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SN/T 1105-2019

Chinese Standard: 'SN/T 1105-2019'
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SN/T 1105-2019EnglishRFQ ASK Days<=3 Detection and identification of Coptotermes curvignathus Holmgren Valid SN/T 1105-2019
SN/T 1105-2019ChineseRFQ ASK <=1-day [PDF from Chinese Authority, or Standard Committee, or Publishing House]  


Standard ID SN/T 1105-2019 (SN/T1105-2019)
Description (Translated English) Detection and identification of Coptotermes curvignathus Holmgren
Sector / Industry Commodity Inspection Standard (Recommended)
Date of Issue 2019-09-03
Date of Implementation 2020-03-01
Older Standard (superseded by this standard) SN/T 1105-2014
Regulation (derived from) Natural Resources Department Announcement No. 7 of 2019

SN/T 1105-2019
(Everyone's quarantine identification method)
ICS 65.002.01B16
People's Republic of China Entry-Exit Inspection and Quarantine Industry Standard
SN/T 11105-2019 instead of Sn/T 1105-2014
Everyone termite quarantine identification method
2020-03-01 implementation
Published by the General Administration of Customs of the People's Republic of China
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This standard was drafted in accordance with the rules given in GB/T 1.1-2009.
This standard replaces Sn/T 1105-2014 "Everybody's termite quarantine and identification method".
Compared with SN/T 1105-2014, the main technical changes of this standard are as follows.
--- Removed normative references;
--- Modified terms and definitions;
--- Modified equipment and reagents, integrated morphology and molecular biology;
--- Increase the content of on-site quarantine to facilitate the detection of termites;
--- Increased identification feature measurement locations;
--- Added the host and distribution of termites in Appendix A;
--- Added Appendix B to the research overview of the world termite, and clarified the synonym of the termite genus. Delete the original standard approximation
Synonyms of termites, Philippine island termites are synonymous with Gerst's stage termites, and Sinabon's termites are synonymous with termites
--- Modified the search form for appendix C termites and their approximate species;
--- Added Appendix E color pictures of termites;
--- Added Appendix F color pictures of similar termites.
This standard is proposed and managed by the General Administration of Customs of the People's Republic of China.
This standard was drafted. Xiamen Customs of the People's Republic of China, Gongbei Customs of the People's Republic of China.
The main drafters of this standard. Pang Jin, Huang Pengying, Fang Zhipeng, Fang Yuanwei, Lin Zhenji, Yao Xiangrong, Dong Wenguang, Zhang Weidong, Liao Li.
The release of the version replaced by this standard is.
--- SN/T 1105-2014.
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Everyone termite quarantine identification method
1 Scope
This standard applies to the quarantine and identification of termites.
2 Terms and definitions
The following terms and definitions apply to this document.
The termite is recessed or protruding at the confluence of the seam of the head cover, which is the opening of the forehead.
Round opening in the middle of the forehead.
Soldier and worker ants refer to the widest part of the head, and winged adults refer to the width of the head including compound eyes.
The maximum width of the pronotum.
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3 Basic information on termites
Spreading of bedding materials such as sawn wood and wood packaging spread. The host, distribution and biology are shown in Appendix A. Overview of World Termite Research
See Appendix B. Because there are not many similar species of milk termites in the Americas, Africa, and Oceania, they are generally easier to identify. There are many types of milk termites in Asia, which are difficult
distinguish. See Appendix C for the retrieval tables of termites and other important milk termites in Asia.
4 Method steps
According to the damage of termites, you can obtain eggs, soldier ants, worker ants, or adults at the quarantine site or where suspected termites are. Present
If field quarantine finds soldier termites with termites, they are identified based on morphological characteristics. If termite soldier ants are not found, but have eggs, worker ants or winged adults
When you wait, the conventional PCR detection method and real-time fluorescent PCR detection method of the termite-specific amplification primers of everyone can be accurate and fast.
Quickly distinguish everyone from termites, Gerst milk termites, Taiwan milk termites, and South Asia milk termites.
5 Equipment and reagents
5.1 Equipment
Microscope, dissecting mirror, bark shovel, handsaw, chainsaw, iron cone, screwdriver, small axe, scalpel, dissecting needle, forceps, petri dish, writing brush,
Worm tube, finger tube, micrometer; PCR amplification instrument, real-time fluorescence PCR instrument, electrophoresis instrument, gel imaging system, centrifuge, desktop centrifuge, nuclear
Acid protein analyzer, oven, autoclave, electronic balance, mortar, shaker, water bath, vortex shaker, refrigerator, micro-sampler
(0.5 μL, 2 μL, 10 μL, 20 μL, 100 μL,.200 μL, 1000 μL), PCR reaction tube (0.2 mL, 0.5 mL).
5.2 Reagents
75% ethanol, double distilled water, liquid nitrogen, proteinase K (20mg/mL), RNA digestion enzyme, Tris saturated phenol, PremexExTa
(2 ×), chloroform, isoamyl alcohol, isopropanol, 75% ethanol, 95% ethanol, 10 × PCR buffer, deoxyribonucleoside triphosphates (dNTPs),
Tazyme, DL2000 Marker, agarose, ethidium bromide (EB), primer probe, insect genome DNA extraction kit. Extract A. 1% SDS, 50 mmol/L Tris · Cl (PH8.0), 25 mmol/L NaCl, 25 mmol/LEDA (PH 8. 0).
6 On-site inspection
6.1 Check for holes
Tap the ends and middle of the log or wood with an iron cone to hear if there is a hollow echo and find out if there are holes. If it makes an unusual sound,
The hole should be pried open and carefully checked for termites.
6.2 Checking the exposed traces of termites
Check whether the quarantine materials such as logs and wood cubes have termite sludge (ie, termite excrement and soil accumulation of ants), fly holes (this hole is winged into
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Insects breeding fly holes, which are often sealed by soil) and vent holes (ie, pinpoint-shaped holes with a diameter of about 1mm outside the ant nest, usually more than ten
There are dozens of holes, sometimes with fine soil and guards by soldiers and ants). Use a saw to cut the wood to check the signs of danger, if necessary,
Sampling this paragraph back to indoor quarantine. When the log has bark, you need to lift the bark with a shovel or screwdriver and observe whether the inside of the bark and the xylem of the log
There are termite hazards or bugs.
6.3 Finding the Nest
On the scene, carefully search the nests of suspicious woods with exposed termite activity. If termite activity is found, collect specimens in time to
Prepare for laboratory identification.
7 Laboratory identification
7.1 Specimen processing
The specimen was placed in a petri dish and washed with 75% ethanol, and the wood chips and dirt attached to the insect body were gently removed with a fine brush. Stereomicroscopy
Use a dissecting needle to straighten the worm body under the microscope, so that the main characteristics of the identification are on a plane, especially the soldiers need to separate the upper jaw after the worm body softens.
7.2 Morphological characteristics measurement
Specimen bodies were observed with a stereo microscope with a micrometer, and important identification characteristics were measured.
10 (except for a small number of specimens). Soldier ant specimen measurements include. head to palate, head widest, head height, left palate length, long hole diameter, short hole
Diameter, widest posterior condyle, narrowest posterior condyle, width of pronotum, prostrate length, and number of antennae. Winged adult specimen measurements include body length (not
With wings), head to upper lip, head to upper jaw, head wide with eyes, compound eyes long diameter, compound eyes short diameter, compound eyes to head lower edge, monocular long diameter, monocular short
Diameter, fore-wing length (without scales), pronotum length, pronotum width, hindfoot tibial length, and number of antennae nodes. Units are millimeters (mm).
7.3 Records and calculations
Record all measured features, and convert the measured values according to the multiples of the stereo microscope, uniformly converted to millimeters
(Mm). At the same time, the average of all measured individuals is calculated.
7.4 Morphological identification
7.4.1 Identification of Milky termites Morphological characteristics of soldier ants
The head is ovate, and the anterior segment is obviously narrowed. The ridge is located at the front of the head and extends as a short tube.
The tooth shape is notched, the inner edge is smooth and toothless, the antennae are 13 to 17 knots, and the tip of the upper lip swallows. The pronotum is flat and narrower than the head. Morphological characteristics of winged adults
The head is broadly ovate, the base of the rear lip is extremely short and flat, and the antennae are 18 to 25 knots. The pronotum is flat, narrower than the head, and the front wings
The scales are larger than the hind wing scales and cover the hind wing scales. The veins of the wings have a very shallow reticulate pattern, and the wings have mask hairs.
The veins emerge from the base of the tibial vein, and the midrib is very close to the elbow.
7.4.2 Identification of termites Morphological characteristics of worker ants
The body length is from 4.10mm to 5.05mm. The head is nearly round, light brown, with a head width of 1.30mm to 1.75mm, and a head length of 0.85mm to
1.05mm, antennae 14 to 15 knots. The width of the pronotum is 0.60mm ~ 0.85mm, the leading edge is slightly raised, there is a nick in the center.
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Belly is milky white, sparsely yellowish short-haired. Morphological characteristics of soldier ants
The head shell is dark yellow, the upper jaw is dark brown, and the chest, abdomen and feet are pale yellow. The head shell is sparsely hairy, with a pair of end hairs on the upper lip end, one on each side of the palate
1 bristle, there is also a thick bristle between the palate and the antennae, about 20 short hairs in the middle of the pronotum. Oval head shell, widest behind the head
Half, long and thin palate, saber-shaped, strong end of the jaw, a deep recess at the base of the left upper jaw, with a small dent in the front, tongue-shaped upper lip, slightly longer than
Wide, slightly pointed at the ends, translucent at the ends of the lips, large, round calyx holes, oblique openings on the side, and the upper part slightly backward. Antennae 15 knots to 16 knots, generally 16
Sections 3, 4 are short and almost equal, and section 2 is slightly longer than or equal to section 3. Prosthetic dorsum plate ingot-shaped, wider than long, with central anteroposterior edge
Slightly concave, rounded on the front side, and obliquely trailing on both sides (see Appendix D, Appendix E, and Appendix F). Morphological characteristics of winged adults
The total length is 16mm ~ 17mm, the wing length is 13mm ~ 14mm, the body length (excluding wing) is 7.5mm, the head is nearly round, dark brown, and the head is long
1.15mm. The tentacles are yellow-brown, 21 knots, and the second, third, and fourth knots are shorter than the other knots, and the third knot is the shortest. Compound eyes large, near-circular, monocular oval
shape. The distance between the antennae and the compound eye is smaller than the distance between the antennae and the compound eye, and the distance between the single eye and the compound eye is less than the width of the single eye. The front edge of the pronotum
Concave, the center of the trailing edge is recessed forward. Pronotum brown, densely yellow-brown long-haired. Fore-wing scales are larger than hind-wing scales.
It is colored hairy, with the forelimb midline protruding independently from the shoulder seam, and the elbow vein having 4 to 10 branches. Hind wing midribs are from the base of the radial veins, and the elbow veins have 8
There are 10 to 10 branches, and the wing veins vary greatly among different individuals.
7.5 Molecular biology identification
7.5.1 Isolation of Termite Genomic DNA
Take one termite (worker ants, soldier ants, winged adults, etc.) and rinse it in double-distilled water, blot it with absorbent paper, and remove the attachments on its surface under a dissecting microscope.
And abdominal contents to prevent protozoa contamination. Place it in a 1.5 ml Eppendorf tube, add 40 μL of extraction solution A, put it in a -70 ° C refrigerator, remove it after 4 min, mash it with a toothpick, homogenize, add an equal volume of 3 mmol/L potassium acetate (PH7.2), and ice Place on top for 1h, add
Add an equal volume of phenol/chloroform, mix, centrifuge at 2,000 g for 10 min, take the supernatant, add 2 times the volume of frozen absolute ethanol, mix and place in the refrigerator for 1 h
Precipitate DNA; centrifuge at 1,500 g for 15 min, discard the supernatant, add 1 mL of 75% ethanol to wash the pellet, centrifuge at 2,000 g for 1 min, and dry the ethanol
Then, the precipitate was dissolved with 100 μL double-distilled water, and the concentration and purity of DNA were measured by a microspectrophotometer.
Store at -20 ° C until use. Genomic DNA can also be extracted by kit method.
7.5.2 DNA quality check
The purity and concentration of DNA were measured with a nucleic acid protein analyzer, and the absorption values at 260 nm and 280 nm were obtained, respectively.
Purity and concentration, the calculation formula is as follows.
DNA purity = OD260/OD280
DNA concentration = 50 × OD260 μg/mL
The OD260/OD280 ratio of the PCR grade DNA solution should be 1.7 to 1.9.
7.5.3 Routine PCR detection Primers
The sequence of the PCR primers for termites is shown in Table 1.
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Table 1 Primer sequences
Type primer name primer sequence amplified fragment
Forward primer
Reverse primer
5'-CCCATCTCTCGCGCGCCCGCGCGCT-3 '254bp Reaction system
See Table 2 for the conventional PCR reaction system.
Table 2 Reaction system
Reagent volume/μL
10 × PCR buffer (with Mg2 +) 2.5 μL
dNTP mixture (10 mmol/L) 2 μL
Forward primer and reverse primer (10 μmol/L) each 1 μL
Tazyme (5U/μl) 0.2μL
Template DNA 5μL
ddH2O supplemented to a total volume of 25 μL Reaction conditions
Pre-denatured at 94 ° C for 5 min; denatured at 94 ° C for 1 min, annealing extended at 72 ° C for 1 min, 30 cycles; finally, extended at 72 ° C for 5 min. Setting of negative control, positive control and blank control
The DNA templates of other species of Pterotermes were used as negative controls, and the DNA of every termite was used as a positive control, and sterilized deionized water was used.
For blank control, make 2 replicates each. Agar gel electrophoresis detection
The size of the amplified target fragments was checked by agarose gel electrophoresis on conventional PCR products. Take 5μL of PCR product, add 1μL of 10 × PCR buffer, and use DL2000 Marker as molecular weight marker, electrophoresis on a 1.5% agarose gel containing DNA staining agent for 30min (90V), check whether the amplification is on the gel image analyzer A target segment of the expected size is taken, and the results are recorded and recorded.
7.5.4 Real-time fluorescent PCR detection Primers and probes
The real-time fluorescent PCR primers and probe sequences of the termites are shown in Table 3.
Table 3 Primer and probe sequences
Type primer name primer sequence amplified fragment
Forward primer
Reverse primer
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The real-time fluorescence reaction system of termites is shown in Table 4.
Table 4 Reaction system
Reagent volume/μL
PremExTaz (2 ×) 12.5 μL
Forward primer and reverse primer (10 μmol/L) each 0.5 μL
Probe (10 μmol/L) 0.25 μL
Template DNA 2μL
ddH2O supplemented to a total volume of 25 μL Reaction conditions
Pre-denatured at 94 ° C for 1 min; denatured at 94 ° C for 15 s, and annealed at 60 ° C for 1 min, 40 cycles. Setting of negative control, positive control and blank control
Same as
8 result judgment
8.1 Morphological methods
Based on the morphological characteristics of soldier ants, the identification characteristics of winged adults can be used as a reference, and they can be judged as termites if they conform to
8.2 Molecular biology methods
8.2.1 Routine PCR detection
Electrophoresis imaging results, if the positive control sample and the test sample both show a specific DNA product band of about 254 bp, and the negative
There is no DNA product band in the control sample. The above experiment was repeated two or more times. If the results are consistent, it can be determined that the type of sample is everyone.
8.2.2 Real-time fluorescent PCR detection
When the results of the positive control, negative control, and blank control are normal, then.
--- The detection Ct value is less than or equal to 36, the test result is positive and the sample type is everyone termites;
--- The detection Ct value is greater than or equal to 40, and the test result is negative to determine that the sample type is not a termite;
--- The detection Ct value is between 36 and 40, it is necessary to redo the real-time fluorescence PCR.
9 Specimen preservation
Specimens collected for morphological identification should be stored in wide-mouth bottles containing 75% ethanol, and the source of the goods should be indicated.
Master, collection time, place and collector, storage period is 6 months to 1 year; specimens used for molecular biological identification are soaked with 100% ethanol,
Store in -70 ℃ refrigerator for future use.
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Appendix A
(Informative appendix)
Everyone termite host, distribution and damage
A. 1 host
Albizia, Soybean, Cashew, Araucaria, Jackfruit, Kapok, Olive, Jibe, Eucalyptus, Hevea, Twin Leaf Bean, Mango,
Wood butterfly, Pinus, Caribbean pine, Sumatra pine, Salix, Red eucalyptus, Salo, benzoin, mahogany, teak, etc.
A. 2 Distribution
Asia. Singapore, Malaysia, Thailand, Myanmar, Vietnam, Indonesia, Cambodia, Brunei, India, Philippines, Papua New Guinea
Asia, Papua New Guinea, South Korea.
Europe. Germany.
Africa. South Africa
A. 3 harm
Everyone termites are social polymorphic insects. The same nest group has different grades of differentiation, including ants, queens, worker ants, soldier ants, and reproduction.
They are divided into ants, and each has their own responsibilities. Among them, soldier ants have strange changes and have obvious discriminative value in taxonomy. The worm is a terrestrial termite that can
Nesting in wood, you can also nest in the soil. The hidden lifestyle, the occurrence of various grades and the development of the entire nest group are in the nest.
Nesters in the wood are mostly in the hollow of the wood. Small holes, vent holes or openings for worker ants are sometimes visible on the surface of the damaged wood, and are common
There are excreta or dirt.
Termites invaded from the roots of the ground, eating stems. The ant can cause severe damage to a variety of economic forest trees, can damage live trees, and erode
Ground cables, instruments and equipment in the forest, damage to the house and various types of wood packaging.
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Appendix B
(Informative appendix)
Overview of World Termite Research
According to the "World Term Essay Collection" written by Dr. Krishna of the American Museum of Natural History, etc.
There are 70 species in the genus, including 4 fossil records. Of the 70 species, 8 are in the Americas, 7 in Africa, 11 in Oceania, and 44 in Asia.
There are currently 20 species of intercepted milk termites (Wang Mingsheng,.2015).
Divided by different geographical regions, there are 8 species of American termites in America, including 2 species intercepted by China's quarantine department, including South American termites.
More consistent).
狌 狊 狌 狊.
Synonyms. Ding Junjie (2013) collected 10 species of Chinese termites for molecular biology research. It is speculated that there may be only two species of Chinese termites in China, namely
Taiwan milk termites and Gerst milk termites. With the exception of Chinese termites, there are 24 species of other termites in Asia, including Taiwanese termites.
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Appendix C
(Informative appendix)
Everyone termite and its approximate species retrieval form (Soldier)
1. Pore hair 2 or more 2
Pore hair 1 to 3
The head shell is wide pear-shaped, with the widest head 1.30 to 1.34; antennae 15 to 16 knots; saber-shaped upper jaw, slightly curved jaw body;
Clearly cut in the center, shallowly concave at the trailing edge; upper lip spear-shaped; translucent end-pointed triangular, with a pair of end hairs
3. Oval head shell 4
Head shell wide pear-shaped 6
4. Antennae 14 to 15 knots, knot 2 is slightly longer than knot 3, knots 3 and 4 are equal in length, saber-shaped upper jaw, thin and curved tip, obvious front edge of pronotum
Tentacles 13 to 14 knots, hair in middle of pronotum 5
5. The front edge of the pronotum is deep indented, the front edge is narrow and round, the middle part has a horizontal set of bristles, and the periphery has sparse upright bristles
6. The antennae is 15 knots, with the tip of the upper jaw being thin and curved;
Antennae 15 to 16 knots, head with a wide pear-shaped shell, near the back of the head at its widest point, saber-shaped upper jaw, strong bending of the jaw end; center of the front and back edges of the pronotum
It is shallowly concave, with narrow frontal corners and straight oblique rear edges on both sides. The upper lip is blunt-shaped, slightly longer than wide, and the translucent ends are nearly straight.
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Appendix D
(Informative appendix)
Everyone termite soldier ant scale
Table D. 1 Unit of measurement for termites and soldiers (mm)
Original record of Project Holgren (1913)
Ahanad (1965)
According to Thai specimen
Ping Zhengming (1992)
According to Malaysian specimens
Head to palate 1.48 to 1.56 1.65 to 1.76 (1.69) 1.45 to 1.65 (1.55)
Head width (base of jaw) 0.78 to 0.83 0.86 to 0.90 (0.87) 0.72 to 0.83 (0.78)
The widest head is 1.35 to 1.43 1.40 to 1.63 (1.53) 1.25 to 1.37 (1.33)
Head height 0.93 to 0.98 0.97 to 1.11 (1.06) 0.85 to 0.93 (0.99)
Left upper jaw length-1.08 to 1.22 (1.14) 0.90 to 1.10 (0.99)
Long hole diameter-0.15 to 0.21 (0.18) 0.21 to 0.25
Short diameter of countersunk hole-0.15 to 0.18 (0.16)-
Houyi length-0.97 to 1.11 (1.08) 0.90 to 1.000 (0.94)
Houyi width 0.39 to 0.44 0.43 to 0.46 (0.44) 0.37 to 0.45 (0.41)
Houya narrow-0.23 to 0.30 (0.26) 0.22 to 0.25 (0.24)
The length of the pronotum is 0.48 0.54 to 0.63 (0.57) 0.47 to 0.52 (0.51)
The width of the back of the chest is 0.93 to 1.01 1.04 to 1.18 (1.12) 0.87 to 0.98 (0.93)
Note. The value of each item in parentheses is the average value of the item.
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Appendix E
(Informative appendix)
Everyone termite soldier ant feature map
(A. back of head; b. side of head; c. left upper jaw; d. right upper jaw; e. antennae; f. posterior condyle; g. pronotum
Figure E. 2 Front view of everyone's termite soldier ant head E. 3 Everyone's termite soldier side view
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Figure E. 4 Everybody termite soldier ant pronotum E. 5 Everyone termite palate
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Appendix F
(Informative appendix)
Morphological characteristics of termites and their approximate species
Figure F. 1. Characteristic map of termites and soldiers. 2 Taiwan milk termite soldier ant characteristic map
Figure F. 3. Characteristic map of Gerst milk termite soldier F. 4 Characteristic map of milk termites in South Asia
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[1] Chen Lanfen, Lai Xuewen. Milking termites in curved jaws were intercepted at Tianjin Port [J]. Plant Quarantine,.1992, 6 (4). 269-270.
(2). 225 to 232.
[3] Ding Junjie. Study on the Revision of the Milky Termite Classification in China. Master's Degree Thesis of Jiangxi Agricultural University [M]..2013.
[4] Huang Yuanda. Introduction to Chinese Termite Science [M]. Wuhan. Hubei Science and Technology Press,.2001, 149-153.
[5] Jiang Jinlin, Tang Lianfei, Huang Ping, etc. Identification of termites and Taiwan milk termites by PCR method, Plant Quarantine,.2009, 23 (3).
28 ~ 30.
[6] Wang Lizhong, Du Guoxing, Zhou Minghua. Five descriptions of Papua New Guinea and Solomon Islands [J]. Phytosanitary,.2002, 16
(5). 284 to 287.
[7] Wang Mingsheng, Zhou Pei, Army, etc. Overview of World Milk Termite Research [J]. Jiangsu Agricultural Science,.2015, 43 (3). 124-126.
[8] Wang Xingguo, Wang Dingguo, Liang Fan, et al. Morphological study of eight species of milk termite soldier ants [J]. Journal of Zhongkai Agricultural Engineering College,.2015,
28 (1). 1 ~ 7.
[9] Yu Daojian, Chen Zhizheng, Kang Lin. Adjacent termites intercepted in imported logs [J]. Plant Quarantine,.2001, 15 (1). 34 ~ 35.
[10] Yu Daojian, Chen Zhihuan. Two termites intercepted in imported Indonesian logs [J]. Plant Quarantine,.2001, 15 (4). 227-228.
[11] Zeng Lingzhao, Chen Weixiong, Xue Kuiwei, etc. Intercepted lucid termites from logs imported from Papua New Guinea [J]. Phytosanitary
Epidemic Disease,.1998, 12 (5). 290-291.
[12] Zheng Jianzhong, Ye Jun, Jin Yan. Termites intercepted by Chinese ports at imports from Southeast Asia [J]. Phytosanitary,.1999, 13 (4).
224 to 231.
[13] Thomas Bouguignon, ZvesRoisin. Revisioftermitifamiliy Rhinotermitida
(Isoptera) inNewGuinea. Yookeys [M],.2011, 148. 55 ~ 103.
[14] TraceM. Jenkins, SusanC. Jones, Cow-Zangle Lee. etal. Phyloguegrahythyminiumes
Materaloringinxoxic Coxtocopymesistroi (Isoptera. Rhinoceritia) [J]. MolcularPhyloge-
nettric Evolution,.2007, 42. 612-621.
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Related standard:   SN/T 5122-2019  SN/T 5123-2019
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