Overview of ARS On-farm Research · S. cubana T10 S. cubana T8 S. mbandaka T38 S. knatum T39 S. mbandaka T36 S. mbandaka T37 S. havana T33 S. havana T34 S. havana T35 S. newport T18

Overview of ARS OnOverview of ARS On--farm farm ResearchResearch

J. Allen Byrd

Food Animal Protection Research LaboratoryFood Animal Protection Research LaboratoryUSDAUSDA--ARSARS

College Station, Texas, USACollege Station, Texas, USA

Production Cost

Selling Price

Consumer Demands

Animal Welfare

Environmental Regulations

Food Safety

21st Century Decision Tree

Food Animal

Agriculture

Preharvest Intervention Preharvest Intervention StrategiesStrategies

VaccinationVaccinationChemical or Biological Litter Treatment Chemical or Biological Litter Treatment Competitive Exclusion CultureCompetitive Exclusion CultureMedicationMedicationFeed WithdrawalFeed Withdrawal

Areas of ConcernAreas of Concern

Feed millFeed mill

BreedersBreeders

HatcheryHatchery

Growout Growout

Feed millFeed mill

Pelleting temperaturesPelleting temperaturesPest control Pest control Housekeeping Housekeeping SanitationSanitationLittle (if any) research is performed by Little (if any) research is performed by ARS ARS

BreedersBreeders

Pest controlPest controlHouse SanitationHouse SanitationEgg Room ManagementEgg Room ManagementWater QualityWater QualityDocumentationDocumentationGeneral Practices General Practices Research by ARS Research by ARS ––Kogut, FFSRUKogut, FFSRU

Background & HypothesisBackground & HypothesisCurrent selection Current selection Single pathogen resistanceSingle pathogen resistance

The hostsThe hosts’’ response in the earliest stage is critical in response in the earliest stage is critical in determining resistance or susceptibility.determining resistance or susceptibility.

The first week postThe first week post--hatch, poultry are highly hatch, poultry are highly susceptible to pathogens common in poultry facilities.susceptible to pathogens common in poultry facilities.

Our HypothesisOur Hypothesis•• Innate response drives acquired responseInnate response drives acquired response•• BroadBroad--spectrum resistancespectrum resistance•• Improve existing linesImprove existing lines

Dr. Kogut’s group, Avian Immunology

Summary of Lines A & BSummary of Lines A & B

In vitro In vitro heterophil functionheterophil functionA > BA > B

In vivo In vivo challenges challenges ((SalmonellaSalmonella & & EnterococcusEnterococcus))A more resistant > BA more resistant > B

Genomic profile (innate immunity) Genomic profile (innate immunity)

A > BA > B

Dr. Kogut’s group, Avian Immunology

Significance & ImpactSignificance & Impact1.1. Identified potential selection parametersIdentified potential selection parameters

Line levelLine levelChicks from a single damChicks from a single damIndividual adult breedersIndividual adult breeders

2.2. Select for high responders in lowerSelect for high responders in lower--responding lines responding lines

3.3. Development of new immunologically Development of new immunologically competent lines competent lines

4.4. Improve valuable existing linesImprove valuable existing lines

Dr. Kogut’s group, Avian Immunology

Where are we now?

parental lines improve line B

screen progeny

in vitroHΦ function identify

high/low sires

innate immunity profile

in vivochallenges

screen line B sires

Dr. Kogut’s group, Avian Immunology

HatcheryHatchery

SanitationSanitationPest ControlPest ControlBiosecurityBiosecurityWater ControlWater ControlChick Box Pads ManagementChick Box Pads ManagementHatchery Cabinet InterventionsHatchery Cabinet InterventionsARS ResearchARS Research-- Athens GroupsAthens Groups

GrowoutGrowout

Pest Control Pest Control House SanitationHouse SanitationFarm BiosecurityFarm BiosecurityWater Quality/TreatmentsWater Quality/TreatmentsARS Research ARS Research ––Fayetteville, Athens, & Fayetteville, Athens, & College StationCollege Station

Poultry Production and Product Safety Poultry Production and Product Safety Research Unit, Fayetteville, ARResearch Unit, Fayetteville, AR

Salmonella Salmonella ResearchResearch--

Identify mechanisms by which Identify mechanisms by which stressors suppress immunity to stressors suppress immunity to pathogens in turkeyspathogens in turkeys

Determine if joint and bone Determine if joint and bone infections are an overlooked infections are an overlooked source of product and source of product and processing plant contaminationprocessing plant contamination

Development of alternative approaches to antibiotics for controlling bacterial pathogens in poultry

Salmonella Salmonella ResearchResearch--

Bacteriophage therapyBacteriophage therapylaboratory studies demonstrate reduction of laboratory studies demonstrate reduction of SalmonellaSalmonella post harvestpost harvest

ProbioticsProbioticsin vitroin vitro selection for selection for in vivoin vivo competition against competition against pathogenspathogens-- discovery licensed, 100 million turkeys discovery licensed, 100 million turkeys and chickens treated internationally last yearand chickens treated internationally last year

Soil AmendmentsSoil AmendmentsAlum (aluminum sulfate) 10 patents related to improving Alum (aluminum sulfate) 10 patents related to improving poultry productionpoultry production

Recovery of Recovery of SalmonellaSalmonella from Commercial turkey from Commercial turkey Carcasses rinsed with Control or 72 Bacteriophage Carcasses rinsed with Control or 72 Bacteriophage

Isolates Amplified in SE Host or Field IsolateIsolates Amplified in SE Host or Field Isolate

0102030405060708090

100

Control SE S14 SE + S14

20/30

3/30

a

b

Perc

ent %

2/305/30

bb

Salmonella host used to propagate Bacteriophagea-b Different superscripts indicate significant (p<0.05) differences in incidence.

Impact of Alum ResearchApproximately 600 million chickens are grown using alum/year

Ammonia reduction and pathogen control are the primary reasons poultry producers are using alum although the initial goal was controlling P runoff.

Saves poultry industry $5-10 million per year

NR

NO3- NO2

-

ClO3 ClO2

Intracellular

Extracellular

UQ

1010

1011

1012MPN controlMPN chlorate

Rum

en

Cec

um

Rec

tum

Tota

l Cul

tura

ble

Ana

erob

es(M

PN/g

inte

stin

al c

onte

nts)

Chlorate effects in continuous Chlorate effects in continuous cultureculture

pHpHA:P ratioA:P ratioTotal culturable Total culturable anaerobic bacteriaanaerobic bacteriaTotal VFATotal VFANHNH33 productionproductionCHCH44 productionproductionAll unchangedAll unchanged1

100

104

106

108

1010

1012

0 50 100 150 200

Con

cent

ratio

n C

FU/m

l

Time (h)

Beneficial anaerobes

E.coli O157:H7

Pure culture Mixed fecal culture

10

1000

105

107

109

1011

10 mM Chlorate0 mM Chlorate

0 50 100 150 200

933

CFU

ml-1

a

E. c

oli

Time (h)

1

100

104

106

108

1010

1012

0 50 100 150 200

MPN/10 mM ChlorateMPN/0 mM Chlorate

M

PN m

l-1 T

otal

cul

tura

ble

anae

robe

sTime (h)

b

-1 o

rE.

col

iE.

Col

i93

3 C

FU m

l-1or

(Continuous culture)Time (h) Time (h)E. C

oli O

157:

H7

933

CFU

ml-1

10 mM Chlorate

Experimental Chlorate Product Experimental Chlorate Product (ECP)(ECP)

DayDay--ofof--hatch Broiler chickshatch Broiler chicks–– 1x ECP (48 hour)1x ECP (48 hour)

Reduced cecal Salmonella 99.99% (32,400 Reduced cecal Salmonella 99.99% (32,400 2)2)

Preharvest BroilersPreharvest Broilers–– 1x ECP (48 hour)1x ECP (48 hour)

Reduced crop Salmonella 95% (36.7% Reduced crop Salmonella 95% (36.7% 1.7%)1.7%)Reduced cecal Salmonella 40% (53% Reduced cecal Salmonella 40% (53% 31.7%)31.7%)Reduced cecal E.coli 1 Log (6 Reduced cecal E.coli 1 Log (6 5)5)

Byrd et al, 2002

Evaluation of CE and Evaluation of CE and ECPECP on Nonon Non--Challenged Broiler ChicksChallenged Broiler Chicks --

CONTACTSCONTACTS

Exp. Groups (+/total) Log 10 ST

Control 33/39 (84.6%)A 3.94 (8710)A

CE 13/39 (33%)B 1.31 (20)B

ECP – 1x 7/40 (15%)B 0.42 (3)C

ECP- 1x + CE 1/40 (2.5%)C 0.07 (1)C

Effect of 38 Hour Experimental Chlorate Product (ECP) Treatment Provided Prior to Feed Withdrawal on Salmonella Ceca Colonization in Commercial Turkeys

Treatment Log10Salmonella

Salmonella(Pos/Total)

Control 14/40 (35%)A

4/40 (10%)B

0/40 (0%)B

3x ECP 0.14 + 0.11B 3/40 (7.5%)B

1x ECP

2x ECP

0.95 + 0.31A

0.27 + 0.19B

0.00 + 0.00B

Twelve Hour Feed Withdrawal, Ten Hour Water Withdrawal *P< 0.05

Evaluation of Evaluation of ECPECP as a Feed Supplement for 7 as a Feed Supplement for 7 days on days on SalmonellaSalmonella ceca colonization in broiler ceca colonization in broiler

Treatment Salmonella Pos per total

Log10Salmonella/mL

Control 14/20 (70%)a 2.27 + 1.69ab (186)

0.3% ECP 11/20 (55%)a 1.76 + 1.73b (57)

3.1% ECP 13/20 (65%)a 2.15 + 1.73ab (141)

11.1% ECP 4/20 (20%)b 0.47 + 1.01c (3)

6.2% ECP 5/20 (25%)b 0.63 + 1.17c (4)

**Significantly different from control at P< 0.05

Residue in Broiler TissueResidue in Broiler Tissue

David J. Smith, Fargo, ND, 2006

Salmonella SerotypingSalmonella Serotyping

Conventional methods to identify Salmonella serotypes are time consuming, labor intensive, and expensive.

We are examining the utility of 16S/23S ribosomal DNA spacer region polymorphisms (Bakshi et al., 2002) to identify Salmonella serotypes among poultry isolates.

S. kentucky T66 S. kentucky T67 S. kentucky T26 S. kentucky T27 S. albany T43 S. soerenga T51 S. soerenga T52 S. 6,8,1,2 monophasic S. bredeney T48 S. hadar B97 S. 3,10,1 monophasic S. unknown T54 S. kentucky B94 S. untypable T64 S. senftenberg T11 S. senftenberg T12 S. cubana T9 S. cubana T10 S. cubana T8 S. mbandaka T38 S. knatum T39 S. mbandaka T36 S. mbandaka T37 S. havana T33 S. havana T34 S. havana T35 S. newport T18 S. tennesseeT20 S. cerroT6 S. barranquilla T4 S. senftenberg T13 S. oranienburg T42 S. typhimurium K75 S. anatum T1 S. anatum T2 S. anatum T3 S. dublin T29 S. copenhagen T31 S. quakumT40 S montevideo T14 S. montevideo T16 S montevideo T15 S. cerro T30 S. barranquilla T5 S. copenhagen T32 S. tennessee T21 S. tennessee T22 S. oranienburg T41 S. newport T17 S. newport T19 S. meleagridis T23 S. cerroT7 S. lillie K88 S. beita K89 S. copenhagen T28 S. heidelberg B91 S. senftenberg B93 S. hadar K83 S. heidelberg K81 S. heidelberg K82 S. infantis K76 S. braenderup K77 S. 4,5,12:i monophasi S. 4,5,12:i monophasi S. dublin T46 S. dublin T47 S. untypable T62 S. enteritidis B95 S. typhimurium B96 S. 4,12,2 monophasic S. untypable T60 S. untypable T61 S. give T68 S. give T69 S. give T70 S. ohio K73 S. taksony K90 S. agona K72 S. 3,10,1 monophasic S. untypable T63 S. agona T71 S. worthington K84 S. adelaide K87 S. untypable T65 S. schwarzengrund K86 S. oranienburg K74 S. unknown T55 S. bredeney T49 E. coli H1A E. coli H5A S. montevideo B92 S. soerenga T50 S. albany T45 Clostridium perfringen 1 Clostridium perfringen 4 VRE H1A VRE H5A C. coli C. coli C. jejuni

10080604020

FlavomycinFlavomycin

Used in chickens and swine to promote Used in chickens and swine to promote growthgrowthPhosphoglycolipid antibiotic Phosphoglycolipid antibiotic Active against Gram positive bacteriaActive against Gram positive bacteriaIs not absorbed from the gutIs not absorbed from the gutNothing similar to flavomycin is used in Nothing similar to flavomycin is used in human medicinehuman medicineNo known mechanism of resistance No known mechanism of resistance

In vitroIn vitro conjugation with Flavomycinconjugation with Flavomycin

0

12

345

6

7

8

0 8 16 24 32 40 48 56 64 72Flavomycin concentration mg/L

Log

10 C

FU/m

l

Pre onlyPre+conjConj. only

Proposed FFSRU Salmonella Proposed FFSRU Salmonella Poultry ResearchPoultry Research

Identify environmental and management Identify environmental and management influences that contribute to the influences that contribute to the colonization of foodcolonization of food--borne pathogens.borne pathogens.–– Newly constructed broiler houseNewly constructed broiler house

Where FFSRU is headed in Poultry Where FFSRU is headed in Poultry Research ContinuedResearch Continued

Identify prebiotics and symbiotics Identify prebiotics and symbiotics –– LactoseLactose–– ChlorateChlorate–– CottonseedCottonseed–– Quorum sensingQuorum sensing

Identify environmental and management influences that Identify environmental and management influences that contribute to the antibiotic resistancecontribute to the antibiotic resistance–– FlavomycinFlavomycin

Characterize the complex interaction between the Characterize the complex interaction between the immune and endocrine systems and their role in food immune and endocrine systems and their role in food safety. safety. –– Endocrine assayEndocrine assay–– Endocrine in vitro model (epithelial cells)Endocrine in vitro model (epithelial cells)

PrePre--harvest Research at ARSharvest Research at ARS

A broad research area being addressed by a A broad research area being addressed by a diverse group of scientists in many locations.diverse group of scientists in many locations.

Many others in ARS are studying preMany others in ARS are studying pre--harvest harvest epidemiology and developing intervention epidemiology and developing intervention strategies in poultry. The knowledge gained in strategies in poultry. The knowledge gained in these studies may impact the overall health of these studies may impact the overall health of poultry as well as food safety issues that may poultry as well as food safety issues that may influence future applications in the preinfluence future applications in the pre--harvest harvest poultry research.poultry research.