Pennsylvania Animal Diagnostic Laboratory System
PADLS User Guide Tests Bacteriology/Mycology
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9.2 Bacteriology/Mycology

Test/ Agent	Site	Procedure	Tissue/ Specimen	Container	Shipping*	Setup	Turn- Around	Remarks
Acid fast stain	All	Acid fast stain	Tissue Swab	Sterile container Culturette	Overnight on gel packs	M-F	1 day	Specimens to be cultured for mycobacteria other than M. paratuberculosis may be referred to NVSL.
Acid fast stain	All	Acid fast stain	Heat-fixed slide	Slide mailer	None	M-F	1 day
Aerobic culture (includes Listeria)	All	Aerobic culture	Swab	Culturette or Amies transport media (latter preferred for Actinobacillus and Haemophilus)	Overnight on gel packs	M-F	2-5 days	Take specimens from the margin of lesions. If tissue and fluid specimens are not protected by transport medium, ship overnight. Uterine swabs should be taken with a shielded swab system, such as Kalajian, Acu-cul-shure, or Tiegland swab. Listeria isolation may require up to 1-3 months.
Aerobic culture (includes Listeria)	All	Aerobic culture	Tissue, fluid	Sterile container	Overnight on gel packs	M-F	2-5 days
Anaerobic culture (e.g. fusobacterium spp)	All	Anaerobic culture	Tissue 3.0 cm³	Sterile container, anaerobic transport medium	Samples should not be refrigerated	M-F	4-10 days	Anaerobic bacteria should be suspected if the infection involves tissue that is normally sterile, a site of recent trauma, or a poorly vascularized area. Contact laboratory to obtain transport media.
			Fluid aspirate	Vacutainer (>= 3 ml), anaerobic transport medium
			Swab	Anaerobic transport medium, Acu-cul-shure swab system
Antibiotic residue	PSU	LAST	Urine, 5ml		overnight on gel packs	M-F	1 day	Detection of antibiotic residue excreted in urine prior to shipping for slaughter
Antibiotic susceptibility	All	Minimum inhibitory concentraton (MIC) or Disk diffusion	Bacterial isolate derived from PADLS culture work Characterized bacterial culture	N/A Medium slant with growth	N/A	M-F	3-5 days	-Susceptibility reports will be released to veterinarians only. -Susceptibility test reports contain in vitro test results only and do not constitute recommendations for treatment. -Charges may reflect reisolation and identification by PADLS.
Black Leg Screen (Clostridium chauvoei, Cl. novyi, Cl. septicum, Cl. sordellii)	PSU PVL	Fluorescent antibody assay	Deep muscle tissue	Sterile container	Overnight on gel packs	M-F	1-3 days	Specimens for Clostridium FA testing must arrive at the laboratory within 24 hours of collection. Cl. septicum is a frequent post-mortem invader.
Blood Culture	PSU NBC	Blood bottle culture (aerobic and anaerobic)	7-10 ml whole blood	SPS Vacutainer or blood collection bottle	Overnight on gel packs	M-F	3-7 days	Indicate if patient is on antimicrobial therapy, define drug and dose. Collect specimen following complete surgical preparation. Preferred specimen collected prior to initiation of antimicrobial therapy.
Borrelia burgdorferi	NBC	PCR	1 ml synovial fluid or urine	Purple top EDTA tube	Overnight on gel pack for fluid & urine, ship M-Th	M-F	7 days	Refrigerate samples immediately upon collection.
Borrelia burgdorferi	NBC	PCR	Synovial membrane, kidney	Sterile tube	Preserve all tissues in 95% ethanol or s paraffin embedded formalin-fixed sections	M-F	7 days	Tissues sent in ethanol should be about 1 cm x 1 cm x 1 cm in size. Paraffin-embedded formalin fixed sections should be 10 micron in thickness and curly. Change blades between tissue blocks. For best results, do not leave tissues in formalin longer than 24 hours prior to paraffin embedding.
Brucella	All	Brucella culture	Bovine female: milk (all 4 quarters 20 ml ea.)	Sterile capped vial	Overnight on ice/gel packs	M-F	7-14 days	Include foremilk in samples for Brucella culture, but not routine cultures. Aseptic collection technique is extremely important. If milk samples cannot arrive at the laboratory within 24 hours of collection, they can be frozen, provided that the freezer is not a "frost free" unit.
			Supramammary lymph nodes	Sterile container
			Bovine male: Semen (2ml)	Sterile capped vial
			Superficial inguinal lymph nodes	Sterile container
			Other species: tissue	Sterile container
			Swab	Ames transport media
Campylobacter culture (Enteric) Campylobacter coli, Campylobacter jejuni culture	All	Microaerophilic culture	Feces, rectal swab: Amies or Cary Blair transport media	Sterile wide-mouth container	Overnight on ice/gel packs	M-F	3-5 days
Clostridium perfringens screen	All	Anaerobe culture PEA and EYA plating	Feces or intestinal loop	Sterile container	Overnight on ice/gel packs	M-F	2-3 days
Clostridium perfringens toxin typing Toxin Detection	PSU PVL	PCR	Feces or intestinal loop	Sterile container	Overnight on ice/gel packs	Th	2-8 days
	PSU PVL	PCR	Isolated organism	Anaerobic BA in AnaPak	Overnight on ice/gel packs	Th	2-8 days
	NBC	ELISA	Feces	Fecal cup	Keep feces frozen	W/Sa	1 day
Darkfield microscopic exam	All	Darkfield microscopic exam	Fluid specimen	Sterile container		M-F	1 day	Protect samples for trichomonas examination from temperature extremes. Refrigerate all other samples. Direct examination is a rapid but insensitive procedure. Culture procedures are recommended to confirm the presence or absence of a particular organism.
Dermatophilus exam	All	Geimsa Stain and/or culture	Tissue	Sterile container	Overnight on ice/gel packs	M-F	1-7 days
Dermatophilus exam	All	Geimsa Stain and/or culture	Swab	Culturette	Overnight on ice/gel packs	M-F	1-7 days
E. coli typing (0+H)	PSU	Serotyping	Ligated loop of intestine or colon, feces or isolate	Sterile container Overnight on gel packs M-F	Overnight on ice/gel packs	M-F	3-14 days	Submission of tissue/feces will include culture fees
E. coli virulence testing	PSU	Adhesin and toxin PCR	Fresh feces or Bacterial isolate	Sterile container Overnight on gel packs M-F	Overnight on ice/gel packs	M-F	3-14 days	Submission of tissue/feces will include culture fees
Erhlichia risticii*	NBC	PCR	2 ml whole blood	Purple top EDTA tube	Overnight on ice/gel packs	M-F	7 days	Refrigerate samples immediately upon collection.
Erhlichia risticii*	NBC	PCR	Cecum, right dorsal colon, colon	Sterile tube	Preserve all tissues in 95% ethanol or as paraffin embedded formalin-fixed sections	M-F	7 days	Tissues sent in ethanol should be about 1 cm X 1 cm X 1 cm in size. Paraffin-embedded formalin fixed sections should be 10 micron in thickness and curly. Change blades between tissue blocks. For best results, do not leave tissues in formalin longer than 24 hours prior to paraffin embedding.
Gram morphology	All	Gram stain	Heat-fixed slide	Slide mailer	None
Johne's disease (Mycobacterium paratuberculosis)	PVL	Mycobacterial culture (Liquid ESP or Agar culture) PCR*	Feces (10 g)	Wide-mouth screw-top container, child-resistant prescription vial	Overnight on ice/gel packs	M-W	42-112 days	Animal identification should be clearly written on submission form. For correct billing it is important to provide information about program participation. Withhold silage or moldy feed 48 hrs prior to fecal collection. Obtain samples directly from the rectum if possible. Ship samples to arrive at the laboratory within 36 hours of collection. Contact the laboratory for specimen containers: samples received in open containers of gloves will be refused. *Contact PVL for availability of PCR
			Ileocecal valve, mesenteric lymph node	Sterile container	Overnight on ice/gel packs
			Feces, tissues, paraffin embedded blocks	Sterile container	Overnight on ice/gel packs
Leptospira	All	FA Microscopic exam PCR	Fetal fluids, urine, kidney, placenta, spleen, liver	Sterile container Lepto transport medium Kidney impression slides	Overnight on ice/gel packs	M-W	1-3 days	Fetal fluids and tissues should be refrigerated but should not be frozen Urine collection-see below
Leptospirosis urine collection	For best recovery potential, inject with furosemide IV or IM. Clean vulva/prepuce and wait 10 min or second urination. Collect 10ml urine midstream in a red top or other sterile container.
Mastitis culture	All	Aerobic milk culture	Quarter sample	Sterile capped vial Freeze immediately (minimum of 10 ml for bulk tank)	Overnight on ice/gel packs	M-F	2-5 days	Please see below for sample collection; call laboratory for additional information. Bulk tank kits available from PSU
Mastitis culture	All	Aerobic milk culture	Quantitative bulk tank sample		Overnight on ice/gel packs	M-F	2-5 days
Mastitis culture sample collection	Wash the udder and teats and allow to air dry. Clean the teat orifice with alcohol pledgets. Sample in reverse order to reduce contamination. For routine culture, the first stream of milk should be discarded. Hold the collection vial as nearly horizontal as possible, and collect 2-3 ml of milk. When collecting composite samples, include an equal volume of milk from each quarter. Freeze samples immediately after collection.
Mycology	All	Fungal culture and/or microscopic exam	Tissue, fluid	Sterile container	Overnight on ice packs	M-F	1-42 days	If dermatophytes are suspected, specimens should not be transported in a sealed container and should not be refrigerated.
Mycology	All	Fungal culture and/or microscopic exam	Skin scrapings	Paper envelope	None	M-F	1-42 days
Mycoplasma spp.	PSU, PVL	Mycoplasma culture	Bovine milk bulk tank sample	Sterile-capped vial	Overnight on ice packs	M-F	7-28 days	Avoid the use of wood-shaft swabs; they interfere with the detection of Mycoplasma in culture.
		Speciation with PCR (MG, MS, M. bovis, etc)	Tissues (1.5 in³), aspirates	Sterile container
		Speciation with PCR (MG, MS, M. bovis, etc)	Swabs	Amies transport medium w/o charcoal
Quantitative bacterial culture	All	Dilution Plating Count	Water	1 L sterile container	Overnight on ice/gel packs	M-F	2-5 days
			Bedding or litter	1 qt Ziplock bag
			Raw semen	Sterile container				Raw semen samples must arrive at the laboratory within 24 hours of collection in order for examination to be considered valid.
			Processed semen	Semen straw	Ship frozen on ice/gel packs
			Urine	Sterile container	Ship frozen on ice/gel packs			Urine samples collected by cytocentesis or from catheterized animals are preferred, however, mid-stream collections are suitable. Samples must arrive at the laboratory on the same day as collection in order for the examination to be considered valid.
Salmonella	All	Salmonella culture	Feces (10 g)	Wide-mouth screw-top container, child resistant prescription vial	Overnight on ice/gel packs for all except feed	M-F	Presumptive id: 3-4 days	Samples should be obtained from the rectum of animals whenever possible. Specimens received in open containers or gloves will be refused.
			Tissue (Ligated ileum, mesenteric lymph node, etc)	Sterile container		M-F	3-4 days
			Exudate Swab	Culturette, Amies or Cary-Blair transport medium		M-F	3-4 days
			Environmental swab	Condensed evaporated skim milk		M-Th	3-4 days
			Swiffers	Sterile Whirlpak bag		M-F	3-10-days
			Eggs	Sterile Whirlpak bag		M-F	7-15 days
			200 gm feed	Sealed bag		M-F	7-15-days
Salmonella sp. Serotyping, molecular delineation	NBC	Molecular assays as described	Bacterial isolate	Contact laboratory	Overnight on ice/gel packs	M-F	1-10 days	Other tests available on bacterial isolate: Pulsed-Field Gel Electrophoreses, Phage Typing Contact laboratory.
Serpulina spp.	PSU	Serpulina culture	Ligated spiral colon Ceca	Sterile container	Overnight on ice/gel packs	M-F	5-14 days	Tissue specimens must be shipped to arrive overnight. Contact laboratory prior to collection.
Tritrichomonas	All	Tritrichomonas culture Microscopic exam	Cow: vaginal wash/cervical mucus Bull: preputial wash	Sterile capped vial Diamond's media or Lactated Ringers Solution (LRS) In-pouch media for culture	Overnight on ice/gel packs (see shipping note below) LRS-hand carry within 4 hours	M-F	5-10 days	Contact lab prior to collection for instructions and transport media Samples collected in Clark's/Weybridge media for venereal campylobacterosis cannot be tested for Trichomoniasis
Tritrichomonas	All	Tritrichomonas culture Microscopic exam	Semen Fetal fluids	Sterile, capped vial		M-F	5-10 days
Trichomonas Container and Shipping Information	Ship overnight on ice/gel packs if using Diamonds media. If using Lactated ringer's, hand carry at room temperature to reach laboratory in 4 hours
Ureaplasma spp.	PSU, PVL	Ureaplasma culture	Fetal fluids (stomach, per-icardial, & thoracic fluids	Sterile capped vial	Overnight on gel/ice packs	M-F	4-21 days	Wood shaft or cotton-tipped swabs should not be used; these may obscure the detection of Ureaplasma in culture. Shielded swabs should be used when collecting specimens from the female reproductive tract. Ureaplasmas may be a component of normal flora in the posterior vagina, particularly around the urethra.
			Swab	Amies transport medium w/o charcoal
			Raw semen (2 ml)	Sterile capped vial
Venereal Campylobacter spp.	PSU, PVL	Venereal Campylobacter culture	Bovine female: vaginal wash, cervical mucus	Clark's transport medium, Vacutainer, Weybridge transport media	Should preferably arrive same day or Overnight on gel packs	M-F	7-17 days	Direct aspiration of cervical/vaginal mucus or preputial material is the best method of specimen collection from adult bovines. Collect into a Vacutainer containing USP Lactated Ringer's solution. This material must arrive at the laboratory within 4 hours of collection. Clark's transport medium (available from Colorado State University) can be inoculated with this material; however, specimens inoculated into transport medium are not suitable for direct examination. Submitters requiring volume testing, e.g. for export, must make arrangements 5 days in advance. Contact the laboratory for specific instructions.
			Bovine male: preputial wash	Same as above	Should preferably arrive same day or Overnight on gel packs
			Semen	Semen straw, cryovial	Frozen
			Abortions: tissues	Sterile container	Overnight on gel packs
			Fluids	Clark's transport medium, sterile capped vial	Overnight on gel packs
Yersinia enterocolitica or Y. pseudotuberculosis	All	Yersinia culture	Intestinal, loop, feces	Sterile container	Overnight on ice packs	M-F	1-3 weeks	Cold enrichment and selective plating

*If shipping to PSU, do not use the US Postal Service

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