Hematological And Biochemical Values For Chinese Alligator And Bacterial Pathogeny Identification

Hematologic and serum biochemical values are significantly important for the health management of Chinese alligator species and never have been reported. During deep and late hibernation period, we drew blood samples respectively from30healthy captive Chinese alligators of three age groups (adults, sub-adults, and juveniles) in the Anhui Research Center of Chinese Alligator Reproduction (ARCCAR). Hematologic and serum biochemical measurements were performed.In deep hibernation, serum cholesterol, uric acid, globulin concentration among three age groups of Chinese alligators had a significant difference(p<0.05), indicating serum biochemical parameters affected by age. Compared with deep hibernation period, in late hibernation period there was a significant difference (p<0.05) for three age groups in some hematological indices (thrombus cell count, the percentage of neutrophils, monocytes, lymphocytes, basophils in the total number of white blood cells) and serum biochemical indices(total bilirubin, direct bilirubin, indirect bilirubin, lactate dehydrogenase, alkaline phosphatase, aspartate aminotransferase activity, calcium ions). These changes indicated that hematologic and serum biochemical values were affected by hibernation stage. In late hibernation, uric acid, globulin concentrations and white blood cell count of sub-adult and juveniles were significantly higher (p<0.001), the ratio of calcium and phosphorus, albumin concentration decreased significantly (p<0.001). Based on the comprehensive statistic analysis, sub-adults and juveniles were presumed that kidney had injury in artificial hibernation environment.When Chinese alligator eggs were hatched artificially, the embryo is most likely to suffer from death in early phase. Our experiment intended to provide some useful preparations for the connection between embryonic death likely occurred in earl stage with bacterial pathogens. In ARCCAR, we collected28dead embryonated eggs,16unfertilized eggs and4amniotic fluid of newly hatched young alligator for bacterial isolation and toxicity test to chicken embryos.26/28dead embryos isolated bacteria. Germ-carrying rate of Pseudomonas, Stenotrophomonas, Citrobacter, Enterobacter, Aeromonas, Proteus, Klebsiella in the dead embryos respectively was57.1%(16/28)、28.5%(8/28)、28.5%(8/28)、21.4%(6/28)、21.4%(6/28).10.7%(3/28)、10.7%(3/28). All of16unfertilized eggs isolated bacteria. The carrying rate of Proteus, Enterobacter, Citrobacter, Pseudomonas, Aeromonas in unfertilized eggs respectively was43.75%(7/16)、37.5%(6/16)、25%(4/16)、12.5%(2/16、6.25%(1/16), the carrying rate of Stenotrophomonas, Klebsiella, Pantoea, Serratia bacteria in the unfertilized egg was all18.75%(3/16). All of4amniotic fluid isolated bacteria. The carrying rate of Enterobacter, Citrobacter, Stenotrophomonas, Klebsiella in amniotic fluid respectively was100%(4/4),50%(2/4),25%(1/4),25%(1/4).A total of seven bacterial strains isolated from kidney, rectum, blood of two years old dead Chinese Alligator and lesions ovarian and lung of two adult killed Chinese Alligator. All this7bacteria strains were identified as Proteus vulgaris. Meanwhile, previous10strains of Proteus sp. isolated from dead embryos and unfertilized eggs were also identified as Proteus vulgaris by specific PCR. In this study, all this17strains of Proteus vulgaris were tested for pathogenicity in mice, ERIC-PCR genotyping, OMP-typing and medicines sensitive test. The LD50of XX2, XS2, XS1, WL16, WL9, SP1, SP11, SP13strains was2.5×10-6、5.0×10-6、5.0×106、1×109、6.8×108、5.5×108、6.8×108、5.5×108, respectively.17strains of Proteus vulgaris all showed good polymorphic fingerprinting map by both methods. Two to six bands could be amplified between100bp to3000bp by ERIC-PCR, typing17strains into4groups and6classes. The result of SDS-PAGE of outer membrane protein (OMP) showed3to6bands between20kD and130kD, typing17strains into5groups and7classes Two kinds of typing methods have no correlation and are not able to separate strains into different virulence. Resistance rates of17strains of Proteus vulgaris to ceftriaxone, imipenem, aztreonam, cephalosporins, cefepime, gentamicin, ticarcillin+clavulanic acid was0. Resistance rates to Cefazolin, amoxicillin, ampicillin, ticarcillin was100%. Resistance rates to carbenicillin, nitrofurantoin, co-trimoxazole was41.1%,58.8%,70.5%, respectively. Resistance rates to kanamycin, ciprofloxacin, streptomycin, piperacillin, amoxicillin+clavulanic acid, norfloxacin was lower, respectively was17.6%,11.7%,11.7%,11.7%,5.8%,5.8%.