point-of-care estrus detection for improved dairy cattle

Point-of-care Estrus Detection for Improved Dairy Cattle Farming in

Kenya

Dickerson, Matthew W.*; Milkey, Kendall1; Wong, Michelle1; Nakami, W. N., Odede, O. Rezin;Tsuma, Victor; Kipkemoi, K Nickson, and Osano, Odipo

Diagnostics For All (DFA).840 Memorial Dr Cambridge, MA 02139, USASidai Kenya LTD

University of Nairobi, Faculty of Veterinary Medicine, P. 0. Box 29053, NairobiUniversity of Eldoret, P. O. Box 1125 Eldoret, 30100

University of Nairobi

Introduction

• Diagnosis of stages of estrous cycle at the point of care – on the farms has a potential to improve dairy farming.

• Challenges farmers face that are related to estrous detection:– onset of estrous cycles in heifers,

– Pregnancy diagnosis in the first trimester,

– early pregnancy losses,

– cystic ovarian conditions

– infection-related abortions.

– Abortions due to manipulative pregnancy diagnoses during the early stages of pregnancies

– Misdiagnoses of pregnancies

– Silent estrous

Traditional means of estrous detection

• Human error prone

• reliance on assistants

• involves moving around the animal pens and chasing around animals that are lying down.

• Observation of typical behaviors of animals on heat.

• Others– Use of aid like chalk on tail heads

– Use of detector animals.

Diagnoses of stages of estrous by use of hormonal assays

• These offer alternatives to physical examination.

• Progesterone assays in blood and milk offer 95% accuracy in non – pregnancy cases and

• 80% accuracy of pregnancy diagnoses.

• Pregnancies can also be verified with ultrasonography.

DFA kit

• 1st rapid progesterone test designed for whole blood.

• Cheapest

Table 1: Bovine Estrus Test Competitive Landscape

Company Name of test Type of test

Sale price per test kit Sample

Reagent warm-up time (min)

Sample prep time (min)

Run & read time (min)

On-Farm Notes

DFA Bovine Estrus Test lateral flow < $3*whole blood

0 0 20* yes*estimated parameters: assay still in development, price includes sample collection kit

SPAN Biotech

Cow Pregnancy Rapid Test

lateral flow $3.00+ serum 0 20 10 yes

+ Sample Collection Kit cost not included; DFA tested with two samples: worked in serum, whole blood makes illegible. Also reported to work in saliva/hair/urine/milk

SPAN Biotech

Cow Ovulation Rapid Test lateral flow $3.00+ milk 0 20 10 yes+ Requires centrifuge. Tests for luteinizing hormone instead of progesterone

Ridgeway P4 Rapid dipstick $4.40 milk 0 0 10 yes

Biovet OVUCHECK Rapid Wellrapid well ELISA

$3.12 milk 180 0 30 yes

BioMetallics Target Bovine CL Chekrapid well ELISA

$4.40 milk 180 0 30 yes

Biolab Hormonost Farmertest ELISA $4.21+ milk 15 0 30 yes+ Requires $1028 Hormonostmicrolab

BiovetOVUCHECK Bovine Plasma

ELISA $11.84+ plasma 180 20 60 no+ Requires sample collection kit, centrifuge & $995 plate reader

Biovet OVUCHECK Bovine Milk ELISA $9.54+ milk 180 0 60 no + Requires $995 plate reader

Process of testing with DFA kit

• Affordable: the DFA kit is positioned to be lower price than any other kit on market. Unlike other kits, the low price includes sample collection supplies.

• Convenient: the DFA kit has everything needed to go from blood draw to results in less than 20 minutes. Blood test can be performed at any time; no need to wait for milking.

• On-Farm: a true cow-side test; no equipment required.

• Appropriate: test any cow; even heifers or cows who may have dried off; a particular challenge in low resource settings.

Requirements

• Obtain 1 ml of blood from the coccygeal vein, jugular, ear lobe or even tip of the tail.

• Transfer into heparinized tube

• Label the tube appropriately (i.e cow id, date of collection)

• Run point of care DFA test

35 µl Whole blood

Insert test strip

35 µl Diluent 75 µl ChaseProvided in the kit

Incubate for 10 minthen remove strip

Test set upNote that 1 ml of blood could be adequate for the test.

Read test strip against read chart provided in kit.

Total 20 minutes required to carry out Diagnosis.

PREGNANTNOT PREGNANT

LINENO LINE

Adopted from http://research.vet.upenn.edu/Dairy/Reproduction/EstrousCycle/EstrousCycleFollicularWaves/tabid/3965/Default.aspx

How To: Heat Detection with DFA Kit

Farmer Observes Signs of Heat

AI Provider Draws Blood

Farmer Calls AI Service Provider

AI Provider Visits Farm

AI Provider Runs Test

AI Provider Discusses Results with Farmer

Provide AI More Milk per Year and a High Quality Calf Earlier

NegativeTest

Result

Positive Test

Result

Expected Benefit of Improved Heat Detection

Heat detection with DFA kit can lead to shorter calving intervals.Today Average Breeding Scenario: Smallholder Farmers, Kenya

Calving Interval 21 months

Lactation 12 months

Annual % Days Lacatation 57%

Calves/year 0.57

Month 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21

Lactating 1 1 1 1 1 1 1 1 1 1 1 1 0 0 0 0 0 0 0 0 0

Pregnant 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 1 1 1 1

Attempts at AI AI wait to confirm AI wait to confirm confirmed!

Average Case w/DFA kit: Smallholder Farmers, Kenya


Lactation 12 months


Calves/year 0.71

Month 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17

Lactating 1 1 1 1 1 1 1 1 1 1 1 1 0 0 0 0 0

Pregnant 0 0 0 0 0 0 0 0 1 1 1 1 1 1 1 1 1

Attempts at AI 1 Dx AI+Dx wait 1 Dx AI+Dx wait to confirm confirmed!

Median & Shortest Case w/DFA kit: Smallholder Farmers, Kenya


Lactation 9 months


Calves/year 1.09

Month 1 2 3 4 5 6 7 8 9 10 11

Lactating 1 1 1 1 1 1 1 1 1

Pregnant 0 0 1 1 1 1 1 1 1 1 1

Attempts at AI AI+Dx wait to confirm confirmed!

Shorter calving intervals yield higher annual net farmer income.

Assumption:DFA Kit performs with clinical sensitivity of 75% and specificity of 75%.Specificity of behavioral heat detection is 85% (van den Berg, 2014)Conception rate is 61% for cows given AI during low progesterone. (R.D. Smith, 1984: IRD-10)Additional assumptions and references on Smallholder ROI Farmer Slide.

Net

An

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al In

com

e p

er C

ow

Figure 2: Net Farmer Income by Calving Interval with Heat Detection using DFA Kit

Calving Interval (months)

$-

$50.00

$100.00

$150.00

$200.00

$250.00

$300.00

$350.00

$400.00

11 13 15 17 19 21 23 25

Avg net income with DFA Kit

Avg net income today is $231

Figure 1: % Cows Pregnant Per Calving Interval

with Heat Detection Using DFA Kit

% o

f C

ow

s p

regn

ant

Calving Interval (months)

0.0%

5.0%

10.0%

15.0%

20.0%

25.0%

30.0%

35.0%

40.0%

45.0%

11 12 14 16 18 19 21 22 24

Average calving interval with DFA Kit

Average calving interval today is

21 months58% 1 AI

24% 2 AI 10%

3 AI 8% > 3 AI

Expected Benefit of Improved Heat Detection

The ROI for Smallholder Farmers

Heat detection with DFA kit can provide an average 9X ROI.

[1] Average calving interval among smallholder farmers in Kenya is 646 days. (Odima et al., 1994; Waithakaka et al. 2002)[2] Friesian cows dry off naturally around 12 months after calving. (Meyn et al., 1972)[3] On average, smallholder farmers in Kenya sell 4 L milk per day per cow. (Technoserve 2008)[4] Farm gate price for milk is 30ksh per liter at Brookside Dairy, the largest buyer of milk. (Daily Nation 2014)[5] Average AI service call costs 937 ksh for smallholder farmers in Kenya. Average two AI/year/cow (Makoni et al. 2015)[6] Assumes DFA kit will cost <$3 inclusive of sample collection supplies, and full heat detection services $5.25 including DFA kit. (final pricing depends on partner agreements)Photo courtesy Luigi Guarino

Table 2: Return on Investment of Heat Detection Services: Smallholder Farmers in Kenya

Average Today Average Case w/DFA Kit Median Case w/DFA Kit

Calving interval (months) [1] 21 17 11

Annual % Days Lactation [2] 57% 71% 82%

Income from Selling Milk at Farmgate

Additional annual per cow income [3,4] $ - $ 58.56 $ 105.24

Expenses for Reproductive Health

# of annual AI service calls per cow 2 1.1 1.1

# of annual heat detection service calls per cow 0 1.6 1.1

Additional annual per cow expense [5,6] $ - $ 5.79 $ 2.80

Return on Investment

ROI for smallholder farmer per cow 912%

The Additional ROI for FarmersGet calves and heifers milking earlier; weed out problem cows.

• New Calves Earlier: a new calf every 17 months instead of every 21 months means more cash flow earlier.

• Heifer to Cow Earlier: One high progesterone reading indicates your heifer is cycling. Watch for signs of heat in the next 2 weeks to get the animal pregnant and then milking earlier.

• Manage Non-Cycling Animals: one test per week for three weeks shows whether an animal is cycling at all. Fix the problem or cull the animal instead of continuing to sink expenses.

• Identify Unsuccessful Pregnancy Earlier: In the cases of precise record keeping, get your cow milking earlier by identifying false pregnancy. The DFA kit can be used at 21 and 24 days after AI as confirmation.

Photo courtesy Amy the Nurse

Farmer Willingness to Pay

Heat detection service call must cost less than AI service call.

Table 3: Total Cost of AI Service in Kenya [1]

Table 4: Estimated Cost of Heat Detection Service in Kenya

# Item/Description Cost (Ksh) Cost (USD) # Item/Description Cost (Ksh) Cost (USD)

1 Bull semen 400 $ 3.92 1

Estrus Detection Kit including sample collection supplies < 300 $ < 3.00

2 AI Service 350 $ 3.43 2 Estimate of Service Charge 100-275 $ 1.00-2.75

3 Transport 127 $ 1.25 3 Transport 127 $ 1.25

4 Lubricant 29 $ 0.28 4 $ -

5 Glove 10 $ 0.10 5 $ -

6 Liquid Nitrogen 8 $ 0.08 6 $ -

7 Depreciation 7 $ 0.07 7 $ -

8 Sheath 6 $ 0.06 8 $ -

Total 937 $ 9.19 Total 525-700 $5.25 - $7.00

• In interviews with ten smallholder farmers in Kenya, farmers expressed willingness to pay between USD$3 and $10 for heat detection services.

• Farmers were not taught anything about the value proposition. Final price for services will depend on commercial partner agreements for kit pricing and distributor and service provider agreements for heat detection service pricing.

• [1] Makoni et al 2015, Table 5

The Case for Support

[1] Statistics for # cows in Kenya receiving AI per year and projected annual AI market growth come from: Makoni et al, April 2015.[2] Average 1.6 heat detection service calls per cow per year, and average $53 per cow per year increased farmer income.

Over 5 years in Kenya, current investment could achieve a 27X ROI.

Table 5: Return on Investment of Heat Detection Services: Investing in the Market in Kenya

Annual cows receiving AI

DFA kit market share

DFA kit annual market Cumulative impact

Cumulative increased farmer income

(# cows) (% share) (# cows) (# cows) (USD)

Year 1 (launch) 980,000 5% 49,000 49,000 $ 2,585,647.08

Year 2 1,155,000 10% 115,500 164,500 $ 11,266,033.70

Year 3 1,330,000 15% 199,500 364,000 $ 30,473,697.71

Year 4 1,505,000 20% 301,000 665,000 $ 65,564,622.35

Year 5 1,680,000 20% 336,000 1,001,000 $ 118,385,698.38

Foundation Investment 1022219: milk contamination, aflatoxin, bovine estrus/pregnancy $ 1,988,318.00

DFID Investment 1022219: milk contamination, aflatoxin, bovine estrus/pregnancy $ 1,000,000.00

Foundation Investment 1078804: bovine estrus $ 1,267,971.00

Total Investment To-Date $ 4,256,289.00

ROI Factor To-Date 2681%

Thank You

point-of-care estrus detection for improved dairy cattle

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