concluded that some property of biosolids reduces aerosolization of microorganisms relative to groundwater. Additional experiments utilizing a novel air sampling protocol showed that the duration of bioaerosol exposure immediately (2 m) downwind of biosolids spray application is brief and the plume of bioaerosols generated is discrete. Additional air samples showed that aerosolization of coliphages and coliform bacteria after liquid biosolids have been applied to land does not occur at detectable levels.     

包装饮用水生产工艺中微生物风险分析

本研究在A、B、C、D四个市的包装饮用水产工艺中的微生物进行调查,获得微生物污染的基础数据,评价企业生产工艺中微生物污染控制系统的有效性。全面对四个工厂生产工艺过程中原水、过程水及成品水进行微生物检测,利用传统和分子方法对优势菌进行菌种鉴定。结果显示,33份样品霉菌和酵母计数、大肠菌群检出值均1 CFU/mL,砂滤水和碳滤水的菌落总数均有检出,且检出值较高。过程水中的产气荚膜梭菌均未检出,其中碳滤水有检出粪链球菌,反渗透(Reverses Osmosis,RO)水有检出铜绿假单胞菌。共分离到34株细菌,优势菌包括食酸菌属、假单胞菌属、分枝杆菌属和芽胞杆菌属。包装饮用水企业生产工艺对微生物的控制较为有效,微生物危害风险较低。议包装饮用水企业建立生产现场微生物识别系统,加强微生物监控,掌握生产过程中的微生物风险,为合理有效地控制生产工艺中微生物风险提供科学依据。     

Male-specific coliphages as an additional fecal contamination indicator for screening fresh carrots

The objective of this study was to evaluate the efficacy of male-specific (F+) coliphages as a fecal-contamination indicator for fresh carrots. The prevalence of specific pathogens and indicator organisms on the surface of carrots obtained from a farm, truck, and processing shed was studied. Twenty-five carrot samples collected from each of these locations were washed, and aliquots of the wash were analyzed for the presence of F+ coliphages, Escherichia coli, Salmonella, and Shigella. Additionally, the Salmonella isolates were genotyped using pulsed-field gel electrophoresis (PFGE). Our studies detected the presence of F+ coliphages, E. coli, and Salmonella on carrots. All samples, however, tested negative for Shigella. Although none of the carrot samples from the field were positive for E. coli, one sample was positive for Salmonella, and another was positive for F+ coliphages. From the truck, two carrot samples (8%) were positive for Salmonella, four (16%) were positive for F+ coliphages, and four (16%) were positive for E. coli. None of the carrot samples from the processing shed were positive for Salmonella. However, 2 carrot samples (8%) were positive for E. coli, and 14 carrot samples (56%) were positive for F+ coliphages. The PFGE results suggest that there were three distinct Salmonella genotypes among the carrot samples from the truck and that the Salmonella isolates identified on carrot samples from the field and truck locations were different. Microbiological screening of fresh produce such as carrots (which can be exposed to fecal contaminants in soils and water) should ensure the detection of both viral and bacterial contaminants. Overall, in this study, F+ coliphages were detected in 25% of the carrot samples, compared to E. coli (8%), Salmonella (4%), and Shigella (0%). The results suggest F+ coliphages can serve as a conservative indicator of fecally associated viruses on carrots. This suggests that in addition to E. coli screening, F+ coliphages should be included when produce such as carrots that are vulnerable to fecal contaminants are screened. Since the detection of specific enteric viral pathogens is expensive, screening for viral indicators of fecal contamination using F+ coliphages can be an economical approach to providing an additional level of assurance about the microbiological quality of fresh carrots.     

Phylogenetic analysis implicates birds as a source of Cryptosporidium spp. oocysts in agricultural watersheds

Cryptosporidium parvum and C. hominis are protozoan parasites responsible for cryptosporidiosis, an acute gastrointestinal illness that can be life-threatening for immunocompromised persons. Sources and genotypes of Cryptosporidium oocysts were investigated in two agricultural areas within the Wachusett Reservoir watershed, a drinking water source for Boston, Massachusetts. Two brooks (denoted Brook SF and Brook JF, respectively), each downgradient from a dairy farm, were chosen as sample sites. For one year, Brooks SF and JF were sampled monthly; oocysts were detected in 6 (50%) out of 12 samples from Brook JF, and no oocysts were detected in Brook SF. Oocyst genotypes from agricultural surface waters were compared to oocyst genotypes from Genbank, as well as fecal samples of cattle and birds, using phylogenetic analysis of a hypervariable region of the 18S rRNA gene by both neighbor-joining and parsimony methods. Results show extensive heterogeneity among Cryptosporidium spp. 18S rRNA sequences, and also suggest that birds are an oocyst source in this watershed. Principal components analysis showed oocyst presence correlating strongly with seasonal factors, and oocysts in surface waters were only detected in the summer through late fall, co-incident with the presence of migratory birds in this watershed. If birds are confirmed to be an important source of oocysts infectious to humans, the data suggest that protection of raw drinking water supplies in some agricultural areas may depend upon management and control of resident and migratory bird populations.