Final Report for LS02-134
Several standard varieties of turkeys and a commercial strain were compared in range-based production systems, in DNA analysis, and for immune system response. Although the commercial variety reached market weight in fewer days and grew to a larger size, the standard varieties had lower mortality and better immune response. DNA micro-satellite analysis showed some standard varieties are only distantly related to the commercial strains, providing valuable genetic diversity essential for the long- term sustainability of turkeys. Increasing market demand over the past 5 years has supported increasing populations, rescuing standard turkey varieties from extinction, and providing new sustainable enterprises for farmers across the nation.
Tables, figures and photographs referred to in the following text are available from SARE.
Objective 1: Define range-based turkey production systems as the term will be applied in this project.
Objective 2: Identify similarities and differences of specific standard varieties and industrial turkey stocks in range-based, on-farm settings by measuring health status, weight gain, morbidity/ mortality, and feed conversion.
Objective 3: Identify similarities and differences of standard varieties and industrial turkey stocks by measuring response to immunologic tests and biochemical assays, including lymphocyte isolation, lymphocyte proliferation, and flow cytometric analysis
Objective 4: DNA fingerprint standard turkey varieties. This information documents the genetic differences and similarities of the turkey genomes.
Objective 5: Correlate immune response, DNA fingerprint and production characteristics to support the promotion of standard varieties for range-based production.
Objective 6: Inform farmers interested in range-based turkey production, the poultry science community, and consumers about project results.
Objective 7: Evaluate project effectiveness at meeting each objective and define next steps.
While crop genetics is nearly always an important consideration for both conventional and sustainable cropping systems, appropriate genetics are rarely considered in the decision for sustainable livestock and poultry production. Yet genetics cannot be overlooked as an important factor in the development of successful sustainable systems.
A breed or variety of livestock or a variety of a plant is the expression of its genome – the combination and configuration of the genes that it carries. Breeds and varieties often carry unique genomes which have been selected for specific habitats (e.g. pasture-based systems or confinement systems). Genomes cannot be reconstructed despite the wonders of current technology. Genes can be manipulated but they cannot be created. If a genome becomes extinct an opportunity to increase and improve our food supply and to modify our production system also is lost.
There is little current research on standard turkey varieties, but anecdotal information from small scale producers indicates that they possess valuable attributes for range production. Before specific turkey varieties can be promoted for use in ranged-based systems the immunological health, productivity and biological fitness (hardiness, reproductive health, and foraging activity) of standard varieties compared to the industrial stocks needs to be assessed and evaluated in extensive production systems. With the support of a SSARE Planning Grant (RD309-036/1789727) a group of experienced turkey breeders, farmers, poultry scientists, an industry representative, program personnel from non-profit organizations with expertise in livestock conservation and in educational outreach met in July 2001 to establish a collaborative group. The group articulated the questions to be researched and the approaches to be taken for gathering the needed information on standard varieties of turkeys. Our hypothesis is that standard varieties of turkeys have superior immuno-competence and perform better in range-based production systems than industrial stocks. To test this hypothesis the project will document the immuno-competence of turkey genomes and their performance in range-based production, providing farmers with valuable information needed to make thoughtful decisions about genome selection, conserve the genetic diversity these varieties represent, and provide further evidence that genetic biodiversity is essential to prevent the catastrophic collapse of the turkey industry.
Range-based production is economically, environmentally, and socially far more sustainable than current confinement turkey production. Range-based production more closely resembles the turkey’s natural habitat making it a more humane system. It is not capital intensive as large, expensive housing is not required. Well managed, range-reared turkeys effectively spread their manure over the pasture, recycling consumed nutrients. Consumers are demanding more naturally and humanely raised poultry, creating an important niche for sustainable farmers. As recently as the 1980s range-reared turkeys were an important farming enterprise. Turkeys foraged for much of their own food at low cost to the farmer in both time and money, and served as an active partner in pest control. At season’s end turkeys often gleaned harvested fields before going to market. The purebred standard varieties used did not require artificial insemination, giving farmers the freedom to propagate their own replacement stock.
Modern farmers are increasingly interested in diversifying their agricultural enterprises, capturing the new and emerging markets, and rediscovering the benefits of integrated livestock and cropping systems. But finding purebred standard turkeys – production birds of their day – is no longer easy. The 1997 ALBC turkey census found only 1335 breeding birds of the seven recognized varieties of standard turkeys, (Standard Bronze, Bourbon Red, Narragansett, White Holland, Slate, Black, and Royal Palm). A few additional varieties exist, but because of their limited numbers and closely held populations, they do not effectively participate in breeding programs and genetic conservation efforts. These historically significant genomes are disappearing before our very eyes.
The National Turkey Federation estimates that nearly 300 million turkeys are produced annually. Several southern states lead the nation in the industrial turkey production. Industrial turkeys have been selected for rapid growth, broad breasts, white feathers, and ability to produce in confinement systems. Intensive selection has resulted in a highly efficient and consistent production of meat. Toms can be ready for slaughter at 35-40 lbs at only 20 weeks. This strategy has, however, led to an increase in health problems such as compromised immune systems, skeletal failures, hypertension, and ruptured aortas. Survival characteristics such as disease resistance have not been a priority (Christman, 1999). As a result, significant resources are necessary to prevent disease through routine use of prophylactic antibiotics, anthelmentics, and other medications. Artificial insemination is a necessary standard practice since mature toms have such broad breasts and short legs as to be unable to mount the hens.
Why does this matter? Such narrow genetics makes the whole population vulnerable to disease in much the same way that past blights were catastrophic for genetically uniform potato, wheat, and corn crops. Genetic resources were the solution to these crop disasters. The greatest problem facing the industry may be the narrow genetic foundation of industrial turkeys since only a few strains of the Large White turkey variety are used commercially. Conservation of standard turkeys will provide the diverse genetics needed to stem such a disaster.
The National Turkey Federation estimates that 276 million turkeys were produced in 2000. (National Turkey Federation). Several southern states, including North Carolina, Virginia and Arkansas, lead the nation in the commercial production of turkeys. (USDA-NASS). In spite of these numbers turkeys are the most genetically eroded of all livestock species. Modern methods of poultry production seek to isolate birds from disease and are antagonistic to the natural processes which enable genetic selection for disease resistance. In fact, such practices are likely allowing susceptibility to disease to increase in breeding populations (Lamont, 1994). This is compounded by the fact that only a few strains of the Large White turkey variety are used commercially and only three primary turkey breeder companies produce genetic stock for commercial use worldwide. These birds have been selected for broad breasts, white feathers, and the ability to grow rapidly in high-input confinement systems. The American Livestock Breeds Conservancy (ALBC) conducted a census of turkeys in 1997. Only seven varieties of standard turkeys, (purebred, non-industrial turkey varieties), and a total of 8,212 breeding females were found. (Christman, 1999). When adjusted for the Broad Breasted Bronze population, a variety not currently considered “heritage,” a mere 1182 breeding females were found. (ALBC 1997 Turkey Census files) Additional varieties exist but were not readily available to the public because of their very limited numbers and closely held populations. This isolation and anonymity only serves to perpetuate their decline. All of these genomes are disappearing through lack of attention and a viable market.
For this reason, maintaining and characterizing a large gene pool and facilitating improved disease resistance through genetic selection is a desirable long-term objective. To this end we propose to characterize the immuno-competence and genetic diversity of several historic and commercial breeds of turkeys.
Why does this matter? As ALBC learned while researching turkeys, “The greatest problem facing the industry … may be the narrow genetic foundation of industrial turkeys. Intensive selection of a few strains of Large Whites has resulted in a highly efficient and consistent production of meat. At the same time, this strategy has led to the increase in health problems such as compromised immune systems, reduced fertility, joint and bone problems, high blood pressure, and ruptured aortas. Survival characteristics, such as disease resistance, have not been a priority.” (Christman, 1999) Artificial insemination is now standard practice since mature toms have such broad breasts and short legs that they are unable to mount the hens. These industrial strains are generally biologically unsuitable for small scale, sustainable agriculture ventures.
Such narrow genetics makes the whole population vulnerable to disease and infection, in much the same way that past blights were catastrophic for genetically uniform potato and corn crops. The turkey industry directs significant resources toward preventing disease and infection through the extensive use of vaccines, prophylactic antibiotics, anthelmentics, and bio-security. “With cheap and effective vaccines available for many virus diseases, there is little incentive for [commercial] breeders to become involved in complex selection programmes for genetic resistance to them.” (Hunton, 1993). Should industrial turkey strains experience an outbreak, disease not controlled through present genetic resistance or medical and management interventions, the consequences would be disastrous. “The extent of disruption which the industry might experience, as a result of a new disease with no solution other than the introduction of new genotypes, is almost impossible to calculate. Assuming that resistant genotypes existed among breeders’ stocks, a minimum of three to five years would be necessary for multiplication [of healthy stock], and distribution [of production flocks].” (Hunton, 1993). Sustainable systems would be safe from such catastrophe only if the immuno-competence, genetic diversity and production attributes of other genomes had been characterized, were readily available, and were already used by producers. In the end, the lack of genetic diversity among industrial strains may prove to be the undoing the turkey industry. For this reason, maintaining and characterizing a large gene pool and facilitating improved disease resistance through genetic selection is a desirable long-term objective. To this end we propose to characterize the immunocompetence and genetic diversity of several historic and industrial breeds of turkeys.
SARE has supported three other projects which explicitly evaluated turkeys in sustainable systems:
- "Minor Breed Turkeys - Growth Rate and Eating Qualities" (Project number: FNE94-038) conducted in 1994 by Anne Bossi of Maine for $980. In Bossi's experience the "minor breed” turkeys were not good production birds but they were hardier and more disease resistant than the industrial varieties.
- "Evaluating Hoophouses for Rotationally Grazed Turkeys" (Project number: FNE94-051) conducted in 1994 by John Hayden of Vermont for $705. Hayden found that the turkeys raised in hoophouses showed greater feed efficiency than those raised in conventional confinement warehouses.
- "Evaluate Standard Turkeys for the Small Scale Producer" (Project number: ENE01-374) was funded in 2001. David Fritz, the project coordinator, resides in Maryland and received $751. Fritz evaluated the Wishard strain of the Standard Bronze turkey. He describes these birds as "smart, self-sufficient, good foragers and pleasant to work with. At 8 months birds dressed for the holiday market ranged from 15 - 20 pounds and were very tasty, though slightly tough because of high level of activity. He was very pleased with the result and intends to evaluate other varieties.
Genetic resources were the solution to crop disasters. Only through the conservation of standard turkeys, “the natural resource upon which [turkey] production depends” (1990 Farm Bill), will diverse genetics be available to stem such a disaster.
Until the 1950s all poultry were raised outdoors. Turkeys were raised on range into the 1980s in some areas. In a 1983 Poultry Science article the economics of raising turkeys in confinement and on range in Georgia were compared. It was found that both tom and hen turkeys could be raised to heavier weights more efficiently on range during the summer, fall, and early winter (Lance, 1983).
Today’s poultry industry is vertically integrated, with only a few large companies owning most stages of production, processing, and marketing. By contrast, range-based production is a grassroots movement that focuses on farm-scale production and direct marketing. It has been developed from the ground up by hundreds of family farms, and is driven by consumers seeking an alternative product. Some producers use standard turkey varieties but most use industrial strains developed for confinement rearing. Marketing is usually direct to customers and advertising is often word-of-mouth. Producers report more demand than they can supply (Fanatico, 2001).
The U.S. pastured poultry movement was begun by pioneer Joel Salatin and his book Pastured Poultry Profits (Salatin, 1996). Other leaders in pastured poultry include Andy Lee (The Chicken Tractor, 1998) an early advisor to this project, and Herman Beck-Chenoweth (Free-Range Poultry Production and Marketing, 1996). SARE has supported several projects by pastured poultry producers as well as some landmark initiatives by nonprofit organizations including:
- A partnership between Heifer Project International (HPI) and the National Center for Appropriate Technology (NCAT) that helped limited-resource farmers in the South test small-scale pastured poultry field pens (Polson, 1996). NCAT published a booklet, Pastured Poultry: An HPI Case Study Booklet (Fanatico, 1999).
- Another HPI/NCAT partnership is helping farmers interested in expanding range poultry businesses by developing mobile processing units that meet government inspection (Muntz, 1999). NCAT is developing a range poultry entrepreneurial Toolbox (Fanatico, 2001).
- The Center for Integrated Agricultural Systems at the University of Wisconsin led a project to examine economics and quality of life for small pastured poultry producers, nutritional qualities of the meat, sensory/microbial analysis, and marketing analyses (Stevenson, 1997).
Current outdoor poultry production systems include both “contained” and “uncontained” systems. In contained systems, the foraging of the birds is contained - and protected - by a fence, pen, or netting, allowing a high level of management. Contained systems include yarding, field pens like the "chicken tractor" (Salatin, 1996, Lee, 1998) and moveable or stationary net range systems. In uncontained systems, the foraging of the birds is not contained. Birds range during the day — usually in a pasture — and return to a portable house at night. The house is moved regularly to a fresh site. Uncontained systems include portable houses on skids, colonies (Fanatico, 2001), and herding.
In avian species, antibody-mediated immunity has been studied extensively (Dunnington , 1992; Hovi , 1978; Kaspers, 1993; Scott, 1991; Thiel and Burkhardt, 1984). By comparison, much less is known about the avian cell-mediated immunity. Like mammals, it is believed that the cell-mediated immune response plays a prominent role in the ability to resist/survive an infectious insult. In order to better understand the immunological potential of turkeys, we have developed tools that enable evaluation of cell-mediated immune function (Barta , 1992; Gogal , 1997, Caldwell , 2001). Combined with existing humoral function tests, it is now possible to compare on a quantitative basis, multiple immunological traits.
DNA fingerprinting i.e., Restriction Fragment Length Polymorphism analysis (RFLP) has been shown to be useful in determining the genetic relationships among and within populations of chickens (Dunnington, 1991; Dunnington , 1994; Kuhnlein , 1991; Siegel , 1992). Although not new technology, the technique is sensitive enough to distinguish between breeds, selected lines within breeds, and selected lines and their F1 crosses (Dunnington, 1991) as well as highly divergent populations such as jungle fowl and domestic chickens (Siegel, 1992). The relationship between RFLP pattern and quantitative morphological / physiological traits has also been demonstrated (Dunnington, 1990; Kuhnlein, 1991). This technology has only recently begun to be exploited in turkeys (Ye, 1998; Zhu, 1996).
(The following three paragraphs have been excerpted from a publication by Smith, et al., 2005, which was funded by this project.)
Molecular approaches have made it possible to rapidly and reliably determine genetic diversity or variation within and among livestock species. This determination often can be useful for the various processes used in the genetic improvement of livestock and poultry. In particular, the ability to identify existing variation correctly can influence the efficiency of selection, a major genetic improvement approach in the livestock and poultry industries. Molecular approaches that have been used for the determination of genetic variation in poultry include the techniques based on polymerase chain reaction (PCR), including randomly amplified polymorphic DNA (RAPD) and microsatellites or simple sequence repeats. Methods based on Southern analysis, including DNA fingerprinting using a variable number of tandem repeat DNA sequences as probes, have also been used. Recently, for example, Ponsuksili et al. (1998) used microsatellites as probes to evaluate genetic variation within and among 12 chicken lines. Using aminisatellite-specific probe, both Dunnington et al. (1994) and Kuhnlein et al. (1990) used DNA fingerprinting to evaluate genetic variation within chicken lines divergently selected for specific traits over several generations. Several PCR-based genetic marker systems have also been used to evaluate variation between and within chicken lines including RAPD-PCR (Sharma et al., 2001) and microsatellites (Zhou and Lamont, 1999). These molecular methods have also been used by a variety of groups to evaluate genetic variation within and among commercial turkey populations. Using a multilocus probe, Zhu et al. (1996) and Ye et al. (1998) evaluated both diversity within and among selected turkey lines and commercial populations. In another study, Smith et al. (1996) used the RAPD procedure to distinguish among commercial turkey strains. Although these analyses have been informative, they were limited, as the populations and strains evaluated were primarily commercial strains with an expected narrow genetic base.
Efforts by others (Reed et al., 2000) and us (Huang et al., 1999) have led to the development of turkey-specific DNA marker systems. These DNA marker systems are essential for increasing our understanding of the turkey genome and the relatedness among turkey populations. Recently, Latch et al. (2002) and Mock et al. (2002) used microsatellites developed by Huang et al. (1999) to evaluate genetic variation and differences among subspecies of turkeys across both geographical and historical ranges. In the present work, the use of these molecular systems, including RAPD, microsatellites, and EST-based SNPs, was investigated to distinguish among five turkey varieties—Blue Slate (BS), Bourbon Red (BR), Narragansett (N), Royal Palm (RP), and Spanish Black (SB). Additionally, a novel PCR-RFLP assay was developed to genotype birds for one of the four SNPs detected.
In conclusion, scientific investigation employing the aforementioned tools will be of help in determining the usefulness of historic breeds in sustainable agriculture and their value as a genetic repository for immunologic traits.
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Literature was reviewed. Information gathered from experts, specifically Frank Reese, Jr. and Anne Fanatico. Discussion and agreement on definition occurred at planning meeting.
Eight farmers from across the country participated in the project. Each brought a unique set of skills to this effort. They were:
- Pam Marshall, Amenia, NY. A small scale turkey breeder & farmer; committed steward of standard varieties, with access to the New York City market.
New England Heritage Breeds Conservancy, Richmond, MA, Heather Bean Ware, Associate Director of Agriculture Education. An educational facility dedicated to the conservation of rare breeds of livestock.
Gail & Harry Groot, Hiwassee, VA. Small scale, multi species, farmers who strongly prefer standard turkeys that are naturally mating, brood own young, with good growth potential and economic return.
Gerry Cohn, Snow Camp, NC. A small scale, multi species farmer who buys poults to raise for direct sales to consumers at Thanksgiving. He prefers standard varieties with good growth potential and economic return.
Center for Environmental Farming Systems, North Carolina State University, Goldsboro, NC, Paul Mueller, Professor, Crop Science, Sustainable Agriculture Coordinator. CEFS is research farm dedicated to sustainable agriculture, including multi-species and integrated crop & livestock systems. Scientists research and demonstrate production systems that are environmentally and economically self-sustaining.
Glenn Drowns, Calamus, IA. A committed steward of hundreds of breeds and varieties of poultry, including fifteen varieties of standard turkeys, as well as hundreds of heirloom vegetables. He is dedicated to the rescue of standard turkey genetics from the brink of extinction and their long term conservation. He is a breeder and operates a moderately sized, seasonal hatchery. He does not raise birds for the processing market.
Frank Reese, Jr., Lindsborg, KS. A committed steward of standard varieties of turkeys. He is dedicated to maintaining breed standard, carcass quality in naturally mating turkeys. A significant breeder and producer of standard turkeys for the Thanksgiving market.
Paula Johnson, Las Cruces, NM. A committed steward and breeder of standard varieties of turkeys. She dedicated to the rescue of standard turkey genetics from the brink of extinction and their long term conservation.
Bourbon Red turkey poults from Privett Hatchery and a commercial strain of medium white turkey poults from British United Turkeys of America (BUTA) were shipped to each farm on either May 15 or 29, 2002. Glenn Drowns also had a combined flock of Black Spanish and Blue Slates turkeys that were from Privett. Paula Johnson had a flock of a commercial strain of large whites turkeys from Nicholas Hatchery. Each farmer received shipments of a few more than 30 poults of each variety, the extras to allow for some loss due to shipping stress. Gerry Cohn, Paul Mueller, Pam Marshall each received 60 Bourbon Red poults. Paula Johnson received 15 of each. While each farm was different, their production systems all complied with the definition of "range-based", as defined by this collaboration. Each farm provided the following, using forms developed specifically for the project:
(1) a detailed description of their production systems and a general overview of their farm:
(2) weights taken on a random subset of 5 birds once a week through slaughter;
(3) final live weights and dressed weights of all birds;
(4) morbidity and mortality information, with necropsy information provided when possible;
(5) weight of feed fed to each flock;
(6) weather data; and
(7) observations of behavior.
Significance and Justification
Presently, commercial or industry turkeys have been selected based on their growth rates, meat potential (i.e. broad breasts), white coloring and their ability to artificially reproduce in highly controlled commercial environments. The outcome of this selection is a bird with high feed efficiency and growth characteristics. However, there are costs to this type of management system. The turkey gene pool has narrowed considerably. Natural breeding can not occur in these commercial birds due to the anatomical characteristics of these genetically altered lines. Other problems encountered include hypertension, ruptured aortas, skeletal failure of weight support and immune dysregulation. As a result, the industry has had to make significant financial investment to control disease. The long-term outcome does not appear favorable for the industry bird. One obvious remedy would be to expand the gene pool.
The standard varieties of turkey have been in existence in range-based farms for years with some lines pre-dating the industry lines. Minimal research has been conducted on these lines. However, the fact that these lines have remained in existence for many of these years primarily in outdoor systems would seem to indicate that their immune system and production indices would warrant evaluation.
Standard varieties of turkeys possess genetic characteristics that code for optimal immune response and may potentially serve as a useful gene pool for enhancing the commercial lines.
Aim 1: To compare select production indices in standard and commercial turkey lines at various stages of growth.
Commercial turkeys have been selectively bred for their production performance traits at the expense of the birds’ other resources. The current commercial bird’s gene pool is so narrow that there is no room to genetically manipulate this line. Aim 1 assessed, of the 5 standard varieties studied, if any of the lines were comparable to the commercial line in production indices.
Five varieties of standard turkey were evaluated: Bourbon Red, Blue Slate, Narragansett, Royal Palm, and Black Spanish. Twenty birds of each of variety, 11 males and 9 females were used in the experiment.
One commercial turkey strain was evaluated: British United Turkey of America (BUT). Twenty birds were used, only males were available.
All varieties were maintained under identical management. Live weight was measured at 9, 11, and 13 weeks of age utilitzing an electronic kilogram scale.
Egg Hatch and Infertility Analysis
Five varieties of standard turkeys were evaluated: Bourbon Red, Blue Slate, Narragansett, Royal Palm, and Black Spanish. Seven hens of each were inseminated, and eggs collected over a period of seven weeks. Data collected included the number of eggs each hen laid, infertile eggs (evaluated through candling), the number that hatched, and hatchability (the number that did not hatch). Inseminated hens from the commercial strain were not available for evaluation.
Aim 2: To employ an immune testing panel of assays to evaluate immunophenotypes of the standard turkey lines to the chosen commercial line.
Commercial turkeys have been shown to be relatively immunologically incompetent. As a result, the industry has had to create, at great costs, clean environments in which these commercial birds can be raised.
This study compared the following immunological and production characteristics of five standard varieties of turkeys (Blue Slates, Black Spanish, Royal Palms, Bourbon Reds, and Narragansetts) one commercial breed (British United Turkeys of America): lymphocyte recovery, leukocyte proliferation in response to Concanavalin A at 5 µg/ml, 50 µg/ml and phorbol 12-myristate 13-acetate (PMA) 0.02 µg/ml with ionomycin 0.40 µg/ml, T-cell subset flow cytometric analysis, packed cell volume (PCV), total protein (T.P.), and five point differentials.
Blood was transferred to hematocrit tubes and placed in an Hematocrit Centrifuge for 6 minutes then measured.
Total Protein was measured using a refractometer.
The five-point differential was conducted using bloods smears. Specific cells included lymphocytes, basophils, heterophils, monocytes, and eosinophils which were analyzed under a light microscope. One hundred leukocytes were enumerated per slide.
Blood collection/Lymphocyte Isolation
- Peripheral blood (8 ml) was collected from jugular vein with a 23-g needle attached to a pre-heparinzed 10-ml syringe.
Blood transferred to vacuum tubes were slow centrifuged at 3x at 25 degrees C, 50 xg for 10 min.
Buffy Coat Layer (lymphocyte-rich layer) was collected by gentle "swirl" technique with a 1-ml sterile glass pipette.
Buffy Coat collection repeated after each centrifugation; all suspensions collected were combined and chilled on ice.
Washed buffy coat suspensions 3x with RPMI 1640 media.
First wash: 12 degrees C, 1200 rpm, 10 minutes.
Second and third wash: 12 degrees C, 1200 rpm, 7 minutes.
Resuspended pellet in 3-ml RPMI 1640 media.
Stained lymphocytes and other cells with Natt-Herrick’s Stain.
Lymphocytes were identified and enumerated with a hemacytometer and adjusted to 5 x 106 cells/ml in RPMI 1640 media with 10% FBS.
Lymphocyte Proliferation Assay
- To 96-well round bottom tissue culture plates, 100 ul of 5 x 106 lymphocytes/ml were added to quadruplicate wells containing 100 ul of medium alone and concanavalin A (Con A 5-100 mg/well)
Incubated cells at 37oC with 5% CO2
20 ul of Alamar Blue dye added to each will at 24 hours.
24 hours after dye added, fluorescent changes of plate were measured using a CytoFluorII fluorescence Multi-well Microplate reader.
Flow Cytometric Analysis
- Lymphocytes stained with monoclonal antibodies (PE-CD4, FITC-CD8)
Analyzed by Epics XL flow cytometer and Immuno-4 software program
Packed Cell Volume & Total Protein
- After collection, blood was transferred to hematocrit tubes and placed in an Hematocrit Centrifuge for 6 minutes.
After centrifugation, packed cell volume was determined utilizing a Circular Micro-Capillary Tube Reader.
Total Protein was subsequently determined utilizing a Refractometer.
Five Point Differential
- After making blood smears, specific cells to include: lymphocytes, basophils, heterophils, monocytes, and eosinophils were analyzed under a light microscope. One hundred leukocytes were enumerated per slide.
I. Endogenous Production of Ascorbic Acid in Historic Turkeys: Physiology and Applications
The antioxidant ascorbic acid has been the subject of extensive scientific investigation since its discovery by Albert Györgyi in the 1920s. Ascorbic acid (AA) has been shown to be physiologically multifunctional, enhancing function of the innate immune system, modulating gene expression, acting as a co-factor in enzymatic reactions, and protecting organisms from free radical damage during oxidative stress. The fact that avian species have retained their ability to synthesize AA has generated much interest into the possible benefits this ability affords birds. Previous studies have suggested that in times of stress (ie. disease, heat, and environmental stressors) endogenous AA may not provide adequate protection. Some studies have shown benefits in supplementation of domestic avian species with dietary AA.
The purpose of this study was to modify a reduction-based spectrophotometric method to specifically quantify AA levels in avian tissue and plasma samples. The goal was to develop an assay that was rapid, specific for AA, accurate and reproducible. A secondary aim of this study was to use this assay to determine if endogenously produced AA levels varied between varieties and between sexes of the same variety. The assay presented here is novel in that it has never been attempted in avian species and it provides a method for not only specific measurement of AA in plasma but in tissue samples as well. The experiments performed were designed to evaluate the efficacy and reliability of this assay in plasma and/or tissues of different avian species. The results section contains the data from a submitted manuscript that reviews the standard turkey varieties.
(The following was excerpted from a paper entitled “Specific Determination of Plasma and Tissue Ascorbic Acid in Avian Species” by Robert Gogal, F. William Pierson, and Cal Larson Virginia Tech, Blacksburg, VA and submitted to Poultry Science.)
Blood was collected with aseptic technique from the jugular vein of each bird. A 22-g needle was used with a 10 mL syringe. Blood was then transferred to a heparin-containing vacuum tube with an 18 g needle. Plasma was separated from the whole blood by centrifugation a 1300 X g for 15 min. The resulting plasma was transferred in 500 µL aliquots to 1.5 mL microcentrifuge tubes. The plasma was immediately snap-frozen in a dry ice/ethanol slurry and then stored at –45º C until needed.
Preparation of Ascorbic Acid Assay Reagents
An 8mM 2,2’-dipyridyl solution was made with 313 mg of 2,2’-dipyridyl (Sigma Chemical, St.Louis, MO, USA ) dissolved in 250 mL of a 36 mM HCl solution. (770 µL of concentrated HCl in 250 mL water) The dipyridyl solution showed a shelf stability of 3 days at room temperature. The 0.3 M Sodium Acetate buffer was made with 3.1 g of sodium acetate (Fisher Chemical, Fair Lawn, NJ, USA) dissolved in 900 mL of water. 16 mL of glacial acetic acid (Sigma Chemical) was then added and the pH adjusted to 3.6. The final volume was brought up to 1 L with deionized water. This solution was found to be stable for 1 month at room temperature. The ascorbate oxidase (ao) solution was prepared with 1 mg of lyophilized ascorbate oxidase (Sigma Chemical) (102.3 units/mg solid) dissolved in 20 mL of deionized water. Twenty, 1 mL aliquots, each containing approximately 5 units of enzyme were frozen at –45º C until needed. A 20 mM ferric chloride solution was prepared with 540 mg of FeCl3 6 H2O (Sigma Chemical) dissolved into 100 mL of deionized water. This solution was found to be stable at room temperature. A 2.84 mM ascorbic acid stock solution was made with 50 mg of L-Ascorbic Acid (Sigma Chemical) dissolved in 100 mL of deionized water. This solution was made daily and placed on ice and shielded from light until needed. The reduction reagent was prepared as a mixture of the dipyridyl solution, acetate buffer, and ferric chloride solution. The ratio used for the mixture was 1.25 mL sodium acetate: 0.15 mL dipyridyl: 0.1 mL ferric chloride, such that 1.5 mL of the reagent was required for each sample tested. The mixture was made daily during the incubation step in the assay procedure.
Ascorbic Acid Assay
Ascorbic acid standards were prepared as follows: A 200 µL aliquot of the ascorbic acid stock solution was added to a tube containing 800 µL of water. The tube was vortexed and 500 µL of the resulting solution was transferred into a new tube with 500 µL of deionized water and vortexed. This serial 1:2 dilution was continued to obtain ascorbic acid standards of 0,6.25,12.5,25,50 and 100 µg/mL. Deionized water served as the 0 µg/mL standard. Plastic culture tubes were labeled as follows: #1 ao+, #1 ao-, etc. up to #6 ao+#6 ao-. To each numbered tube, 100 µL of the corresponding standard or experimental sample was added. Next, to each tube labeled ao-, 50 µL of water was added, and to each tube labeled ao+, 50 µL of the ascorbate oxidase solution was added. All tubes were incubated in a 37º C water bath for 15 min. While the samples were incubating, the reduction reagent mixture was prepared. After incubation, 1.5 mL of the reagent mixture was added to each tube and immediately transferred to a 96 well plate. Approximately 150 µL of each sample was pipetted into each well (in triplicate) and read 525 nm on a UV spectrophotometer (Molecular Devices VERSAmax, Sunnyvale, CA ). The plate was read 5 min from the addition of reagent mixture.
Ascorbate Oxidase allows specificity for ascorbic acid using reduction-based spectrophotometry. Blood samples were collected from eight historic turkeys, five males and three females. Plasma was separated and snap-frozen immediately. At the time of assay, samples were thawed at room temperature. The assay for ascorbic acid was run as previously described. Data averages from the enzyme negative group were compared to averages obtained by subtraction of the enzyme positive group from the enzyme negative group.
One-way analysis of variance (ANOVA) was performed on the data sets using the IGOR-PRO statistical analysis software. Statistical significance was set at p<0.05.
II. Comparison of Immunological Profiles between Commercial Line and Standard Varieties of Turkeys Following challenge with HEV and E. coli
The purpose of this study was to compare immunologic profiles of commercial turkey to a number of standard varieties of turkeys. Peripheral blood was collected every two days from flocks of age-matched birds consisting of a commercial line and standard varieties (Royal Palm, Narragansett, Bourbon Red, Blue Slate, and Black Spanish). At six week-of-age, the turkeys were challenged with HEV (Hemorrhagic Enteritis Virus). Blood was collected on days 0, 2, 4, and 6, before they were exposed to E. coli at 108 CFU/mL on the seventh day. Immune endpoints measured included: lymphocyte recovery enumeration and sizing, lymphocytic function using in vitro mitogen proliferation assays with Alamar Blue™, and morphologic analysis from cytospins.
(This following section is excerpted directly from Biochemical Genetics, Vol. 43, Nos. 1/2, February 2005 (©2005)
Molecular Analysis of the Relatedness of Five Domesticated Turkey Strains
Edward J. Smith,1,3 Tuoyu Geng,1 Elizabeth Long,1 F. William Pierson,2
D. Phillip Sponenberg,2 Cal Larson,2 and Robert Gogal2
1 Comparative Genomics Lab, Department of Animal and Poultry Sciences.
2 Department of Biomedical Sciences & Pathobiology, Virginia Tech, Blacksburg, Virginia.
3 To whom correspondence should be addressed at 2250 Litton-Reaves Hall,Virginia Tech, Blacksburg, Virginia 24061-0306; e-mail: firstname.lastname@example.org.)
Abbreviations: EST, Expressed Sequence Tag; SNP, Single Nucleotide Polymorphism.
The birds that provided DNA samples were maintained as true-breeding strains at the Virginia Tech Turkey Research Center. The parents were obtained from a retailer who originally acquired foundation stock from a few breeders and then subsequently maintained each strain as a closed and pure breeding population. Privett Hatcheries provided Virginia Tech the birds which were produced from random matings based on pedigree records. ALBC maintains a collection of the strains and has initiated a multi-institution project, including the study described here, to characterize noncommercial domesticated turkeys. To our knowledge, there is no peer-reviewed publication that describes the general, external, and functional characteristics of these turkey strains. There is limited historical information about these and other turkey strains in Dohner (2001). In the initial among-strain molecular analysis, two DNA pools, each containing samples from five birds and made according to Smith et al. (2002), were used. The rationale for using DNA pools in the initial analysis was to determine whether differences within strains were lower than among strains. Analyses of individual samples involved a total of 94 birds, including 23 from Blue Slate (BS), 15 from Bourbon Red (BR), 19 from each of Narragansett (N) and Royal Palm (RP), and 18 from Spanish Black (SB). An aliquot of 50 mL of whole blood collected in EDTA from the birds using standard techniques was used to isolate genomic DNA according to Smith et al. (1996).
2. STS-Based Analysis
Primers specific for a chicken EST, accession number AW328799, previously described as TUCEST200 (Smith et al., 2001), were used in the sequence tagged site (STS)-based analysis. The chicken EST as well as the turkey STS derived from it does not match any database gene sequences. The sequences of the forward and reverse primers were 5_-TGGGCTGAAAAATAAAAGCA-3_ and
5_-GCCATGGAAAATGCATCAGT-3_, respectively. The amplification, processing of PCR products, sequencing, and sequence analysis to detect and validate SNPs were also according to Smith et al. (2001). Although sequences of amplicons produced using these primers are identified by the prefix TUCEST200, the SNPs detected have the prefix VPIcest200SNP.
3. Restriction Fragment Length Polymorphism (RFLP) Analysis and Conditions
Two naturally occurring Msl1 sites in the consensus turkey DNA sequence, accession number AF268673, were used as a basis for RFLP-based genotyping of the turkey strains for an informative SNP detected from the comparative sequence analysis. The restriction digestion of the amplified products was according to the manufacturer’s (New England Biolabs) recommended protocol.
4. RAPD Analysis
Five primers were used in the RAPD-PCR-based screening (Table I). The primers were chosen on the basis of previous analysis of commercial turkeys (Smith et al., 1996). The amplification reaction was in a final volume of 10 μL containing 10 mM Tris-HCl (pH 8.3), 50 mM KCl, 1.5 mM MgCl2, 0.001% gelatin, 100-ng template DNA, 40-ng primer, 200 μM of each dNTP, and 1.5 units of Amplitaq DNA polymerase (Roche). Amplification was carried out in a PTC100 thermal cycler using the following cycling conditions: initial denaturation at 95◦C for 5 min, followed by 34 cycles of denaturation at 94◦C for 1 min, annealing at 35◦C for 45 s, and extension at 72◦C for 2 min. A final extension at 72◦C for 5 min was also carried out. Amplified products were analyzed on agarose gels and stained with ethidium bromide. Stringent controls, to ensure repeatability of each primer, were included in each PCR including blanks (water for negative control), two DNA pools, and duplicates of two random samples from two strains. Only distinct and reproducible fragments were scored.
5. Microsatellite Analysis
Three new primer sets, each unique for a novel microsatellite from previously described enriched turkey DNA libraries (Huang et al., 1999), were used. Each forward primer was tagged with an M13 forward primer as suggested by Oetting et al. (1995). The rationale for using tailed primers is the reduced cost for genotyping a large number of individuals using a labeled universal tail primer and not using either the labeled forward or reverse for each primer pair. Additionally, the tailed primer permits the efficient use of multiplexing, though not necessary in the current work. The sequence ID and sequences of the primers tested were TUM261, accession number AY170649, and containing (CA)12 microsatellite repeat,
5_-ACCTTCAAGCTGTCTTAGC-3_, 5_-CACCCTTGGTAATTTAGAATAAATGTT-3_; TUM312, accession number AF111596, and containing (TG)8: 5_-ACCCAT TAACAGTATGGGG-3_, 5_-AAAAATATGCATTTCCTA-3_; TUM940, accession number AF111668, and containing (CCT)3 microsatellite repeat: 5_-ACAGTTATCTTCCTTCTTT-3_, 5_-ATGATGGGTTTACAGGAGGG-3_. Each primer pair was optimized to yield a single fragment in cycling and reaction conditions that included an additional primer, a fluorescently taggedM13 forward (Oetting et al., 1995).
6. Statistical Analysis
Birds were scored for each informative RAPD fragment as either present (1) or absent (0). In general, conventional assumptions for RAPD-based genetic analyses were made, including the correlation of fragment sizewith locus and independence of different fragments. The biallelic SNPs and microsatellite alleles were analyzed by direct count within each population. For each locus within each marker system, heterozygosity was computed according to Hartl and Clark (1997). Because of the small sample sizes and assuming presence and absence of a band as dominant and recessive alleles, respectively, heterozygosity from RAPD data was calculated using the unbiased approach of Lynch and Milligan (1994). For the heterozygosity estimates, genotype frequencies were assumed to be in Hardy–Weinberg equilibrium. Using the RAPD data, genetic distance D_ was estimated using the statistics suggested by Lynch (1991). The method assumes genetic distance to be a function of the average band-sharing frequency between any two strains. Band-sharing frequency was computed as previously described (Smith et al., 1996), including adjustments for within-strain similarity. The genetic distance estimates were used to construct a dendrogram using the UPGMA approach (Sneath and Sokal, 1973).
2006 follow-up genetic study
The genetic research was expanded upon between 2003 and 2006. Using resected liver samples, Dr. Robert Gogal and his colleagues used identifying molecular markers to distinguish relatedness of several varieties of domestic turkeys. These included: Jersey Buff (4 samples from 1 source), Bourbon Red (17 samples from 2 sources), Black (48 samples from 6 sources), Chocolate (3 samples from 1 source), White Holland (8 samples from 1 source), Standard Bronze, Kardosh strain (7 samples from 2 sources), Midget White (8 samples from 2 sources), Narragansett (13 samples from 4 sources), Royal Palm (7 samples from 2 sources), Sweet Grass (3 samples from 1 source), and Slate (3 samples from 1 source). Microsatellite analysis on up to 5 loci was conducted on 139 of the 189 samples.
DNA was extracted from the samples using either a salting-out protocol or a Qiagen Tissue DNA Extraction kit, and the quantity and size range of the extracted DNA was assessed using a 0.7% agarose gel stained with ethidium bromide and visualized under a UV transilluminator.
Results and trends from objectives 2, 3 and 4 were compared.
Articles, press releases, information packets, collaborations with other organizations, and presentations were all vehicles for disseminating project results.
Evaluation was conducted by the collaborators ability to meet the objectives, and the level of response to promotional information. Additionally, the number of breeding birds was counted in 2003 and 2006 as a means of measuring adoption rate. An increase in breeding stock, breeders, and hatcheries directly correlates with demand. This became the project’s primary measure of effectiveness.
Results for the American Livestock Breeds Conservancy 2006 Turkey Census show that total numbers of heritage turkeys are increasing. The increase in breeding populations of turkeys is the direct result of the rise in market demand for these birds. Many organizations and individuals have contributed to this success, and the total effort over the past 10 years of engaging farmers, chefs, and consumers has generated and maintained momentum. Sustained momentum has led to a much more secure status for heritage turkeys.
The precarious position of naturally mating turkeys began to receive public attention beginning in 1997 with the American Livestock Breeds Conservancy’s (ALBC) census of standard varieties of turkeys maintained by hatcheries (Christman & Hawes, Birds of a Feather). Censuses conducted by the Society for the Preservation of Poultry Antiquities (SPPA) from 1998 through 2000 of both standard and non-standard varieties of turkeys held by small scale breeders continued to bring attention to the near extinction of naturally mating turkeys.
ALBC was able to use two grants, one from the Bernice Barbour Foundation and a second from the Southern Sustainable Agriculture Research and Education Program (SSARE), to generate information that led to securing a significant research grant from SSARE to study the immunologic health of standard and industrial varieties of turkeys and their suitability for range production. ALBC partnered with Virginia Tech and turkey producers around the country for this effort. ALBC was awarded a subsequent SSARE grant to develop an educational program and materials on the production of heritage turkeys. Concurrently, Slow Food USA recognized a number of rare turkey varieties on the Ark of Taste, its rare American foods list, and initiated a project to produce and distribute heritage turkeys. This project effectively connected an ALBC conservation priority with consumers. The success of this project initiated growth in the marketplace.
In 2002, the Slow Food USA project spun off as a for-profit company called Heritage Foods. Frank Reese, Jr., the owner of Good Shepherd Turkey Ranch, supplied turkeys to Slow Food and then Heritage Foods, and has continued to develop new market outlets. Frank has served as a close advisor to ALBC’s heritage turkey conservation project. All of the people associated with these organizations, plus many others, have played invaluable roles in the success of turkey conservation. The positive trend seen in this census is the result of the combined time, effort, money, and good will of all involved in this important project for a period of several years.
A CENSUS OF TURKEYS
Turkeys produce many offspring in a single year, but the vast majority of this production ends up on our tables and never passes genes to another generation. Because only a select few turkeys produce the next generation ALBC monitors breeding populations – rather than total populations – to determine which breeds and varieties are numerically secure as well as which are endangered and to what degree. The ALBC Conservation Priority List is derived from this information.
A census of poultry is a challenging undertaking. It is in many ways more difficult, and potentially less complete, than a census of four-footed livestock. A census of purebred livestock can usually be conducted by contacting a breed registry and obtaining information on the number of animals registered in a given year. Registration numbers indicate the size of active breeding populations. Registries, however, are not maintained for poultry. To census poultry, owners must be identified and surveyed individually to determine (1) who is maintaining breeding stock, and (2) how many breeding birds of each breed and/or variety each breeder is retaining for the current breeding season. Additionally, participation in the census is voluntary, which means that not all flocks are counted. While both owner identification and voluntary participation leave room for omissions, the compiled data provide a valid indicator of population size and trends.
During 2006 ALBC contacted hatcheries and breeders of heritage turkeys throughout the United States. A list of 447 names was compiled using a number of different sources: participants in a previous turkey census conducted by the ALBC and SPPA; ALBC and SPPA members who self-identified as raising turkeys; turkey producers listed on ALBC’s holiday turkey producers list and the Slow Food USA website.
The survey was distributed by US Post or email in early 2006 to those on the list. Additionally, the survey form was included in the ALBC News and posted on the ALBC website. Notices of the census were printed in a number of farming and poultry publications, inviting additional participation. Phone calls were made to a few known key breeders to assure their flocks were included in the census.
At the planning meeting held in July 2001, participants agreed to the following definition for range-based turkey production: (1) birds will have daily access to outdoor range, (2) birds will have daily access to forage, shelter and roosting locations, and (3) systems will NOT be 24 hour, 7 day per week housing, the single yard model (implying no range), or field pens or “chicken tractors,” too small for normal turkey behavior such as roosting, spreading wings, and exercise.
In January 2005 a definition for “Heritage Turkey” was developed. Over the past 5 years “heritage turkey” has become the market identity for standard turkeys. Unfortunately, some producers have sold broad breasted turkeys, or crosses thereof, as “heritage,” getting premium dollar for an inauthentic product. Through the leadership of Good Shepherd Turkey Ranch a definition for heritage turkeys was developed that appears to have reduced this practice. This definition has been embraced by the National Turkey Federation and has also educated retailers and consumers.
Heritage turkeys are defined by the historic, range-based production system in which they are raised. Turkeys must meet all of the following criteria to qualify as a Heritage turkey:
1. Naturally mating: the Heritage Turkey must be reproduced and genetically maintained through natural mating, with expected fertility rates of 70-80%. This means that turkeys marketed as “heritage” must be the result of naturally mating pairs of both grandparent and parent stock.
2. Long productive outdoor lifespan: the Heritage Turkey must have a long productive lifespan. Breeding hens are commonly productive for 5-7 years and breeding toms for 3-5 years. The Heritage Turkey must also have a genetic ability to withstand the environmental rigors of outdoor production systems.
3. Slow growth rate: the Heritage Turkey must have a slow to moderate rate of growth. Today’s heritage turkeys reach a marketable weight in about 28 weeks, giving the birds time to develop a strong skeletal structure and healthy organs prior to building muscle mass. This growth rate is identical to that of the commercial varieties of the first half of the 20th century.
(See http://www.albc-usa.org/cpl/turkdefinition.html or contact ALBC for the complete definition.)
The following data was have been collected:
(1) Days to Market Weight: Commercial birds performed better, as expected, attaining marketable weight in fewer days
(a) The commercial strain (BUTA Medium White Males) attained a marketable weight in 110 – 144 days (3.6 – 4.8 months)
(b) The Bourbon Red (Privett strain) attained a harvest weight in 162 – 190 days (5.4 – 6.3 months)
(2) Average Final Live Weight. Commercial birds grew larger, weighing one-third more both at harvest and dressed, as expected. The Bourbon Red turkeys have not been selected in recent years for meat production, including those sold through hatcheries like Privett. A few independent breeders have continued to select for meat production as well as the breed standard for both color and conformation for the variety. These strains, however, are not readily available to the public.
(a) Commercial strain – Toms: Ranged from 20 – 26.3 pounds. The median was 22.2 pounds.
(b) Bourbon Red – Toms: Ranged from 12 – 16.4 pounds. The median was 15 pounds.
(c) Bourbon Red – Hens: Ranged from 8.8 – 12 pounds. The median was 10.26 pounds
(3) Average Dressed Weight.
(a) Commercial strain – Toms: Ranged from 14.92 – 19.59 pounds dressed. The median was 17.52 pounds.
(b) Bourbon Red – Toms: Ranged from 9.17 – 11.95 pounds dressed. The median was 11.3 pounds.
(c) Bourbon Red – Hens: Ranged from 6.29 – 7.45 pounds dressed. The median was 7.38 pounds
(4) Average Percent Dress Out. Percent dress out was comparable between the commercial strain and the Bourbon Red. However, the smaller final live weight for the Bourbon Reds translated into smaller dressed turkeys, many under 10 pounds. This is problematic for the producers because of the economics (feed consumption & lower poundage for sale), and for the consumers because many have come to expect larger turkeys with more breast meat.
(a) Commercial strain: Ranged from 67% - 85% dress out. The median was 75%.
(b) Bourbon Red: Ranged from 70% - 79%. The median was 75%.
(5) Feed conversion. The Bourbon Red turkeys required 20% more feed per pound of weight gain than the commercial strain. Again, the commercial strain has been rigorously selected for feed efficiency and rapid weight gain. This may come at the expense of skeletal development, expressing itself as joint problems, especially in older breeds. The standard turkey varieties, like the Bourbon Red, develop their skeletal and internal organs prior to adding significant muscle mass.
(a) Commercial strain: Feed conversion ranged from 3.49 to 6.32 pounds of feed per pound of weight. The median was 4.99 pounds of feed per pound of weight. (Adjusted for mortality)
(b) Bourbon Red: Feed conversion ranged from 5.36 to 7.06 pounds of feed per pound of weight. The median was 6.03 pounds of feed per pound of weight. (Adjusted for mortality)
(6) Mortality. Mortality was higher among the commercial strain. Dr. Bill Pierson, Poultry Immunologist at Virginia Tech, had anticipated an even greater mortality. Mortalities occurred at three distinct times. Shipping stress was a significant cause of loss, especially among the commercial birds. We experienced a particularly cold spell when we were shipping birds to participants. This combined with problems resulting from a change in the US Postal Service contractor for expedited shipping contributed increased mortality. Brooder mortalities were also high. Improved management in the brooder, including preventing predation by rodents and snakes, will reduce these mortalities. Predation, especially in the final weeks of growth took a significant toll among the Bourbon Reds. Early preparation for preventing predation is necessary to curtail this expensive loss.
(a) Commercial strain: Participants lost between 13% and 93%. The median was 46%
(b) Bourbon Red: Participants lost between 15% and 31%. The median was 21%.
(7) Morbidity. All participants reported heat stress related behaviors in the commercial strain, including panting, reduced activity, standing in water, and “flushing”(watery manure as a result of increased water intake). One participant had most of his commercial strain succumb to a respiratory disease. Dr. Pierson said the lung and cardiac to body mass ratio is no longer in balance, making it very difficult for the birds to tolerate stressors. This imbalance can also result in spontaneous death. While autopsies were not performed, Dr. Pierson suspected that the mortalities in the commercial strain reported by the one participant were due to failure of the lung immune system caused by either a mycoplasma or E. coli.
(8) Economic analysis. When health related mortality is considered, economics is fairly similar. Caring for the birds to reduce brooder deaths and to prevent predation will enhance the economics of both the commercial and the standard varieties significantly.
(a) Production cost, adjusted for mortality. Production cost is calculated as cost per poult divided by the survival rate, plus cost for feed, plus the cost of processing. This sum was divided by the total average dressed weight.
(i) Commercial strain tom: $1.96/pound
(ii) Bourbon Red tom: $2.66/pound
(b) Pounds of meat produced, adjusted for health related mortality.
(i) Commercial strain tom: 9.46 pounds/poult started.
(ii) Bourbon Red tom: 8.95 pounds/poult started.
Other producers, who have begun raising standard turkeys following this on-farm phase of the study, and who experienced fewer mortalities, have reported that the industrial turkey cost ~$1.00/pound to raise, and the standard varieties averaged ~ $2.00/pound in production costs, less as volume is increased and the standard varieties are selected for production attributes.
The actual price per pound charged by participants in this study varied widely. One participant had no access to processing facilities or to markets. She gave her birds away. Others charged between $1.25 and $4.00/pound depending on their market. Recent data collected from new turkey producers show standard varieties selling for $2.50 – 4.50 per pound from the farm, and commercial strain turkeys selling for $2.00 - $2.50/pound. Some very high end markets retail the standard varieties for $6.00/pound.
Aim 1: To compare select production indices in standard and commercial turkey lines at various stages of growth.
The commercial turkeys gained more than twice the amount of weight than any of the standard varieties between nine and thirteen weeks of age (Figure 1). Based on Dr. Gogal’s observations, the weight gain of the turkeys was inversely related to the amount of the “flight or fight” response: as the bird’s “flight or fight” response increased the amount of weight that they gained decreased.
Aim 2: To employ an immune testing panel of assays to evaluate immunophenotypes of the standard turkey lines to the chosen commercial line.
The British United Turkeys (commercial strain) consistently had a lower Packed Cell Volume and total proteins values than the standard varieties at nine, eleven and thirteen weeks of age. For all three time points the heterophil/lymphocyte ratio decreased with one exception - an increase in the B.U.T at thirteen weeks of age. Trends in the average lymphocyte recoveries were variable within the standard and commercial varieties. During all three time points that were evaluated the commercial turkeys had a much lower mitogen response supporting the conclusion that they are functionally weaker to non-specific stimulation.
Egg Hatch and Infertility Analysis
Hatchability of the standard varieties was as a whole lower than what has been reported for the commercial line. However, the British United Turkey eggs were not evaluated at the same hatchery. Within the standard varieties, the Blue Slates, Bourbon Reds, and Narragansetts had the highest hatchability. The mean number of infertile eggs was lower for Royal Palms and Narragansetts (Table 1).
Packed Cell Volume and Total Protein Analysis
When compared to the standard varieties, commercial turkeys consistently had lower packed cell volumes and total proteins at nine, eleven and thirteen weeks of age (Table 2). This translates to a reduced cardiovascular capacity by commercial birds compared to the standard varieties. In short, the commercial birds would be far less mobile in a field management system. The low protein could reflect a lower overall antibody level (which was not evaluated) in the commercial strain, which is not favorable if the birds were exposed to an infectious agent.
Five Point Leukocyte Differential Analysis:
Average heterophil/lymphocyte numbers declined in all six varieties of birds during all three weeks, except for the commercial birds which increased in week thirteen (Figure 2). Bourbon Reds, Royal Palms and Blue Slates had the lowest mean heterophil/lymphocyte ratios of the six lines.
At nine, eleven and thirteen weeks of age, the Black Spanish, Bourbon Reds and Narragansetts had a consistent statistically significant decrease in the number of basophils. Royal Palms were consistent in their basophil numbers at nine and eleven weeks of age, but had statistically significant decrease at thirteen weeks of age (Figure 3). Blue Slates and commercial birds had a statistically significant decrease between nine and eleven weeks, then had a statistically significant increase between eleven and thirteen weeks of age.
Blue Slates had consistent monocyte numbers at nine and eleven weeks of age, but the percentage decreased at thirteen weeks of age. Black Spanish, Royal Palms, commercial birds and Narragansetts decreased in percentage of monocytes between nine and eleven weeks of age, then decreased at thirteen weeks of age. Bourbon Reds decreased in percentage of monocytes between nine and eleven weeks, then remained constant between eleven and thirteen weeks-of-age. (Figure 4)
In general, a decrease in percentage of leukocytes in the peripheral blood infers that an animal would be more susceptible to an infectious agent. A review of the trend differences in leukocyte expression, although statistically significant in some cases, did not reveal any biologic significance.
Average lymphocyte recovery increased between nine and eleven weeks of age, then decreased between eleven and thirteen weeks of age in the Blue Slates, Black Spanish, and commercial turkeys (Data not shown). Royal Palm average lymphocyte recovery decreased each for all three weeks of the experiment (Data not shown). Bourbon Reds, and Narragansetts average lymphocyte recovery increased each week for all three time points (Data not shown).
None of the trends in lymphocyte recovery showed significance across the varieties. In fact, the patterns of changes were consistent with what is expected during avian growth.
When compared to Blue Slate, Black Spanish, Royal Palm, and Bourbon Red, the commercial turkeys had a statistically significantly lower proliferation response to Concanavalin A (Con A) at a concentration of 5 µg/ml at nine, eleven and thirteen weeks of age. When compared to Royal Palms, the commercial turkeys had a higher proliferation response to Con A at a concentration of 5 µg/ml at nine weeks, the same amount of response at eleven weeks and a lower response at thirteen weeks of age. All five standard varieties had a higher proliferation response to Con A at a concentration of 50 µg/ml for all three time points. The commercial turkeys’ proliferation response to phorbol 12-myristate 13-acetate 0.02 µg/ml with ionomycin (PMA/I) at 0.40 µg/ml was lower than the Blue Slates, Royal Palms, and Bourbon Reds for all three time points. Black Spanish and Narragansetts had a higher proliferation response to PMA/I at nine weeks of age, and a lower response at eleven and thirteen weeks of age, when compared to the commercial turkeys. (Figures 5, 6, and 7).
These observations seem to support a functional inhibition or suppression of the lymphocytes of the commercial birds. Once again, this would seem support a general deficiency in the commercial birds’ immune system.
Analysis of the lymphocyte subsets from peripheral blood did not show any significant differences in the cell marker expression across all 6 of the varieties (data not shown).
I. Endogenous Production of Ascorbic Acid in Historic Turkeys
Assay of standard turkeys for breed and sex differences in endogenous ascorbic acid production. Plasma ascorbic acid concentrations were analyzed in samples from each of the 96 standard turkeys in the study. Fourteen birds out of the 96 were excluded in order to age-match the study. The 14 that were excluded were older than the remaining 82. It was found that there were statistically significant differences between the breeds (Figure 8) when analyzed with ANOVA (p-value < 0.01). The Narragansett turkeys had the lowest average ascorbic acid concentration (0.54) - approximately half that of the Black Spanish turkeys, which had the highest average plasma AA concentration overall. These results show that there are breed differences in endogenous ascorbic acid production in historic turkeys as measured by plasma ascorbic acid concentration.
The projected interpretation is that a higher level of basal ascorbic acid levels would be more protective against certain infectious agents that utilize reactive oxygen species as a mechanism for pathogenesis.
Analysis was conducted with 82 age-matched standard turkeys across 5 different varieties. Plasma AA concentrations averaged for each variety are shown here graphically. Statistical analysis showed significant difference in plasma AA concentration between breeds (p<0.01). The Narragansett turkeys showed an average plasma [AA] approximately half that of the Black Spanish turkeys, with the other 3 breeds falling in between.
When gender differences were analyzed across breeds, a statistically significant difference was found between males and females (Figure 9), with a p-value of 0.011. Within breeds, only the Blue Slate and the Royal Palm turkeys showed a statistically significant gender difference in plasma AA concentrations.
A statistically significant difference was found between gender across varieties while only the Blue Slate and Royal Palm standard turkeys showed significant intra-varietal gender differences in plasma [AA]. That is, AA levels were different across genders becoming statisically significant only in the Bourbon Red, Blue Slate and Royal Palm.
First, the study of plasma levels of AA in standard turkeys showed significant breed differences in average plasma AA. The birds in this study did not receive AA supplementation, thus the plasma AA concentration was solely the result of endogenous production. Differences in endogenous AA production in breeds may point to an adaptive mechanism originally acquired to protect against environmental stresses present in certain groups during breed divergence. Future genetic studies are required to validate such a hypothesis. A definitive study would include comparison of gene expression for enzymes in the ascorbic acid synthetic pathway (i.e. L-gulono lactone oxidase) among breeds.
Physiology and Applications
Analysis of Ascorbic Acid (Vitamin C) levels in the standard and commercial lines was a fortuitous study. Ascorbic acid (AA) has been shown to be physiologically multifunctional, enhancing function of the innate immune system, modulating gene expression, acting as a co-factor in enzymatic reactions, and protecting organisms from free radical damage during oxidative stress. Animals having high levels of ascorbic acid would be at an increased benefit. A unique feature with birds is that they synthesize their own ascorbic acid. Although this was not part of the original proposal, we decided to include the turkeys from this study in our ascorbic acid study. An exciting finding was that the same lines that had the more robust immune responses also had the higher mean ascorbic acid levels.
II. Comparison of Immunological Profiles between Commercial Line and Standard Varities of Turkeys Following challenge with HEV and E. coli
On day 0, the commercial birds exhibited a high proliferative response to the T-cell mitogen Concavalin A (100μg/mL) the Blue Slates and Royal Palms responded comparably. However, by day 6, post-challenge, the BUTAs exhibited the lowest proliferative response to mitogens compared to the standard varieties, Blue Slates, Royal Palms, and Black Spanish had the highest proliferative response to Con A. Additionally, the peripheral blood lymphocyte recovery increased for the standard varieties, but remained low for the BUTAs. (Figure 10). Following exposure to E. coli on day 7 of the study, the BUTAs demonstrated a negative response to the mitogens, and all but two had died: One died by day 10, one died on day 11. In contrast, the majority of the standard turkeys survived past three days of bacterial infection surviving to the study termination. (Figure 11).
These results indicate a significant difference in immunologic response exists between the BUTA production and historic turkeys. These observations support our theory that select lines of the standard turkey could serve as a contributing gene pool to the commercial lines, expanding its gene pool and allowing for other management options of these birds.
Examining all the data collected in the HEV/ E.coli challenge study, it is clear that the production BUT-8 line of turkey was the least immuno-competent breed studied. It adapted least to bacterial or viral infection. Conversely, as projected, the Blue Slates, Black Spanish and Bourbon Red breeds possessed the most competent immune systems, as they both had the highest survival rates when infected with E. coli, and had the highest lymphocyte responses of all the breeds during the HEV and E. coli phases of the experiment.
The following is excerpted directly from Biochemical Genetics, Vol. 43, Nos. 1/2, February 2005 (©2005)
Molecular Analysis of the Relatedness of Five Domesticated Turkey Strains
Edward J. Smith,1,3 Tuoyu Geng,1 Elizabeth Long,1 F. William Pierson,2
D. Phillip Sponenberg,2 Cal Larson,2 and Robert Gogal2
1 Comparative Genomics Lab, Department of Animal and Poultry Sciences.
2 Department of Biomedical Sciences & Pathobiology, Virginia Tech, Blacksburg, Virginia.
3 To whom correspondence should be addressed at 2250 Litton-Reaves Hall,Virginia Tech, Blacksburg, Virginia 24061-0306; e-mail: email@example.com.)
Abbreviations: EST, Expressed Sequence Tag; SNP, Single Nucleotide Polymorphism.
Tables & figures referred to in this section can be found in the published paper
In addition to the five SNPs detected within AY327574 using Consed, four SNPs were detected within AF268673 (Table IV). Three SNPs, including VPIcest200SNP1, VPIcest200SNP3, and VPIcest200SNP4, were previously detected in commercial turkey populations (Smith et al., 2001). The two Msl1 sites in the sequence of the amplified turkey fragment (AF268673) corresponding to nucleotide positions 259 and 578 were the basis of the PCR-RFLP method developed to genotype turkeys for the polymorphism detected among the strains at this locus (Fig. 2). The presence of a C allele at nucleotide position 259 created an Msl1 site in addition to the naturally occurring but monomorphic Msl1 site at position 578.
In total, six alleles, including two for each locus evaluated, were detected from the microsatellite analysis (Table V). Allelic frequencies at the microsatellite loci, as well as for the two sequence-based analyses, are also presented. The chart in Fig. 2 shows the allelic frequency from the PCR-RFLP genotyping of individual birds for the SNP identified as VPIcest200SNPl (Table IV). The VPIcest200SNPl frequency data suggest significant genetic differences among the strains, especially between RP and all of the other strains. The UPGMA tree implies genetic relatedness between the Royal Palm and Narragansett and a separation from the other three strains (Fig. 3). These data, however, were not entirely supported by the comparisons based on the three microsatellite loci used in the analyses (data not presented).
In the present work, we have described resources that may be useful in resolving an old question about the genetic composition of domesticated turkeys. Traditionally, and as reflected in the American Poultry Association’s Standard of Perfection, all turkey strains are classified as a single breed. To support this traditional classification, molecular evidence is needed as to the genetic relatedness among varieties. The molecular approach may, as implied in the current work, divide the traditional varieties into a few subgroups. The Royal Palm and Narragansett form one such group, and the Blue Slate, Spanish Black, and Bourbon Red form another group. The relatedness of the Royal Palm and the Narragansett is consistent with unpublished observations (at Web sites of poultry fanciers) that the two varieties may have a common ancestry. It is assumed, for example, that the color pattern of the Royal Palm is consistent with the assumption that it is derived from matings of Narragansett and another variety, as the Narragansett color mutation is a component of the final Royal Palm color. While these two varieties may be distinct strains, the sharing of common genotypes at many different loci suggests that they may not be distinct breeds in the traditional sense or the analyses are detecting common genotypes that underlie common phenotypes, such as the shared color genotype that characterizes these two strains. It should be noted that the Royal Palm is much smaller than all the other strains and may have only color as a common phenotype with the other varieties. The markers developed here may provide resources needed to address these important population genetics questions, including the question of whether genetic diversity is correlated with the ability of the different turkey strains to adapt to common or different environments.
The RAPD-, microsatellite-, and STS-based combinations of analyses also suggest a closer relationship among BR, BS, and SB strains. These data appear to support unpublished speculation based on historical records by Ms. Laura Phillips (www.natureofanimals.com/article1016.html) that BS is a product of matings that involved SB and other unknown turkey strains. According to Ms. Phillips, several groups including the Society for the Preservation of Poultry Antiquities and the American Livestock Breeds Conservancy consider BS a critically rare strain (www.albc-usa.org/slate.htm). knowledge of its genetic relatedness and origin may help biologists develop adequate plans for this strain’s preservation.
We used three different DNA marker systems to evaluate the genetic relatedness of five domesticated turkey strains based on polymorphisms observed at multiple loci. The systems, in combination and separately, were able to differentiate unambiguously the RP population from the other four turkey strains. Though not supported by SSR- and SNP-based analyses, the RAPD marker system also showed genetic differentiation of the Narragansett from BR, BS, and SB strains. The differences revealed by these marker systems, however, may not be indicative of adaptive divergence of the strains that may have arisen from emphasis on plumage color phenotypes by different breeders. The DNA markers in combination with the turkey strains established and maintained at different locations by ALBC will be useful in addressing questions of adaptive diversity among noncommercial turkeys. Though preliminary, results of the SNP analysis using two different fragments indicate a high density (a frequency of approximately 1 SNP in 100 bases) and pattern of sequence variation in the noncommercial turkeys similar to that previously observed in a commercial turkey population (Smith et al., 2001). The novel markers described could also be useful for other genetic studies, including being a resource for building a turkey genetic map. The PCR-RFLP method, for example, can be useful as both a marker and a population genetic tool. Other resources developed include three new microsatellite markers and SNPs within an RAPD-amplified fragment. The American Poultry Association lists in its Standard of Perfection eight turkey varieties, five of which were included in the current analysis. Although the data do not establish firm genetic relatedness, they provide resources that can be used to develop further molecular data that will complement the use of physical characteristics for classification. Such information can be complementary to phenotypic and other genotypic data required to validate the evolutionary relationships among turkey strains.
2006 Follow-up Genetic Study
Microsatellite data at these 5 loci for these samples was used to estimate relative similarities among varieties/strains using two approaches: (1)an individual-based approach using principle coordinates analysis (PCA), and (2) a group-based approach using UPGMA clustering. The primary finding in this preliminary analysis is one of low statistical signal (low bootstrap values, weak clustering within variety/strain). This means that, based on this set of samples, data within a variety did not cluster, indicating relatedness within that variety. Instead data scattered more randomly. This low signal could be the result of (1) low sample size for each variety/strain, (2) low number of loci and low polymorphism of some loci, (3) the effect of missing data for some of the samples, or (4) actual lack of signal due to interbreeding and common and/or recent derivation of these varieties/strains. Without additional samples and data it is impossible to distinguish among these possibilities.
To differentiate among varieties/strains and to assess genetic diversity within varieties/strains future studies would need (1) a large collection of pedigreed samples, to the extent possible, of each variety, and (2) genetic assessment using additional microsatellite loci (possibly up to 12 loci) or single nucleotide polymorphisms (SNPs).
This research has not been published. A more complete description of this work, including loci used, and tables and graphs of data may be obtained from ALBC or through SARE.
The hypothesis of the study was that standard varieties of turkeys are superior to commercial strains in immuno-competence (disease resistance) and perform better in range based production systems.
In the Field: Production attributes
The field trial results were reported in the 2003 annual report. To recap, the field trial compared a commercial turkey line, the BUT-8, a medium sized commercial strain, with a Bourbon Red turkey, the Privett strain, on 8 farms across the United States. The commercial strain performed better than the Bourbon Red in rate of gain attaining market weight in 110 – 144 days (3.6 – 4.8 months). The Bourbon Reds attained market weight in 162 – 190 days (5.4 – 6.3 months). The commercial birds also grew larger than the Bourbon Red, weighing one-third more both at harvest and dressed. Commercial strain toms ranged from 20 – 26.3 pounds with a median of 22.2 pounds. (Hens were not available for this project.) Bourbon Red toms ranged from 12 – 16.4 pounds with a median of 15 pounds. Bourbon Red hens ranged from 8.8 – 12 pounds with a median of 10.26 pounds. These results were not unexpected. The Bourbon Red turkeys have not been selected in recent years for meat production, including those sold through hatcheries like Privett. A few independent breeders have continued to select for production attributes as well as for breed standard for both color and conformation for the variety. These strains, however, are not readily available to the public.
Mortality in the field tests demonstrated superior immune function in the Bourbon Red turkey, the standard variety used in the study. Mortalities for the Bourbon Red turkeys averaged 21%, with a range of 15 – 31%. Mortalities were attributed primarily to lack of proper brooder management. BUT-8 line mortalities averaged 46%, with a range of 13-93%. The mortalities were attributed to shipping stress which resulted in the complete loss of several batches, lack of proper brooder management, heat stress, and respiratory illness.
The field study confirmed the hypothesis that the standard varieties of turkeys had superior immuno-competence in pasture based setting by its lower rate of mortality.
In the Laboratory: Immunophenotyping
Immune function studies were reported in the 2003 annual report. To recap the results all the studies support that select lines of standard turkeys possess genetic traits that code for enhanced immunity and thus, allow them to perform better under social and pathogenic challenge compared to the commercial lines. However, the BUT commercial line outperformed all the standard varieties in terms of weight gain only. In this production index, none of the standard lines came even close in weight gain although Blue Slates, Black Spanish and Bourbon Reds had the highest mean weights of the standard varieties. Egg Hatch and Infertility analysis obtained from the Avian Medicine Veterinary Research Center’s Hatchery showed that the Royal Palms and Narragansetts had the lowest mean number of infertile eggs. However, Royal Palms had the lowest percentage hatchability. The BUT eggs were not evaluated.
The immunologic data generated in the first year of the study clearly identified differences among the lines during their growth time-points. The blood smear cytology showed comparable leukocyte numbers during the 9 and 11-week period. However, by thirteen weeks the BUT birds had a much higher heterophil/lymphocyte (H/L) ratio than the standard lines. The H/L ratio is an indication of a stress response. The higher the ratio the greater the stress response, which is a negative factor in bird health. Lymphocyte proliferation assays were performed to assess the functional response of the birds’ lymphocytes to non-specific stimulants. At all time periods analyzed, the standard lines out performed the commercial line. Of the standard lines, the Bourbon Red, Royal Palm and Blue Slates had the stronger response to the stimulants. Packed red blood cell and total proteins analyses were higher in all of the standard lines compared to the commercial line which are all favorable endpoints. In summary, results from the immunophenotyping of the standard lines suggested that the Bourbon Red, Royal Palm and Blue Slates were stronger from an immunologic assessment.
A challenge study conducted tested the hypothesis that the high immune responders would have a better outcome when challenged. A viral-bacterial challenge experiment was included in the second year of the study. Examining all the data collected in the HEV/ E.coli challenge study, it is clear that the production BUT-8 line of turkey was the least immuno-competent breed studied as none of these birds survived the challenge. The BUT-8 line turkey is adapted least to bacterial or viral infection. Conversely, as projected, the Blue Slates, Black Spanish and Bourbon Red breeds possessed the most competent immune systems, as they both had the highest survival rates when infected with E. coli, and had the highest lymphocyte responses of all the breeds during the HEV and E. coli phases of the experiment. The Narragansett and Royal Palms suffered greater mortalities than the other standard varieties but fewer than the commercial line.
Analysis of Ascorbic Acid (Vitamin C) levels in the standard and commercial lines was a fortuitous study. Ascorbic acid (AA) has been shown to be physiologically multifunctional, enhancing function of the innate immune system, modulating gene expression, acting as a co-factor in enzymatic reactions, and protecting organisms from free radical damage during oxidative stress. Animals having high levels of ascorbic acid would be at an increased benefit. A unique feature with birds is that they synthesize their own ascorbic acid. The same lines that had the more robust immune responses also had the higher mean ascorbic acid levels.
The cumulative results of the studies suggest that there were significance differences in the standard turkey lines when compared to the commercial line in bird immunity. Further, some of these lines had stronger immune responses than others. Analysis of the ascorbic acid (Vitamin C) blood levels paralleled the line with the stronger immune response. Thus, if the industry wished to increase or expand the gene pool of the commercial/production turkey, serious consideration is warranted to the Blue Slates, Black Spanish and Bourbon Red based on their strong immunological performance. Evaluating other historic lines of birds may locate additional potential donors to the gene pool.
The results of the genetic analysis, performed by Dr. Ed Smith, supported the laboratory and field data. The Narragansett and Royal Palms appeared to be the most genetically similar. This proved to be an equally similar conclusion drawn from the results of the immune, Vitamin C and challenge studies.
The results from the follow-up DNA analysis conducted by Dr. Gogal and colleagues used a different analytical protocol. Results from this study did not segregate along varietal lines. The single nucleotide polymorphisms protocol may enable differentiation.
Inquiries were fielded throughout the years from farmers, media (radio, TV, newspaper, periodicals), and consumers. All were interested in standard varieties of turkey, though the area of interest and the depth of information requested varied. Significant on-going advise, council, and moral support has been provided to breeders and grow-out producers as they struggle with the challenges associated with being on the leading edge of a new and appealing enterprise in the context of a highly industrialized agricultural and competitive market environment. The lack of an infrastructure, specifically processing, that is independent of corporately held systems threatens to undermine producers at every turn. The lack of genetic lines selected for production attributes while maintaining biological fitness also thwarts progress. Growers across the country are exploring many models, including cooperatives and contractual relationships, to meet their urgent needs. Constraints due to processing alone could undermine this emerging market and slow farmers return to diversified livestock, poultry and crop farms.
A listing of publications and outreach events can be found in the Publication/Outreach section.
Turkey populations have risen substantially over the past ten years, which is most encouraging. An extensive survey of turkey breeders and hatcheries in the United States was completed in 2006. In 1997 the total population of naturally mating turkeys was 1335 breeding birds (male and female). In 2003, the total population had more than tripled, rising to 4412. In 2006, the population had more than doubled again, reaching a total of 10,404 breeding birds. (Table 3).
Individual varieties have at least doubled in number. (Table 4) Despite of this fabulous growth, all standard turkey varieties remain endangered and significant increases are needed to move them into the Recovering category and secure this valuable genetic resource, ALBC’s ultimate goal. The Standard Bronze made the most significant progress, increasing 439% from 441 in 2003 to 1,938 in 2006. The bronze color pattern is the image that most often comes to mind when people think about what a turkey looks like.
The Beltsville Small White and the White Holland turkeys both continue to languish. Both began with very small populations, so the road to recovery is longer. Both are white-feathered, and because “heritage turkeys” have been associated with colored-feather turkeys they have been less popular than the colored varieties.
Non-standard varieties of turkeys have increased by 350% between 2003 and 2006, growing from 106 breeding birds to 867. (Table 5) While this is good news, the population of non-standard varieties as a group and by individual variety continues to teeter on the brink of extinction. The Midget White and the Chocolate are the only exceptions. The Midget White is a known and named variety and has been listed on the ALBC Conservation Priority List (CPL) since 2000. This degree of visibility has supported its population growth. The population of the chocolate colored variety has increased for very different reasons. It is not listed on the CPL except insofar as it is implied in the listing of “Other Naturally Mating Varieties.” The name itself, Chocolate, referring to the brown color of the feathers, may have a real market appeal, connecting it with our fondness for chocolate confections and the popular Mexican mole sauce that contains Mexican chocolate.
The marketplace plays an increasingly important role in the success of standard turkey conservation. Since 2002 the market for “heritage turkeys” has approximately doubled annually, based on reports from producers and distributors in the ALBC network. For example, Good Shepherd sold:
- In 2002, 800 birds
In 2003, 1200 birds, a 50% increase;
In 2004, through a cooperative venture with a group of Kansas farmers, sold 4400 birds, a 260% increase, and
In 2005, 8900 birds, 102% increase.
In 2006, 6000 birds. A decision to double the size of the breeding flocks resulted in this decrease of 33%.
Educational & Outreach Activities
Mid-summer – fall 2002. Sunrise Valley Farm held a field day. CEFS/NCSU and Matzah Rising Farms were hosts for the SARE national conference. All three presented the objectives of the project to visitors. The other farms were closed to protect them from increasing disease threats.
Summer/Fall 2002. Four producers held field days. (1) Harry & Gail Groot held a farm field day for friends and potential customers. They presented the purpose of the research and their experience. They held a taste testing. While the results showed no clear preference for the standard or the commercial variety, their customers showed their preference for the standard turkeys with their purchases. (2) Gerry Cohn held a farm open house for customers, sharing the purpose of the research and his experience. (3) New England Heritage Breeds Conservancy is open to the public. Their flocks were easily viewed by the public. A taste testing did not reveal significant difference. (4) NCSU’s Center for Environmental Farming Systems hosted the SARE National Conference in Oct 2003. Attendees visited the flocks and learned about the research objectives.
March, 2003 NCSU posts an excellent documentation of their research results, including coop design. Go to http://www.cefs.ncsu.edu/frsu.htm. Click on "CEFS 2002 Pastured Poultry Project" or go direct to http://chatham.ces.ncsu.edu/growingsmallfarms/CEFSTurkey2002Report.pdf
June 23-24, 2003 American Association of Animal Science, Phoenix, AZ. Presentation to National Animal Germplasm Program about the issues of turkey genetic conservation.
September 7-8, 2003 Seeds and Breeds Summit in Washington, DC. Support for public funding of breeding programs for crops & livestock, including naturally-mating turkeys.
Oct 1 – 2, 2003 Turkey Breeder Clinic, Hutchison, KS. Brought together twenty breeders of standard turkeys from across the US and Canada attended a two day clinic on how to select birds for the breeding flock. Clinician: Frank Reese, Good Shepherd Turkey Ranch. Moderated by Marjorie Bender
Oct 3, 2003 ALBC Conference, Wichita, KS. “Recovery of the Heritage Turkey: Rediscovering Their Place in the Market.” Presented research data, future directions, market opportunities to a national audience. Marjorie Bender & Frank Reese. This was followed by a break-out discussion group for more in-depth discussion of standard turkey conservation and production.
Sept – Nov 2003 Marjorie Bender and Don Bixby fielded calls from journalists and reporters from across the country about standard turkeys. ALBC’s press file sports 38 articles we were able to collect. Such high profile publications as the USA Today, Chicago Tribune, LA Times, and Mother Earth News featured standard turkeys. ALBC’s publications – ALBC News and the Snood News – have included multiple articles on this research.
October 18, 2003 Don Bixby spoke at the Bioneers Conference in San Francisco. More than 500 farmers and supporters attended a presentation on the necessity of genetic conservation. ALBC’s turkey research was highlighted.
October 20, 2003. Don Bixby visited Slow Foods USA representatives in California, including their president. Discussion included ALBC’s turkey research & SF’s market promotion.
November 12, 2003 Slow Food USA’s NC Triangle convivium sponsored a promotional dinner at Panzanella featuring Matzah Rising Farm’s turkeys.
December, 2003 Developed “Heritage Turkey Information Packet.” Outlines the basic questions potential producers need to address to be successful. Includes a resource list and hatchery list. Distributed on request at no charge. As of April 2004 ALBC had distributed over 300 packets. Distribution continues.
January 25, 2004 Southern Sustainable Ag Working Group Conference, Gainesville, FL. “The Recovery of Heritage Turkeys” by Marjorie Bender and range production by Mike Walters, Walters Hatchery, Stilwell, OK. Approximately 40 farmers attended. Animated discussion of the emerging market for range-reared, standard turkeys was raised in other sessions. This is a hot topic!
February 8-9, 2004 Don Bixby attended the National Turkey Federation Annual Conference in Savannah, GA to better understand promotion, marketing, husbandry, and genetic issues in the turkey industry.
March 13, 2004 Organic Growers School, Flat Rock, NC, “The Recovery of Heritage Turkeys” by Marjorie Bender and range production by Alex Hitt, Peregrine Farm, Graham, NC. Approximately 40 farmers attended. Again, other poultry presentations discussed turkeys with great interest.
April 5, 2004 Central Carolina Community College Sustainable Agriculture Program, Pittsboro, NC. “The Recovery of Heritage Turkeys” by Marjorie Bender and range production by Alex Hitt, Peregrine Farm, Graham, NC. 12 students are enrolled in the class. Cooperative Extension Agents and reporters invited.
April 2004 ALBC staff visited Mike Walters Farm, Stilwell, OK Mike has been an active breeder and producer of standard turkeys. He has been an important outlet for poults and for dressed turkeys.
April 2004 Renewing America’s Food Traditions. ALBC is collaborating with 6 other non-profits in a project called Renewing America’s Food Traditions (RAFT). Other participants include Slow Food, USA; Chefs’ Collaborative; Seed Savers Exchange; Native Seed SEARCH; Cultural Conservancy; and the Center for Sustainable Environments at Northern Arizona University. The purpose of the collaboration is to draw attention to and promote traditional food and food processes such at the turkey breeds ALBC has been studying and promoting. The Cedar Tree Foundation has recently funded the collaboration for $750,000 over three years. The first RAFT effort was the publication “Renewing America’s Food Traditions: Bringing Cultural and Culinary Mainstays from the Past into the New Millennium”, edited by Gary Paul Nabhan and Ashley Rood. The book features a handsome Narragansett Bronze turkey on the cover and includes listing of the ten most endangered foods and ten foods already making a comeback because of conservation efforts. The heritage turkeys are listed in the comeback group, in part because of the support of SSARE research to understand the characteristics of these varieties for sustainable production.
July 2004 Seed Savers’ Exchange Conference. Don Bixby spoke at the Seed Savers’ Exchange summer conference in Decorah, IA, on integrating poultry into gardening. Information from ALBC research on turkeys was presented. A pairs of Narragansett turkeys was on display. Over 250 people attended.
July 2004 National Animal Germplasm Program Poultry Committee. Don Bixby attended the National Animal Germplasm Program Poultry committee meeting in conjunction with the American Society of Animal Scientists in St. Louis, MO. Since the turkey is the most genetically compromised of the industrial livestock species ALBC has been advocating for developing technique for and long term storage of turkey DNA in the national gene bank. Technology has yet to be developed for cryopreserving turkey semen, ova, or embryos.
July 2004 Elementary School Teacher In-service Training. Marjorie Bender presented information to 18 North Carolina teachers attending an in-service training on Noah’s Ark Today (NAT) conducted by ALBC in Pittsboro, NC. NAT is an educational program on biodiversity and livestock and poultry breed conservation developed for Kindergarten through fifth grade. Marjorie’s presentation on turkeys, based on SSARE funded research, helped to illustrate the value of genetic conservation among all livestock and poultry species.
July 2004 ALBC Special Event. Marjorie Bender presented research, marketing and census information to 35 people at an ALBC Special Event hosted by Hamilton Rare Breeds Foundation, Hartland, VT. Attendees included the Vermont Commissioner of Agriculture, and a reporter from the Wall Street Journal. Ensuing discussion focused on how the turkey conservation efforts can and should serve as a model for conservation, and effective partnerships that promote niche market development.
August 2004 Processing Forum. Processing continues to be the most tenuous link in the success of any independent poultry enterprise. A USDA inspected independent processing plant located in Chatham County, North Carolina, is the only one in a four state region. An area food company purchased this processing plant. The sale raised huge concerns for all independent poultry producers in the region. ALBC and North Carolina Cooperative Extention held a forum to inform farmers of changes in access and procedures. Without independent processors, all of the fruits of our efforts over the past several years would vanish.
September 2004 “How-to Produce Standard Turkeys on Range” and “Selecting Breeding Stock” Workshops. Marjorie Bender, Frank Reese and Bill Yockey conducted a two day workshop for the Pennsylvania Association for Sustainable Agriculture. Twenty breeders and producers attended this intensive clinic on range production of standard turkeys and breeder selection. This provided the template for ALBC’s recently funded Professional Development Program.
September – December, 2004 Fielded calls from media seeking information on standard turkeys for the holiday market. Coverage appeared in local, regional and national publications, most accompanied by interviews with local producers and photos.
November 2004 The Discover Conference in Cheyenne, WY. The Discover Conference is an intermittent gathering, by invitation only, sponsored by the American Dairy Science Foundation, for the purpose of uncensored intellectual exploration of livestock agriculture issues. The National Animal Germplasm Program of the USDA-Agricultural Research Service was implemented as a result of the 1999 Discover Conference. The 2004 conference marked the 5-year anniversary of the NAGP. Don Bixby’s presentation covered all the livestock and poultry species, and included discussion about the erosion of genetic diversity in the commercial turkey lines and the need for preserving turkey DNA.
November 2004 ALBC’s Annual Meeting, was held in Florida. Turkey research updates were provided to the members.
November 2004 Panzanella Restaurant held a benefit dinner for ALBC, in recognition of standard turkeys. Marjorie Bender and Don Schrider, ALBC’s communications director, attended and shared information about ALBC’s conservation efforts with attendees.
January 2005 Quivira Coalition meeting, Albuquerque, NM. The Quivira Coalition is a collaboration of ranchers, environmentalists, and managers of public lands, all of whom recognize that they have more in common than in conflict. Their work has been recognized and applauded for the progress made in restoring degraded range to economic production. Don Bixby presented at the meeting on integrating other species into the managed grazing systems. Turkeys were emphasized. There is a strong historical precedent for turkeys in the native American culture of the Southwest.
January 2005 Good Shepherd Turkey Ranch Summit, Tampa, KS. Frank Reese, owner of Good Shepherd, and the farmers he works with raised 5000 (4400 were salable) turkeys for the high-end Thanksgiving and Christmas market for 2004. The purpose of the meeting was to introduce the farmers, who are raising turkeys and chickens for Frank, to his team of advisors. Marjorie Bender and Chuck Bassett, ALBC’s executive director, traveled to the meeting. This was an excellent opportunity to share ALBC’s efforts and information about the turkeys with active producers, and researchers and extension specialists at the University of Kansas, and national marketers. Others are beginning to carry this work forward.
February 2004 Pastured Poultry Conference, North Carolina Cooperative Extension, Chatham County. ALBC and North Carolina Cooperative Extension – Chatham County, cooperated to present a full day conference on pastured poultry to farmers. Turkeys, chickens (broilers & layers), ducks and geese were all covered. Producers presented information on production how-to’s, ALBC staff presented information on appropriate genetics for outdoor production. The turkey research figured prominently in this conference. The capacity crowd of 130 included, as attendees, university extension specialists and agents, USDA and NCDA personnel, commercial producers, experienced independent farmers, and others exploring farming. (For many of the handouts and powerpoint presentations see www.ces.ncsu.edu/chatham/ag/SustAg. Visit the sections on Web Resources/Small-scale Livestock and also the Grower Resource List/Livestock)
February 2005 Virginia Biological Farming Association Conference. Don Bixby presented a workshop entitled “Beyond Pastured Poultry,” addressing the many issues of turkey and chicken production that is beyond the movable confinement cages. For example, and a primary lesson, was that confinement of turkeys to the low-profile, densely stocked “chicken tractors” borders on inhumane, not allowing the birds room to engage in healthy turkey behaviors.
March 2004 ALBC hosted two Swedish visitors who traveled to North Carolina to learn about the Center for Environmental Farming Systems, NCSU’s research and education related to sustainable agriculture, and their partners. These two professors were specifically interested in ALBC’s successful turkey research and conservation efforts.
March 2005 The American Grassfed Association is a maturing organization of producers dedicated to raising all livestock on pasture and forage, with ruminants raised on 100% forage. Field and laboratory research continues on the health and nutritional benefits to humans of eating grassfed meat and dairy products. Don Bixby conducted a workshop on the integration of turkeys and chickens into ruminant grazing systems with an emphasis on the seasonal production of high value turkeys for the holiday market.
April 2004 The Lantern Restaurant of Chapel Hill hosted a benefit dinner for ALBC. While the focus of the meal was the Ossabaw Island pig, the ALBC staff had opportunity to present short educational pieces needed to help this group of consumers understand the value breed conservation. Because of the success of the turkeys, this story was told to help people understand how conservation is done from the ground up.
January 2005 “Heritage Turkeys” defined. “Heritage turkey” has become the market identity for standard turkeys. Unfortunately, some producers have sold Broad Breasted turkeys, or crosses thereof, as “heritage,” getting premium dollar for their inauthentic product. Through the leadership of Frank Reese, a definition for heritage turkeys was developed that appears to have stopped this practice. It has also educated consumers. Heritage turkeys are defined as having long productive lives (5-7 years for hens, 2-3 years for toms), slow rate of growth (reaching market weight in 26-28 weeks), and reproducing through natural mating with a 70% or greater fertility rate. This definition has been embraced by the National Turkey Federation.
April 2005 ALBC awarded a Professional Development Grant from SSARE to develop a series of pamphlets and educational programming for humane, range-based production of standard turkeys. The pamphlets are currently being written.
July 2005 9th Continental Bioregional Congress, Black Mountain, NC Don Bixby was an invited speaker, providing information about integrating turkeys and other poultry and livestock in to an environmentally balanced agriculture.
July 2005 American Society of Animal Scientists, Cincinnati, OH Don Bixby represented the ALBC at the annual meeting of the species committees of the USDA-National Animal Germplasm Program. The poultry committee held significant dialog on characterizing turkey genetics as well as the research needed for the preservation of turkey genetic materials.
September 2005 Seeds and Breeds Conference, Ames, IA This conference was convened to develop a position statement for the upcoming farm bill. The goal is that the farm bill would include increased public support for the development of improved crops and livestock for feeding an increasing global population faced with a changing market, limited resources and environmental factors. Don Bixby was invited to present the issues of genetic characterization and conservation.
October 2005 Pastured Poultry PDP Workshop, Perryville, AR. Marjorie Bender attended this educational event by invitation, serving as a resource person for pasture turkey production and genetic conservation.
November 2005 ALBC Annual Conference, Greeley and Fort Collins, CO. Several varieties of standard turkeys were on display at the conference. Standard turkeys were served at the awards banquet. The keynote speaker was Dr. Vincent Amanor-Boadu, an assistant professor of Agricultural Economics at Kansas State University. He has advised the nation’s largest standard turkey producer, Frank Reese, in the marketing and branding of his product. In his typical charismatic manner, he shared his perspectives on niche marketing, punctuating it with many turkey examples.
November 2005 ALBC Special Event at Ayrshire Farm, Upperville, VA. Approximately 70 people gathered to celebrate rare breeds of livestock and poultry through creative cuisine, understanding that “we have to eat them to save them.” A fast-emerging trend in food consumption is the eating of rare and hard to find foods that have been sustainably raised. Epicureans are finding that meat flavor and texture varies by breed. This event demonstrated the wonderful and complex flavors represented in rare breeds of livestock and poultry. Standard varieties of turkeys are quickly being discovered for the rich flavor they bring to the table.
February 2006 Pastured Poultry Workshop, Pennsylvania Association for Sustainable Agriculture, State College, PA. ALBC organized and implemented a full day workshop on pastured poultry. Most were from Pennsylvania and surrounding states, but the conference drew people from across the country. Turkeys, chickens (broilers & layers), ducks and geese were all covered. Producers presented information on production how-to’s, ALBC staff presented information on appropriate genetics for outdoor production. The turkey research figured prominently in this conference. A marketing panel, that included chefs, a retailer, and a market developer, shared their experiences and needs when buying poultry directly from producers. The state veterinarian for the Pennsylvania Department of Agriculture reviewed biosecurity, an especially important topic as concerns for highly pathogenic Avian Influenza are increasing. The capacity crowd of 100 included university extension agents, PDA personnel, experienced independent farmers, and others exploring farming. A manual of the presentations and supportive material was distributed. A poultry listserv was established by PASA, at the request of ALBC to serve the attendees.
February 2006 Virginia Association for Biological Farming Conference, Lynchburg, VA. Marjorie Bender coordinated a workshop on the production of standard turkeys. Again, this research was presented. Harry Groot, a collaborator in the SSARE on-farm research, provided how-to production information. Harry and his wife, Gail, breed and raise Narragansett turkeys. They are selecting for survival attributes and broodiness. Dr. William Pierson, a research collaborator, presented information on diseases of turkeys, disease management and prevention. Biosecurity featured prominently. An enthusiastic 40+ farmers attended.
February 2006 Virginia Professional Development Program Annual Meeting, Lynchburg, VA. At the invitation of state sustainable ag coordinator Andy Hankins, Marjorie Bender & Dr. William Pierson presented research results on standard turkey comparison, and diseases of turkeys, disease management and prevention, and biosecurity. Approximately 20 extension and natural resource personnel attended.
August 2006 National SARE Conference, Milwaukee, WI. This project was invited to present both a poster and workshop at the National SARE conference. Marjorie Bender, Frank Reese, and Robert Gogal shared information from the research. Approximately 20 attended the workshop.
November 2006 Turkey Breeder Selection Clinic, Indianapolis, IN. This two day clinic on selecting breeding stock was attended by 25 people prior to the ALBC conference. It featured Good Shepherd Turkey Ranch (GSTR) owner and breeder Frank Reese, Lindsborg, KS and GSTR Board Member, breeder, and grower Danny Williamson, Tampa, KS. This clinic continues to be in high demand as many stewards are new to breeding. Fewer than 6 people in the US are master turkey breeders who learned at the feet of the previous generation. Hands-on education is critical if these skills are to be effectively passed to others.
January 2007 Turkey production clinic, Louisville, KY. Jeannette Beranger, ALBC Technical Program Manager, Mike Walters, Walters Hatchery, Stilwell, OK, and Holly Borne, Economics Specialist, ATTRA, conducted a 2 day clinic on standard turkey production at the SSAWG Conference. 20 people attended.
Smith, Edward J.,Tuoyu Geng, Elizabeth Long, F. William Pierson,
D. Phillip Sponenberg, Cal Larson, and Robert Gogal. “Molecular Analysis of the Relatedness of Five Domesticated Turkey Strains.” Biochemical Genetics, Vol. 43, Nos. 1/2, February 2005
Endogenous Production of Ascorbic Acid in Historic Turkeys Robert Gogal, F. William Pierson, Cal Larson, Virginia Tech. Status: A manuscript was submitted to Poultry Science in the fall of 2003. It was rejected not on the merits of the work, but rather the reviewers felt that the manuscript would be more appropriate in a nutrition-oriented journal. The manuscript was submitted in the spring of 2005 to a Nutrition journal. It was rejected based on the omission of HPLC analysis as a gold standard for measuring ascorbic acid. We are re-writing this manuscript with less focus on technique of AA identification and more focus on differences observed across bird and plan submission in the spring of 2007.
Immunologic Evaluation, by Dr. Robert Gogal, Virginia Tech. This paper is in draft form and is currently undergoing revisions. It is to be submitted for publication to Avian Diseases.
Comparison of Immunological Profiles between Commercial Line and Standard Varieties of Turkeys Following challenge with HEV and E. coli by Dr. Robert Gogal, Virginia Tech. This paper is in draft form and is currently undergoing revisions. It is to be submitted to Avian Diseases.
Heritage Turkeys in North America: The 2006 Census of Turkeys. By Marjorie Bender and D. Phillip Sponenberg. It is in the final stages of writing and will be published by ALBC upon completion.
Summary article drawing together the results of the on-farm research & the laboratory work by Marjorie Bender, ALBC. This document is considered the final summation of the research.
The number of breeder birds across the nation has increased over 600%, creating a significantly more secure future for these important genetic resources. (See results section for Objective 7)
The number of breeders (including hatcheries) maintaining breeding flocks has increased. According to the 2006 census a total of 81 breeders are breeding flocks,13 of which are hatcheries. This is a 44% increase since 2003.
A more extensive network of breeders and producers has developed which provides greater security (through distribution and increased participation) for these genetic resources.
A national demand for the flavorful and historic heritage turkey has developed. A new niche market has developed with the listing of several varieties on the Slow Food USA’s Ark of Taste and a subsequent presidium that has connected producers to consumers. This has literally breathed life back into heritage turkeys by giving them back their job. While this project did not track grow-out producers of heritage turkeys, the number of small and mid-sized producers across the nation has increased dramatically. Access to independently managed, USDA inspected processing facilities, and high quality breeding and grow-out stock remain the primary obstacles to long term adoption and success.
The cumulative results of the laboratory studies would suggest that there were significance differences in the standard turkey lines when compared to the commercial line in bird immunity. Further, some of these lines had stronger immune responses than others. Analysis of the ascorbic acid (Vitamin C) blood levels paralleled immune response. Thus, if the industry wished to increase or expand the gene pool of the commercial/production turkey, serious consideration is warranted to the Blue Slates, Black Spanish and Bourbon Red based on their strong immunological performance. Evaluating other historic lines of birds may locate other potential donors to the gene pool. Additionally, it is important to match a genetic resource to a production system. These results demonstrate that the heritage turkeys are better suited for outdoor production where incidental disease exposure may be greater. Therefore farmers could expect both lower morbidity and mortality when using the heritage varieties.
ALBC presentations have coupled the genetic and research information with farmer-led production information. This has been a great success. Additionally, ALBC and Frank Reese have developed a clinic on how to select breeding stock, a knowledge and skill set that has largely been lost through consolidation and industrialization of the turkey industry. Both of these educational programs are now regularly requested as this information is not otherwise available in print or in other programs. (See Publications and Outreach for listings)
Spin off projects have occurred:
SSAWG produced a brief video/DVD on raising heritage turkeys with SSARE funding. It featured Mike Walters, Walters Hatchery, Stilwell, OK.
SSARE PDP has funded the development of educational materials and clinics of the pasture-based production of heritage turkeys. ALBC is the PI.
SSARE has funded a sensory analysis of heritage turkeys. Anne Fanatico, ATTRA/NCAT is the PI.
Numeric information is not available, but it is clear from the niche market development and the corresponding increase in breeders and breeding stock that farmers have been adopting these practices and utilizing this information.
Areas needing additional study
Continued promotion of standard turkeys to potential breeders and the discriminating consumer.
Work with hatcheries to recover production attributes of the strains within the varieties to secure genetic diversity. Presently, Good Shepherd Turkey Ranch’s lines are the only ones known to meet both growth expectations and meet breed standards. Hatcheries, producers and consumers would benefit from this effort.
Provide education to individuals on managing breeding flocks and a hatchery and distributing poults.