PRELIMINARY REPORT: Epidemiologic Analysis of HPAI-Affected Turkey Flocks. May 1, 2015

Transcription

1 United States Department of Agriculture Animal and Plant Health Inspection Service Veterinary Services PRELIMINARY REPORT: Epidemiologic Analysis of HPAI-Affected Turkey Flocks May 1, 2015

2 I. Descriptive analysis of epidemiologic investigation of infected flocks Project Background The purpose of the survey and analysis is to assess potential pathways of initial introduction of highly pathogenic avian influenza (HPAI) viruses onto commercial poultry operations and potential transmission routes of HPAI viruses from infected premises to non-infected premises. The survey was designed as an assessment tool to provide an in-depth review of the current biosecurity and management practices and exposure risks on an infected farm. Project Status Data collection and analyses continue. This document represents an initial preliminary descriptive analysis. Methods A survey instrument (Appendix I) is being administered by State and Federal animal health officials in areas affected by HPAI strain H5N2. Survey administrators are requesting that respondents be individual(s) most familiar with the farm s management and operations. Instructions request responses be provided for the two week period prior to HPAI detection. Investigators have been asked to complete the investigation within one week of detection. Additionally, for each survey completed for an infected barn/farm, at least one non-infected barn/farm within the same complex or as near as possible to the infected flock should be completed. Completed questionnaires are sent via to USDA-APHIS-Veterinary Services (VS). Analytical epidemiologists located at VS, Center for Epidemiology and Animal Health (CEAH), Fort Collins, Colorado, are responsible for questionnaire review, data entry, and analysis. The questionnaire (Appendix I) includes both closed- and open-ended questions focused on the following categories: premises description, farm biosecurity, farm help/workers, farm equipment, litter handling, dead bird disposal, farm visitors, and wild birds. Additionally, respondents have been asked to provide mortality data (charted over the duration since placement of turkeys in a barn), a copy of the most recent biosecurity audit or assessment if available, and a farm diagram. Preliminary Findings Preliminary findings for questionnaires analyzed to date are given in Table 1. All three control farms had fewer than five barns compared to half of case farms. All three control farms (versus half of case farms) had three or more employees. Although all farms used footbaths, two of three control farms used a dry type. This was rarely used on case farms. All three control farms (and only 39% of case farms) used fly control. All three case farms disposed of dead birds via burial pit. None had an uncovered rendering bin. Wild birds were generally seen less often on and around control farms. 2

3 Table 1 - Percent case farms, control farms, and control barns on case farms by characteristics Premises characteristic Level Case farm N=23 Control farm N=3 Control barn N=10 Number of barns Total number of employees < Foot pans Yes, in use Footbath type Dry Fly control Dead bird disposal Burial pit Render Yes, no bin cover Yes, bin cover not routinely closed Yes, bin cover routinely closed No rendering Wild bird characteristic Level Case farm N=23 Control farm N=3 Control barn N=10 Wild birds around farm Waterfowl Gulls Small perching Other water birds Other birds Any Birds year round Seasonality Bird location Away from facilities On farm, not in barns On farm, in barns Any In addition to directly responding to survey questions, all respondents included written statements to elaborate on or to qualify a response, and most questionnaires included a written narrative. Several themes have emerged while reviewing these written responses. Many respondents indicated that they have changed farm activities and protocols (i.e., improvements in biosecurity) over the last four to six weeks as more farms have been confirmed as HPAI positive in the United States. Frequently, comments have been included with details of wild bird observations on or around the farm. These comments include the frequency of bird sightings in and around the houses and on the premises as well as the types of birds being observed. Several respondents described the level of security of carcass bins in relation to the presence of scavenger species. Finally, several respondents mentioned wind conditions around the time of presumed infection. 3

4 Copies of mortality data were requested for case and control barns/farms. Figure 1 shows the high variability in daily flock mortality (number of dead birds/ 1000) for six barns with confirmed or presumed HPAI infection. Mortality rates are displayed over the period from placement to the date of disease confirmation. Figure 1. Mortality rates (number of dead birds per 1,000) since placement for six barns with presumed or confirmed HPAI infection. 4

5 Interpretation and Limitations These results are preliminary, and several limitations should be recognized. The numbers of infected and non-infected farms available for the initial descriptive analysis presented in this document were small. Additionally, 10 of the 13 controls were non-infected barns on infected farms and 3 controls were non-infected farms (i.e., non-infected farm with similar characteristics located near an infected flock). Many of the analysis variables were collected at the farm level (e.g., other animals located on the farm, premises biosecurity, dead bird disposal), and therefore will be the same for both infected and noninfected farms/barns. CEAH analysts will continue to review, enter and analyze surveys and update results regularly. Variables (Appendix II) will continue to be evaluated in more detail as more data become available, and results could represent areas of focus for a more rigorous follow-up study to evaluate risk factors for virus introduction and transmission. Information collected in the written responses and mortality data will also continue to be evaluated in conjunction with the analysis for purposes of hypothesis generation and to inform the next steps of the investigation. II. Comparison of General Wind Direction and Direction of HPAI Spread in One Cluster of HPAI in Minnesota Project Background This portion of the spatial analysis investigates the hypothesis that HPAI (EA/AM-H5N2) in MN is spread by air. To test this hypothesis we compared a directional analysis of positive premises in one cluster of positive HPAI premises in MN using Cluster Seer software with a generalized compass rose based on weather stations in the area. The results suggest very little alignment of general wind direction to disease spread direction although the data and methods used were very limited. Data and Methods - Generalized Wind Rose The generalized wind rose was developed based on wind direction and speed from the four weather stations found in Stearns, Meeker, and Kandiyohi counties, Minnesota. We chose to group wind direction for the four stations to get a view of how wind behaves across the area of interest used in the analysis. Combining would also reduce localized variations that could affect the directional analysis across the larger area of infections. Dates used to create the generalized wind rose were March 23, 2015 through April 2. These data are collected through the Automated Surface Observing System (ASOS). The data used were downloaded from the Iowa Environmental Mesonet website: 5

6 Figure 1. Location of weather stations used to create wind rose and resulting wind rose integrating data from all four stations. Data and Methods - ClusterSeer Analysis ClusterSeer [1] is a software package developed for spatio-temporal analysis of disease. Within ClusterSeer we used the direction method to evaluate the direction of disease spread in one area of clustered HPAI cases in Minnesota. The Direction Method tests for a space-time interaction and calculates the average direction of disease spread. A relative model was used, which connects each case 6

7 to all subsequent cases. This method was chosen since each positive case had the potential to infect all subsequent cases throughout the period of time for the cluster (approximately 3 weeks). The null hypothesis is that cases following (in a temporal sense) a given case are located in a random direction. The alternative hypothesis is that subsequent cases are located in a specific direction. ClusterSeer provides the following results: a significance test for the above hypothesis, the average direction of disease spread, and a measure of the variance in the angles between connected cases. Case data for the ClusterSeer analysis were extracted from the APHIS EMRS (Emergency Management Response System) and imported into ArcGIS software. The spatial locations of all confirmed positive premise were validated using geocoding and aerial imagery interpretation to ensure accuracy of the locations using ArcGIS software. Next, we identified a cluster of 35 cases in Kandiyohi, Stearns, and Meeker counties. The start date of the premise status represents the date premises were confirmed positive by NVSL and these dates were used for ClusterSeer analysis. The selected set of 35 cases were exported from ArcGIS as a text file and then prepared for input to ClusterSeer. Results Based on the ClusterSeer directional test, subsequent cases typically occurred in the southwest direction ( degrees) to previous cases. The analytic results were statistically significant (p = 0.001), and the results were weakly consistent (ClusterSeer concentration value of 0.35, with 0 being randomly spread and 1.0 being strongly consistent in directional spread.) The generalized wind rose shows wind direction during this time window to be predominantly in the west-northwest direction but highly variable throughout the period. Based on this comparison, the two do not match and suggest a simple wind movement of infection based on predominant wind direction during this time window does not explain the spread of Avian Influenza in this cluster of positive cases in Minnesota. 7

8 Figure 2. Positive premises used in ClusterSeer analysis and direction of spread as reported by ClusterSeer. Limitations and Future Work The evidence suggests that there are likely multiple routes of disease spread for HPAI. Possible routes of disease spread include direct and indirect contacts between premises, such as movement of trucks, feed, people, and equipment. Movement of wild birds carrying HPAI can spread the virus to new areas and interactions between wild and domestic birds can cause infection. This analysis does not account for these methods of disease spread. The potential for HPAI to be spread by air is dependent on the period of viral shedding and the distance that HPAI can travel on dust particles and survive in the atmosphere. Detailed information on the survival characteristics of EA/AM-H5N2 HPAI may not be available at this time. The generalize approach to measuring wind direction over the entire period of a cluster of cases used here makes it difficult to identify a predominant wind direction. A large scale case by case analysis of disease spread and wind patterns using commonly employed plume models would enable a shorter time period of wind data to be used and highlight predominant wind directions. The large scale case by case analysis would also enable more accurate temporal modeling of virus shedding and periods of 8

9 infectivity. This approach has been used by other researchers to evaluate wind-borne spread of HPAI between farms [2]. Future work for HPAI geospatial analysis will investigate corn fields proximate to premises as a potential epidemiologic factor for infection, as well as further investigating ice out timing and clustering along highways 71, 12, and 4. Sustained wind events will be investigated for associations with spikes in cases 5-7 days after the wind events to see if sustained winds of a certain velocity are an epidemiologic factor for HPAI spread. Depopulation events will be investigated as a potential factor for HPAI spread. Wind breaks will be investigated as a protective effect for uninfected barns versus infected barns. References: Ssematimba A, Hagenaars TJ, de Jong MCM (2012) Modelling the Wind-Borne Spread of Highly Pathogenic Avian Influenza Virus between Farms. PLoS ONE 7(2): e doi: /journal.pone III. Initial Examination of Geospatial Risk Factors for Infection with HPAI in Minnesota Methods The premises location and species of birds present on poultry farms in Minnesota were obtained from the Minnesota Poultry Laboratory. Latitude, distance to the nearest water body, distance to the nearest river, distance to the nearest of three highways (71, 12, and 4), and the amount of surface water located within 0.5, 1, or 3 km were calculated and examined as possible risk factors for infection with HPAI. Factors which were unconditionally associated with the outcome at p value<0.20 were selected for further analysis. Multivariable ordinal logistic regression models were constructed using a forward stepwise selection approach. Variables with a p value<0. 05 based on Z-statistics or Wald tests for factors with multiple levels were considered significant and retained in the model. Results Being a turkey operation, latitude of the premises, and nearness to one of three highways were identified as risk factors. Controlling for latitude and distance to the nearest highway, the odds of being an infected flock are 6.4 times greater in turkey operations. Additionally, a band between the latitudes of and degrees (Figure 1, green band) was identified as being an area of higher risk (odds ratio 2.1). Being a farm located above this latitude range confers a protective effect. Distance to nearest highway (p=0.077) was forced into the model in order to compare predicted probabilities of infection. For turkey operations located in the high risk latitude band and within 30 km of any of the examined highways, predicted risk of infection was 17.6%. This decreased to 8% for turkey operations located greater than 30km from the highways but still in the high risk band. For turkey operations located outside of the high risk latitude band and greater than 30 km from the examined highways, the predicted probability of infection decreases to 2%. Being within 30 km of an examined highway outside of the high risk latitude band only slightly increases the predicted risk of infection to 4-5% depending on latitude. The final model explains approximately 17% of the variation between infected and non-infected farms. 9

10 Interpretation Results suggest that geography is playing a role in disease risk. This analysis identified broad categories of risk factors using readily available data, which may be used to help guide future analysis. Consistent with the outbreak dynamics seen so far in Minnesota, being a turkey operation was the most significant risk factor identified. The risk of infection also increased within a distinct band of latitude in southern Minnesota, where the majority of cases have appeared. Since highway 12 runs east to west across this band, farms located close to this highway may be at increased risk. The effect of being near a highway decreases outside of the high risk latitude band. A follow up analysis of farms just within the high risk band may yield additional clues to the effect of highway proximity. Additional data collected from surveys of cases and controls may also provide greater insight into the factors driving this effect. Figure 1. Latitude bands identified as having decreased odds of becoming a case, the non-shaded band at the bottom is the comparison group, while the upper three bands represent areas of decreased risk. 10

11 Limitations The geographic factors examined are most likely proxies for underlying risk factors and so represent only a first look at possible risk factors. Further data collection and analysis will aid in identifying underlying factors driving the effects identified in this analysis. Further work is needed to examine the fit of the model, and the identification of additional infected farms could change the effect of factors identified. Because the model used is nonlinear, the magnitude of the change in the infection probability that is associated with a given change in one of the risk factors depends on the levels of all of the risk factors examined. Adding additional risk factors to the analysis could change how these risk factors affect the probability of infection occurring. IV. HPAI Air Sampling Project (University of Minnesota in collaboration with USDA, APHIS, Veterinary Services) Project background This project was designed to investigate the ability to detect HPAI viruses in the air of poultry barns and in their immediate environments. This project was designed as a proof of concept and further investigative work is required. Methods Three turkey flocks were chosen to participate in the study. Flock 1 The flock was a turkey layer confirmed positive on Monday 20 of April, and had 4 barns (about 6,000 to 7,000 birds/each). Barn 2 is where infection started and had an estimated mortality of 70-80% on the day of sampling. Infection in farm 1 (April 24) was suspected since there was elevated mortality (~20 birds) the morning of sampling. There was no suspicion of HPAI in barns 3 and 4. Air samples were collected in the interior of barn 1, interior barn 2 and outside of the barn about 3 yards from the exhaust fan. Flock 2 The flock was a turkey grow-out confirmed positive on April 25, and had 6 barns. Two of the barns (barn 3 and 7) had between 30 to 40% mortality on the day of sampling (April 27). Only barn 3 was sampled. There were samples collected inside, at the exhaust point and at ~ 150 yards from the barn. Air samples were collected from 150 yards, at the exhaust point outside the barn and inside the barn (in this order). Flock 3 The flock was a male breeder confirmed positive on April 26 and samples were collected on April 28. Clinical signs of HPAI were only observed in one barn. The Affected barn had about 30% mortality. Air samples were collected at 150 yards downwind from barn, at the exhaust point outside the barn and inside the barn (in this order). Only one type of air samplers was used. Air samples were collected using a cyclonic air sampler (Microtek, Midwest) for 30 min. Estimated air sampler capacity is 200 l/min. Results using this air sampler are shown in Tables 1, 2. Samples were tested using the matrix PCR against influenza and if positives, they were re-tested using the specific H5 and H7 PCR. 11

12 Results Tables 1 and 2 and 3 show results obtained using the air cyclonic collector. This air sampler does not discriminate based on particle size. Table 1. Summary of results from air samples obtained by matrix RT-PCR No.samples (%) Positive 10 37% Suspect 11 41% Negative 6 22% Total % Table 2. Summary of results from air samples by sampling location obtained by matrix RT-PCR Inside Exhaust 150 yards Positive 5 (42%) 5 (56%) 0 (0%) Suspect 4 (33%) 4 (44%) 3 (50%) Negative 3 (25%) 0 (0%) 3 (50%) Total 12 (100%) 9 (100%) 6 (100%) Conclusion Influenza virus genetic material could be detected in air samples collected inside and outside of poultry facilities with birds acutely infected with HPAI. Viral genetic material was found in samples collected with the three air samplers. The quantity of virus present in the air could only be semi-quantified and it was not considered very high (only Ct values above 30 were obtained). Samples positive or suspect were obtained inside and outside at the exhaust point, and only suspect results were obtained at 150 yards. In conclusion, we showed that HPAI can be aerosolized from infected facilities. However, the implications of these findings in terms of understanding the transmission of HPAI between flocks needs further investigation. Future work needs to include sampling of more flocks, detection at various distances from infected premises and the assessment of environmental contamination outside infected facilities. 12

13 Appendix I. Questionnaire. Animal and Plant Health Inspection Service Veterinary Services HPAI Investigation Questionnaire INSTRUCTIONS The purposes of these investigations are to assess potential pathways of initial introduction of HPAI viruses onto commercial poultry operations and potential lateral transmission routes of HPAI viruses from infected premises to noninfected premises. Following confirmation of an HPAI virus introduction into a commercial flock, an investigation should be initiated as soon as possible, no later than 1 week following detection. The investigator(s) assigned should be integrated into other response activities but their primary focus is on completion of the introduction investigation. The investigation form provided is a guide for conducting a systematic and standardized assessment of potential pathways of initial virus movement onto the farm and potential movement of the virus off the farm. All sections of the form should be completed through direct conversation with the individual(s) most familiar with the farm s management and operations and questions are to be answered for the period 2 weeks prior to the detection of HPAI. Where applicable, direct observation of the biosecurity or management practice asked about should be conducted. This is not a box-checking exercise but an indepth review of the current biosecurity and management practices and exposure risks on an affected farm. For example, direct observation of the farm employee donning and doffing procedures and compliance with company biosecurity practices is more important than checking the box on the form indicating that workers wear coveralls into the poultry houses. Investigators are encouraged to take notes and include them with the investigation form when completed. An investigation form should be completed for the infected house or farm and at least one noninfected house or farm within the same complex as near as possible to the index infected flock. 13

14 Date: date Interviewer name/organization: intrname Interviewee name/organization: intename A. PREMISES INFORMATION Farm name: frmname Farm address: frmadd Farm (premises) ID: frmid County: frmcty Township: frmtshp Range: frmrng Section: frmsec Is facility enrolled in NPIP?... npip 1 Yes 3 No B. PREMISES CONTACT INFORMATION 1. Contact name: h201 Phone: h202 Cell phone: h203 h Contact name: h205 Phone: h206 Cell phone: h207 h Contact name: h209 Phone: h210 Cell phone: h211 h Flock Veterinarian: h213 Phone: h214 Cell phone: h215 h216 14

15 C. PREMISES DESCRIPTION 1. Poultry type: 1 Broiler 2 Layer 3 Turkey 4 Other (specify: ) h301/h301oth 2. Production type: 1 Meat 2 Egg 3 Breeding 4 Other (specify: ) h302/h302oth 3. Age: 1 Multiple age 2 Single age h Sex: 1 Hen 2 Tom 3 Both h Flock size:... h305 # birds 6. Facility type: [Check all that apply] Brood Grow Other (specify: ) h308oth Both brooder & grower houses are present on the same premises Breeder Commercial 7. If brooder and grower houses are present on the same premises, are there multiple stages of management (brooding and growing), in the same house?. h312 h306 h307 h308 h309 h310 h311 1 Yes 3 No 8. a. Farm capacity... h313 # birds b. Number of barns... h314 # barns c. Barn capacity... h315 # birds 9. What is the primary barn type/ventilation: [Check one only.] h316 1 Curtain sided 2 Environmental control 3 Side doors 4 Other (specify: ) h316oth 10. Are cool cell pads used?... h317 1 Yes 3 No If Yes, what is the source of water for these pads? h Distance in yards of closest body of water near farm:... h319 yd 15

16 12. Water body type: [Check all that apply.] Pond Lake Stream River Other (specify: ) h324oth h320 h321 h322 h323 h What other types of animals are present on the farm? a. Beef cattle... h325 1 Yes 3 No b. Dairy cattle... h326 1 Yes 3 No c. Horses... h327 1 Yes 3 No d. Sheep... h328 1 Yes 3 No e. Goats... h329 1 Yes 3 No f. Pigs... h330 1 Yes 3 No g. Dogs... h331 1 Yes 3 No h. Cats... h332 1 Yes 3 No i. Poultry or domesticated waterfowl... h333 1 Yes 3 No j. Other (specify: ) h334oth... h334 1 Yes 3 No 14. What is the primary water source for poultry? [Check one only.] h335 1 Municipal 2 Well 3 Surface water (e.g., pond) 4 Other (specify: ) h335oth 15. Is water treated prior to delivery to poultry?... h336 1 Yes 3 No If Yes, how is it treated and with what? h337 16

17 D. FARM BIOSECURITY 1. Is there a house with a family living in it on the property?... h401 1 Yes 3 No 2. Is there a common drive entrance to farm and residence?... h402 1 Yes 3 No 3. Do you have signage of no admittance or biosecure area on this property? h403 1 Yes 3 No 4. Is there a gate to this farm entrance?... h404 1 Yes 3 No 5. Is the gate secured/locked?... h405 1 Yes 3 No If Yes, what hours is it secured? h Is the farm area fenced in?... h407 1 Yes 3 No 7. How frequently is vegetation mowed/bush hogged on the premises?... h408 times/month 8. Is facility free of debris/clutter/trash piles?... h409 1 Yes 3 No 9. Is there a wash station/spray area available for vehicles?... h410 1 Yes 3 No If Yes, what disinfectant is used? 10. Is there a designated parking area for workers and visitors away from the barns/pens?... h412 h411 1 Yes 3 No 11. Is there a changing area for workers?... h413 1 Yes 3 No Do they shower?... h Do workers don dedicated laundered coveralls before entering each house on the premises?... h415 1 Yes 3 No 1 Yes 3 No 13. Do worker wear rubber boots or boot covers in poultry houses?... h416 1 Yes 3 No 14. Are the barn/pen doors lockable?... h417 1 Yes 3 No Are they routinely locked?... h418 1 Yes 3 No 15. Are foot pans available at barn/pen entrances?... h419 1 Yes 3 No Are they in use?... h420 1 Yes 3 No 16. Are foot baths dry (powdered or particulate disinfectant)?... h421 1 Yes 3 No 17. Are foot baths liquid disinfectant?... h422 1 Yes 3 No 17

18 18. Frequency foot pan solutions are changed?... h423 times/month What disinfectant is used? h Is there an entry area in the barns/pens before entering the bird area?... h425 1 Yes 3 No 20. What pest and wildlife control measures are used on this farm? a. Rat and mouse bait stations... h426 1 Yes 3 No b. Bait stations checked at least every 6 weeks... h427 1 Yes 3 No c. Fly control used... h428 1 Yes 3 No If Yes, type and frequency: d. Houses are bird proof... h430 1 Yes 3 No e. Wild birds seen in house... h431 1 Yes 3 No If Yes, type, number and frequency: f. Raccoons, possums, foxes seen in or around poultry houses... h433 1 Yes 3 No g. Wild turkeys, pheasants, quail seen around poultry... h434 1 Yes 3 No 21. Are biosecurity audits or assessments (company or third party) conducted on this farm?... h435 If Yes, when was the last audit or assessment conducted? (Obtain a copy of the result of the audit or assessment if available.) h429 h432 1 Yes 3 No 22. Has this farm been confirmed positive for HPAI?... h437 1 Yes 3 No h436 E. FARM HELP/WORKERS 1. Total number of persons working on farm... h501 # 2. Number of workers living on the farm premises who are: a. Family... h502 # b. Nonfamily... h503 # 3. Workers are assigned to: [Check one only.] h504 1 Entire farm 2 Specific barns/areas 4. Do the workers have a common break area?... h505 1 Yes 3 No If Yes, location: h506 18

19 5. Are workers employed by other poultry operations?... h507 1 Yes 3 No 6. How often are training sessions held on biosecurity for workers?... h508 times/year 7. Are family members employed by other poultry operations or processing plants? h509 1 Yes 3 No If Yes, poultry operation or processing plant: h Do part-time/weekend help and other extended family members on holidays and vacations?... h Are workers (full & part-time) restricted from being in contact with backyard poultry?... h512 How is this communicated? 1 Yes 3 No 1 Yes 3 No h513 F. FARM EQUIPMENT Is the equipment used on this premises farm specific, under joint ownership that remains on this premises, or under joint ownership and used on other farm premises? A list of equipment follows. 1. Company vehicles/trailers: Farm specific?... h601 If No, by whom is equipment jointly used: Dates: 2. Feed trucks (excess feed): Farm specific?... h604 If No, by whom is equipment jointly used: Dates: 3. Gates/panels: Farm specific?... h607 If No, by whom is equipment jointly used: Dates: 4. Lawn mowers: Farm specific?... h610 If No, by whom is equipment jointly used: 1 Yes 3 No h602 h603 1 Yes 3 No h605 h606 1 Yes 3 No h608 h609 1 Yes 3 No h611 19

20 Dates: h Live haul loaders: Farm specific?... h613 1 Yes 3 No If No, by whom is equipment jointly used: Dates: 6. Poult trailers: Farm specific?... h616 1 Yes 3 No If No, by whom is equipment jointly used: Dates: 7. Pre-loaders: Farm specific?... h619 1 Yes 3 No If No, by whom is equipment jointly used: Dates: Describe pre-loader cleaning and disinfection procedures: h614 h615 h617 h618 h620 h621 h Pressure sprayers/washers: Farm specific?... h623 1 Yes 3 No If No, by whom is equipment jointly used: Dates: 9. Skid-steer loaders: Farm specific?... h626 1 Yes 3 No If No, by whom is equipment jointly used: Dates: 10. Tillers: Farm specific?... h629 1 Yes 3 No If No, by whom is equipment jointly used: Dates: h624 h625 h627 h628 h630 h631 20

21 11. Trucks: Farm specific?... h632 1 Yes 3 No If No, by whom is equipment jointly used: Dates: h633 h Other equipment: h635 Farm specific?... h636 1 Yes 3 No If No, by whom is equipment jointly used: Dates: h637 h638 G. LITTER HANDLING 1. Litter type: h Supplier/source: h Is a litter shed present?... h703 1 Yes 3 No 4. Do you do partial cleanouts?... h704 1 Yes 3 No If Yes, give dates of last partial cleanout: h Date of last cleanout:... h706 date Frequency of cleanout:... h707 times/month 6. Who does the cleanout? [Check one only.] h708 1 Grower 2 Contractor If contractor, name and location h Litter is disposed of: [Check one only.] h710 1 On farm 2 Taken off site If taken offsite, name and location: h711 21

22 H. DEAD BIRD DISPOSAL 1. Approximate normal daily mortality... h801 # birds 2. How is daily mortality handled? [Check all that apply.] On-farm: Burial pit/incinerator/composted/other (specify: ) h802oth Off-farm: Landfill/rendering/other (specify: ) h803oth Off-farm disposal performed by: Owner/employee/other (specify: ) h804oth h802 h803 h804 If burial or compost pits are used, are carcasses covered with soil on a daily basis?... h805 1 Yes 3 No 3. Contact name of company or individual responsible for disposal: h806 If rendering is used, include location of carcass bin on the farm map. 4. What is the pickup schedule? h Does the carcass bin have a cover?... h808 1 Yes 3 No Is it routinely kept closed?... h809 1 Yes 3 No I. FARM VISITORS 1. How many visitors do you have on a daily basis?... h901 # 2. Is there a visitor log to sign in?... h902 1 Yes 3 No Is it current?... h903 1 Yes 3 No 3. Do you provide any outer clothing to visitors entering the farm?... h904 1 Yes 3 No If Yes, identify items of clothing provided: 4. Mark the following services that were on the farm when this flock was on the farm. List date of service and name of person (or contract company) and if they had contact with the birds. Service Dates Name Contact? Service person h906 Yes No h907 h908 h909 1 Yes 3 No Vaccination crew h910 Yes No h911 h912 h913 1 Yes 3 No h905 22

23 Service Dates Name Contact? Moving crew (moving from brood to grow, or pullet house to layer house) h914 Yes No h915 h916 h917 1 Yes 3 No Processing plant load out h918 Yes No h919 h920 h921 1 Yes 3 No Load-out crew (positive flock) h922 Yes No h923 h924 h925 1 Yes 3 No If load-out took more than one night, was returning crew the same crew?... h926 1 Yes 3 No Truck #/# s Trailer #/# s What plant did flock go to? Load-out crew (flock previous to positive flock) h930 Yes No h931 h932 h933 1 Yes 3 No If load-out took more than one night, was returning crew the same crew?... h934 1 Yes 3 No Truck #/# s h935 Trailer #/# s h936 What plant did flock go to? h937 Poult delivery h938 Yes No h939 h940 h941 1 Yes 3 No Rendering pickup h942 Yes No h943 h944 h945 1 Yes 3 No Litter services h946 Yes No h947 h948 h949 1 Yes 3 No Cleanout services h950 Yes No h951 h952 h953 1 Yes 3 No Equipment shared/rented/loaned/borrowed (each of the categories of visitor is likely to be accompanied by equipment of some sort or another) h954 Yes No h955 h956 h957 1 Yes 3 No Feed delivery h958 Yes No h959 h960 h961 1 Yes 3 No 5. Who makes sure covers are closed after delivery? h Are feed covers kept closed?... h963 1 Yes 3 No J. WILD BIRDS 1. Do you see wild birds around your farm?... h Yes 3 No h927 h928 h929 If Yes, what type of birds? [Check all that apply.] Waterfowl Gulls Small perching birds (sparrows, starlings, swallows) Other water birds (egrets, cormorants) Other h1006oth h1002 h1003 h1004 h1005 h Do you see birds all year round?... h Yes 3 No 23

24 If Yes, what type of birds? h Is there seasonality to the presence of some types of birds?... h Yes 3 No If Yes, what type of birds and what seasons do you see them? h Where are wild birds seen in relation to the farm? [Check all that apply.] On adjacent habitats away from facilities and equipment (identify location of habitat on photos) On the farm but not in the barns (identify facilities or equipment birds have contact with) On the farm and sometimes in the barns (identify facilities or equipment birds have contact with) h1011 h1012 h

25 K. NARRATIVE/COMMENTS h

26 FARM DIAGRAM -Attach a download from satellite imagery if possible. In addition, draw a simple schematic map of the farm site centering with the poultry houses/pens. Identify where the HPAI positive flocks were housed. Also include: fan banks on houses, residence, driveways, public roads, bodies of water, feed tanks, gas tanks, out buildings, waster dumpsters, electric meters, dead bird disposal, parking areas, other poultry sites. Digital photographs, if allowed, are excellent supporting documentation. North 26

27 Appendix II. Variables continuing to be evaluated from the survey. Premises variables of interest being evaluated. Premises Characteristic Production type Age Sex Flock size Facility type Brooder & grower same house Farm capacity Number of barns Ventilation Cool cell pads Water source for cool cell pads Closest body of water (yards) Water body type Other animals Level Meat Egg Breeding Other Multiple age Single age Hen Tom Both <20,000 birds 20,000 + Brood Grow Other Both Breeder Commercial <50,000 birds 50,000 + Curtain sided Environ. Control Side doors Other NA < 350 yards Pond Lake Stream River Other Beef cattle Dairy cattle Horses Sheep 27

28 Water source Water treated Goats Pigs Dogs Cats Poultry or domestic waterfowl Other Municipal Well Surface Other Farm biosecurity variables of interest being evaluated. Biosecurity Characteristics House with family on property Signage Gate to farm entrance Farm area fenced in Freq. vegetation mowed (per month) < Facility free of debris/trash Wash/spray area for vehicles Designated parking workers/visitors Changing area for workers Dedicated coveralls Rubber boots or boot covers Barn doors locked Foot pans Footbath type Level Yes, common drive Yes, no common drive No Yes, locked Yes, not locked No Yes, shower Yes, no shower No Yes, Routinely locked Yes, not routinely lock No Yes, in use Yes, not in use No Dry Liquid None 28

29 Ante area Rodent bait station Yes, checked every 6 weeks Yes, checked less freq. No Fly control Houses bird proof Wild birds in house Raccoons, possums, foxes Wild turkeys, pheasants, quail Biosecurity audits Farm help/worker variables of interest being evaluated. Employee Characteristics Total number < 3 3+ Any nonfamily Worker assigned to: Level Workers employed by other poultry op. Biosecurity training sessions per year 0 1+ Family members employed by other poultry op. Part-time/weekend help Restrict contact with backyard poultry Entire farm Specific barn/area Farm equipment variables of interest being evaluated. Equipment Characteristic Farm specific Level Company trucks Feed trucks Gates/panels Lawn mowers Live haul loaders Poultry trailers Pre-loaders Pressure sprayer/washer Skid-steer loader 29

30 Tillers Trucks Other Litter handling variables of interest being evaluated. Litter Characteristic Type Shed Partial cleanouts Who does cleanout Litter disposal on-farm Level Wood shavings Other Grower Contractor Dead bird disposal variables of interest being evaluated. Disposal Characteristic Dead bird disposal Render Level On-farm, covered with soil On-farm, not covered with soil Off-farm Yes, no bin cover Yes, bin cover not routinely closed Yes, bin cover routinely closed No rendering Farm visitor variables of interest being evaluated. Visitor Characteristic Level Number of daily visitors All =0 Visitor log Outer clothing provided Visitor: Service person Vaccination crew Moving crew Yes, bird contact Yes, no bird contact No Yes, bird contact Yes, no bird contact No Yes, bird contact Yes, no bird contact No 30

31 Poult delivery Rendering pickup Litter service Cleanout service Equipment shared Feed delivery Feed covers kept closed Yes, bird contact Yes, no bird contact No Yes, bird contact Yes, no bird contact No Yes, bird contact Yes, no bird contact No Yes, bird contact Yes, no bird contact No Yes, bird contact Yes, no bird contact No Yes, bird contact Yes, no bird contact No Wild bird variables of interest being evaluated. Wild Bird Characteristic Wild birds around farm Birds year round Seasonality Bird location Level Waterfowl Gulls Small perching Other water birds Other birds Any Away from facilities On farm, not in barns On farm, in barns 31