Chapter 5 - Sampling and Testing

5.8 Sample Submission Procedures

5.8.1 Laboratory Sample Tracking System (LSTS)

There are four components to the LSTS which are of importance for field staff.

5.8.1.1 Sample Submission Form generator (LSTS/SSF)

5.8.1.1.1 Introduction

Samples for CFIA laboratories should be entered directly into the LSTS electronically. The Sample Submission Form generator is the tool used for this purpose.

The CFIA is in the process of introducing new software to handle the LSTS. During the transition period, both old and new systems are in use. The system to use depends on the type of sample being submitted.

The following sample submission forms are on the old (Citrix Metaframe) system.

Meat Inspection Sampling (CFIA/ACIA 5258)
Food Product Sampling (CFIA/ACIA 5164)

The following sample submission forms are on the new (Web-based) system:

Animal Health Disease Control (DC) Specimen Submission (CFIA/ACIA 5439), for tuberculosis, granulomas, pathology, cysticercosis, and suspected diseases other than TSE & Rabies
Animal Health Rabies (RAB) Specimen Submission (CFIA/ACIA 2908)
Animal Health Transmissible Spongiform Encephalopathy (TSE) Specimen Submission (CFIA/ACIA 5420)
Animal Health Import, Export and Artificial Insemination (IEA) Specimen Submission (CFIA/ACIA 5473)

5.8.1.1.2 Sample submission forms (metaframe)

If you have never used this application before, you will have to apply for access, by completing form CFIA/ACIA 5190, Request for System Access, and submitting it to your Regional Director for approval. Under "System requested", select "Input Application - PRODUCTION (LSTS)". Under "database", select "Prod". You may also wish to request access to the test/training database, to gain familiarization with the application.

Once access has been granted to run the application, first log on to the network. Then go to the Start menu on your computer, and select Start > All programs -> CFIA Applications > Lab sample tracking system (LSTS) > Submission forms - Metaframe (LSTS).

For testing of samples collected at slaughter, select "Food Products", then select "Meat Inspection Sampling". This will lead you through the generation of a form CFIA/ACIA 5258 (Meat Inspection Sample Submission Form).

For samples of imported or processed product, select "Food Products", then select "Food Product Sampling". This will lead you through the generation of a form CFIA/ACIA 5164 (Food Product Sampling Submission Form).

For samples for histopathology or parasitology, select "Pathology". This will lead you through the generation of a form CFIA/ACIA 1528 ("Pathology Specimen Submission").

Once you have "submitted" the form, print off two hard copies, one to accompany the shipment and one for your records. You must also affix a form CFIA/ACIA 1461 - Specimen Identification Tag to the sample, giving the job system number and other applicable information.

5.8.1.1.3 Sample submission forms (Web-based)

To log into LSTS and generate a submission form, click the "Start" button at the bottom left corner of your screen, and choose All Programs > CFIA Applications > Lab Sample Tracking System (LSTS) > LSTS User Services (2006). The Login page will be displayed. Enter Username and Password to enter the application.

If this is the first time that you are logging in, click on "First time User" at the bottom left of the page. Click the link in the e-mail and follow the online prompts. Your password and user name will be e-mailed to you.

Select English or French. The Landing page, showing the Web forms menu and the "other utilities" menu, is displayed. Use this page to create Web submission forms, perform a search, print a blank form, and to request Report of Analysis notification through PENMan or find test reports.

The first time you log in to LSTS User Services, you must select the submission forms that you want to work with. This is done by clicking on the "my preferences" link under "My LSTS". To add a form, select it in the "Default list" and click the "Add" arrow. The form is moved to the "User Selection" list. To remove a form, select it in the User Selection list and click the "Remove" arrow. Click Save to update your changes. The forms added to your User Selection list remain selected until you remove them. The forms that you select are then displayed on the Landing page and in the left-hand navigation tree on other pages. You can change your preferences at any time.

5.8.1.2 Reports of Analysis

The Reports of Analysis application is the tool for viewing test results. It is located on Merlin under LSTS User Services at: http://webapp/lsts/login.asp. The use of Merlin hyperlinks refer to an internal CFIA Intranet site for staff use only. A password is required to access this application.

If a notice of report of analysis is received electronically, the email message will contain a hypertext link which will take you directly to the result. Otherwise, you can search for a result by the reference number or system ID number, which are on the hard copy of the submission form.

5.8.1.3 Submission Status Inquiry

The Submission Status Inquiry tool is the tool for checking on a sample that you have submitted, and for which you have not yet received a result. It is located on Merlin under LSTS User Services at: http://webapp/lsts/login.asp. The use of Merlin hyperlinks refer to an internal CFIA Intranet site for staff use only. Select "Food Meat Inspection", then enter your search criteria. If you submitted the sample electronically using the Sample Submission Form generator, then you will have a "System no." from the hard copy of the submission form (CFIA/ACIA 5439, CFIA/ACIA 5164 or CFIA/ACIA 5258), and this will be the easiest way to locate the correct sample.

5.8.1.4 Personal Email Notification Manager (PENMan)

The Personal Email Notification Manager is the tool which you can use to have LSTS automatically notify you of test results submitted by you or your establishment. It has not yet been implemented for samples from the meat program.

If you enter your name on the sample submission form and you have an individual email account, a report of analysis notification is sent to you automatically by LSTS without requiring the use of PENMan.

5.8.2 Recording of livestock identification

When submitting samples collected at slaughter to CFIA laboratories, ensure that the animal identification number is recorded on the sample submission form, to permit traceback if necessary. On either form CFIA/ACIA 5258 (Meat Inspection Sample Submission Form) or form CFIA/ACIA 5439 (Disease Control Specimen Submission), enter it in the field marked "eartag" or "tattoo", as applicable. Also indicate the type of tag: Canadian Cattle Identification Agency (CCIA), Agri-Traçabilité Québec (ATQ), Canadian Sheep Identification Program (CSIP), etc.

For samples collected under the National Chemical Residue Monitoring Program, the animal identification number must be recorded on the page in the sampling plan booklet.

5.8.3 Samples for chemical residue analysis

5.8.3.1 Collection and preservation

Each tissue (skeletal muscle, liver, kidney, fat, etc.) for a particular sample must be in a separate bag, to prevent any residue which may be present from diffusing from one subsample into another.

Samples must be frozen and shipped in an insulated container with freezer packs, unless the product is shelf-stable.

5.8.3.2 Documentation

Each bag must be clearly identified using a form CFIA/ACIA 1461 Specimen Identification label. See section 5.8.1 for detailed instructions.

If the sample is from a carcass sampled during slaughter, and being submitted to a CFIA lab, the sample must be accompanied by form CFIA/ACIA 5258 Meat Inspection Sample Submission Form, which is generated when you enter the samples into LSTS.

If the sample is from portions or processed product not traceable to a single animal or place of origin, and being submitted to a CFIA lab, the sample must be accompanied by form CFIA/ACIA 5164 Food Product Sampling Submission Form, which is generated when you enter the samples into LSTS. See section 5.8.1.1.

In each case, a hard copy must accompany the sample. Alternatively, if the sampling submission form has been electronically generated and submitted, you can affix a form CFIA/ACIA 1461 - Specimen Identification Tag to the sample, giving the job system number and other applicable information. This will allow the lab to match the sample to the corresponding sample submission information in LSTS.

Samples submitted to private labs under the domestic National Chemical Residue Monitoring Program are not currently tracked in LSTS, and therefore do not require a CFIA sample form; the lab will match the sample to the sampling plan based on the sample number affixed to the sample. Because there is no accompanying submission form, it is critical that each bag have a CFIA/ACIA 1461, fully completed with NCRMP sample number, Establishment registration number, actual date sampled, sampling plan, species or production class, name of the inspector, and an identification of the tissue sampled ("Sample of").

5.8.3.3 Submission

Only the laboratory which is scheduled to receive the sample will have all the necessary information to complete the testing. It is therefore absolutely essential to submit the sample to the correct laboratory. The name and address of the correct lab is provided on every sheet in the sampling plan booklet. One establishment may make use of several different labs, so check each sheet carefully and ensure that the sample goes to the correct lab.

Ship the sample by courier for overnight delivery. Do not ship on a Friday unless you have verified that someone will be at the lab to receive the sample the next day. Product which arrives off condition will not be analyzed.

5.8.4 Samples for antibiotic confirmation

5.8.4.1 Collection and preservation

Each tissue (skeletal muscle, liver, kidney, fat, etc.) for a particular sample must be in a separate bag, to prevent any residue which may be present from diffusing from one subsample into another. Each bag must be clearly identified using a form CFIA/ACIA 1461 Specimen Identification label. See section 5.8.1 for detailed instructions. Ensure that the sample number or held tag number has been included.

Do not submit injection sites. Levels measured at the injection site cannot be used to determine carcass disposition, and the sample can act as a source of contamination for other tissues and for the lab.

The sample must be frozen and shipped in an insulated container with freezer packs, unless the product is shelf-stable.

5.8.4.2 Documentation

The sample must be accompanied by form CFIA/ACIA 5258 Meat Inspection Sample Submission Form, which is generated when you enter the samples into LSTS. See section 5.8.1.1.

Alternatively, if the sampling submission form has been electronically generated and submitted, you can affix a form CFIA/ACIA 1461 - Specimen Identification Tag to the sample, giving the job system number and other applicable information. This will allow the lab to match the sample to the corresponding sample submission information in LSTS.

If the carcass is held, ensure that "under detention" is selected under "sample priority" on the submission form.

Whether the carcass is detained or not, when the sample is sent to the lab, ensure that the owner's name and address are included under "submitter comments". This will simplify traceback in the event that the laboratory confirms a violative result.

5.8.4.3 Submission

Samples for antibiotic residue analysis should be shipped to the Centre for Veterinary Drug Residues, Saskatoon.

5.8.5 Samples for compositional analysis

5.8.5.1 Collection and preservation

The sample must be frozen and shipped in an insulated container with freezer packs, unless the product is shelf-stable.

5.8.5.2 Documentation

Samples must be accompanied by a form CFIA/ACIA 5164 Food Product Sampling Submission, which is generated when you enter the samples into LSTS. See section 5.8.1.1. In the box marked "Sampling plan no.", enter the number of the applicable sampling plan, prefixed by the fiscal year and an underscore, for example "2009_M104".

The form must be completed in sufficient detail to permit a rapid and efficient identification of the production lot sampled. For imported products the Import Inspection Report number must also be entered in the "identification code" field.

A hard copy must accompany the sample. Alternatively, if the sampling submission form has been electronically generated and submitted, you can affix a form CFIA/ACIA 1461 - Specimen Identification Tag to the sample, giving the job system number and other applicable information. This will allow the lab to match the sample to the corresponding sample submission information in LSTS.

In the case of an unsatisfactory laboratory result, the submission form must provide Ottawa with all necessary product information to initiate a recall or follow up investigation and in the case of imported products, to advise the foreign country.

5.8.5.3 Submission

Ensure that samples for compositional analysis are sent to the correct lab, as indicated on your sampling plan. Other labs will likely not be set up to perform the requested assay.

5.8.6 Samples for microbiological analysis

5.8.6.1 Collection and preservation

Micro-organisms from the environment, hands, clothing, sample containers, sampling devices, etc., may lead to erroneous analytical results. Stringent requirements for microbiological sampling are necessary. Use of aseptic sampling techniques and clean, sanitized equipment and supplies are of utmost importance.

Sterile gloves should be used for collecting samples. The only items which may contact the external surface of the glove are the exposed sample being collected and the sterile sample utensil. Keep in mind that the outside surfaces of the sample container are not sterile. Do not handle the inside surface of the sterile sample containers. Do not touch anything else.

Sampling and packaging instructions are to be found in the individual sections for each program. If special requirements should arise, they will be communicated in writing to all inspectors concerned.

Samples should be cooled down to refrigeration temperatures (4 - 8°C) as quickly as possible, which can be facilitated by using pre-chilled shipping containers and packing material. Samples in an individual bag or container can be put inside a second bag filled with insulation and ice or other coolant. Under no circumstances should the samples be subjected to freezer temperatures! Ice bags or freezer-packs are recommended to maintain a low enough temperature during shipping.

5.8.6.2 Documentation

A CFIA/ACIA 5164 Food Product Sampling Submission Form should be completed by the inspector and accompany the sample submission. The box marked "Sample plan no." must contain the sampling plan number (e.g. 2009_M200) and the "analysis requested" box must also be filled in. If completing a form manually for samples collected under MASS, this section should be filled in with the word "Bacteriology". If using the Sample Submission Form application, click on the button marked "Load per plan". The receiving laboratory will automatically subject the sample submission to the necessary test procedures called for by the program, based on the sampling plan number.

Two copies of the form should accompany the sample, and one copy should be retained by the submitter. Alternatively, if the sampling submission form has been electronically generated and submitted, you can affix a form CFIA/ACIA 1461 - Specimen Identification Tag to the sample, giving the job system number and other applicable information. This will allow the lab to match the sample to the corresponding sample submission information in LSTS.

The submitter should have an individual GroupWise email account to receive email notifications when the electronic Reports of Analysis (results) are available.

5.8.6.3 Submission

All samples, except cans, should be shipped refrigerated. The internal temperature should not exceed 10°C upon arrival at the laboratory.

Samples should arrive at the designated laboratory within 24 hours accompanied by the proper form. Several courier and express companies offer an overnight service. Stickers or labels of "Overnight Delivery By Air" should be put on cardboard boxes. These samples do not fall under the Transport of Dangerous Goods Regulations.

5.8.6.4 Acceptance criteria - CFIA food laboratories

Food samples submitted to CFIA laboratories for microbiological examination must be transported in a manner that will maintain the stability of sample characteristics (e.g. microflora, pH, moisture content, etc.) from the point of collection to reception by the laboratory.

5.8.6.4.1 Foods stored at room temperature

Foods usually stored or sold at room temperature (e.g. canned food, dried herbs and spices, etc.) should be sent using appropriate procedures to prevent any temperature abuse that could affect sample characteristics; the temperature should not fall below 0°C or exceed 35°C.

This type of food may include:

canned foods
dry mixes (soups, sauces, etc.)
spices (dried)
products without a "Keep refrigerated" declaration

"Dry cured" meats or other similar products that are stable at room temperature should be sent using the procedure for "Refrigerated Foods", below.

5.8.6.4.2 Refrigerated foods and environmental samples

Refrigerated foods include all commodities that are stored, shipped or presented to the consumer under refrigerated conditions.

Refrigerated samples, except shellfish and shellstock should be cooled and transported in a sample chest with suitable refrigerant capable of maintaining the temperature of the sample between 0°C and 7°C until arrival at the laboratory.

This type of food may include:

raw meats; or
processed meats.

Do not freeze refrigerated products unless otherwise specified. Frozen environmental samples will not be accepted for lab analysis.

5.8.6.4.3 Frozen foods

Do not allow sample units that are usually frozen to thaw during shipment; keep frozen samples solidly frozen at all times. Transport samples in a suitable container using ice packs or dry ice, where necessary to keep product in a frozen state.

This type of product may include:

frozen dairy
frozen fruits and vegetables
frozen dinners
ice (pre-packaged)
all types of frozen fish

5.8.6.4.4 Special samples

In some cases where the sample has a "high priority" status and it cannot be re-sampled, (e.g. food poisoning incidents, limited availability of imported product), and the above conditions are not met, the laboratory supervisor shall inform the submitter of the situation (e.g. frozen sample thawed, refrigerated sample higher than 7° or 10°C or in a frozen state, etc.) and agree on the action to be taken by the laboratory.

Any sample submitted to the laboratory that does not comply with the above guidelines shall be declared unfit for testing and the inspector notified.

5.8.7 Samples for BSE

5.8.7.1 Collection and preservation

Brain samples for BSE surveillance may be collected using the spatula technique or the spoon technique. The flushing technique is considered unacceptable because of the risk of generating a contaminated aerosol.

These techniques are described in the Bovine Spongiform Encephalopathy Manual of Procedures, Appendix 1 (Section 6.1).

Samples derived from BSE surveillance cases are classified as "Biological Samples - Non Regulated", and can be packaged and shipped accordingly. The availability of the rapid tests allows for submission of fresh tissue samples. Do not fix BSE surveillance samples in formalin. Do not freeze tissue samples.

Inner packaging
Samples should be submitted inside a leak-proof primary container such as a Whirl-pak™ bag. The primary container is to be placed inside a secondary receptacle containing sufficient absorbent material to absorb the entire liquid contents of the primary receptacle(s). The secondary receptacle could also be a Whirl-pak™ bag. Attach animal identification to the outer Whirl-pak™ to identify the specimen. Record this identification on the CFIA/ACIA 5420. The dry submission form and any other paperwork should be submitted outside the immediate sample container.
Outer packaging
The inner packaging and, depending on the number of samples, one or two freezer packs is placed in an outer packaging to protect its contents from outside influences such as physical damage and heat while in transit. The green metal "rabies can" may be used as an outer packaging if desired and available. A styrofoam cooler is also acceptable.

5.8.7.2 Documentation

Enter the sample information in the Laboratory Sample Tracking System (LSTS), by clicking on CFIA applications->Lab sample tracking system (LSTS)->User services 2006. Then select the "Create" link under "TSE". Under "Reason for test", select "Surveillance." Under "Disease category", select "Bovine Spongiform Encephalopathy." Then click on "Load all".

In all cases, ensure that all the animal's identifiers have been recorded on the laboratory submission form in order to facilitate traceability of the animal to the farm of origin, if required. In the "Samples" tab, under "National animal ID", record the CCIA tag or ATQ tag, as applicable. If there is a Health of Animals eartag, enter this number under "Health of Animals tag". If there is an auction or sale backtag, enter this under "Other ID". Any other identifying information, such as a Dairy Herd Improvement tag, herd management tag, or brand, should also be recorded under "Other ID".

Report the clinical abnormalities observed on ante mortem examination in the "Sample comments" section for that sample. Also indicate the category of risk animal that has been sampled (found dead, non ambulatory, emergency slaughter, ante mortem suspect). Categorizing the sample according to the risk population is important to allow verification that the high-risk populations of adult cattle are being included in the surveillance program.

Attach the LSTS Transmissible Spongiform Encephalopathy Specimen Submission form and any additional documentation to the outside of the outer packaging. This will ensure that the laboratory personnel are aware of the package contents upon receipt and prior to opening the outer packaging.

The shipper's waybill or form GC47 Government Bill of Lading documentation must accompany the shipment. Under "description of articles", indicate "Biological Sample - Non Regulated".

5.8.7.3 Submission

Routine BSE surveillance samples may be sent to any of the following CFIA laboratories: Lethbridge, Winnipeg, St-Hyacinthe. Select the laboratory to which you can most quickly ship samples.

The outer package must indicate the name, address, and phone number of the shipper and consignee.

In addition, the following statement should appear:
Emergency 24 Hour Number: 613-239-4604

These samples are not considered infectious substances (section 2.4.1 of the Transportation of Dangerous Goods Regulations) if the contents do not exceed 10 kg in weight.

5.8.8 Samples for scrapie

5.8.8.1 Collection and preservation

Brain samples for scrapie surveillance may be collected using the spoon technique as described above for BSE. Sample collection technique is described in more detail in Appendix 1A of the Animal Health Scrapie manual of procedures and on the training CD titled "Transmissible Spongiform Encephalopathies: Surveillance and Specimen Collection" published by the CFIA and distributed to all CFIA District Offices during the summer of 2005.

Place each sample in a Whirl-Pak™ bag, and attach a CFIA/ACIA 1461 Specimen Identification Tag. The four tissues from one animal may be placed in a single bag. Place this inside a second bag along with sufficient paper towel, cotton, or other absorbent material to hold any liquid which escapes from the inner bag.

These samples must be frozen prior to shipping.

5.8.8.2 Documentation

In all cases, ensure that all the animal's identifiers have been recorded on the laboratory submission form in order to facilitate traceability of the animal to the farm of origin, if required. In the "Samples" tab, under "National animal ID", record the CCIA tag or ATQ tag, as applicable. If there is a Health of Animals eartag, enter this number under "Health of Animals tag". Any other identifying information, such as a herd management tag, should also be recorded under "Other ID".

Report any clinical abnormalities observed on ante mortem examination in the "Sample comments" section for that sample.

5.8.8.3 Submission

Place the samples in a styrofoam container along with a freezer pack.

Submit the sample to the Ottawa Laboratory, Fallowfield. These samples do not fall under the Transport of Dangerous Goods Regulations. Follow instructions for shipping an "exempt animal specimen" published in the Shipping Biological Samples Guidebook.

Samples must be shipped by courier for overnight delivery. Samples must arrive at the designated laboratory within 24 hours accompanied by the proper form. Several courier and express companies offer an overnight service. Stickers or labels of "Overnight Delivery By Air" should be put on cardboard boxes.

5.8.9 Samples for parasitology

5.8.9.1 Collection and preservation

Specimens should be individually wrapped in a plastic bag and marked. They should be chilled to 4-8°C immediately. Do not freeze!

Samples should be shipped in bulk with sufficient ice or other coolant and insulation to ensure that samples remain in a fresh state while in transit. A double bag system is recommended. Samples in an individual bag or container can be put inside a second bag filled with insulation and ice or other coolant.

5.8.9.2 Documentation

Use form CFIA/ACIA 5439, Disease Control Specimen Submission. Ensure that the sampling date and animal identification for each specimen are reported. For farm trace-back, it is essential that the animal's identification, such as a tattoo or CCIA ear tag number, be recorded. If positive cases are found, the Veterinarian in Charge will be contacted in order to get the complete address of the affected producer for follow-up action at the farm. Traceability is critical for follow-up action.

A hard copy of the form must accompany the shipment. Alternatively, if the sampling submission form has been electronically generated and submitted, you can affix a form CFIA/ACIA 1461 - Specimen Identification Tag to the sample, giving the job system number and other applicable information. This will allow the lab to match the sample to the corresponding sample submission information in LSTS.

5.8.9.3 Submission

Samples should arrive at the Centre for Food-Borne and Animal Parasitology in Saskatoon within 24 hours accompanied by the proper form. Several courier and express companies offer an overnight service. Stickers or labels of "Overnight Delivery By Air" should be put on cardboard boxes. Specimens must be shipped to the laboratory according to the sampling schedule. These samples do not fall under the Transport of Dangerous Goods Regulations.

5.8.10 Samples for histopathology

5.8.10.1 Collection and preservation

Specimens for histopathology are normally forwarded in fixative. Specimens should be collected into wide-mouthed glass, or preferably sturdy plastic, jars with tight fitting lids, with fixative. As a rule, a jar should contain 20 volumes of fixative to one volume of tissue. Specimens should be collected as soon as possible after death of the animal and contain all stages of the lesion. For quick fixation, each specimen should be no thicker than 1 cm. The exception to this is brain, which should be placed entire and intact in formalin, while maintaining the ratio of 10 to 20 parts of fixative for one part of tissue.

A 4% formaldehyde (10% formalin) solution may be prepared as follows:

Ingredient	Volume of ingredient
Commercial formalin (40% formaldehyde in water)	50cc
Water	450cc
Table salt	3.8g

Containers must be leakproof. The cartons containing the samples must contain sufficient absorbent material (cotton, absorbent paper) to prevent leakage of contents in case of breakage.

Formaldehyde is exempt from the provisions of the Transport of Dangerous Goods Regulations, provided the quantity is less than 50 kg (Schedule VIII Table III) and the concentration is less than 25% (Schedule II List II). The package does not have to bear any special marking, but the waybill/bill of lading shall be marked with the statement: "Preserved Material, Non Hazardous, Unregulated".

If the volume of fixative is too large, tissues can be fixed locally for 24 hours, removed from formalin, and wrapped in paper towel previously soaked in formalin, then placed in a Ziplok® type of bag and send along with the CFIA/ACIA 5439 form to the lab. Do not use this procedure for whole brains; adequate fixation for brains may take up to 1 week, and premature removal from formalin for shipping may create artifacts which could affect the histology.

5.8.10.2 Documentation

Identify the submission "for histopathology" on the outside of the package, so that it will be sent directly to the pathology unit on receipt.

Samples must be accompanied by form CFIA/ACIA 5439, Disease Control. Under "Reason for test", select "Meat inspection pathology". Enter the history and gross pathological description under "Submission Comments/History." Alternatively, if the sampling submission form has been electronically generated and submitted, you can affix a form CFIA/ACIA 1461 - Specimen Identification Tag to the sample, giving the job system number and other applicable information. This will allow the lab to match the sample to the corresponding sample submission information in LSTS.

5.8.10.3 Submission

Samples from Western Area should be shipped to the Animal Disease Research Institute, Lethbridge. Samples from Ontario, Quebec, and Atlantic Areas should be shipped to the Laboratoire d'Hygiène Véterinaire, St.-Hyacinthe.

Note that samples in formalin can be shipped immediately, since they will continue to fix while in transit. It also does not matter if samples are shipped on a Friday, since they will not deteriorate in transit.

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