Haemophilus parasuis, streptococcus suis and actinobacillus pleuropneumoniae triple inactivated vaccine and application thereof
A technology of Haemophilus suis and Streptococcus suis, which is applied in the field of veterinary biological products, can solve the problems of lack of immune cross-protection, and achieve good immunogenicity, good safety, and long-term immunity
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Embodiment 1
[0045] Example 1: Isolation and identification of strain HNHPS1
[0046] 1.1 Collection of disease data
[0047] The disease material came from a 2-month-old nursery pig that died of severe pneumonia, dyspnea and neurological symptoms in a large pig farm in Ruyang County, Henan Province in November 2016.
[0048] 1.2 Preparation of medium
[0049] TSB liquid medium: Dissolve 30 g of TSB broth powder (Tryptic Soy broth, tryptone soybean broth) in 1000 mL of ultrapure water, sterilize at 115 °C for 15 min, add 50 mL of fetal bovine serum, sterile 1% NAD (Nicotinamide adeninedinucleotide, nicotinamide adenine dinucleotide, coenzyme Ⅰ) 1 mL;
[0050] TSA solid medium: Dissolve 40g of TSA agar powder (Tryptic Soy Agar, tryptone soybean agar) in 1000mL ultrapure water, sterilize at 115°C for 15min, add 50mL of fetal bovine serum, 1mL of sterile 1% NAD, in 4 Store at ℃ for later use.
[0051] 1.3 Isolation and culture of bacteria
[0052] Aseptically collect the brain tissue of ...
Embodiment 2
[0072] Example 2: Isolation and Identification of Bacterial Strain HN1570
[0073] 2.1 Culture medium preparation
[0074] TSB liquid medium and TSA solid medium were prepared according to the method in Example 1.
[0075] 2.2 Isolation and cultivation of strain HN1570
[0076] In 2017, the applicant collected the heart blood of nursery pigs with typical polyserositis under aseptic conditions, inoculated them on TSA solid medium containing NAD, cultured them in a constant temperature incubator at 37°C for 36 hours, and picked suspected colonies for passage Purify and culture for another 24-48 hours to observe the colony morphology of Haemophilus parasuis, and grow consistent needle-shaped, colorless, transparent, smooth, moist colonies with a diameter of 1-2 mm (see Figure 4 ); while purified and inoculated on NAD-free TSA solid medium, it cannot grow on NAD-free TSA medium;
[0077] Pick a single purified colony for Gram staining and microscopic examination, and observe t...
Embodiment 3
[0091] Example 3: Isolation and identification of strain HNSS1
[0092] 3.1 Culture medium preparation
[0093] TSB liquid medium and TSA solid medium were prepared according to the method in Example 1.
[0094] 3.2 Isolation and cultivation of strain HNSS1
[0095] The disease material came from a 2-month-old nursery pig that suffered from severe pneumonia, dyspnea and neurological symptoms and died in a large pig farm in Jun County, Henan Province in November 2016. The brain tissue of the diseased pig was aseptically collected and inoculated on TSA solid culture containing NAD On the base, cultured in a constant temperature incubator at 37°C for 24 hours, a uniform needle-like size, a single colony of 0.5-1 mm on the blood plate, off-white translucent, round, smooth, and neatly edged α-hemolyzed colonies (see Figure 7 ). After purification and inoculation, carry out Gram staining. The bacteria are Gram-positive bacteria, round or oval, arranged in chains or in pairs, and...
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