BTSF. Better Training for Safer Food. Initiative. Sampling protocols and biosecurity measures. Geneviève LIBEAU. Peste des Petits Ruminants (PPR)

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1 Better Training for Safer Food Sampling protocols and biosecurity measures Geneviève LIBEAU Initiative Peste des Petits Ruminants (PPR) This presentation is delivered under contract with the Consumers, Health, Agriculture and Food Executive Agency ( The content of this presentation is the sole responsibility of Opera S.u.r.l., the Istituto Zooprofilattico Sperimentale Lombardia e Emilia Romagna and the State Food and Veterinary Service of Latvia and it can in no way be taken to reflect the views of the Consumers, Health, Agriculture and Food Executive Agency or any other body of the European Union. The Consumers, Health, Agriculture and Food Executive Agency or any other body of the European Union will not be responsible under any circumstances for the contents of communication items prepared by the contractors. BTSF Sofia, Bulgaria 6 th -8 th June 2018

2 Training Workshop Peste des Petits Ruminants Title: Sampling protocols and biosecurity measures Name: Geneviève LIBEAU CIRAD Astre Unit, Montpellier FRANCE FAO, OIE and EU reference laboratory 6 th -8 th June 2018 Sofia, Bulgaria 2

3 Collection of samples for the purpose of PPR diagnosis Confirm the current or retrospective infection of animals with PPR Collect the suitable samples to confirm PPR infection Ensure biosecurity is practiced during clinical investigation and onward Respect procedures for transport and shipment

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5 Virus shedding Incubation Clinical signs in acute cases Period Erosive Phase Diarrhea Convalescence 2-10 days multiplication of the virus Prodromal phase 4-8 days epithelial oral lesions become necrotic erosive Secretions become muco-purulent In mild cases oral erosions are reduced; convalescence One week days C inappetence, marked depression somnolence Starting of the secretions 4-8 days Diarrhea T drops In peracute cases the clinical signs are limited to high fever, severe depression and death; first occurrence in naïve pop.;

6 Procedure for specimen collection Truong et al.,plos One Jan 30;9(1) Uninfected/unvaccinated Infected Recovered/vaccinated Post-mortem Specimen to collect Tests to perform Test results Serum Antibody ELISA VNT NEGATIVE Absence of Ab Conjunctival, buccal cavity, nasal and rectal swabs, whole blood in EDTA Isolation VERO-SLAM LN and affected sections of the alimentary tract omasum, duodenum, ileum, caecum, lung, spleen* Gene and Antigen RT-PCR, ICE, field test, Immunostaining POSITIVE by One or several tests Serum Antibody ELISA VNT POSITIVE Presence of Ab

7 Principles of collection and dispatching PPRV specimen Sample collection: Conventional sampling on live animals (e.g. eye, mouth, rectal swabs. Serum for serology). The swab stick is cut to release the cotton end of the swab into a cryovial Non-invasive sampling, faeces/droppings (both with rectal swabs will be related to one animal and to the other samples taken from this animal). Post-mortem, spleen, lymph nodes, pieces of intestine, lungs, liver and tonsil. Kept cold during sampling 7

8 Principles of collection and dispatching PPRV specimen Sample preservation for shipment: The vial with the specimen is then well secured with the cap, labeled, surface decontaminated, recorded (excel). It is advised to keep specimens cold from the time they are collected until they arrive at the laboratory Specimens arriving in the laboratory in a decomposed state will not be processed, since processing and culture of these tissues lead to meaningless or erroneous results. 8

9 Principles of collection and dispatching PPRV specimen Dispatching PPRV specimen to the testing laboratory: The tubes containing specimen are wrapped in absorbent material and put in the secondary container The secondary container should be fluid-tight (with a screw cap and a rubber washer) Specimens should then be dispatched to the testing laboratory in insulated containers with carbide ice or a sufficient number of ice packs to last 48 to 96 hours and with correct external label (urgency, address, T ) The specimen inventory is dispatched in the same package

10 Sampling protocols and biosecurity measures Biosecurity practiced during clinical investigation exercise: Disinfectant Foot Dip Risk for disease transmission may be associated with personnel especially during surveillance and farm visits. Biosecurity is a set of practices designed to prevent the spread of the disease-causing agent. It is vital that disease investigators lead examples by observing biosecurity procedures because it will be difficult then to persuade other staffs and farm visitors. 10 dirty areas

11 Sampling protocols and biosecurity measures Biosecurity practiced during clinical investigation exercise: During PPR investigations sampling and epidemiological information collated while maintaining biosecurity: Wear disposal personal protective equipment (PPE), gloves, overboots etc.. Carry out cleansing and disinfection before and after visiting any farm Strict segregation between dirty and clean area are essential Quarantine period for the persons entering the dirty area should be observed (at least 5 days for PPRV). Containers, vehicles coming from the dirty area where the clinical team is, should be decontaminated before exiting this area. Disinfectant Foot Dip 11 dirty areas

12 The International Air Transport Association (IATA) Dangerous Goods Regulations Responsibility and definitions underlying IATA 6.2 regulations Transportation of infectious material is subject to international regulations applicable to any mode of transport and based upon technical instructions of the ICAO* and IATA Dangerous Goods Regulations. Transportation of infectious material is the responsibility of the biologist sending the material. Thus the sender is responsible of his parcel for the duration of the transport. He must therefore ensure with its partners, with a prior agreement that the samples will be forwarded as quickly as possible and in accordance with both regulatory and technical requirements. *International Civil Aviation Organization

13 The International Air Transport Association (IATA) Dangerous Goods Regulations Responsibility and definitions underlying IATA 6.2 regulations Animal biospecimens: Are considered as dangerous goods, which are substances capable of posing risk to health, as they may contain pathogens (microorganisms or prions) known to cause infectious diseases in humans and / or animals, to safety and environment. 9 UN hazard classes; Class 6. is relevant for biological specimen; division 6.2, for infectious substances. Infectious substances are divided into: - Infectious Material, affecting human (category A) - Biological Material (category B) - Exempt (human or animal samples) - Live infected animals

14 The International Air Transport Association (IATA) Dangerous Goods Regulations Transport planning check list: The sender must ensure Correct designation, labelling and documentation related to all infectious diagnostic specimen Classify infectious material (generally B) Select the packaging according to the regulations in force Choice of shipping conditions (packaging, shipping temperature, accompanying documents and means of transport) Inform the receiver laboratory so that the material is transported safely and arrives on time and in good conditions

15 The International Air Transport Association (IATA) Dangerous Goods Regulations Transport planning check list: The laboratory must ensure Obtaining the necessary authorizations from national authority Good coordination between the sender and the carrier Arrange with the carrier the most timely and efficient collection at departure and arrival Provide the carrier with the import permit and other documents Help the sender for designation, classification of the infectious material and choice of the carrier and shipping conditions

16 The International Air Transport Association (IATA) Dangerous Goods Regulations Biological substance, category B Samples of animal (or human) origin transported for diagnosis or research purposes, not a pure bacterial of viral culture such as tissue samples An infectious substance which does not meet the criteria of category A, Eg : Bacillus anthracis (samples taken from an animal) EXEMPLE D IMAGE Highly pathogen Avian Influenza Virus (samples taken from an animal) Infectious substances are assigned to UN shipping numbers according to their nature: UN 3373 / Biological material Class B 16

17 Packing Instructions 650 (P650) Triple packaging system is used for biological materials, category B EXEMPLE D IMAGE 17

18 Accompanying documentation: Biological substance, Category B No Dangerous Goods Declaration. A proforma invoice. Air waybill "(LTA) The Import Authorization (obtained by CIRAD with the French Veterinary Services) if necessary Weight and volume Limitations for shipping (excluding over packs) for Category B: Maximum 4 litres or 4 kg per package for passenger aircraft or cargo flights Maximum 1 litre per primary container for passenger aircraft or cargo flights 18

19 Exempt human / animal specimen Samples that do not contain: Or dry ice / infectious substances / Or biological substance - Class B / Or genetically modified organisms (fedex domestic) Low probability of pathogens in the sample (serum samples for PPR) No UN number but marked "Exempt animal specimen" (except animal sample) EXEMPLE Packaging must include three elements: the leak proof primary receptacle; a watertight secondary packaging; D IMAGE and an outer packaging of sufficient strength. 19

20 Exempt packaging requirements Specimen Primary container leakproof or siftproof Absorbent and cushioning material Secondary container leakproof or siftproof (eg. Sealed plastic bag) EXEMPLE D IMAGE Outer packaging 20

21 Classify infectious material Situation Categorization Proper Shipping Name UN Number Animal presents with suspected case of PPR, swabs samples are sent to a reference laboratory Category B Biological substance Affecting humans or animals UN 3373 Sending a culture of - Mycobacterium tuberculosis to a reference laboratory - PPRV Category A Infectious substance, Affecting humans Affecting animals UN 2814 UN 2900 Ebola virus has been found in bats and samples which have not been cultured are being shipped Category A Infectious substance, affecting humans UN 2814 A vet declares that samples collected for serosurveillance (seromonitoring) of PPR have only minimal likelihood of containing pathogens (or PPRV) Exempt Exempt animal specimen -

22 Infectious material classified in category A

23 Team involved 23 Arnaud Bataille PPR expertise, evolutionist, reverse genetics methodology Renata Servan de Almeida sirna methodology, animal challenges Geneviève Libeau PPR expertise, ELISA and vaccine development Olivier Kwiatek PPR expertise, Lab techniques Emmanuel Albina Virology expertise Tetiana Kwan Tat Intellectual Property CIRAD (ASTRE, UMR117) Thank you

24 This presentation is delivered under contract with the Consumers, Health, Agriculture and Food Executive Agency ( The content of this presentation is the sole responsibility of Opera S.u.r.l., the Istituto Zooprofilattico Sperimentale Lombardia e Emilia Romagna and the State Food and Veterinary Service of Latvia and it can in no way be taken to reflect the views of the Consumers, Health, Agriculture and Food Executive Agency or any other body of the European Union. The Consumers, Health, Agriculture and Food Executive Agency or any other body of the European Union will not be responsible under any circumstances for the contents of communication items prepared by the contractors. Thank you! OPERA Viale Parioli 96, Roma - Italy Tel / Fax info@opera-italy.it; Better Training for Safer Food BTSF European Commission Consumers, Health and Food Executive Agency DRB A3/042 L-2920 Luxembourg