Persistence of F-Specific RNA Coliphages in Surface Waters from a Produce Production Region along the Central Coast of California

F+ RNA coliphages (FRNA) are used to source-track fecal contamination and as surrogates for enteric pathogen persistence in the environment. However, the environmental persistence of FRNA is not clearly understood and necessitates the evaluation of the survival of prototype and environmental isolates of FRNA representing all four genogroups in surface waters from the central coast of California. Water temperature played a significant role in persistence–all prototype and environmental strains survived significantly longer at 10°C compared to 25°C. Similarly, the availability of host bacterium was found to be critical in FRNA survival. In the absence of E. coli F_amp, all prototypes of FRNA disappeared rapidly with a D-value (days for one log reduction) of <1.2 d from water samples incubated at 25°C; the longest surviving prototype was SP. However, in the presence of the host, the order of persistence at 25°C was QB>MS2>SP>GA and at 10°C it was QB = MS2>GA>SP. Significant differences in survival were observed between prototypes and environmental isolates of FRNA. While most environmental isolates disappeared rapidly at 25°C and in the absence of the host, members of genogroups GIII and GI persisted longer with the host compared to members of GII and GIV. Consequentially, FRNA based source tracking methods can be used to detect phages from recent fecal contamination along with those that persist longer in the environment as a result of cooler temperatures and increased host presence.     

Fecal Shedding of Zoonotic Food-Borne Pathogens by Wild Rodents in a Major Agricultural Region of the Central California Coast

Recent outbreaks of food-borne illness associated with the consumption of produce have increased concern over wildlife reservoirs of food-borne pathogens. Wild rodents are ubiquitous, and those living close to agricultural farms may pose a food safety risk should they shed zoonotic microorganisms in their feces near or on agricultural commodities. Fecal samples from wild rodents trapped on 13 agricultural farms (9 produce, 3 cow-calf operations, and 1 beef cattle feedlot) in Monterey and San Benito Counties, CA, were screened to determine the prevalence and risk factors for shedding of several food-borne pathogens. Deer mice (Peromyscus maniculatus) were the most abundant rodent species trapped (72.5%). Cryptosporidium species (26.0%) and Giardia species (24.2%) were the predominant isolates from rodent feces, followed by Salmonella enterica serovars (2.9%) and Escherichia coli O157:H7 (0.2%). Rodent trap success was significantly associated with detection of Salmonella in rodent feces, while farm type was associated with fecal shedding of Cryptosporidium and Giardia. Seasonal shedding patterns were evident, with rodents trapped during the spring and summer months being significantly less likely to be shedding Cryptosporidium oocysts than those trapped during autumn. Higher rodent species diversity tended to correlate with lower fecal microbial prevalence, and most spatiotemporal pathogen clusters involved deer mice. Rodents in the study area posed a minimal risk as environmental reservoirs of E. coli O157:H7, but they may play a role in environmental dissemination of Salmonella and protozoa. Rodent control efforts that potentially reduce biodiversity may increase pathogen shedding, possibly through promotion of intraspecific microbial transmission.     

Diversity of Salmonella Isolates from Central Florida Surface Waters

Identification of Salmonella serotypes is important for understanding the environmental diversity of the genus Salmonella. This study evaluates the diversity of Salmonella isolates recovered from 165 of 202 Central Florida surface water samples and investigates whether the serotype of the environmental Salmonella isolates can be predicted by a previously published multiplex PCR assay (S. Kim, J. G. Frye, J. Hu, P. J. Fedorka-Cray, R. Gautom, and D. S. Boyle, J. Clin. Microbiol. 44:3608–3615, 2006, http://dx.doi.org/10.1128/JCM.00701-06). Multiplex PCR was performed on 562 Salmonella isolates (as many as 36 isolates per water sample) to predict serotypes. Kauffmann-White serogrouping was used to confirm multiplex PCR pattern groupings before isolates were serotyped, analyzed by pulsed-field gel electrophoresis, and assayed for antimicrobial susceptibility. In 41.2% of the Salmonella-positive water samples, all Salmonella isolates had identical multiplex PCR patterns; in the remaining 58.8%, two or more multiplex PCR patterns were identified. Within each sample, isolates with matching multiplex PCR patterns had matching serogroups. The multiplex patterns of 495 isolates (88.1%) did not match any previously reported pattern. The remaining 68 isolates matched reported patterns but did not match the serotypes for those patterns. The use of the multiplex PCR allowed the number of isolates requiring further analysis to be reduced to 223. Thirty-three Salmonella enterica serotypes were identified; the most frequent included serotypes Muenchen, Rubislaw, Anatum, Gaminara, and IV_50:z4,z23:−. A majority (141/223) of Salmonella isolates clustered into one genotypic group. Salmonella isolates in Central Florida surface waters are serotypically, genotypically, and phenotypically (in terms of antimicrobial susceptibility) diverse. While isolates could be grouped as different or potentially the same using multiplex PCR, the multiplex PCR pattern did not predict the Salmonella serotype.     

BMI Compared With Central Obesity Indicators as a Predictor of Diabetes Incidence in Mauritius

The aim of the study was to compare BMI with waist circumference (WC), waist‐to‐hip ratio (WHR), and waist‐to‐stature ratio (WSR) as a predictor of diabetes incidence. A total of 1,841 men and 2,104 women of Mauritian Indian and Mauritian Creole ethnicity, aged 25–74 years, free of diabetes, hypertension, cardiovascular disease, and gout were seen at baseline in 1987 or 1992, and follow‐up in 1992 and/or 1998. At all time points, participants underwent a 2 h 75 g oral glucose tolerance test. Hazard ratios for diabetes incidence were estimated applying an interval‐censored survival analysis using age as timescale. Six hundred and twenty‐eight individuals developed diabetes during the follow‐up period. Multivariable adjusted hazard ratios for diabetes incidence corresponding to a 1 s.d. increase in baseline BMI, WC, WHR, and WSR for Mauritian Indians were 1.49 (1.31–1.71), 1.58 (1.38–1.81), 1.54 (1.37–1.72), and 1.61 (1.41–1.84) in men and 1.33 (1.17–1.51), 1.35 (1.19–1.53), 1.39 (1.24–1.55), and 1.38 (1.21–1.57) in women, respectively; and for Mauritian Creoles they were 1.86 (1.51–2.30), 2.07 (1.68–2.56), 1.92 (1.62–2.26), and 2.17 (1.76–2.69) in men and 1.29 (1.06–1.55), 1.27 (1.04–1.55), 1.24 (1.04–1.48), and 1.27 (1.04–1.55) in women. Paired homogeneity tests showed that there was no difference between BMI and each of the central obesity indicators (all P > 0.05). The relation of BMI with the development of diabetes was as strong as that for indicators of central obesity in this study population.     

Health Risk Assessment of Urban Surface Waters Based on Real-Time PCR Detection of Typical Pathogens

Typical enteric pathogens including enteroviruses, Salmonella typhi, Shigella spp., and Eschierichia coli were selected and monitored during a 1-year period in urban surface waters using the real-time polymerase chain reaction (PCR) method. By considering two routes of human exposure to urban surface waters (i.e., drinking water and involuntary intake), and supposing that the dose–response relation may follow either an exponential model or the Beta-Poisson model, health risk assessment was conducted to estimate the safety under a given acceptable risk level upon exposure to each water and to evaluate the required level of pathogen inactivation for safeguarding human health. As a result, it was found that human health risk due to enteroviruses is often greater than that due to bacterial pathogens, and greater removal of enteroviruses would be required for safeguarding at the same acceptable risk level.     

Salmonellae as an Index of Pollution of Surface Waters

Screening enrichments of surface water specimens by means of a polyvalent fluorescent antibody reagent for the salmonellae yielded approximately 60% more positive specimens than was obtained by cultural procedures. It is not known what fraction of the excess of fluorescent antibody-positive over culturally positive specimens represents staining of non-salmonellae or non-arizonae as opposed to the staining of non-cultivatable organisms of these two genera. Cotton gauze and rayon-polypropylene fiber swabs were equally sensitive for collecting salmonellae from the streams examined. Tetrathionate enrichment incubated at 41.5 C appeared to be superior to selenite-cystine for isolation of salmonellae from surface waters. Twenty-eight serotypes of Salmonella and two serotypes of Arizona were identified in the 121 positive specimens. In water rated moderately polluted, 65% of all specimens tested were positive; in minimally polluted waters, 38% were positive; and in unpolluted streams, 44% were positive.     

New Species of Lankesteria (Apicomplexa,Eugregarinida) from Ascidians on the Central California Coast1

The following species of the gregarine genus Lankesteria were found in a study of ascidians from Monterey Bay, on the central California coast of the Pacific Ocean: L. abbotti n. sp. in Clavelina huntsmani, L. aplidii n. sp. in Aplidium solidum, L. ascidiae in Ciona intestinalis, L. diaphanis n. sp. in Archidistoma diaphanes, L. euherdmaniae n. sp. in Euherdmania claviformis, L. montereyensis n. sp. in Archidistoma molle, L. pescaderoensis n. sp. in Ritterella rubra, L. pittendrighi n. sp. in Ascidia ceratodes, L. psammii n. sp. in Archidistoma psammion, L. ritterellae n. sp. in Ritterella pulchra, L. ritterii n. sp. in Archidistoma ritteri, L. synoici n. sp. in Synoicum parfustis, and Lankesteria sp. in Botrylloides sp. No gregarines were seen in the following ascidians: Aplidium californicum, A. propinquum, Botryllus tuberatus, Diplosoma macdonaldi, Perophora annectens, and Polyclinum planum.     

The Presence of Clostridium botulinum in Indonesian Waters

S ummary . In initial surveys of Indonesian waters and marine animals, Clostridium botulinum , predominantly of types A and C, was detected in 10% of all enrichment cultures tested.     

hypD as a Marker for [NiFe]-Hydrogenases in Microbial Communities of Surface Waters

Hydrogen is an important trace gas in the atmosphere. Soil microorganisms are known to be an important part of the biogeochemical H₂ cycle, contributing 80 to 90% of the annual hydrogen uptake. Different aquatic ecosystems act as either sources or sinks of hydrogen, but the contribution of their microbial communities is unknown. [NiFe]-hydrogenases are the best candidates for hydrogen turnover in these environments since they are able to cope with oxygen. As they lack sufficiently conserved sequence motifs, reliable markers for these enzymes are missing, and consequently, little is known about their environmental distribution. We analyzed the essential maturation genes of [NiFe]-hydrogenases, including their frequency of horizontal gene transfer, and found hypD to be an applicable marker for the detection of the different known hydrogenase groups. Investigation of two freshwater lakes showed that [NiFe]-hydrogenases occur in many prokaryotic orders. We found that the respective hypD genes cooccur with oxygen-tolerant [NiFe]-hydrogenases (groups 1 and 5) mainly of Actinobacteria, Acidobacteria, and Burkholderiales; cyanobacterial uptake hydrogenases (group 2a) of cyanobacteria; H₂-sensing hydrogenases (group 2b) of Burkholderiales, Rhizobiales, and Rhodobacterales; and two groups of multimeric soluble hydrogenases (groups 3b and 3d) of Legionellales and cyanobacteria. These findings support and expand a previous analysis of metagenomic data (M. Barz et al., PLoS One 5:e13846, 2010, http://dx.doi.org/10.1371/journal.pone.0013846) and further identify [NiFe]-hydrogenases that could be involved in hydrogen cycling in aquatic surface waters.     

Distribution and Diversity of Pathogenic Leptospira Species in Peri-domestic Surface Waters from South Central Chile

BackgroundLeptospirosis is a neglected zoonosis affecting animals and humans caused by infection with Leptospira. The bacteria can survive outside of hosts for long periods of time in soil and water. While identification of Leptospira species from human cases and animal reservoirs are increasingly reported, little is known about the diversity of pathogenic Leptospira species in the environment and how surveillance of the environment might be used for monitoring and controlling disease.ConclusionsThis study reports the presence of pathogenic Leptospira in the peri-domestic environment of households in three community types and the differences in Leptospira diversity at the community level. Systematic environmental surveillance of Leptospira can be used for detecting changes in pathogen diversity and to identify and monitor contaminated areas where an increased risk of human infection exists.     

Methanobrevibacter ruminantium as an Indicator of Domesticated-Ruminant Fecal Pollution in Surface Waters

A PCR-based assay (Mrnif) targeting the nifH gene of Methanobrevibacter ruminantium was developed to detect fecal pollution from domesticated ruminants in environmental water samples. The assay produced the expected amplification product only when the reaction mixture contained DNA extracted from M. ruminantium culture, bovine (80%), sheep (100%), and goat (75%) feces, and water samples from a bovine waste lagoon (100%) and a creek contaminated with bovine lagoon waste (100%). The assay appears to be specific and sensitive and can distinguish between domesticated- and nondomesticated-ruminant fecal pollution in environmental samples.     

Evaluating the Performance of Chemical Control in the Presence of Resistant Pathogens

Resistance to chemical control is a major impediment to combating many socially and economically important diseases. Theoretical and experimental studies have shown that reducing the intensity of treatment can slow, or even prevent, the invasion of resistance, yet reducing treatment levels also results in a net increase in disease severity. Clearly there is a need to identify control strategies that balance the conflicting aims of resistance management and disease suppression. Using a mathematical model for the dynamics of fungicide resistance in crop pathogens, we present a broadly applicable measure of the performance of chemical control in the presence of resistant pathogen strains. We illustrate how to optimise fungicide performance with respect to the intensity of treatment as a function of the duration of treatment and the fitness of the resistant strain. We find that in the short term, fungicide performance is optimised at high levels of treatment despite rapid selection for resistance, while the long-term optimum performance is achieved when treatment renders the fungicide-sensitive and fungicide-resistant pathogens equally fit. We further present evidence that under prescribed conditions, the ratio of dose size and frequency, and the fungicide mode of action, can have a significant effect on fungicide performance.     

The Eco-epidemiology of Pacific Coast Tick Fever in California

Rickettsia philipii (type strain “Rickettsia 364D”), the etiologic agent of Pacific Coast tick fever (PCTF), is transmitted to people by the Pacific Coast tick, Dermacentor occidentalis. Following the first confirmed human case of PCTF in 2008, 13 additional human cases have been reported in California, more than half of which were pediatric cases. The most common features of PCTF are the presence of at least one necrotic lesion known as an eschar (100%), fever (85%), and headache (79%); four case-patients required hospitalization and four had multiple eschars. Findings presented here implicate the nymphal or larval stages of D. occidentalis as the primary vectors of R. philipii to people. Peak transmission risk from ticks to people occurs in late summer. Rickettsia philipii DNA was detected in D. occidentalis ticks from 15 of 37 California counties. Similarly, non-pathogenic Rickettsia rhipicephali DNA was detected in D. occidentalis in 29 of 38 counties with an average prevalence of 12.0% in adult ticks. In total, 5,601 ticks tested from 2009 through 2015 yielded an overall R. philipii infection prevalence of 2.1% in adults, 0.9% in nymphs and a minimum infection prevalence of 0.4% in larval pools. Although most human cases of PCTF have been reported from northern California, acarological surveillance suggests that R. philipii may occur throughout the distribution range of D. occidentalis.     

Occurrence of Klebisiella pneumoniae in Surface Waters

The occurrence of Klebsiella pneumoniae in surface waters was not found to be ubiquitous. When it was isolated, Escherichia coli could also be found. The fecal coliform to fecal streptococci ratio suggest that its origin could be human animal, or mixed sources.     

Diversity and Transport of Microorganisms in Intertidal Sands of the California Coast

Forced by tides and waves, large volumes of seawater are flushed through the beach daily. Organic material and nutrients in seawater are remineralized and cycled as they pass through the beach. Microorganisms are responsible for most of the biogeochemical cycling in the beach; however, few studies have characterized their diversity in intertidal sands, and little work has characterized the extent to which microbes are transported between different compartments of the beach. The present study uses next-generation massively parallel sequencing to characterize the microbial community present at 49 beaches along the coast of California. In addition, we characterize the transport of microorganisms within intertidal sands using laboratory column experiments. We identified extensive diversity in the beach sands. Nearly 1,000 unique taxa were identified in sands from 10 or more unique beaches, suggesting the existence of a group of “cosmopolitan” sand microorganisms. A biogeographical analysis identified a taxon-distance relationship among the beaches. In addition, sands with similar grain size, organic carbon content, exposed to a similar wave climate, and having the same degree of anthropogenic influence tended to have similar microbial communities. Column experiments identified microbes readily mobilized by seawater infiltrating through unsaturated intertidal sands. The ease with which microbes were mobilized suggests that intertidal sands may represent a reservoir of bacteria that seed the beach aquifer where they may partake in biogeochemical cycling.     

The Rise of Pathogens: Predation as a Factor Driving the Evolution of Human Pathogens in the Environment

Bacteria in the environment must survive predation from bacteriophage, heterotrophic protists, and predatory bacteria. This selective pressure has resulted in the evolution of a variety of defense mechanisms, which can also function as virulence factors. Here we discuss the potential dual function of some of the mechanisms, which protect against heterotrophic protists, and how predation pressure leads to the evolution of pathogenicity. This is in accordance with the coincidental evolution hypothesis, which suggests that virulence factors arose as a response to other selective pressures, for example, predation rather than for virulence per se. In this review we discuss some of those environmental factors that may be associated with the rise of pathogens in the marine environment. In particular, we will discuss the role of heterotrophic protists in the evolution of virulence factors in marine bacteria. Finally, we will discuss the implications for expansion of current pathogens and emergence of new pathogens.