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Prevalence and Genetic Characterizationon of Cryptosporidium Isolates from Domestic Rabbits in Henan Province

Author: ShiKe
Tutor: NingChangShen;ZhangLongXian
School: Henan Agricultural University
Course: Preventive Veterinary Medicine
Keywords: Rabbit Cryptosporidium Epidemiological investigation Animal infection test Nested PCR-RFLP 18SrRNA actin COWP Phylogenetic
CLC: S858.291
Type: Master's thesis
Year: 2009
Downloads: 47
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Abstract


Rabbit as economic animals and pets, intimate relationship with humans. Reported in the literature rabbits susceptible to zoonotic Cryptosporidium rabbit genotype is a potential threat to human health. Domestic molecules very little information about the rabbit Cryptosporidium, Henan Province so far no complete rabbit Cryptosporidium echinococcosis epidemiological data, epidemiological data to master the Henan Province rabbit cryptosporidiosis and rabbit Cryptosporidium species and genotypes, the epidemiological investigation of Henan Province, parts of rabbit cryptosporidiosis from morphological, animals, and check to Cryptosporidium infection experiment, oocyst law and the small ribosomal subunit (18SrRNA) gene, muscle actin (actin) gene and oocyst wall protein (COWP) gene amplification and bioinformatics analysis aspects of the study of species identification and biological characteristics, identified in Henan rabbit, species and genotypes of Cryptosporidium popular evolutionary relationships determine Henan rabbit Cryptosporidium isolates and other types of Cryptosporidium between molecular species, and for the prevention and control of man and beast, suffering from cryptosporidiosis provides a theoretical basis. Parasitology detection methods to understand of Henan Province rabbit intestinal parasites infection repeated water washing precipitation, saturated sugar flotation technique, modified acid-fast staining and Lugol iodine solution staining in eight large-scale rabbit farms collected a total of 1081 fecal samples parasite examination. 5 eggs and oocysts were detected, including protozoa (coccidia, Cryptosporidium, Giardia lamblia), nematodes Class 2 (class round worm and rabbit tied filaria). The survey found 732 stool samples positive for parasites, parasite infection rate was 67.72% (732/1081); the 8 Tuchang different types of parasites average infection rate: 3.42% of Cryptosporidium, ball worms 64.11%, 5.27%, Giardia, rabbit bolt filaria 1.39%, 8.97% stercoralis. The survey found that rabbit parasites mixed infections are quite serious, including two parasites mixed infection is more prevalent. Saturated sucrose solution flotation method was to master the Henan Province of rabbit cryptosporidiosis insects epidemiological data, and modified acid-fast staining of 8 rabbit farms in five regions in Henan Province of the 1081 rabbit dung samples to check, and rabbit Cryptosporidium infection risk factors to assess. The results show that: identified 37 positive samples, Cryptosporidium total positive rate of 3.42% (37/1081); 1-3 month old rabbit and less than 1 month old unweaned rabbits highest infection rate amounted to 10.93% and 4.11%. Oocysts isolated from 37 positive samples in the size and morphology consistent the oocyst size is 4.426-5.163μm × 3.934-4.672μm, an average of 4.91μm × 4.28μm oocyst shape index is an average of 1.15. Cryptosporidium oocysts morphological analysis results will be found in the rabbit Cryptosporidium oocysts preliminary identification of Cryptosporidium parvum (Cryptosporidium parvum) or Cryptosporidium rabbit genotype (of Cryptosporidium rabbit genotype) specific pending molecular identification. Rabbit Cryptosporidium infestation also found that there are age differences. Animal infection tests showed that the 30-day-old rabbit, rabbit Cryptosporidium can infect immunosuppressed infected immune normal 30-day-old meat rabbits, just Cryptosporidium infection after the incubation period, the 2d and 8d; infection 2 peak of oocyst after all, but the first peak of the less obvious; the oocyst duration (exposed period) is also different, 5d and 8d. Published Cryptosporidium 18SrRNA gene actin gene and COWP gene sequence to establish the PCR method, in the domestic first rabbit Cryptosporidium isolates 18SrRNA, actin gene and COWP gene was successfully amplified and sequenced, the sequencing of sequence Blast in NCBI Nucleotide Sequence Database search for homologous sequences, then Lee Clustal Ⅹ 1.81, Phylip3.64 and DNAstar4.0 biology software sequence comparison construct molecular phylogenetic tree, and homology analysis. Analysis of evolutionary relationships in the the 18SrRNA gene locus, actin gene loci and the COWP gene locus germline Henan rabbit infected with Cryptosporidium Cryptosporidium rabbit genotype (Cryptosporidium rabbit genotype). Molecular characterization of rabbit Cryptosporidium insects, few reports at home and abroad. At home and abroad for the first time, the study of Cryptosporidium rabbit genotype (Cryptosporidium rabbit genotype) GP60 gene PCR amplification and sequence analysis showed that the Henan Province rabbit saphenous Cryptosporidium isolates found a subtype of the family, two different sub type.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Livestock, poultry, wildlife diseases > Livestock > Rabbit
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