In 2006, a deadly Escherichia coli O157:H7 outbreak in bagged spinach was traced to California's Central Coast region, where >70% of the salad vegetables sold in the United States are produced. Although no definitive cause for the outbreak could be determined, wildlife was implicated as a disease vector. Growers were subsequently pressured to minimize the intrusion of wildlife onto their farm fields by removing surrounding noncrop vegetation. How vegetation removal actually affects foodborne pathogens remains unknown, however. We combined a fine-scale land use map with three datasets comprising ∼250,000 enterohemorrhagic E. coli (EHEC), generic E. coli, and Salmonella tests in produce, irrigation water, and rodents to quantify whether seminatural vegetation surrounding farmland is associated with foodborne pathogen prevalence in California's Central Coast region. We found that EHEC in fresh produce increased by more than an order of magnitude from 2007 to 2013, despite extensive vegetation clearing at farm field margins. Furthermore, although EHEC prevalence in produce was highest on farms near areas suitable for livestock grazing, we found no evidence of increased EHEC, generic E. coli, or Salmonella near nongrazed, seminatural areas. Rather, pathogen prevalence increased the most on farms where noncrop vegetation was removed, calling into question reforms that promote vegetation removal to improve food safety. These results suggest a path forward for comanaging fresh produce farms for food safety and environmental quality, as federal food safety reforms spread across ∼4.5 M acres of US farmland.agriculture | biodiversity | disease ecology | E. coli | foodborne pathogens

bRecent 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.

A year-long study was conducted to determine the fecal prevalence of Escherichia coli O157:H7 in three sheep ranches. Strain diversity and persistence were compared with multiple-locus variable-number tandem repeat analysis and pulsed-field gel electrophoresis. Ranch C, a feedlot, consisted of young sheep raised predominantly on a high-grain diet. The other two sites consisted of sheep raised on native pasture and a combination of native and irrigated pasture. Forty fecal samples were collected every month from each ranch. Samples were examined for E. coli O157:H7 by immunomagnetic separation and culture of the magnetic beads onto selective media. Detection of virulence markers in positive isolates was determined by PCR. E. coli O157:H7 was isolated from 100 (22.7%) of 440 fecal samples collected from ranch C. On ranch B, 9 (1.9%) of the 480 fecal samples were positive for the pathogen, while none of the samples from ranch A were positive. On ranch C, the odds of detecting E. coli O157:H7 was 3.2 times greater during the warmer months compared with the cooler months of the year. There was no association between days spent in the feedlot and fecal prevalence of the pathogen (P = 0.62). Most multiple-locus variable-number tandem repeat analysis types were isolated only once from ranch C (14 of 23), but several strains were isolated over 4 to 6 months, often in many intervening negative months. This study revealed that the prevalence of E. coli O157:H7 can be high in some sheep ranches in California, especially in feedlots where young sheep are fed predominantly high-grain rations.

In endemic African areas, such as Tanzania, Brucella spp. cause human febrile illnesses, which often go unrecognized and misdiagnosed, resulting in delayed diagnosis, underdiagnosis, and underreporting. Although rapid and affordable point-of-care tests, such as the Rose Bengal test (RBT), are available, acceptance and adoption of these tests at the national level are hindered by a lack of local diagnostic performance data. To address this need, evidence on the diagnostic performance of RBT as a human brucellosis point-of-care test was reviewed. The review was initially focused on studies conducted in Tanzania but was later extended to worldwide because few relevant studies from Tanzania were identified. Databases including Web of Science, Embase, MEDLINE, and World Health Organization Global Index Medicus were searched for studies assessing the diagnostic performance of RBT (sensitivity and specificity) for detection of human brucellosis, in comparison to the reference standard culture. Sixteen eligible studies were identified and reviewed following screening. The diagnostic sensitivity (DSe) and specificity (DSp) of RBT compared to culture as the gold standard were 87.5% and 100%, respectively, in studies that used suitable “true positive” and “true negative” patient comparison groups and were considered to be of high scientific quality. Diagnostic DSe and DSp of RBT compared to culture in studies that also used suitable “true positive” and “true negative” patient comparison groups but were considered to be of moderate scientific quality varied from 92.5% to 100% and 94.3 to 99.9%, respectively. The good diagnostic performance of RBT combined with its simplicity, quickness, and affordability makes RBT an ideal (or close to) stand-alone point-of-care test for early clinical diagnosis and management of human brucellosis and nonmalarial fevers in small and understaffed health facilities and laboratories in endemic areas in Africa and elsewhere.

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Background Many ecologically important plants are pollinated or have their seeds dispersed by fruit bats, including the widely distributed African straw-colored fruit bats ( Eidolon helvum ). Their ability to fly long distances makes them essential for connecting plant populations across fragmented landscapes. While bats have been implicated as a reservoir of infectious diseases, their role in disease transmission to humans is not well understood. In this pilot study, we tracked E. helvum to shed light on their movement patterns in Tanzania and possible contact with other species. Methods Tracking devices were deployed on 25 bats captured in the Morogoro Municipal and Kilombero District area near the Udzungwa Mountains of Tanzania. Nightly flight patterns, areas corresponding to foraging bouts and feeding roosts, and new day roosts were determined from bat movement data and characterized according to their proximity to urban built-up and protected areas. Sites for additional environmental surveillance using camera traps were identified via tracking data to determine species coming in contact with fruits discarded by bats. Results Tracking data revealed variability between individual bat movements and a fidelity to foraging areas. Bats were tracked from one to six nights, with a mean cumulative nightly flight distance of 26.14 km (min: 0.33, max: 97.57) based on data from high-resolution GPS tags. While the majority of their foraging locations were in or near urban areas, bats also foraged in protected areas, of which the Udzungwa Mountains National Park was the most frequented. Camera traps in fruit orchards frequented by tracked bats showed the presence of multiple species of wildlife, with vervet monkeys ( Chlorocebus pygerythrus ) observed as directly handling and eating fruit discarded by bats. Conclusions Because we observed multiple interactions of animals with fruits discarded by bats, specifically with vervet monkeys, the possibility of disease spillover risk exists via this indirect pathway. With flight distances of up to 97 km, however, the role of E. helvum in the seed dispersal of plants across both protected and urban built-up areas in Tanzania may be even more important, especially by helping connect increasingly fragmented landscapes during this Anthropocene epoch.

Deer mice ( Peromyscus maniculatus ) are abundant and widely distributed rodents in North America that occupy diverse habitats, including agricultural landscapes. Giardia and Cryptosporidium are common parasites in wildlife including deer mice, which may play a role in on-farm contamination of produce. An important step in assessing the risk of produce contamination by Cryptosporidium and Giardia shed by deer mice is to determine the prevalence, levels, and genotypes of (oo)cysts in mouse feces. A total of 63 (30.3%) and 53 (25.5%) of 208 deer mice trapped on 12 farms on the California Central Coast were positive for Cryptosporidium and Giardia, respectively. Of these mice, 41 (19.7%) contained both parasites. The odds of Cryptosporidium shedding were 2.5 to 5 times higher for mice trapped in autumn than for mice trapped in summer or spring. Female mice had a higher prevalence and two- to threefold higher levels of Cryptosporidium and Giardia compared with male mice. Female adults and female juveniles had the highest rates of contamination of the environment with Cryptosporidium and Giardia, respectively. We estimated that 20 infected deer mice inhabiting 1 ha of a typical leafy green produce farm in the study region could shed approximately 5.3 × 10 Cryptosporidium and 10.5 × 10 Giardia, respectively, per day into the environment. The small-subunit rRNA gene loci from a subset of protozoan isolates were sequenced and compared with existing sequences in GenBank. Multiple genotypes of Cryptosporidium and Giardia were found, and BLAST analyses suggest that Giardia and the majority of Cryptosporidium genotypes in deer mice circulate within various rodent populations, but some Cryptosporidium isolates possess zoonotic potential.

Infection with Borrelia miyamotoi in California, USA, has been suggested by serologic studies. We diagnosed B. miyamotoi infection in an immunocompromised man in California. Diagnosis was aided by plasma microbial cell–free DNA sequencing. We conclude that the infection was acquired in California.