Materials and reagents

Reagents

1. Cresol solution (Shanghai Hengyuan Biochemical Reagent, catalog number: 7784-24-9)

2. Potassium aluminum sulfate (Shanghai Hengyuan Biochemical Reagent, catalog number: 7784-24-9)

3. Arsenic trioxide (Yunnan Tin Co, catalog number: 1327-53-3)

4. Camphor powder (Shanghai Anpu Experimental Technology, catalog number: 464-49-3)

5. Xylene (Shanghai Aladdin Biochemical Technology, catalog number: 1330-20-7)

6. Distilled water

7. Canadian balsam (Shanghai Macklin Biochemical Technology, catalog number: 8007-47-4), neutral balsam (Shanghai Macklin Biochemical Technology, catalog number: 96949-21-2), or fir balsam (Aibixin Biotechnology, catalog number: 8016-42-0)

8. Pomegranate oil (Xi’an Rongbai Biological Technology, catalog number: 84961-57-9) or clove oil (Jiangxi Baicao Pharmaceutical, catalog number: 8000-34-8)

9. Arabic gum (Shanghai Yiji Industrial, catalog number: 9000-01-5)

10. Formaldehyde (Beijing Qinling Pharmaceutical Technology, catalog number: 79083-29-7)

11. Glycerin (Shanghai Aladdin Biochemical Technology, catalog number 56-81-5)

12. 10% sodium hydroxide (Shanghai Macklin Biochemical Technology, catalog number 1310-73-2) or potassium hydroxide (Shanghai Aladdin Biochemical Technology, catalog number 1310-58-3) solution

    
    

1. 5%–10% cresol soap solution (Lysol) (see Recipes)

2. Alum arsenic preservative (see Recipes)

3. Hoyer’s solution or Berlese’s solution (see Recipes)

4. Different concentrations of ethanol (30%, 50%, 70%, 80%, 90%, 95% and 100% anhydrous ethanol)

   The different concentrations of ethanol can be prepared by diluting anhydrous ethanol with distilled water. Anhydrous ethanol is a kind of very commonly used reagent with a wide choice of catalog numbers and manufacturers. We use anhydrous ethanol from Wuhan Jisi Instruments & Equipment (catalog number: 64-17-5).

   
   

Recipes

1. 5%–10% cresol soap solution (Lysol)

   5 mL of cresol solution + 95 mL of distilled water for making 5% cresol soap solution.

   10 mL of cresol solution + 90 mL of distilled water for making 10% cresol soap solution.

   In a beaker, mix the cresol and water and shake or stir until the cresol is completely dissolved in the water.

   
   

2. Alum arsenic preservative

   Potassium aluminum sulfate (powder, accounting for 40% or 70%) + arsenic trioxide (powder, 20%) + camphor powder (40% or 10%). Mix them together.

   
   

3. Hoyer’s solution or Berlese’s solution

   1. Hoyer’s solution (or Hoyer’s medium)

      50 mL of distilled water, 30 g of Arabic gum, 200 g of formaldehyde, 20 mL of glycerin.

   2. Berlese’s solution (or Berlese medium)

      20 mL of distilled water, 15 g of Arabic gum, 160 g of formaldehyde, 20 mL of glycerin.

      
      

Laboratory supplies

1. Collection and specimen preparation of small mammal hosts

   1. Fieldwork equipment (field tent, backpack, raincoat, non-slip shoes, flashlight)

   2. Disinfection and protection materials: 70% or 75% ethanol (Wuhan Jisi Instruments & Equipment, catalog number: 64-17-5), protective clothing and hat, medical latex gloves, mask

   3. Trapping materials

      1. Rat cage (18 cm × 12 cm × 9 cm, Jiangxi Guixi Li’s Rat Trap Equipment Co., Ltd.)

      2. Food bait (peanuts, corn, cooked meat, fried dough sticks, steamed buns)

      3. Disposable white cloth bag

   4. Recording materials (pen, marker, pencil, collection information record book)

   5. Specimen preparation materials for small mammals [lancet, surgical scissors, tweezers, steel ruler or tape measure, electronic scale, degreased cotton, bamboo sticks, wooden sticks, suture needles, specimen labels, beakers, alcohol lamps (for boiling small animal skulls), ether, alum arsenic preservative]

      1. Host tissue sample collection and preservation materials: Cryopreservation tubes and 95%–100% alcohol

         
         

2. Collection and fixation of ectoparasites

   1. White square plate (large size: 565 mm × 400 mm, small size: 385 mm × 285 mm)

   2. Lidded centrifuge tubes

   3. Ophthalmic forceps

   4. 70% or 75% ethanol

   5. Marking pens

      
      

3. Specimen-making of flea and sucking lice

   1. Glass slides

   2. Coverslips

   3. Baking plate or baking rack

   4. Specimen box

   5. Marker pen

   6. Label paper

      
      

4. Specimen-making of gamasid mites and chigger mites

   1. Petri dish

   2. Brush

   3. Slide

   4. Coverslip

   5. Marker pen

   6. Label paper

      
      

5. Specimen-making of ticks

   1. Wide-mouth bottle

   2. Cover glass

   3. Sealing wax

Equipment

1. Stereomicroscope (Leica, model: DM3000)

2. Baking oven (Shanghai Yiheng Scientific Instrument, model: DHG-9420A)

Procedure