Public Health and Environmental Laboratories

The Rabies Unit of the Public Health and Environmental Laboratories (PHEL) facilitates human prophylaxis programs AND supports rabies prevention and control programs in animal populations.  It is the only rabies TESTING laboratory in New Jersey.

The Rabies Unit is responsible for providing safe, accurate and timely laboratory testing of the rabies virus in animal (not human) tissue.  

The Rabies Unit works closely with the Communicable Disease Service (CDS) as well as Local Health Departments (LHD), veterinarians and animal control officers across the state. PHEL is required to report all positive rabies results directly to the appropriate LHD and to CDS. 

If you see an animal that you suspect may have rabies, 
do one or more of the following:

 1) Contact your Local Health Department

 2) Contact an Animal Control Officer

 3) Call Non-emergency police contact

Signs of rabies virus in animals may vary by species and stage of the disease.

The most obvious signs and behaviors associated with rabies in animals (wild or domestic) is abnormal behavior, such as:

• Nonspecific symptoms including lethargy, fever, vomiting, paralysis, seizures, anorexia, difficulty breathing or swallowing
• Wild animals that seem unusually tame or unafraid of an approaching person or domestic animal 
• Nocturnal animals such as: skunks, foxes, or bats, that are out and active during daylight hours
• Animals behaving strangely such as moving in circles, excessive salivation, aggression, air biting or self-mutilation
• Bats that are unable to fly or have been caught by a domestic animal (for example a domestic dog or cat)
• Animals that have difficulty walking, eating or drinking

Fortunately, rabies post exposure prophylaxis (PEP) with human rabies immune globulin (if given in a timely manner following exposure) and human rabies vaccine can be effective in treating and preventing rabies, respectively.  

Following an exposure, rapid and accurate laboratory testing of the infected animal is essential to enable appropriate preventive treatment in humans.

Rabies testing and diagnosis is done exclusively using postmortem animal brain tissue. The PHEL Rabies Unit follows the CDC’s National Standard for the Laboratory Diagnosis of Rabies and disposal of specimen materials. The diagnostic procedure followed is the Direct Fluorescent Antibody Testing https://www.cdc.gov/rabies/pdf/rabiesdfaspv2.pdf  

Antigen detection by "dFA"

The dFA test is based on the observation that animals infected by rabies virus will have rabies virus proteins (antigens) present in the brain tissue.  The test uses a fluorescent marker-labeled anti-rabies antibody. When labeled antibody is incubated with rabies-suspect brain tissue, it will bind to the rabies antigens. Any unbound antibody is then washed away and areas where antigens are present in the brain tissue can be visualized as specific patterns of apple-green cell staining using a fluorescent microscope (see below, left). If rabies virus is absent there will be no staining (see below, right).  There are unique patterns of staining and the interpretation of findings requires extensive training, routine proficiency testing, CDC training and concurrent interpretations of results between qualified "readers".

    

IMPORTANT information regarding specimen submission

Note:  Once submitted, specimens and associated materials are managed as regulated biological waste and MUST be incinerated (see NJ regulatory guidance).  The rabies laboratory processes many specimens each day, each may contain or become contaminated with highly pathogenic organisms (including several pathogens currently classified as "select agents").  Once accessioned into the rabies Unit any additional manipulation, or handling, places staff at increased risk and increases the likelihood of human error in reporting.  All rabies unit procedures have been Bio-Saftey Risk Assessed to assure proper specimens handling and disposal of contaminated materials.  This protects both staff and the public from potential exposure to dangerous pathogens. 

No tissue, head or carcass can be returned to providers, the public or veterinarians.

Current Rabies Sample Submission Guidelines include the following:

• Only decapitated animal will be accepted for rabies testing, except for bats, which shall be submitted intact.   Even small animals e.g., kittens will need to be decapitated.  Any animal not decapitated will be required to be picked up (WITHIN 24 HOURS) by the sender-provider, for decapitation of the animal, prior to testing. 
• All non-bat, rabies samples (heads) must be drained of blood before packaging for timely shipment to PHEL
• Each sample must be accompanied by a fully completed VIR-16 Form; including the county of exposure. 
  A completed form will assure that testing can be performed promptly and that results are routed correctly.
• When possible, arrange for delivery to PHEL by, or before, Thursday morning, to facilitate prompt testing and reporting.
• Please inform all persons who package specimens to follow the PHEL packaging instructions for rabies specimens.
  
  Compliance with ALL required packaging instructions is mandatory:
  
  https://www.nj.gov/health/forms/vir-16instr_2.pdf  
  
  The following list identifies specific things that can significantly delay testing:

1. In order to protect the Rabies Unit staff, do not wrap live bats in towels.
2. Do not freeze samples, freezing delays testing and increases the chance that samples will be unsatisfactory for testing.
3. Place specimens in a watertight containers such as a leak-proof, coffee can, not something that sharp teeth can puncture, for example a thin plastic bag.
4. During hot weather, pack the outer surface of a specimen container with sealed coolant packs
5. Ensure that a fully completed and legible current (April 2016) VIR-16 form
   ( https://www.nj.gov/health/forms/vir-16.PDF ) accompanies each specimen and is placed in a SEPARATE sealed water proof bag.
6. Samples received without a completed VIR-16 form cannot be processed until a completed VIR-16 form is received by PHEL. This will delay rabies testing.
7. Make sure that your FAX machine is maintained, that you have provided the correct number and it is in working properly.
8. Ensure that all scalpels or instruments used to decapitate are removed from packaging material.

Specimens should not be sent to the PHEL without consulting with the Local Health Department (LHD) where the rabies incident occurred.
A directory of LHDs is posted online: www.localhealth.nj.gov. 

Packaging and Shipping:

1. Specimens accepted for rabies testing: whole bats, animal heads.
   
   
2. BATS

• Whole (alive or dead)
• Must be placed inside a solid, escape proof container less than 1 gallon/128 ounces size (ideally a regular sized coffee can with a tight-fitting lid)
• LIVE BATS

              Must be kept in a container with breathing holes on the lid and clearly labeled “CAUTION LIVE BAT”

CAUTION:

- Do not hinder access with packing material or cloth.

- No wrapping of any kind

- No newspapers, trash, **glue traps

- Do not place on ice, bag, or freeze live bats

- Do not seal, glue, crush or stack live bats

- If you are not sure a bat is dead then treat it as ALIVE

• Dead bats: follow shipping guidelines for All Other Animal Samples

 ALL OTHER ANIMAL SAMPLES

• All animals of any size MUST be decapitated, no exceptions
• Spray heads for ticks/fleas prior to packaging but do not use insecticides that contain Permethrin
• Keep refrigerated or add enough ice packs
• Do not freeze
• If accidental freezing, keep frozen, do not thaw
• NEVER keep at room temperature
• Place 1^st in a watertight container such as a heavy plastic bag tightly secured, or a non-breakable container (NO GLASS) with a tight-fitting lid
• Place this container in a larger, waterproof, non-breakable container (NO GLASS) and in the space between the 2 containers, add coolant packs, do not use DRY ICE or ice in plastic bags
• Leaking or improperly packaged specimens cannot be shipped to other Public Health Laboratories
• Specimens in transit for more than 12hrs must be placed in a secondary insulated container
• Couriers are instructed not to pick up specimens that are improperly packaged
• Avoid delays, submitters must ensure specimens are packaged properly
• If individuals are exposed to fluid, such as leakage from an animal head, or a splash or cut during specimen preparation and packaging, they should
  IMMEDIATELY WASH THE EXPOSED AREA THOROUGHLY WITH SOAP AND WATER. The name and telephone number of the exposed individual(s) should be
  added to the VIR-16 form, along with the type of exposure that occurred

Additional Information:

All persons submitting animals for Rabies testing should be well informed on how Rabies testing will be performed on the animal, including pet and companion animals.

NOTE: Once submitted, contaminated tissues are managed as regulated medical waste and are incinerated, no tissue or part of any sample can be returned once accessioned into the Rabies Unit for testing.  Animal tissues that have been sent for Rabies virus testing may either contain or be contaminated by many types of highly pathogenic organisms, including organisms currently identified as “select agents”, samples may have undergone significant degeneration and consequent bacterial overgrowth.  Additional manipulation or unnecessary handling of discarded material places staff at risk and increases the possibility of cross contamination within the laboratory.   

• Samples not delivered within 24 hours of a high risk exposure
• Improper sample collection and/or submission:
  • Bats in glue traps
  • Live bats wrapped in a towel
  • Inhumane submissions of live bats
  • Damaged samples, e.g., missing areas of the brain, missing or destroyed head, decomposed or dessicated specimens
  • Packaging that exposes all participants in chain of custody
  • Blood not drained
  • Animal specimens (excluding bats) submitted whole and not decapitated
  • Decapitation tools or other sharp objects left in packaging
  • Transport boxes contaminated by biohazard leaks
  • Mislabeled, unlabeled or unidentified specimens
  • Frozen specimens
  • Specimens submitted without ice or with insufficient ice, particularly in warm weather conditions
• Documentation and paperwork:
  • Missing VIR-16
  • Out-of-state forms
  • Incomplete form
  • Illegible or ambiguous information on form

General and Permanent Statutes.

New Jersey Legislature website: https://lis.njleg.state.nj.us/nxt/gateway.dll?f=templates&fn=default.htm&vid=Publish:10.1048/Enu