Spatial scale effects in environmental risk-factor modelling for diseases.

Studies attempting to identify environmental risk factors for diseases can be seen to extract candidate variables from remotely sensed datasets, using a single buffer-zone surrounding locations from where disease status are recorded. A retrospective case-control study using canine leptospirosis data was conducted to verify the effects of changing buffer-zones (spatial extents) on the risk factors derived. The case-control study included 94 case dogs predominantly selected based on positive polymerase chain reaction (PCR) test for leptospires in urine, and 185 control dogs based on negative PCR. Land cover features from National Land Cover Dataset (NLCD) and Kansas Gap Analysis Program (KS GAP) around geocoded addresses of cases/controls were extracted using multiple buffers at every 500 m up to 5,000 m, and multivariable logistic models were used to estimate the risk of different land cover variables to dogs. The types and statistical significance of risk factors identified changed with an increase in spatial extent in both datasets. Leptospirosis status in dogs was significantly associated with developed high-intensity areas in models that used variables extracted from spatial extents of 500-2000 m, developed medium-intensity areas beyond 2,000 m and up to 3,000 m, and evergreen forests beyond 3,500 m and up to 5,000 m in individual models in the NLCD. Significant associations were seen in urban areas in models that used variables extracted from spatial extents of 500-2,500 m and forest/woodland areas beyond 2,500 m and up to 5,000 m in individual models in Kansas gap analysis programme datasets. The use of ad hoc spatial extents can be misleading or wrong, and the determination of an appropriate spatial extent is critical when extracting environmental variables for studies. Potential work-arounds for this problem are discussed.

Incidence and severity of Arcanobacterium pyogenes injection site abscesses with needle or needle-free injection.

Nursery-age pigs (n=198) were used to evaluate the difference in abscess formation at needle-free jet and conventional needle-and-syringe injection sites. Needle-free jet injection was used to administer injections in the neck and ham on one side of the animal whereas needle-and-syringe was used for neck and ham injections on the opposite side. Immediately prior to injection, the injection site surfaces were contaminated with an inoculum of Arcanobacterium pyogenes. Each pig was humanely euthanized 27 or 28 days after injections. Histopathological results showed that needle-free jet injection was associated with more abscesses than needle-and-syringe injection at both neck (P=0.0625) and ham (P=0.0313) injection sites. Out of 792 injection sites, only 13 abscesses were observed, with 12 of those present at needle-free jet injection sites. Needle-free jet injection may increase the occurrence of injection site abscesses that necessitate carcass trimming at pork processing plants.

Package systems and storage times serve as postlethality controls for Listeria monocytogenes on whole-muscle beef jerky and pork and beef smoked sausage sticks.

To validate how packaging and storage reduces Listeria monocytogenes on whole-muscle beef jerky and smoked pork and beef sausage sticks, four packaging systems (heat sealed [HS] without vacuum, heat sealed with oxygen scavenger, nitrogen flushed with oxygen scavenger [NFOS], and vacuum) and four ambient temperature storage times were evaluated. Commercially available whole-muscle beef jerky and smoked pork and beef sausage sticks were inoculated with a five-strain L. monocytogenes cocktail, packaged, and then stored at 25.5 °C until enumerated for L. monocytogenes at 0, 24, 48, and 72 h and 30 days after packaging. The interaction of packaging and storage time affected L. monocytogenes reduction on jerky, but not on sausage sticks. A >2-log CFU/cm(2) reduction was achieved on sausage sticks after 24 h of storage, regardless of package type, while jerky had <2-log reductions for all packaging types. At 48 h, log reductions were similar (P. 0.05) for all types of jerky packaging, ranging from 1.26 to 1.72 log CFU/cm(2); however, at 72 h, mean L. monocytogenes reductions were >2 log CFU/cm(2), except for NFOS (1.22-log CFU/cm(2) reduction). Processors could package beef jerky in HS packages with oxygen scavenger or vacuum in conjunction with a 24-h holding time as an antimicrobial process to ensure a >1-log CFU/cm(2) L. monocytogenes reduction or use a 48-h holding time for HS- or NFOS-packaged beef jerky. A >3-log CFU/cm(2) mean reduction was observed for all beef jerky and sausage stick packaging systems after 30 days of 25.5 °C storage.

Evaluation of two thermal processing schedules at low relative humidity for elimination of Escherichia coli O157:H7 and Salmonella serovars in chopped and formed beef jerky.

Foodborne outbreaks have been linked to jerky produced under insufficient thermal processing schedules. Reduction of Escherichia coli O157:H7 and Salmonella serovars during thermal processing of chopped and formed beef jerky was evaluated under two processing schedules representative of those used by large-scale (LS) and small-scale (SS) jerky production facilities. Fresh chopped and formed all-beef jerky batter was inoculated with 5.8 to 7.3 log CFU of E. coli O157:H7 or Salmonella per g, extruded into strips, and thermally processed by LS or SS schedules. A >or=5.0-log CFU/g reduction of both pathogens occurred with <10% relative humidity and a cumulative process of 44 min at 55.6 degrees C followed by 46 min at 77.8 degrees C into the LS schedule. Additional drying at 77.8 degrees C for 3.5 h was needed to achieve a water activity of 0.67 and a moisture-to-protein ratio (MPR) of 0.77. For the SS process, a >or=5.0-log CFU/g reduction of both pathogens occurred with 15 to 20% relative humidity and a cumulative process of 45 min at 52 degrees C, 60 min at 57 degrees C, 45 min at 60 degrees C, 45 min at 63 degrees C, 90 min at 68 degrees C, and finishing with 30 min at 77 degrees C. After processing for an additional 90 min at 77 degrees C, water activity was 0.60 while the MPR was 0.82. The LS and SS processes for producing chopped and formed jerky provided >or=5.0 log lethality to control E. coli O157:H7 and Salmonella. However, both processes would require additional drying to achieve an MPR of 0.75 to be labeled as jerky.

Ultrastructural and fluorochromatic changes of Anaplasma marginale exposed to oxytetracycline, imidocarb and enrofloxacin in short-term erythrocyte cultures.

Anaplasma marginale causes mild to severe hemoparasitic disease resulting in significant morbidity and mortality in cattle worldwide. In the absence of universally efficacious vaccines, antimicrobial therapy combined with biocontainment and biosecurity strategies are critical to control anaplasmosis. Herein, we compared the effect of oxytetracycline, imidocarb and enrofloxacin on A. marginale isolates in short-term erythrocyte cultures. Electron micrographs detailing antimicrobial-induced changes in rickettsial morphology were scored (0-4) based on ultrastructural changes. These were compared to fluorochromatic changes detected by flow cytometry (FACS) using conversion of hydroethidine (HE) to ethidium bromide (EB) by living organisms to assess viability. A. marginale infectivity in selected cultures was confirmed by subinoculation into susceptible calves. Morphology scores were analyzed using Chi-squared tests and compared to FACS data by ANOVA with isolate, drug and concentration as co-variates in the model. Only the Virginia and Oklahoma isolates exposed to 1.0 microg /ml imidocarb and the Oklahoma isolate exposed to 4.0 microg /ml enrofloxacin were sterilized following antimicrobial exposure. Rickettsia with morphology scores of 0 had significantly more EB positive cells than inclusions with morphology scores of 4 (p=0.039). There was also a significant association between ultrastructural changes and infectivity (p=0.0047). Furthermore, the percent EB positive cells in the antimicrobial exposed cultures was highly predictive of the probability of infectivity (p=0.0026). This is the first study describing ultrastructural changes in A. marginale following exposure to enrofloxacin and imidocarb. These findings demonstrate that FACS and electron microscopy are useful tools to screen new antimicrobials for the use in anaplasmosis chemotherapy.