Phytosanitary and Regulatory Perspective on MLND in Kenya

George Ngundo

Plant Quarantine and Biosecurity Station

Website: www.kephis.org

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Kenya Plant Health Inspectorate Service (KEPHIS )

A state corporation in the ministry of Agriculturemandated to offer regulatory services in theagricultural sector.

VISION:The Lead Regulator And Facilitator Of Globally

Competitive AgricultureMISSION:

To Provide A Science-Based Regulatory Service ByAssuring Quality Of Agricultural Inputs AndProduce To Promote Food Security AndSustainable Development

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Legal FrameworkAgriculture Act, Cap 318

Agricultural Produce (Export) Act, Cap 319

Agricultural Produce Marketing Act, Cap 320

Plant Protection Act, Cap 324

The Suppression of Noxious Weeds Act, Cap 325

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Legal Framework Cont.

The Seeds and Plant varieties Act, Cap 326

The Fertilizers and Animal Foodstuffs Act, Cap 345

The Pest Control Products Act, Cap 346

Biosafety Act, 2009

Supported by a number of International Instruments such UPOV Convention, OECD seed scheme etc

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KEPHIS Activities in relation to MLND

Disease surveillance

Laboratory diagnostics of MLN

- For local and imported/local seed as well as post control testing of maize seed

- Has laboratory capacity to test MLN viruses

Centralized regulation of maize seed imports

Seed Certification process

Training and sensitization of stakeholders

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MLN Disease Surveillance

KEPHIS has been involved in the collaborative field surveys

Surveillance by inspectors from the Plant Quarantine and Biosecurity Station (PQBS)

MLN Surveillance coordinated by KEPHIS Pest Risk Analysis Team (KEPHIS-HQ)

Surveillance by KEPHIS Inspectors in other regional offices

Surveillance Samples analyzed at KEPHIS Plant Quarantine molecular laboratory for MCMV & SCMV

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Laboratory Diagnostics on MLND

Capacity Building on MLND diagnostics done at FERA-UK; January 2013.

Focused on Real-Time PCR assays for seed testing

Conventional PCR protocols for MCMV and SCMV also optimized.

A seed testing protocol developed and adopted

Currently involved in routine testing of MCMV and SCMV for advisory services and seed certification (local and imported seeds)

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Laboratory diagnostics

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Diagnostics for MLND done in the Molecular lab

Centralized regulation of maize seed/grain imports

Also imports of maize grain has been centralized

All Requests to import maize seed are controlled at KEPHIS-HQ Karen.

Maize seed importers required to send a representative sample to KEPHIS for MLND virus Screening prior to importation.

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Seed import/export Regulation

Only registered seed merchants are allowed to import/export seed

An SR14 (notice to import / export) form is filled to notify KEPHIS of the intention to import or export seed.

A Plant Import Permit is issued for the importation.

Seed can also be imported by researchers for experimental purposes

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Seed Companies Dealing In Maize Seed

Currently there are 19 seed companies dealing in maize seed in Kenya. These are

1. Kenya seed co,

2. Pannar seed ,

3. Agriseed co,

4. Western seed co,

5. Freshco,

6. East African seed co,

7. Dryland seed co,

8. Pioneer Hibred,

9. KARI seed unit,

10. Olerai ltd ,

11. Elgon kenya Ltd,

12. Monsanto,

13. Migotiyo plantations,

14. Faida seeds,

15. Pollen Ltd,

16. One Acre fund,

17. Crop Africa,

18. Gicheha and

19. Leldet Ltd.

Import regulation and Phytosanitary measures

MLN is now treated as a regulated non quarantine disease in Kenya. As such, seed samples are delivered to KEPHIS Plant health labs for screening for the two viruses.

A plant Import permit is issued for importation once the lot is declared to be free from the viruses.

The imported seed is subjected to seed testing in the Seed testing lab In KEPHIS, Lanet.

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Seed Certification Measures

Agreement with seed companies to implement recommendations for preventing MLND infection

Field Inspection of Seed fields is based on the minimum thresholds for rejection

Field inspection standards are to be adjusted to 1% at the final inspection, followed by 0 tolerance for lab tests

Seed companies have adopted the dressing of the seed with systemic insecticides e.g Thunder, gaucho, Cruiser.

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Inspectors in the field

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A seed Inspector in the field

A devastated seed field

Seed Certification Measures

Seed treatment with systemic insecticides and fungicides to control the vector and opportunistic fungi for the first 6-8 weeks. (Imidachloprid e.g. Gaucho, Thiamethoxam e.g. Cruiser).

Seed is also dressed with fungicides, eg Maxim and Royalcap

PCPB has been evaluating several systemic insecticides for seed dressing that can protect the maize crop in the field for a longer period

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National performance Trials (NPTs)

Screening of all maize varieties under NPT for

MLN tolerance. This is a collaborative effort with

CIMMYT.

Tolerant varieties to be fast-tracked for release.

One moderately tolerant variety (WE1101) has

been released

Participation in stakeholder awareness programs

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In 2013 and 2014, involved in MLND awareness program coordinated by MOA in collaboration with FAO and others.

Covered all maize production regions in the country

Focused on early detection of symptoms, and the available management options

Stakeholders awareness forums

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Olenguruon Bomet

On average these seed companies

handle 33,000 tonnes of seed maize

per year

of which 28,000 tonnes is locally

produced and

5,000 tonnes is imported.

Seed Production (2014)

Trans-Nzoia Machakos

Uasin-Gishu Thika

Narok Embu

Taveta Nandi

Tana River Bungoma

Baringo Kakamega

Nakuru

The concentration of maize seed production is in the

following areas;

In the year 2014 the average

hactarage of seed maize inspected

was 25,000 ha

of which only 86.2ha was rejected

due to MLND in the field.

This is about 0.34 percent of the total

production area.

Sampling was undertaken upon

harvesting for laboratory testing

Seed Production (2014)

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MLND Testing at KEPHIS -

Takes two weeks

RNA Extraction

Sample Reception

Sampling leaf material

Pre- germination

One Day

Two Days

One Day

qPCR One Day

7 days

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Use of Molecular Biology techniques

● Diagnosis using both conventional and real-time PCR is the

current practice for MLND

● Currently also used for diagnosis of CBSV, CMD, MLND

and Ralstonia spp.

Real time PCR (qPCR)

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Conventional PCR assays

Bv: 1 2T 3 4 5 2A 2A 2A 2A 2A 2A

Sugarcane mosaic virus

also tested using

conventional PCR

Virus specific PCR

primers

Sugarcane mosaic virus also tested

using conventional PCR

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+ve control

Buffer

Healthy

Positive samples

Double Antibody Sandwich (DAS)

ELISA assays

Since December 2014 and April 2015 the KEPHIS has tested a total of 1,005 samples

Two other labs have been trained and equipped to undertake MLND testing (KEPHIS Nakuru and Headquarters)

KEPHIS Nakuru has already started testing all samples from their region

The headquarters lab will commence as soon as a new batch of reagents is delivered

MLND testing at KEPHIS

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THANK YOU