Laboratory Animal Science

Comparative Study on the Laboratory Rabbits and Cats in the Test for Depressor Substances #br#

2020, 37(05): 1.

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Abstract: Objective To explore the feasibility of appling the laboratory rabbits in the test for depressor substances of drugs by comparing the differences of depressor response to histamine between the laboratory rabbits and cats. Method The amount of conventional New Zealand rabbits,Japanese white rabbits,Chinchilla rabbits and healthy cats was respectively 6, half male and half female. After anesthetized with pentobarbital sodium, they were injected intravenously with standard histamine solution by a dose of 0. 05—5. 0 μg / kg ( rabbits ) or 0. 05—1. 0 μg / kg ( cats) and Shengmai injection both by a dose of 0. 5 mL / kg and 1. 0 mL / kg. We try firstly to compare the sensitivity, reproducibility and stability of blood pressure of the depressor responses to the standard histamine solution among three stocks of laboratory rabbits. Then the consistency of the result in the sensitivity test for histamine with the requirement of Chinese Pharmacopoeia( ChP) was also compared, and whether was different for histamine and Shengmai injection that induce depressor effects between the laboratory rabbits and cats. Result The sensitivity and reproducibility of the depressor response to histamine in 3 stocks of rabbits were consistent, without any significant differences, but it was easier to inject by the ear margin veins of New Zealand rabbits, with more stable blood pressure during operating. Comparing with cats, the rabbits were not very sensitive to the depressor effect of histamine, its depressor response was only about 1 / 8 of cats, so that the result of histaminesensitivity test did not meet the requirement of ChP. However, Shengmai injection also had the same significant depressor response to all the laboratory rabbits as the cats, which was about 2 / 3 of that of the cats. Conclusion The laboratory rabbits are not sensitive to histamine and not be currently used for the test for depressor substances, but as being sensitive to Shengmai injection, so that the study also demonstrated that histamine had obviously species selectivity on the depressor response of animals.

Evaluation of the Effectiveness of Canine Parvovirus Immunoglobulin #br#

2020, 37(05): 9.

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Abstract: Objective To explore the effectiveness of canine parvovirus immunoglobulin injection. Method In this experiment, the canines were divided into model control group, symptomatic treatment group and globulin treatment group, and artificial infection of canine parvovirus was carried out. After the onset of the canines, the symptomatic treatment group and the globulin treatment group were treated according to the daily state. The experimental group was injected with canine fine immunoglobulin in addition to the same adjuvant therapy as the symptomatic treatment group. To observe clinical symptoms and score daily, collect anal swabs to detect fecal detoxification with colloidal gold antigen test strips; daily blood collection, perform blood routine and blood biochemical tests, analyze the test result and anatomical result . For analysis, the treatment effects of the other two treatment groups were compared according to the model establishment group. Result The result showed that the incidence rate of all dogs was 100%, the improvement rate of the model control group was 20%, the improvement rate of the symptomatic treatment group was 40%, and the improvement rate of the test treatment group was 80%. Conclusion The result show that the comparison of the treatment effect between the artificial infection model and the treatment group confirms that the canine parvovirus injection has the effect of treating canine parvovirus disease.

Effect of Shenfu Injection on the Expression of PPARγ and TNFα in Lung Tissue of Rats with Endotoxic Shock

2020, 37(05): 14.

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Abstract: Objective To investigate expression of peroxisome proliferator activated receptor gamma ( PPARγ) and tumor necrosis factor alpha ( TNFα) in lung tissue of rats with endotoxic shock and the intervention effect of Shenfu injection. Method SD rats were selected to establish the animal model of endotoxin shock by intraperitoneal injection of lipopolysaccharide ( LPS ) , followed by the treatment with shenfu injection ( low, medium and high doses) . The pathological changes of lung tissue were observed. The transcription and expression of PPARγ and TNFα in lung tissue and secretion of TNFα and IL-1β in plasma were detected. Result There were typical acute lung injury in the rats of LPS group, the transcription and expression of PPARγ in lung tissue were significantly decreased ( P<0. 01) , the expression of TNFα was obviously increased ( P < 0. 01) , and the levels of TNFα and IL-1β in plasma were significantly increased ( P < 0. 01) . However, compared with LPS group, Shenfu Injection relieved pulmonary congestion, edema and inflammatory cell infiltration, increased the transcription and expression of PPARγ in lung tissue in a dose-dependent manner, decreased TNFα expression, and inhibited the levels of inflammatory factors TNFα and IL-1β in plasma as well ( P < 0. 01) . Conclusion Shenfu injection can protect lung tissue of rats with endotoxic shock, and its mechanism may be related to up regulating PPARγ and inhibiting the production of inflammatory mediators.

Pathological Diagnosis and Analysis of Epidermal Tumor in Canine

2020, 37(05): 19.

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Abstract: Objective To investigate the occurrence and pathological characteristics of canine epidermal tumors, the background information and pathological diagnosis result of 39 cases of canine epidermal tumor were analyzed.Method Thirty nine cases of canine epidermal tumors were collected and sectioned for HE staining. Pathological observation and diagnosis were made by light microscope. According to the diagnosis result , the information of tumor types, ages, sexes and sites of the lesion were statistically analyzed. Result The statistical results show that the most common epidermal tumor in canine is squamous cell carcinoma, accounting for 46% of all cases of epidermal tumors, most of which are moderately differentiated squamous cell carcinoma. The second is papillomas with the proportion of 39%, mainly exophytic papillomas. Basal squamous cell carcinoma is rare. Papillomas are more common in 0-3-year-old dogs, squamous cell carcinoma and basal squamous cell carcinoma are more common in dogs over 7 years old. The incidence of male was slightly higher than that of female. In canine, the epidermal tumor most frequently occurs on the head. Conclusion The histological features and epidemic characteristics of canine epidermal tumors are summarized, which provides a reference for the diagnosis and differential diagnosis of this kind of tumors.

The Selection and Preliminary Application of Microsatellite Markers in Outbred Beagle Dogs

2020, 37(05): 24.

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Abstract: Objective In order to select microsatellite loci suitable for genetic detection of the outbred beagle dogs and to primarily adapt to analyze the genetic structure of one outbred population of small beagle dogs. Method We selected 120 microsatellite markers included with 23 loci from beagle dogs and 97 loci from other canine strains. The PCR conditions of these loci were optimized by DNA pool. The loci of better amplification effect were selected to analyze further with fluorescence labeled STR scanning and population genetic structure. Result A total of 73 loci were successfully amplified, within which 49 loci were used to calculate genetic information after STR scanning. It was shown that the small beagle dog population exhibited the effective number of allele 4. 9230, expected heterozygosity 0. 7574, average heterozygosity 0. 7375, Shannon ’ s information index 1. 6625 and PIC 0. 7038, respectively. It showed that the examined population exhibited high and abundant population genetic polymorphism information and high genetic diversity. Conclusion The set of 49 microsatellite loci we optimized could be adapted for genetically monitoring outbred beagle dogs. These loci are valuable to be verify and optimize using more populations.

Real-time PCR Detection and Analysis of the Gut Microbiota in African Green Monkey and Cynomolgus Macaque

2020, 37(05): 31.

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Abstract:Objective As a SIV natural host, African green monkey ( AGM ) shows AIDS resistance. Also, spontaneous hypertension develops in AGM. Different capacities in lipid metabolism between AGM and Cynomolgus macaque ( CM) exist. This study aims to characterize the gut microbiome profiles of AGM and CM. Method Fecal samples were collected for 12 AGM and 12 CM. Real-time PCR detection of the main gut bacteria in AGM and CM was performed and analyzed. Result Our results demonstrated a lower abundance of Bifidobacteria and a higher abundance of Prevotella in AGM. Compared with CM, AGM possessed a higher abundance of the butyrateproducing Faecalibacterium prausnitzii and Eubacteria ( P < 0. 05 ) . Conclusion Compared with CM, the gut microbiota of AGM resembles a trend as found in early SIV infection or chronic HIV infection. AGM possesses a relative higher abundance of the butyrate-producing gut bacteria which correlates with gut homeostasis and mucosal integrity. These findings may provide insight into environmental adaption and AIDS resistance in AGM.

Establishment of Biological Decontamination Hartley Guinea Pig Colonyand Genetic#br# Quality Analysis by Microsatellite Markers

2020, 37(05): 36.

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Abstract: Objective The guinea pig colony was decontaminated by caesarean section. Genetic testing confirms that the decontaminated colony conforms to the genetic characteristics of the closed colony. Method Female and male individuals were widely selected in different breeding units in production groups, mated at 1 ∶ 1. Cesarean section at 70 days after mating. The offspring fed by artificial milk. Breeding F1 generation by cycle mating system. DNA were extracted from 30 individuals ear tissue samples in F1 generation, 18 guinea pig microsatellite markers were used to genetic testing. Result A total of 85 pups were delivered by Caesarean section, with 56 surviving and 47 weaning. Rate of surviving was 65. 9%, rate of weaning was 83. 9%. Eighteen microsatellites loci

shared 81 alleles, the average heterozygosity was 0. 5747, average polymorphism information content ( PIC) was 0. 5216. Fourteen loci kept H-W balance. Three loci diverged significantly from the H-W balance, and 1 locus was homozygous. Conclusion The guinea pig colony was decontaminated by caesarean section and have ideal heterozygosity, belonging to highly polymorphic population, which accorded with the genetic characteristics of closed colony laboratory animals.

Analysis of DNA Polymorphism of Laboratory Red Crucian Carp Exposed to 137Cs-γ by AFLP

2020, 37(05): 42.

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Abstract: Objective To observe the AFLP changes of 137Cs-γ irradiation on the laboratory red crucian carp, and to explore the effect of different doses of γ-rays on the laboratory red crucian carp DNA polymorphism. Method According to the preliminary experimental results , laboratory red crucian carp C1HD was randomly divided into 5 groups, 10 tails / group, namely blank control group, 1 / 16 LD50 , 1 / 8 LD50 , 1 / 4 LD50, 1 / 2 LD50, 4 irradiation groups of 1. 94 Gy, 3. 88 Gy, 7. 76 Gy, 15. 53 Gy, and blank control group were set, and the laboratory red crucian carp was irradiated once and for all by using a biological irradiator. The blood DNA of the laboratory red crucian carp before and after irradiation was extracted and amplified by AFLP-PCR. The polymorphism information content (PIC) , gene heterozygosity (H) and Shannon information index (I) was analyzed. Result Before the laboratory red crucian carp was treated with 137Cs-γ irradiation, the gene polymorphism information content (PIC) , Nei’ s gene diversity (H) and Shannon’ s Information index (I) in normal laboratory red crucian carp is 45. 20%, 0. 19 and 0. 28. After 137Cs-γ irradiation treatment, they are 72. 27%, 0. 28 and 0. 41 respectively, which showed the genetics index rise. Conclusion After 137 Cs-γ nuclear radiation treatment, PIC and H in laboratory red crucian carp are higher. AFLP marker in laboratory red crucian carp C1HD can be used to monitor nuclear contamination

Effect of Anshen Bunao Syrup on Hypothalamic-Pituitary-Adrenal Cortex Axis in Rats with Depressive Sleep Disorder

2020, 37(05): 50.

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Abstract: Objective To investigate the regulatory effect of Anshen Bunao Syrup on hypothalamic-pituitaryadrenal cortex axis in rats with depressive sleep disorder. Method Forty clean SD male rats were randomly divided into 4 groups: normal control group, depressive sleep disorder model group, Anshen Bunao Syrup low dose group and Anshen Bunao Syrup high dose group, with 10 rats in each group. The behavior ( horizontal and vertical movements) of rats were detected by OFT after modeling, 7 and 14 days after drug intervention. The rats were sacrificed under anesthesia and weighed in the hypothalamus. The contents of corticotropin releasing hormone ( CRH) , adrenocorticotropic hormone ( ACTH ) and cortisol ( CORT ) in serum were detected by ELISA. The expression levels of β-endorphin ( β-EP ) and aquaporin 4 ( AQP4) were detected by immunohistochemistry and RT-PCR. Result Compared with the normal control group, the weight of rats in the model group decreased significantly, but the relative weight of hypothalamus increased significantly, the scores of horizontal and vertical activities decreased significantly, while the levels of CRH, ACTH and CORT in serum increased significantly, and the expression levels of β-EP and AQP4 in brain tissue decreased significantly ( P < 0. 05) . Compared with the model group, the weight of rats in low dose group increased, but the difference was not statistically significant ( P> 0. 05) , while the relative weight of hypothalamus decreased significantly ( P < 0. 05) ; the weight of rats in high dose group increased significantly, and the relative weight of hypothalamus decreased significantly ( P< 0. 05) . On the 7th day after treatment, the scores of horizontal and vertical activities in the high dose group were significantly higher than those in the model group ( P< 0. 05) , while the scores of horizontal and vertical activities in the low dose group were significantly higher than those in the model group ( P > 0. 05) . After 14 days of treatment, the horizontal and vertical activities of rats in the two treatment groups were significantly improved compared with those in the model group ( P< 0. 05) . The expression levels of CRH, ACTH and CORT in the 2 treatment groups were significantly lower than those in the model group ( P < 0. 05) , while the expression levels of β-EP and AQP4 in brain tissue were significantly higher than those in the model group, and the expression levels of β-EP and AQP4 in the high dose group were higher than those in the low dose group ( P< 0. 05) . Conclusion Anshen Bunao Syrup can inhibit the hyperfunction of hypothalamus-pituitary-adrenocortical axis in depression sleep disorder rats by regulating the expression of β-EP and AQP4.

Effect of Zhuyu Tongmai Capsule on Cerebral Ischemic Injury-Induced by Chronic Hypoperfusion of Rats

2020, 37(05): 57.

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Abstract: Objective To investigate the effect of Zhuyu Tongmai capsule on cerebral ischemic- injury induced by chronic hypoperfusion in rats. Method The chronic cerebral hypoperfusion injury model was established by permanent ligation of bilateral common carotid arteries in rats. After the model was made, the animals with better condition and better wound healing were divided into six groups: Model control group, Venoruton table group(100 mg / kg) , Buchang Naoxintong capsule group(800 mg / kg) , and three Zhuyu Tongmai capsule groups(200、100、50 mg / kg) ; A sham-operation group was also established. After the administration of 14 days, the rats were carrying out space exploration experiments using morris water maze, and were examined in the learning and memory abilities. At the end of the experiment, the brain of rats were measured in wet weight, brain coefficient and brain tissue water content. Result Compared with the model group, the retention time and travel distance in the effective area of the Zhuyu Tongmai capsule (200 mg / kg) group increased significantly ( P<0. 05) , and also the frequency of passes through the platform ( P < 0. 05) ; the data of rats decreased significantly in wet weight, dry weight, brain coefficient and brain tissue water content ( P < 0. 05) . It showed that Zhuyu Tongmai capsule can improve the spatial exploration ability of rats with chronic hypoperfusion cerebral ischemia and reduce the brainedema. Conclusion Zhuyu Tongmai capsule can improve the learning and memory ability of rats with chronic hypoperfusion cerebral ischemia.

The Method of Establishing Mouse Osteoarthritis Model by Intraarticular Injection of MIA

2020, 37(05): 61.

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Abstract: Objective To observe and analyze the damage on the mice cartilage and bone tissue produced by different dosages of MIA and find out the most suitable MIA dosage for building mouse osteoarthritis models. Method We choose healthy male C57BL / 6 mice randomly divided into 4 groups. The left knee was injected for MIA as treated group, the right knee was injected for PBS as control group. Knee joint width was measured 1,2 and 4 weeks after the injection of MIA. Microscopic and histological scores were submitted to evaluated the damage degree of knee joint. Result We find out that different kinds of pathological change were noticed in experimental groups. The scores of the experimental groups were significantly higher than that of the control group ( P<0. 05) . In the experimental group, there were 2 relative differences between the large dose group ( ≥ 0. 1 mg) and the small dose group ( < 0. 1 mg) . Conclusion We recommended 0. 1 mg as the respective dose for the large dose group, which produce obvious cartilage destruction and inflammatory, and 0. 05 mg as the respective dose for the small dose group which produce mild cartilage tissue damage and reversible inflammatory reaction.

Study on Building Mouse Asthma Model with Self-designed Experimental Animal Atomization Box

2020, 37(05): 66.

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Abstract: Objective To design an experimental animal atomization box with a household medical atomizer and facilitate airway disease research in experimental animals. Method A household atomizer and a separate ventilating squirrel cage were combined and used to transform into an experimental animal atomization box. We built a mouse asthma model with the self-designed atomization box and ovalbumin. Result The atomization box could produce enough fog for 1 - 20 mice experiment rapidly. We successfully built the mouse asthma model with the atomization box. Conclusion We applied the atomization box to build the mouse allergic asthma model and observed significant lung tissue inflammation. This method provides operational guide for experimental animal inhalation experiment with convenience and low cost.

Study on Building Mouse Asthma Model with Self-designed Experimental Animal Atomization Box

2020, 37(05): 71.

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Abstract: Objective To design an experimental animal atomization box with a household medical atomizer and facilitate airway disease research in experimental animals. Method A household atomizer and a separate ventilating squirrel cage were combined and used to transform into an experimental animal atomization box. We built a mouse asthma model with the self-designed atomization box and ovalbumin. Result The atomization box could produce enough fog for 1 - 20 mice experiment rapidly. We successfully built the mouse asthma model with the atomization box. Conclusion We applied the atomization box to build the mouse allergic asthma model and observed significant lung tissue inflammation. This method provides operational guide for experimental animal inhalation experiment with convenience and low cost.

The Application of MRI in the Model of Acute Cerebral Apoplexy in Mice

2020, 37(05): 74.

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Abstract:Magnetic resonance imaging ( MRI) plays an important role in the diagnosis, treatment and prognosis of acute stroke ( AS) . In animal disease models, MRI can better reflect the pathological and physiological changes of acute cerebral apoplexy, which is faster and more intuitive than traditional detection method to show the lesion area and degree, and its non-invasive imaging in vivo has become an important tool for the study of this disease. In recent years, MRI has also been gradually applied to the establishment and evaluation of the mouse AS model. This paper will expound the role of MRI in the evaluation and establishment of mouse AS model and its future development trend.

Risk Assessment of Novel Coronaviruses in Laboratory Animals

2020, 37(05): 78.

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Abstract: Objective Novel coronavirus ( SARS-CoV-2) broke out in December 2019, which spread rapidly around the world, and seriously affected the normal social life. Due to the conservation of SARS-CoV-2 receptor angiotensin converting enzyme ( ACE2) in humans and animals, it is necessary to evaluate the susceptibility of domestic animals and laboratory animals. Methed Based on the amino acid sequence of ACE2, the evolutionary relationship of ACE2 in human, experimental animals and domestic animals were analyzed. According to the binding sites of RBD ( receptor binding domain) and ACE2, the variation between species was compared, and the susceptibility of different species to novel coronavirus was evaluated. Result The binding sites of SARS-CoV-2 and ACE2 were conservative in different species, and the binding sites of ACE2 receptor and SARS-CoV-2 RBD were completely consistent in human, monkey and hamster. Conclusion Based on the binding sites comparison and features of SARS-CoV-2 infected animals, monkey and hamster may be susceptible to novel coronavirus, and they may be preferred as SARS-CoV-2 infected animal models.

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