New York Codes, Rules and Regulations
Title 1 - DEPARTMENT OF AGRICULTURE AND MARKETS
Chapter II - Animal Industry
Subchapter A - Diseases of Domestic Animals; Garbage Feeding
Part 57 - Pullorum And Typhoid Disease
Section 57.9 - Laboratory procedures

Universal Citation: 1 NY Comp Codes Rules and Regs ยง 57.9

Current through Register Vol. 46, No. 39, September 25, 2024

(a) Double strength skim milk. Double strength skim milk shall be prepared utilizing laboratory grade commercial dehydrated skim milk. Reconstitute according to label directions with the exception that the concentration shall be doubled; (i.e., 2x). Since reconstitution will cause foaming, an appropriately large container must be used. The liquid product shall be autoclaved at 121° C, 15 pounds pressure for 15 minutes. Avoid overheating since this can cause carmelization. Cool slowly, protect from contamination and refrigerate at between 2° and 4° C. Do not use if stored for more than two weeks.

(b) Organ culture.

(1) Visceral organs.
(i) Remove aseptically from each bird the apex of heart, piece of liver, gall bladder, spleen, ovary and the upper 4-6 inches of the oviduct. Other visceral organs with pathological lesions shall be included in the culture. The organs obtained from each individual bird shall constitute an organ set.

(ii) The organ set from each bird shall be processed separately. If the evaluation of the birds involves multiple houses on a single farm site, the approved laboratory may pool the organ sets of not more than five birds from all of the houses except one. The organ sets from the birds in the remaining house shall be processed separately. Seronegative birds shall be pooled separately from seropositive birds if organs are pooled.

(iii) Mince or homogenize organ sets in equal volumes of buffered diluent. Transfer 10 mls. of homogenate to 90 mls. of selective enrichment broth (TT Hajna, or tetrathionate brilliant green).

(iv) Incubate samples from tissues for 24 hours at 37° C.

(v) Plate on brilliant-green agar or XLD with 15 mcg/ml of pure novobiocin and a second media such as BG, XLD or XLT-4. Only one of the two media shall contain novobiocin. The selection of the two mediums shall be based on their complimentary ability to select Salmonella of differing biochemical properties.

(vi) Incubate plates for 24 hours at 37° C. Select aminimum of five suspect colonies per plate and transfer to TSI or LIA or select up to nine colonies for screening using the World Health Organization Salmonella polyvalent bacteriophage 01-0E protocol. If either or both develop typical reactions, proceed to serology from TSI slants with polyvalent serum for identification and then serogrouping.

(vii) Select a minimum of five group D isolates per case for serotype and phage type determination and send isolates to National Veterinary Services Laboratory (NVSL) for determination.

(c) Environmental culture.

(1) For samples obtained from litter, manure scraper, egg belts, dead embryos, or meconium:
(i) Place samples in selective enrichment broth at a ratio of 1 part sample to 10 parts broth (TT Hajna or tetrathionate brilliant green).

(ii) No samples shall be pooled after receipt at the approved laboratory.

(iii) Incubate samples for 24 hours at 42° C.

(iv) Plate on brilliant-green agar or XLD with 15 mcg/ml of pure novobiocin and a second media such as BG, XLD or XLT-4. Only one of these two media shall contain novobiocin. The selection of the two mediums shall be based on their complimentary ability to select Salmonella of differing biochemical properties.

(v) Continue incubating selective enrichment broth culture an additional 24 hours after initial plating or hold 5-7 days at room temperature. Transfer 0.25 ml. of all Salmonella-negative original enrichment broth cultures to 10 ml. fresh enrichment broth and incubate the secondary enrichment broth at either 37° C or 42° C for 24 hours.

(vi) Plate secondary enrichment broth as in subparagraph (iv) of this paragraph.

(vii) Incubate plates for 24 hours at 37° C. Select a minimum of five suspect colonies per plate and transfer to TSI or LIA or select up to nine colonies for screening using the World Health Organization Salmonella polyvalent bacteriophage 01-0E protocol. If either or both plates develop typical reactions, proceed to serology OE protocol. If either or both plates develop typical reactions, proceed to serology from TSI slants with polyvalent serum for identification and then serogrouping.

(viii) Select a minimum of five group D isolates per case for serotype and phage type determination and send isolates to National Veterinary Services Laboratory for determination.

(d) The forwarding of Salmonella enteritidis cultures to cooperating research laboratories for further study is recommended.

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