Development of a TEM-cell-integrated CO2 incubator for cell-based transient electromagnetic field bioeffect study.

To facilitate the cell-based experiment for pulsed electromagnetic field biological effect study, a novel TEM-cell-integrated CO2 incubator was developed. The integrated experimental system could simultaneously meet the requirement of standard cell culture condition and the various Transient Electromagnetic Field (TEF) exposure, which made it possible to study the relationship between different electromagnetic pulse exposure and the cellular responses in a reliable way. During the research, a comparison experiment was carried out to evaluate the necessity of the integrated incubator system: firstly, two different types of cell lines, which are the human prostate cancer cell line (PC3) and the pancreatic ß cell line (MIN6) were chosen and exposed in the TEM-cell which located in the open area and the integrated system, respectively, with the same EFT radiation conditions; then, the cells' viability, the cellular ROS level and the mitochondrial membrane potential (MMP) were detected, respectively. The results showed that in the same parameter of the EFT radiation, the processes of the cells had a significant difference and even opposite in the incubator and open area, and all the results could be reproducible. The phenomenon indicated the stability of the TEM-cell-integrated CO2 incubator, and also demonstrated the necessity to strictly control the cell culture condition when carrying out the precise mechanism study of the TEF bioresponse at the cellular levels.

Molecular Detection and Genetic Characterization of Toxoplasma gondii from Horses in Three Provinces of China.

Horse meat and milk are an important source of nutrition for many Chinese. Previous studies have shown a fairly high prevalence of Toxoplasma gondii infection in horses. However, there is no data concerning the molecular characterization of T. gondii in horses in China. The present study tested 231 cervical lymph node samples of horses from northern China (97 from Jilin, 54 from Liaoning, and 80 from Xinjiang) for the presence of the T. gondii B1 gene by seminested PCR. The B1-positive samples were genotyped at nine nuclear loci, that is, SAG1, alternative SAG2, SAG3, BTUB, c22-8, GRA6, c29-2, PK1, and an apicoplast locus, using PCR-restriction fragment length polymorphism technique. A total of 14 (6.06%) out of 231 samples were T. gondii positive. The overall prevalence of T. gondii in the horses was 5.15% (5/97) for Jilin Province, 5.55% (3/54) for Liaoning Province, and 7.50% (6/80) for Xinjiang Uygur Autonomous Region. Of the 14 positive samples, only 2 were successfully genotyped at all loci, 5 were successfully genotyped at 5-8 loci, and all typed samples belong to ToxoDB genotype no. 9. To our knowledge, this is the first molecular characterization of the T. gondii isolates from horses in China.

First report of bovine viral diarrhea virus and Mycobacterium avium subspecies paratuberculosis infection in Tibetan sheep (Ovis aries) in Tibetan Plateau, China.

Bovine viral diarrhea virus (BVDV) and Mycobacterium avium subspecies paratuberculosis (MAP) are important pathogens, which cause serious disease in animals. However, information about BVDV and MAP infection in Tibetan sheep in China is limited. Two thousand one hundred and eighty-seven blood samples were collected from Tibetan sheep between April 2013 and March 2014 from the Tibetan Plateau and tested for BVDV and MAP antibodies using commercial ELISA kits. The overall seroprevalence of BVDV and MAP in Tibetan sheep was 36.7 and 11.29%, respectively. Furthermore, risk factor analysis indicated that the age of sheep was statistically significant associated with BVDV infection and the region was considered as the risk factor of MAP infection in sheep (P < 0.05), gender and season were not considered as risk factors. This is the first report of seroprevalence and risk factors associated with BVDV and MAP infection in Tibetan Sheep in China, which will provide baseline information for controlling BVDV and MAP infection in ruminants in the Tibetan Plateau, western China.

Prevalence and genotypes of Enterocytozoon bieneusi in pigs in southern China.

Enterocytozoon bieneusi is one of the most important causative agents of microsporidiosis, causing diarrhoea the symptoms of enteric disease in humans and animals. Although there is some information on the prevalence and genotypes of E. bieneusi in China, there is still a lack of data in pigs in southern China. In the present study, a total of 396 faecal specimens were collected from pigs in Zhejiang, Guangdong and Yunnan provinces in southern China, and were examined by nested PCR amplification of the ribosomal internal transcribed spacer (ITS) for the prevalence and genotypes of E. bieneusi. The overall prevalence of E. bieneusi in pigs was 31.57% (125/396), forming 15 genotypes, including 9 known genotypes (EbpC, EbpA, D, G, H, PigEBITS5, Henan-IV, KIN-1, CHS5) and 6 novel genotypes (GD1, ZJ1, ZJ2, YN1, YN2 and YN3), which were all clustered into Group 1. Moreover, multilocus sequence typing (MLST) showed that 6, 3, 4 and 5 types were identified in MS1, MS3, MS7 and MS4 loci, respectively, representing four multilocus genotypes (MLGs), designated as MLGs novel-1 to novel-4 in the present study. This is the first detailed study of E. bieneusi using MLST in pigs in southern China, which extended information about the distribution of E. bieneusi genotypes in China.

First Report of Seroprevalence and Risk Factors of Neospora caninum Infection in Tibetan Sheep in China.

Neospora caninum is an intracellular protozoan parasite which can cause abortion and stillbirth in ruminants. However, there is no information on Tibetan sheep N. caninum infection in China. A total of 2187 serum samples were collected from Tibetan sheep in the major production areas of Luqu, Maqu, and Tianzhu in Gansu province, and Nyingchi in southeast Tibet, China. All samples were analyzed for the presence of antibodies to N. caninum using a competitive-inhibition enzyme-linked immunoassay. Of the 2187 serum samples, 184 (8.4%, 95% CI 7.3-9.6) were tested N. caninum seropositive. The N. caninum seroprevalence ranged from 4.4% (95% CI 1.4-7.4) to 11.3% (95% CI 8.2-14.4) among different regions, seasons, ages, and pregnancies, and there was no statistical significance among those groups (P > 0.05). Seroprevalence in male (10.8% 69/638) (95% CI 8.4-13.2) was significantly higher than in female (7.4% 115/1549) (OR =1.51, 95% CI 6.1-8.7) (P < 0.01). To our knowledge, this is the first report of N. caninum seroprevalence in Tibetan sheep in China, which provides baseline data for the prevention and control of N. caninum infection in Tibetan sheep.

Enterocytozoon bieneusi in Minks (Neovison vison) in Northern China: A Public Health Concern.

Enterocytozoon bieneusi is the most important causative agent of microsporidiosis and can infect almost all vertebrate and invertebrate hosts, including minks (Neovison vison). In the present study, a total of 298 feces samples (including 79 from Heilongjiang province, 31 from Hebei province, 67 from Jilin province, 90 from Liaoning province, and 31 from Shandong province, Northern China) were examined by nested PCR amplification of the internal transcribed spacer (ITS) region of the rRNA gene. The overall prevalence of E. bieneusi in minks was 10.1%, with 10.5% in Jilin province, 32.3% in Hebei province, 8.9% in Liaoning province, 0% in Shandong province, and 6.3% in Heilongjiang province. Furthermore, multiple logistic regression analysis revealed that region was only risk factors associated with E. bieneusi infection in the investigated minks. Five E. bieneusi ITS genotypes (three known genotypes, namely D, Peru11, and EbpC; two novel genotypes, namely, NCM-1 and NCM-2) were found in the current study. Importantly, genotypes D, Peru11 and EbpC, previously identified in humans, were also found in minks, which suggested that minks are the potential sources of human microsporidiosis. To the best of our knowledge, this is the first report of E. bieneusi infection in minks worldwide. The results of the present survey have implications for the controlling E. bieneusi infection in minks, other animals and humans.

[The application of acoustic pharyngometry in diagnosis and treatment of OSAHS].

Obstructive sleep apnea-hypopnea syndrome (OSAHS) is a common clinical disease that seriously threatens human health and life. Accurate location of the upper airway obstruction is the key to the diagnosis and treatment of OSAHS. Acoustic pharyngometry uses sound reflection to quickly assess the cross-sectional area and volume of the upper airway. Acoustic pharyngometry represents a simple, quick, non-invasive method for measuring upper airway dimensions which could predict sleep apnea risk. In this article we sought to introduce the application of acoustic pharyngometry in the diagnosis and treatment of OSAHS.

Prevalence and risk factors of Giardia duodenalis in domestic rabbbits (Oryctolagus cuniculus) in Jilin and Liaoning province, northeastern China.

BACKGROUND: Giardia duodenalis is one of the most important intestinal parasites that can infect virtually all animals, including rabbits and humans. However, there is little information regarding the prevalence and genotypes of G. duodenalis in domestic rabbits in China. METHODS: A total of 426 rabbit fecal samples (136 from Shenyang City, 174 from Changchun City, and 116 from Jilin City) were examined by Lugol's iodine staining with microscopy analysis, and the positive samples were genotyped at the triosephosphate isomerase (tpi) and the beta giardin (bg) gene loci using nested PCR. RESULTS: Forty-two (9.86%) out of 426 rabbit fecal samples were G. duodenalis-positive under microscopy analysis, and the highest G. duodenalis infection rate was 23.08% on farm 6. The prevalence of G. duodenalis in rabbits from different cities ranged from 1.47% to 14.37%. Among different age groups, G. duodenalis prevalence in rabbits ranged from 5.41% to 12.58%. The prevalence of G. duodenalis in outdoor rabbits and indoor rabbits was 14.29% and 6.77%, respectively. In the present study, region and farming mode were highly correlated with G. duodenalis infection in rabbits. All 42 G. duodenalis isolates were successfully amplified and sequenced at the tpi and bg loci, and only G. duodenalis assemblage B were identified. CONCLUSION: This study not only further confirmed the dominance of G. duodenalis assemblage B in rabbits, but also further improved the foundation data concerning the distribution of G. duodenalis assemblages in China.

Genetics association study and functional analysis on osteoporosis susceptibility gene BDNF.

To explore the relationship between brain-derived neurotrophic factor (BDNF) gene and bone mineral density (BMD) in Chinese Han population, we performed association analysis of 14 tag SNPs on BDNF gene with hip/spine BMD in 1300 Han Chinese samples from Shaanxi Province. We found that 8 of the 14 SNPs were significantly associated with hip or spine BMD (P < 0.05). Moreover, the SNP rs16917237 was significantly associated with both hip and spine BMD, with significant Bonferroni correlation (P value 0.05/14 = 0.0036) in hip BMD. To further explore the regulatory mechanism of BDNF gene in osteoporosis, we further performed a set of data analyses, including linkage disequilibrium and haplotype analysis, epigenetic annotation, expression quantitative trait locus (eQTL) analysis and metabolic pathway analysis. Further, we have established a mouse pre-osteoblasts differentiation cell model (MC3T3-E1) by recombination human bone morphogenetic protein (rh-BMP2) induction. siRNA- mediated knock down of BDNF in this cell model showed that all 14 SNPs are in the same haplotype block. Strong signals of active histone H3K4me1, H3K4me3, H3K27ac modifications and P300 binding were observed in osteoblasts, in the region surrounding the most significant SNP rs16917237, suggesting that this SNP might have a regulatory function in osteoblasts. Furthermore, analysis of genotype data of rs16917237 and BDNF expression in multiple tissues from GTEx showed that rs16917237 SNP could significantly affect the expression of BDNF in 11 tissues. Through analysis of the various BDNF pathways, we showed that BDNF participates in the MAPK pathway, which is a vital and well-established pathway affecting osteoblasts proliferation and differentiation. siRNA knock down of BDNF significantly decreased the mRNA and protein levels of CREB, which is important in the MAPK pathway in osteoblast differentiation. These findings suggest that BDNF might affect osteoblast differentiation via regulation of CREB expression. In conclusion, our results from combined genetic association and functional analyses show that BDNF is a vital osteoporosis susceptibility gene, which can affect BMD not only in Chinese Han but also likely in other populations.

Prevalence and Genetic Characterization of Cryptosporidium Infection in Java Sparrows (Lonchura oryzivora) in Northern China.

Cryptosporidiosis is a cosmopolitan parasitosis that affects a wide range of hosts including birds. As information concerning Cryptosporidium in birds is limited, the present study examined the prevalence and genotypes of Cryptosporidium in Java sparrows in Beijing and Shangqiu, northern China. Three hundred and fifty fecal samples were collected from Java sparrows (Lonchura oryzivora, 225 white Java sparrows and 125 gray Java sparrows) in Beijing and Shangqiu in October 2015, and the samples were examined by PCR amplification of the small subunit ribosomal RNA (SSU rRNA) gene. The overall Cryptosporidium prevalence is 13.42% (47/350), with 16.44% (37/225) in white Java sparrows and 8.00% (10/125) in gray Java sparrows. Cryptosporidium prevalence was 9.82% (16/163) in Java sparrows from Beijing and 16.58% (31/187) in Java sparrows from Shangqiu. The prevalence of Cryptosporidium in females and males was 40.63% (26/64) and 7.34% (21/286), respectively. The Cryptosporidium prevalence in Java sparrows of different ages varied from 10.47% to 16.33%. Sequence analysis of the SSU rRNA gene revealed that all the samples represented C. baileyi. This is the first report of Cryptosporidium in gray Java sparrows in China, which extend the host range for C. baileyi. These results provide baseline information for further studies of molecular epidemiology and control of Cryptosporidium infection in poultry in China.

First detection and genetic characterization of Toxoplasma gondii in market-sold oysters in China.

Toxoplasma gondii oocysts in the water might be filtered through the gills of shellfish in the process of feeding, and can be concentrated in the digestive glands. Consumers might become infected through ingestion of the shellfish unless they are discharged or inactivated by the shellfish. Thus, the purpose of this study was to evaluate the presence of T. gondii in market-sold oysters in China under natural conditions using a molecular approach. A total of 998 oysters were collected from markets in four cities (Weihai, Qingdao, Yantai and Rizhao) of Shandong province, eastern China. Of these, 26 samples (2.61%) were tested positive by nested PCR amplification of T. gondii B1 gene. Only one of the 26 positive samples was typed completely, and was identified as ToxoDB Genotype #9. This is first report of T. gondii in market-sold oysters in China, suggesting that oysters have the ability to filter and retain T. gondii oocysts in their tissues, which represents a risk to public health because oysters are frequently ingested in nature.

Detection of Enterocytozoon bieneusi in White Yaks in Gansu Province, China.

Enterocytozoon bieneusi, the most common zoonotic pathogen of microsporidiosis, has been found in various animals and humans, but no information is available concerning the prevalence and genotypes of E. bieneusi in white yaks (Bos grunniens). In the present study, 353 faecal samples from white yaks in Tianzhu Tibetan Autonomous County, Gansu Province, Northwestern China, were collected and examined by PCR amplification of the internal transcribed spacer gene to estimate E. bieneusi prevalence and identify their genotypes. Of the 353 faecal samples, 4 (1.13%) were tested E. bieneusi-positive. Sequences analysis revealed that two known genotypes, namely, I (n = 1) and BEB4 (n = 2), and a novel genotype, namely, WCY1 (n = 1), were found in this study. Among them, genotype WCY1 was clustered into Group 1, and genotypes I and BEB4 belonged to Group 2. The present study firstly indicates the existence of E. bieneusi in yaks in Gansu Province, Northwestern China. This is also the first record of E. bieneusi in white yaks. Effective measures should be taken to control E. bieneusi infection in white yaks, other animals, and humans.

Seroprevalence and Risk Factors of Bluetongue Virus Infection in Tibetan Sheep and Yaks in Tibetan Plateau, China.

Bluetongue (BT), caused by bluetongue virus (BTV), is an arthropod-borne viral disease in ruminants. However, information about BTV infection in yaks in China is limited. Moreover, no such data concerning BTV in Tibetan sheep is available. Therefore, 3771 serum samples were collected from 2187 Tibetan sheep and 1584 yaks between April 2013 and March 2014 from Tibetan Plateau, western China, and tested for BTV antibodies using a commercially available ELISA kit. The overall seroprevalence of BTV was 17.34% (654/3771), with 20.3% (443/2187) in Tibetan sheep and 13.3% (211/1584) in yaks. In the Tibetan sheep group, the seroprevalence of BTV in Luqu, Maqu, Tianzhu, and Nyingchi Prefecture was 20.3%, 20.8%, 20.5%, and 19.1%, respectively. The seroprevalence of BTV in different season groups varied from 16.5% to 23.4%. In the yak group, BTV seroprevalence was 12.6%, 15.5%, and 11.0% in Tianzhu, Maqu, and Luqu counties, respectively. The seroprevalence in different seasons was 12.6%, 15.5%, 15.4%, and 9.0% in spring, summer, autumn, and winter, respectively. The season was the major risk factor concerning BTV infection in yaks (P < 0.05). The date of the BTV seroprevalence in Tibetan sheep and yaks provides baseline information for controlling BT in ruminants in western China.

Molecular detection and genetic characterization of Toxoplasma gondii in farmed raccoon dogs (Nyctereutes procyonoides) in Shandong province, eastern China.

Toxoplasma gondii is a successful opportunistic parasite, affecting a wide range of vertebrate animals and humans. Genetic diversity of T. gondii in raccoon dogs (Nyctereutes procyonoides) is of great importance to understand the transmission of T. gondii in the environment. However, no information is available about the distribution of genetic diversity of T. gondii infection in raccoon dogs. This study was conducted to estimate the prevalence and genetic characterization of T. gondii from raccoon dogs in Shandong province, eastern China. A total of 314 brain tissue samples of raccoon dogs were collected and genomic DNA was extracted and assayed for T. gondii infection using semi-nested PCR targeting B1 gene. The positive DNA samples were typed at 10 genetic markers (SAG1, SAG2(5'+3' SAG2, alter.SAG2), SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico) by multiplex multilocus nested polymerase chain reaction-restriction fragment length polymorphism (Mn- PCR-RFLP) technology. Thirty-five (11.15%) of 314 DNA samples were detected positive. Only six samples were completely typed at all genetic loci, and these samples represented ToxoDB genotype#9. Two sample were typed at 9 genetic loci and one was typed at 8 genetic loci, all of them represented Type I. To our knowledge, this is the first report of genetic characterization of T. gondii in raccoon dogs in China. These results revealed the existence of genetic diversity of T. gondii in raccoon dogs in China. These data provided base-line information for controlling T. gondii infection in raccoon dogs.

Occurrence of Enterocytozoon bieneusi in Donkeys (Equus asinus) in China: A Public Health Concern.

Enterocytozoon bieneusi is an important zoonotic parasite. It can infect virtually all animal species and has a global distribution. However, the prevalence of E. bieneusi in donkeys (Equus asinus) has only been reported in Algeria and Spain, and no information is available concerning genotypes of E. bieneusi in donkeys worldwide. In the present study, a total of 301 donkey fecal samples (48 from Jilin Province, 224 from Shandong Province and 29 from Liaoning Province) were collected and examined by PCR amplification of the internal transcribed spacer (ITS) region. The overall E. bieneusi prevalence was 5.3% (16/301), with 6.3% (3/48) in Jilin Province, 4.9% (11/224) in Shandong Province, and 6.9% (2/29) in Liaoning Province. Prevalence in different age groups ranged from 4.2 to 5.5%. E. bieneusi prevalence in donkeys sampled in different seasons varied from 4.2 to 6.5%. Altogether, four E. bieneusi genotypes were identified in this study, with two known genotypes (J and D) and two novel genotypes (NCD-1and NCD-2). Phylogenetic analysis revealed that genotypes D, NCD-1 and NCD-2 belonged to group 1, while the remaining genotype J was clustered into group 2. These findings revealed the occurrence of E. bieneusi in donkeys in China for the first time. Moreover, the present study also firstly genotyped the E. bieneusi in donkeys worldwide. These findings extend the distribution of E. bieneusi genotypes and provide baseline data for controlling E. bieneusi infection in donkeys, other animals and humans.

Toxoplasma gondii Infection in Farmed Wild Boars (Sus scrofa) in Three Cities of Northeast China.

The apicomplexan protozoan parasite Toxoplasma gondii is a widely distributed etiological agent of foodborne illness. This parasite can cause production losses in livestock and serious disease in humans through consumption of contaminated meat. Pig meat is the most likely source of human infection, and wild boars may play a role in the transmission of T. gondii by serving as a reservoir host. This study aimed to investigate the seroprevalence of antibodies to T. gondii among farmed wild boars in China. In an 11-month survey, a total of 882 serum samples were obtained from farmed wild boars from three cities (Jilin City, Siping City, and Baishan City) in Jilin province, Northeast China and were tested for antibodies specific for T. gondii. Using modified agglutination test and a cutoff titer of 1:25, the prevalence of T. gondii infection in the examined samples was 10.0% (88 of 882). The highest seroprevalence was observed in animals from Jilin city (15.3%, 43/281) and followed by Siping (11.4%, 30/263) and Baishan (4.4%, 15/338). Logistic regression analysis revealed a significant correlation between the investigated geographic region and T. gondii infection. In addition, prevalence was higher in females compared to males, and the highest prevalence was detected in piglets. These findings indicate that farmed wild boars may become a source of foodborne toxoplasmosis, posing a food safety threat to the public health in the investigated areas. Implementation of effective measures to control T. gondii infection in farmed wild boars in China may be warranted.

De novo transcriptome sequencing and analysis of the juvenile and adult stages of Fasciola gigantica.

Fasciola gigantica is regarded as the major liver fluke causing fasciolosis in livestock in tropical countries. Despite the significant economic and public health impacts of F. gigantica there are few studies on the pathogenesis of this parasite and our understanding is further limited by the lack of genome and transcriptome information. In this study, de novo Illumina RNA sequencing (RNA-seq) was performed to obtain a comprehensive transcriptome profile of the juvenile (42days post infection) and adult stages of F. gigantica. A total of 49,720 unigenes were produced from juvenile and adult stages of F. gigantica, with an average length of 1286 nucleotides (nt) and N50 of 2076nt. A total of 27,862 (56.03%) unigenes were annotated by BLAST similarity searches against the NCBI non-redundant protein database. Because F. gigantica needs to feed and/or digest host tissues, some proteases (including cysteine proteases and aspartic proteases), which play a role in the degradation of host tissues (protein), have been paid more attention in the present study. A total of 6511 distinct genes were found differentially expressed between juveniles and adults, of which 3993 genes were up-regulated and 2518 genes were down-regulated in adults versus juveniles, respectively. Moreover, stage-specific differentially expressed genes were identified in juvenile (17,009) and adult (6517) F. gigantica. The significantly divergent pathways of differentially expressed genes included cAMP signaling pathway (226; 4.12%), proteoglycans in cancer (256; 4.67%) and focal adhesion (199; 3.63%). The transcription pattern also revealed two egg-laying-associated pathways: cGMP-PKG signaling pathway and TGF-ß signaling pathway. This study provides the first comparative transcriptomic data concerning juvenile and adult stages of F. gigantica that will be of great value for future research efforts into understanding parasite pathogenesis and developing vaccines against this important parasite.

Seroprevalence and Risk Factors of Fascioliasis in Yaks, Bos grunniens, from Three Counties of Gansu Province, China.

The aim of this study was to determine the seroprevalence and risk factors of fascioliasis in yaks, Bos grunniens, from 3 counties of Gansu Province in China. A total of 1,584 serum samples, including 974 samples from white yaks from Tianzhu, 464 from black yaks from Maqu, and 146 from black yaks from Luqu County, were collected and analyzed using ELISA to detect IgG antibodies against Fasciola hepatica. The overall F. hepatica seroprevalence was 28.7% (454/1,584), with 29.2% in white yaks (284/974) and 27.9% in black yaks (170/610). The seroprevalence of F. hepatica in yaks from Tianzhu, Luqu, and Maqu was 29.2%, 22.6%, and 29.5%, respectively. Female yaks (30.9%) had higher F. hepatica seroprevalence than male yaks (23.4%). Also, F. hepatica seroprevalence varied by different age group from 24.1% to 33.8%. Further, the seroprevalence ranged from 21.8% to 39.1% over different seasons. Interestingly, the season and age of yaks were associated with F. hepatica infection in yaks in the investigated areas. These findings provided a basis for further studies on this disease in yaks from 3 counties of Gansu Province in northwestern China, which may ultimately support the development of effective control strategies of fascioliasis in these areas.

Transcriptomic responses of water buffalo liver to infection with the digenetic fluke Fasciola gigantica.

BACKGROUND: Fasciola gigantica, the tropical liver fluke, infects buffaloes in Asian and African countries and causes significant economic losses and poses public health threat in these countries. However, little is known of the transcriptional response of buffaloes to infection with F. gigantica. The objective of the present study was to perform the first transcriptomic analysis of buffalo liver infected by F. gigantica. Understanding the mechanisms that underpin F. gigantica infection in buffaloes will contribute to our ability to control this parasite. METHODS: We challenged buffaloes with 500 viable F. gigantica metacercariae and collected liver samples through a time course at 3, 42 and 70 days post-infection (dpi). Then, we performed gene expression analysis on liver samples using RNA sequencing (RNA-Seq) Illumina technology and confirmed the RNA-Seq data by quantitative RT-PCR analysis. RESULTS: Totals of 496, 880 and 441 differentially expressed transcripts were identified in the infected livers at 3, 42 and 70 dpi, respectively. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that transcriptional changes in the liver of infected buffaloes evolve over the course of infection. The predominant response of buffaloes to infection was mediated by certain pathways, such as MHC antigen processing and presentation, Toll-like receptor 4 (TLR4), transforming growth factor beta (TGF-ß), and the cytochrome P450. Hepatic drug metabolizing enzymes and bile secretion were also affected. CONCLUSIONS: Fasciola gigantica can induce statistically significant and biologically plausible differences in the hepatic gene expression of infected buffaloes. These findings provide new insights into the response of buffaloes to F. gigantica over the course of infection, which may be useful in determining pathways that can modulate host-parasite interaction and thus potentially important for clearance of the parasite.

Prevalence and risk factors of Cryptosporidium infection in farmed pigs in Zhejiang, Guangdong, and Yunnan provinces, China.

Cryptosporidiosis is caused by species of Cryptosporidium protozoa that can infect a wide range of host animals worldwide. However, data regarding Cryptosporidium infection in farmed pigs in subtropical areas in China is limited. Therefore, a total of 396 fecal samples were obtained from farmed pigs from Zhejiang (n = 124), Guangdong (n = 72), and Yunnan (n = 200) provinces, China, and were tested by PCR amplification of the small subunit (SSU) rRNA gene. The overall prevalence of Cryptosporidium in pigs was 17.68% (70/396), with 20.11% (36/179) in male pigs and 15.67% (34/217) in female pigs. Additionally, Cryptosporidium prevalence was 8.33% (6/72) in Guangdong province, 14.52% (18/124) in Zhejiang province, and 23.00% (46/200) in Yunnan province. A DNA sequence analysis of the SSU rRNA gene revealed that all of the isolates represented C. scrofarum. This is the first report of C. scrofarum infection in pigs in Guangdong and Yunnan provinces in subtropical areas of China. The results of the present study provide foundation data for control and prevention of Cryptosporidium infection in pigs in the study areas in China.