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Haemonchosis remains a significant problem in small ruminants. In this study, the assay of recombinase polymerase amplification (RPA) combined with the lateral flow strip (LFS-RPA) was established for the rapid detection of Haemonchus contortus in goat feces. The assay used primers and a probe targeting a specific sequence in the ITS-2 gene. We compared the performance of the LFS-RPA assay to a PCR assay. The LFS-RPA had a detection limit of 10 fg DNA, which was 10 times less compared to the lowest detection limit obtained by PCR. Out of 24 goat fecal samples, LFS-RPA assay detected H. contortus DNA with 95.8% sensitivity, compared to PCR, 79.1% sensitivity. LFS-RPA assay did not detect DNA from other related helminth species and demonstrated an adequate tolerance to inhibitors present in the goat feces. Taken together, our results suggest that LFS-RPA assay had a high diagnostic accuracy for the rapid detection of H. contortus and merits further evaluation.

Toxoplasma gondii infects all warm-blooded vertebrates, resulting in a great threat to human health and significant economic loss to the livestock industry. Ingestion of infectious oocysts of T. gondii from the environment is the major source of transmission. Detection of T. gondii oocysts by existing methods is laborious, time-consuming and expensive. The objective of the present study was to develop a recombinase polymerase amplification (RPA) method combined with a lateral flow (LF) strip for detection of T. gondii oocysts in the soil and water. The DNA of T. gondii oocysts was amplified by a pair of specific primers based on the T. gondii B1 gene over 15min at a constant temperature ranging from 30°C to 45°C using RPA. The amplification product was visualized by the lateral flow (LF) strip within 5min using the specific probe added to the RPA reaction system. The sensitivity of the established assay was 10 times higher than that of nested PCR with a lower detection limit of 0.1 oocyst per reaction, and there was no cross-reactivity with other closely related protozoan species. Fifty environmental samples were further assessed for the detection validity of the LF-RPA assay (B1-LF-RPA) and compared with nested PCR based on the B1 gene sequence. The B1-LF-RPA and nested PCR both showed that 5 out of the 50 environmental samples were positive. The B1-LF-RPA method was also proven to be sufficiently tolerant of existing inhibitors in the environment. In addition, the advantages of simple operation, speediness and cost-effectiveness make B1-LF-RPA a promising molecular detection tool for T. gondii.

Toxoplasmosis is a globally spread zoonosis. The pathogen Toxoplasma gondii can hijack cellular organelles of host for replication. Although a number of important cellular life events are controlled by cell organelles, very little is known of the transcriptional changes of host cellular organelles after infection with T. gondii. Herein, we performed RNA-sequencing (RNA-seq) and bioinformatics analyses to study the global organelle component changes. It was found that many transcripts of the mouse spleen cellular organelle components were altered by acute T. gondii infection with the RH strain (Type I). Most differentially expressed transcripts of mitochondrial components were downregulated, especially those involved in biosynthetic and metabolic processes. Moreover, mitochondria based apoptosis process was downregulated. In terms of cytoskeleton, most differentially expressed transcript of cytoskeleton components were also downregulated, including septin cytoskeleton, cytoskeleton organization, centrosome and myosin. For endolysosomal system, ion transporters were downregulated at mRNA level, whereas the cytolytic components were increased, such as granzymes, Rab27a and perforin1 (Prf1). The main transcripts of Golgi apparatus components involved in sialylation or vesicle-mediated transportation were downregulated, while immune related components were upregulated. For endoplasmic reticulum (ER), posttranslational modification, drug metabolism and material transportation related transcripts were downregulated. In addition, T. gondii antigen cross-presentation by MHC-I complex could be downregulated by the downregulation of CD76 and ubiquitination related transcripts. The present study, for the first time, described the transcriptional changes of the mouse spleen cellular organelles following acute T. gondii infection, which provides a foundation to study the interaction between T. gondii and host cells at the sub-cellular level.

Toxoplasma gondii is a worldwide prevalent pathogen that infects most of the warm-blood vertebrates. To investigate the regulation network of splenic miRNAs altered by acute infection with T. gondii, we herein investigated the changes of miRNA profile in mouse spleen via next generation sequencing and bioinformatics analysis. A total of 379 miRNAs was identified, 131 miRNAs of them were differentially expressed (including 97 upregulated and 34 downregulated miRNAs). 48 differentially expressed miRNAs had validated targets in the miRWalk2.0 database. Gene Ontology (GO) enrichment analysis revealed that the validated targets of differently expressed miRNAs were significantly enriched in gene transcription regulation. It suggested that T. gondii can modulate host gene expression through targeting to trans-regulation factors. The genes involved in apoptosis or anti-apoptosis were both targeted by differentially expressed miRNAs. The change of power balance between the miRNAs targeting host apoptosis genes and those regulating host anti-apoptosis genes contributes to the fate of host apoptosis process. Twelve pathways were significantly enriched in KEGG analysis with most of them being cancer related, including pathways in cancer, pancreatic cancer, colorectal cancer, axon guidance, MAPK signaling pathway, focal adhesion, chronic myeloid leukemia, renal cell carcinoma, prostate cancer, glioma, regulation of actin cytoskeleton, and Wnt signaling pathway. Our study showed a changed miRNA regulation network in mouse spleen infected by T. gondii. These findings will be helpful for better understanding of miRNA regulation network in host-T. gondii interaction, revealing the relationship among T. gondii infection, gene regulation, apoptosis and cancer process alterations in infected spleen.

Toxoplasma gondii is a protozoan parasite infecting almost all warm-blooded animals and humans. There are three infective stages of T. gondii: the tachyzoites, the bradyzoites, and the oocysts. The tachyzoite is a rapidly multiplying stage and the main pathogenic factor. In North America and Europe, T. gondii is consisted of four major clonal lineages (namely Types I, II, III, and Type 12). In this study, we explored the proteomic profiles of different genotypes (Type I-RH strain, Type II-PRU strain, Type II-TgQHO strain, and ToxoDB 9-TgC7 strain) of T. gondii tachyzoites by using 2D DIGE combined with MALDI-TOF MS. Totally, 110 differentially abundant protein spots were selected. Of these, 98 spots corresponding to 56 proteins from T. gondii were successfully identified. These included surface antigen (SAG1), heat shock protein 70 (Hsp 70), disulfide isomerase, coronin, heat shock protein 60 (Hsp 60), pyruvate kinase, receptor for activated C kinase 1, and peroxiredoxin. Gene ontology enrichment analysis revealed that most of the differentially abundant proteins were involved in biological regulation, metabolic process, response to stress, binding, antioxidant activity, and transporter activity. According to the KEGG metabolic pathway maps of T. gondii, some identified proteins were involved in the glycolytic/gluconeogenesis pathway. The present study identified differentially abundant proteins among different genotypes of T. gondii and these findings have implications for the better understanding of the phenotypic differences among the examined T. gondii genotypes, which in turn may contribute to the better control of toxoplasmosis.

A total of 875 serum samples from dairy cattle in six districts in Zhuang Autonomous Region (GZAR) were collected between June 2009 to March 2010, and Chlamydiaantibodies were assayed by indirect hemagglutination test (IHA). The overall seroprevalence of Chlamydia in dairy cattle was 17.83% (156/875) in GZAR. The seroprevalence of Chlamydia in aborting dairy cattle (18.18%) was higher than that in non-aborting dairy cattle (16.69%) (not statistically significant). Dairy cattle of seven year old had the highest seroprevalence (35.42%), and dairy cattle with seven calves had the highest positive level (100%, 1/1). Statistically significant differences were observed between Chlamydia prevalence and ages (P

Roundworms of the genus Ascaris comprise species of medical (A. lumbricoides) and veterinary (A. suum) importance. Despite the economic importance, public health impact, and worldwide prevalence both parasites still have a controversial taxonomy and their proteomes are poorly described in databases. In the present study, high-resolution two-dimensional polyacrylamide gel electrophoresis (2-DE) coupled with MALDI-TOF/TOF MS was used to characterize and contrast proteomic profiles of adult female A. lumbricoides and A. suum. Approximately, 3-fold higher protein spots were obtained in the present study compared to traditional method, and more than 630 and 750 protein spots were reproducibly detected among the 6 gels examined (3 gels for each of A. lumbricoides and A. suum). The peptide mass finger printings (PMFs) were acquired successfully for 17 from 22 differential 2-DE spots, corresponding to 12 unique proteins. Among them, 2 Asu-enriched and 3 Alu-specific proteins were found in A. lumbricoides; and 6 Asu-enriched and 1 Alu-specific proteins were found in A. suum. Functional analysis revealed low level of difference among differentially expressed proteins of the two nematode species, while the 1 species-specific protein of A. suum was different from that of A. lumbricoides. The proteomic patterns of A. lumbricoides and A. suum provide proteomic evidence that these two taxa are very closely related. This work indicates that the combined 2-DE and MALDI-TOF/TOF MS approaches employed in the current study allow the identification of proteins whose expression is different between Ascaris species.

Dicrocoelium chinensis is commonly known as the "lancet fluke" or "small liver fluke" which lives in the bile ducts and gall bladder of domestic and wild ruminants. In this study, we examined sequence variability in two mitochondrial (mt) genes, namely cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) genes, among D. chinensis isolated from domestic yaks in Gansu and Sichuan provinces, China. A portion of the cox1 (pcox1) and nad1 (pnad1) genes was amplified by PCR separately from the individual adult D. chinensis, and the amplicons were subjected to sequencing from both directions. The lengths of the sequences of pcox1 and pnad1 were 358 and 458 bp, respectively. The A+T contents of the sequences were 58.94-59.22% for pcox1 and 60.04-60.48% for pnad1, respectively. The intra-specific sequence variations within D. chinensis were 0-0.84% for pcox1 and 0-0.87% for pnad1. The inter-specific sequence differences among members of the examined trematodes were significantly higher, being 22.9-29.3% and 33.6-41.5% for pcox1 and pnad1, respectively. Phylogenetic analysis utilizing combined sequences of pcox1 and pnad1 using Bayesian inference revealed that all of the D. chinensis isolates grouped together with high statistical support. These findings demonstrated the existence of low intra-specific variation in mitochondrial DNA (mtDNA) sequences among the D. chinensis isolates from domestic yaks in China and had implications for studying the molecular epidemiology and population genetics of D. chinensis.

Cryptosporidium is a widespread protozoan parasite that infects a large number of vertebrate animals, resulting in varying degrees of diarrhea or even death. As dairy cattle feces is an important source of Cryptosporidium spp. infection, development of a handy and accurate detection method via its oocysts in dairy cattle feces would be interesting and necessary. We herein developed a quick detecting method using recombinase polymerase amplification (RPA) combined with lateral flow (LF) strip to detect DNA of Cryptosporidium oocysts in dairy cattle feces. The DNA was released by boiled water with 0.1 % N-lauroylsarcosine sodium salt (LSS). The established method was proven to be of higher sensitivity than normal polymerase chain reaction (PCR) amplification with the lowest detection of 0.5 oocyst per reaction, and specificity with no cross reactivity to other common protozoan species in the intestine of dairy cattle. The diagnostic method established herein is simple, rapid, and cost-effective, and has potential for further development as a diagnostic kit for the diagnosis of cryptosporidiosis of dairy cattle.

Background Schistosoma turkestanicum is a parasite of considerable veterinary importance as an agent of animal schistosomiasis in many countries, including China. The S. turkestanicum cercariae can also infect humans, causing cercarial dermatitis in many countries and regions of the world. In spite of its significance as a pathogen of animals and humans, there is little transcriptomic and genomic data in the public databases. Methods Herein, we performed the transcriptome Illumina RNA sequencing (RNA-seq) of adult males and females of S. turkestanicum and de novo transcriptome assembly. Results Approximately 81.1 (female) and 80.5 (male) million high-quality clean reads were obtained and then 29,526 (female) and 41,346 (male) unigenes were assembled. A total of 34,624 unigenes were produced from S. turkestanicum females and males, with an average length of 878 nucleotides (nt) and N50 of 1480 nt. Of these unigenes, 25,158 (72.7 %) were annotated by blast searches against the NCBI non-redundant protein database. Among these, 21,995 (63.5 %), 22,189 (64.1 %) and 13,754 (39.7 %) of the unigenes had significant similarity in the NCBI non-redundant protein (NR), non-redundant nucleotide (NT) and Swiss-Prot databases, respectively. In addition, 3150 unigenes were identified to be expressed specifically in females and 1014 unigenes were identified to be expressed specifically in males. Interestingly, several pathways associated with gonadal development and sex maintenance were found, including the Wnt signaling pathway (103; 2 %) and progesterone-mediated oocyte maturation (77; 1.5 %). Conclusions The present study characterized and compared the transcriptomes of adult female and male blood fluke, S. turkestanicum. These results will not only serve as valuable resources for future functional genomics studies to understand the molecular aspects of S. turkestanicum, but also will provide essential information for ongoing whole genome sequencing efforts on this pathogenic blood fluke.

The present study examined sequence variation in three mitochondrial DNA (mtDNA) genes, namely cytochrome c oxidase subunit 3 (cox3) and NADH dehydrogenase subunits 1 and 4 (nad1 and nad4), among Ascaridia galli isolates from different geographical localities in China. A portion of cox3 (pcox3), nad1 (pnad1) and nad4 (pnad4) genes were amplified by polymerase chain reaction (PCR) separately from adult A. galli individuals and the amplicons were subjected to sequencing from both directions. The length of the sequences of pcox3, pnad1 and pnad4 were 408 bp, 471 bp and 333 bp, respectively. The intraspecific sequence variations within A. galli were 0–1.7% for pcox3, 0–2.8% for pnad1 and 0–3.4% for pnad4. The A+T contents of the sequences were 67.16–67.65% (pcox3), 67.09–67.94% (pnad1) and 69.91–71.77% (pnad4). The interspecific sequence differences among members of the Ascaridida were significantly higher, being 13.2–30.9%, 12.8–29.0% and 15.1–34.1% for pcox3, pnad1 and pnad4, respectively. Phylogenetic analyses using combined sequences of pcox3, pnad1 and pnad4, with three different computational algorithms (Bayesian analysis, maximum likelihood and maximum parsimony), all revealed distinct groups with high statistical support. These findings demonstrated the existence of intraspecific variation in mitochondrial DNA (mtDNA) sequences among A. galli isolates from different geographical regions in China, and have implications for studying molecular epidemiology and population genetics of A. galli.

This study examined sequence variation in three mitochondrial DNA (mtDNA) regions, namely cytochrome c oxidase subunit 1 (cox1), NADH dehydrogenase subunit 5 (nad5) and cytochrome b (cytb), among Trichuris ovis isolates from different hosts in Guangdong Province, China. A portion of the cox1 (pcox1), nad5 (pnad5) and cytb (pcytb) genes was amplified separately from individual whipworms by PCR, and was subjected to sequencing from both directions. The size of the sequences of pcox1, pnad5 and pcytb was 618, 240 and 464 bp, respectively. Although the intra-specific sequence variations within T. ovis were 0-0.8% for pcox1, 0-0.8% for pnad5 and 0-1.9% for pcytb, the inter-specific sequence differences among members of the genus Trichuris were significantly higher, being 24.3-26.5% for pcox1, 33.7-56.4% for pnad5 and 24.8-26.1% for pcytb, respectively. Phylogenetic analyses using combined sequences of pcox1, pnad5 and pcytb, with three different computational algorithms (maximum likelihood, maximum parsimony and Bayesian inference), indicated that all of the T. ovis isolates grouped together with high statistical support. These findings demonstrated the existence of intra-specific variation in mtDNA sequences among T. ovis isolates from different hosts, and have implications for studying molecular epidemiology and population genetics of T. ovis.

Clonorchiasis caused by Clonorchis sinensis is an important foodborne parasitosis of humans and animals, and is predominantly a hepatobiliary disease. Globally, nearly 35 million people were infected with C. sinensis, with approximately 15 million being in China. Patients would chronically present fatigue, jaundice, abdominal discomfort, along with the increased risk of developing into a form of cholangiocarcinoma that is fatal to humans. Treatment of clonorchiasis by praziquantel has been very successful, but this is dependent on early accurate diagnosis and correct species identification. The present article reviews the current status of knowledge in genomics and functional genomics of C. sinensis, and summarizes the main DNA-based techniques for the specific diagnosis of C. sinensis infection and studies of genetic variation in C. sinensis, and provides perspectives for future studies. The advances in genomics and molecular genetics of C. sinensis shed new sight on our understanding of population structure of C. sinensis as well as the prevention and control of clonorchiasis.

Clonorchiasis caused by Clonorchis sinensis is a fish-borne parasitic disease which is endemic in a number of countries. Using the sequences of the internal transcribed spacers (ITS-1 and ITS-2) of nuclear ribosomal DNA (rDNA) of C. sinensis as genetic markers, a pair of C. sinensis-specific primers was designed and used to establish a specific PCR assay for the diagnosis of C. sinensis infection in humans, cats and fish. This approach allowed the specific identification of C. sinensis after optimizing amplification conditions, with no amplicons being amplified from related heterogeneous DNA samples, and sequencing of amplicons confirmed the identity of the sequences amplified. The detection limit of this assay was 1.03 pg of adult C. sinensis, 1.1 metacercariae per gram of fish filet, and a single egg in human and cat feces. The PCR assay should provide a useful tool for the diagnosis and molecular epidemiological investigation of clonorchiasis in humans and animals.

Toxoplasma gondii, a eukaryotic parasite of the phylum Apicomplexa, can infect almost any nucleated mammalian and avian cells. Micronemes are sub-cellular organelles of apicomplexan parasites which can secrete microneme proteins (MICs) playing a key role in the invasion process of T. gondii and are potential vaccine candidate molecules against toxoplasmosis. In this work, we examined sequence variation in T. gondii microneme protein 13 (TgMIC13) gene sequences among 18 isolates from different hosts and geographical locations by polymerase chain reaction (PCR) amplification and subsequent sequence analysis. The complete TgMIC13 DNA and cDNA sequences were 2506-2507 and 1407 bp, respectively, and they were quite conserved among the T. gondii isolates, with intra-specific variation of only up to 0.84% (21/2507) for genomic DNA and up to 0.71% (10/1407) for cDNA sequences. Phylogenetic analysis using maximum parsimony (MP) revealed that the TgMIC13 DNA sequences were not able to provide an effective molecular marker for intra-species phylogenetic analysis and differential identification of T. gondii isolates from different hosts and geographical locations. However, the results demonstrated that sequence variation in TgMIC13 gene was quite low among different T. gondii isolates, which may be a useful feature as an anti-toxoplasmosis vaccine candidate molecule in further studies. Key words: Toxoplasma gondii, toxoplasmosis, microneme protein 13 (MIC13), sequence variation, phylogenetic analysis

Ascaris lumbricoides and Ascaris suum are parasitic nematodes living in the small intestine of humans and pigs, and can cause the disease ascariasis. For long, there has been controversy as to whether the two ascaridoid taxa represent the same species due to their significant resemblances in morphology. However, the complete mitochondrial (mt) genome data have been lacking for A. lumbricoides in spite of human and animal health significance and socio-economic impact globally of these parasites. In the present study, we sequenced the complete mt genomes of A. lumbricoides and A. suum (China isolate), which was 14,303 bp and 14,311 bp in size, respectively. The identity of the mt genomes was 98.1% between A. lumbricoides and A. suum (China isolate), and 98.5% between A. suum (China isolate) and A. suum (USA isolate). Both genomes are circular, and consist of 36 genes, including 12 genes for proteins, 2 genes for rRNA and 22 genes for tRNA, which are consistent with that of all other species of ascaridoid studied to date. All genes are transcribed in the same direction and have a nucleotide composition high in A and T (71.7% for A. lumbricoides and 71.8% for A. suum). The AT bias had a significant effect on both the codon usage pattern and amino acid composition of proteins. Phylogenetic analyses of A. lumbricoides and A. suum using concatenated amino acid sequences of 12 protein-coding genes, with three different computational algorithms (Bayesian analysis, maximum likelihood and maximum parsimony) all clustered in a clade with high statistical support, indicating that A. lumbricoides and A. suum was very closely related. These mt genome data and the results provide some additional genetic evidence that A. lumbricoides and A. suum may represent the same species. The mt genome data presented in this study are also useful novel markers for studying the molecular epidemiology and population genetics of Ascaris.

The beef tapeworm Taenia saginata infects human beings with symptoms ranging from nausea, abdominal discomfort to digestive disturbances and intestinal blockage. In the present study, microRNA (miRNA) expressing profile in adult T. saginata was analyzed using Solexa deep sequencing and bioinformatics analysis. A total of 15.8 million reads was obtained by Solexa sequencing, and 13.3 million clean reads (1.73 million unique sequences) was obtained after removing reads smaller than 18 nt. Ten conserved miRNAs corresponding to 607,382 reads were found when matching the reads against known miRNAs of Schistosoma japonicum in miRBase database. The miR-71 had the most abundant expression in T. saginata, followed by miR-219-5p, but some other common miRNAs such as let-7, miR-40, and miR-103 were not identified in T. saginata. Nucleotide bias analysis found that the known miRNAs showed high bias and the uracil was the dominant nucleotide, particularly at the first and 11th positions which were almost at the beginning and middle of conserved miRNAs. One novel miRNA (Tsa-miR-001) corresponding to ten precursors was identified and confirmed by stem-loop RT-PCR. To our knowledge, this is the first report of miRNA profiles in T. saginata, which will contribute to better understanding of the complex biology of this zoonotic trematode. The reported data of T. saginata miRNAs should provide valuable references for miRNA studies of closed related zoonotic Taenia cestodes such as Taenia solium and Taenia asiatica.

The seroprevalence of Mycoplasma bovis infection in dairy cows in Guangxi Zhuang Autonomous Region (GZAR) in subtropical southern China was surveyed between June 2009 and March 2010. A total of 455 serum samples of dairy cows were collected from 6 districts in 4 different cities, and examined for M. bovis antibodies with the indirect enzyme-linked immunosorbent assay (ELISA) using a commercially available kit. The overall seroprevalence of M. bovis infection in dairy cows was 7.69% (35/455). Three year-old dairy cows had the highest seroprevalence (15.0%), followed by dairy cows of 4 year-old (11.1%). Dairy cows with the history of 5 pregnancies had the highest seroprevalence (33.3%). However, no statistically significant association was found between M. bovis infection and age or number of pregnancies (p > 0.05). All the aborting dairy cows were negative for M. bovis antibodies, suggesting that bovine abortion may have no association with M. bovis infection in GZAR. These results indicate that M. bovis infection in dairy cows was widespread in GZAR, and integrated strategies and measures should be performed to control and prevent M. bovis infection and disease outbreak.Key words: Mycoplasma bovis, seroprevalence, dairy cows, Guangxi Zhuang Autonomous Region (GZAR), China, enzyme-linked immunosorbent assay (ELISA).

Angiostrongylus cantonensis causes eosinophilic meningitis and eosinophilic pleocytosis in humans and is of significant socio-economic importance globally. microRNAs (miRNAs) are endogenous small non-coding RNAs that play crucial roles in gene expression regulation, cellular function and defense, homeostasis and pathogenesis. They have been identified in a diverse range of organisms. The objective of this study was to determine and characterize miRNAs of female and male adults of A. cantonensis by Solexa deep sequencing. A total of 8,861,260 and 10,957,957 high quality reads with 20 and 23 conserved miRNAs were obtained in females and males, respectively. No new miRNA sequence was found. Nucleotide bias analysis showed that uracil was the prominent nucleotide, particularly at positions of 1, 10, 14, 17 and 22, approximately at the beginning, middle and the end of the conserved miRNAs. To our knowledge, this is the first report of miRNA profiles in A. cantonensis, which may represent a new platform for studying regulation of genes and their networks in A. cantonensis.

Toxoplasma gondii is an obligate intracellular protozoan parasite, which can invade and multiply within the macrophages of humans and most warm-blooded animals. Macrophages are important effector cells for the control and killing of intracellular T. gondii, and they may also serve as long-term host cells for the replication and survival of the parasite. In the present study, we explored the proteomic profile of macrophages of the specific pathogen-free Kunming mice at 24 h after infection with tachyzoites of the virulent T. gondii RH strain using two-dimensional gel electrophoresis combined with matrix-assisted laser desorption ionization time-of-flight (TOF)/TOF tandem mass spectrometry. Totally, 60 differentially expressed protein spots were identified. Among them, 52 spots corresponded to 38 proteins matching to proteins of the mouse, including actin, enolase, calumenin, vimentin, plastin 2, annexin A1, cathepsin S, arginase-1, arachidonate 12-lipoxygenase, and aminoacylase-1. Functional prediction using Gene Ontology database showed that these proteins were mainly involved in metabolism, structure, protein fate, and immune responses. The findings provided an insight into the interactive relationship between T. gondii and the host macrophages, and will shed new lights on the understanding of molecular mechanisms of T. gondii pathogenesis.

Ascaris suum is an important parasite of pigs that causes tremendous economic losses globally to agriculture and animal husbandry annually. RNA interference (RNAi) technology has been described as a successful and useful approach for the elucidation of gene function in parasitic nematodes. In the present study, RNAi was used to silence the expression of a gene encoding enolase in A. suum by soaking infective larvae in double-stranded RNA derived from an EST (representing As-enol-1) selected from an A. suum infective larvae-specific cDNA library. The mRNA levels of RNAi-treated larvae were examined by Reverse-Transcription PCR (RT-PCR) analysis. The survival of RNAi-treated larvae was compared with larvae treated with dsRNA-free culture medium. The effect of enolase depletion on the development of A. suum larvae was assessed by infecting BALB/c mice with RNAi-treated larvae. The results showed that enolase gene expression was silenced completely and the survival rate of the RNAi-treated nematodes was reduced by 20.11% (P

Enzootic pneumonia caused by Mycoplasma hyopneumoniae is a severe disease of pigs, causing significant economic losses to the pig industry worldwide, including the tropical and subtropical regions. In order to obtain the baseline prevalence of M. hyopneumoniae in pigs from intensive farms in southern China, double-sandwich enzyme-linked immunosorbent assay (ELISA) was used to detect M. hyoneumoniae antibodies in 460 pig serum samples collected from 12 administrative cities in China's southern Guangdong province. According to the proportions of the infected animals, among the 12 intensive farms, only two of them showed no infection of M. hyoneumoniae and the seroprevalence ranged from 0% to 90%, with an averaged prevalence of 45.7%. The highest prevalence was found in breeding boars (68.8%), followed by sows (54.5%). These data showed that the infection of pigs with M. hyopneumoniae is severe, and boars might be more important carriers and transfers of M. hyoneumoniae than sows. Integrated strategies and measures should be taken to control the infection of pigs with M. hyopneumoniae in southern China.

Little is known of the molecular detection of Toxoplasma gondii infection in chickens (Gallus domesticus). The objectives of the present study were to determine the suitable tissues of chickens infected with T. gondii for direct polymerase chain reaction (PCR) amplification of T. gondii DNA. Thirty, 35-day-old broiler chickens were divided into three groups of 10 birds (two replications of five chicks). Of these, two groups were experimentally inoculated intravenously with 4.3×10(6) or 4.3×10(7) tachyzoites of the low virulent T. gondii QHO strain. Two inoculated chickens from each of the two groups were killed on days 7, 14, 21, 28, and 35 post-inoculation, respectively, and two uninoculated chickens were also killed at the same time. Sera from chickens were collected for examination of anti-T. gondii antibodies by indirect hemagglutination test (IHAT) and the modified agglutination test (MAT). Brains, hearts, livers, lungs, spleens and eyes of chickens were sampled and DNA from each tissue was extracted as template for PCR assay. Specific anti-T. gondii antibodies were detected in all infected chickens from day 7 to day 35 p.i. with antibody titers between 1:5 and 1:640 by MAT. PCR assay can detect T. gondii DNA in tissues from the day 21 p.i. to day 28 p.i. This study demonstrates that MAT is more sensitive than IHAT for detecting antibodies to T. gondii in chickens, and PCR assay is a specific, speedy, sensitive and cost-effective method for detecting T. gondii DNA in chickens.

The fish-borne clonorchiasis caused by the oriental liver fluke Clonorchis sinensis is endemic in a number of countries with over 35 million people being infected globally. Rapid and accurate detection of C. sinensis in its intermediate host fish is important for the control and prevention of clonorchiasis in areas where the disease is endemic. In the present study, we established a loop-mediated isothermal amplification (LAMP) approach for the sensitive and rapid detection of C. sinensis metacercariae in fish. The specificity and sensitivity of primers designed from the C. sinensis cathepsins B3 gene were evaluated, and specific amplification products were obtained with C. sinensis, while no amplification products were detected with DNA of related trematodes, demonstrating the specificity of the assay. The LAMP assay was proved to be 100 times more sensitive than a conventional polymerase chain reaction for detection of C. sinensis. The established LAMP assay provides a useful tool for the rapid and sensitive detection of C. sinensis in fish, which has important implications for the effective control of human clonorchiasis.

Monascus products are used as both natural colorants and food additives all over the world. However, its safety is really an issue because of the presence of citrinin, which is considered to be hepatotoxic and nephrotoxic. Therefore, the objective of the present study was to develop an approach for using a fragment of pigment-related gene pks1 to replace citrinin-activator gene ctnA. The character of citrinin antibacterial activity to Bacillus subtilis was used for primary screening of transformants based on the foundation that the inhibition zone formed around the mutant colonies with low citrinin products will become smaller. The selected mutants were then further confirmed by polymerase chain reaction and high-performance liquid chromatography methods. During all of the processes, antibiotic markers and vectors were avoided. The results showed that the citrinin products of mutants were reduced to 42%, while the pigment products were improved to 33.9%, respectively, over those of the wild-type strains.

MicroRNA-36 (miR-36) is a recently discovered miRNA family which including at least eight members, and specifically existed in helminths compared with other miRNAs that widely exists in almost all kinds of lives. This paper reviews recent research advances about miR-36 to provide further fundamental information for helminth and miRNA study.

Toxoplasmosis, caused by Toxoplasma gondii, is a parasitic zoonosis with worldwide distribution. The present study investigated the prevalence of T. gondii in dogs in Zhanjiang city, southern China, using both serological and molecular detection. A total of 364 serum samples and 432 liver tissue samples were collected from the slaughter house between December 2012 and January 2013 and were examined for T. gondii IgG antibody by ELISA and T. gondii DNA by semi-nested PCR based on B1 gene, respectively. The overall seroprevalence of T. gondii IgG antibody was 51.9%, and T. gondii DNA was detected in 37 of 432 (8.6%) liver tissue samples. These positive DNA samples were analyzed by PCR-RFLP at 3'- and 5'-SAG2. Only 8 samples gave the PCR-RFLP data, and they were all classified as type I, which may suggest that the T. gondii isolates from dogs in Zhanjiang city may represent type I or type I variant. This study revealed the high prevalence of T. gondii infection in dogs in Zhanjiang city, southern China. Integrated measures should be taken to prevent and control toxoplasmosis in dogs in this area for public health concern.

Rhipicephalus sanguineus is an ectoparasite of medical and veterinary significance, which can transmit a number of pathogens including Babesia canis, Ehrlichia canis, and Rickettsia conorii. MicroRNAs (miRNAs) are recognized as regulators in sex differentiation in dioecious species. We here investigated and compared the miRNA profiles of male and female adults of R. sanguineus by combining Solexa deep sequencing with bioinformatics platform and quantitative real-time PCR. A total of 11.88 and 16.09 million raw reads were obtained from male and female R. sanguineus, respectively. By mapping to the reference genome, 59 and 76 miRNA candidates from the female and male parasite were obtained, with 19 of each consistent with known Ixodes scapularis miRNAs deposited in the miRBase database. Besides, 51 miRNAs were shared by the two genders, and 8 and 25 were female and male specific, respectively. The number of predicted targets of the identified miRNAs ranged from 1 to 383 with an average of 176. Functional analysis showed that a number of predicted targets corresponded to transcription, splicing, and translation factors, elongation factors, and growth factors which were essential for the development of the parasite. Enrichment analysis revealed that some functions of the predicted targets were higher in female than in male, such as antioxidant and electron carrier. The present study firstly described the global profiling of miRNAs in male and female R. sanguineus and identified a number of gender-specific miRNAs, which are likely to participate in the sex differentiation/maintenance process and provide novel resources for better understanding of the biology of the parasite, and may further lead to effective control of the parasite and diseases it causes.

Eurytrema pancreaticum is one of the most common trematodes living in the pancreatic and bile ducts of ruminants and also occasionally infects humans, causing eurytremiasis. In spite of its economic and medical importance, very little is known about the genomic resources of this parasite. Herein, we performed de novo sequencing, assembly and characterization of the transcriptome of adult E. pancreaticum. Approximately 36.4 million high-quality clean reads were obtained, and the length of the transcript contigs ranged from 66 to 19,968 nt with mean length of 479 nt and N50 length of 1094 nt, and then 23,573 unigenes were assembled. Of these unigenes, 15,353 (65.1%) were annotated by blast searches against the NCBI non-redundant protein database. Among these, 15,267 (64.8%), 2732 (11.6%) and 10,354 (43.9%) of the unigenes had significant similarity with proteins in the NR, NT and Swiss-Prot databases, respectively. 5510 (23.4%) and 4567 (19.4%) unigenes were assigned to GO and COG, respectively. 8886 (37.7%) unigenes were identified and mapped onto 254 pathways in the KEGG Pathway database. Furthermore, we found that 105 (1.18%) unigenes were related to pancreatic secretion and 61 (0.7%) to pancreatic cancer. The present study represents the first transcriptome of any members of the family Dicrocoeliidae, which has little genomic information available in the public databases. The novel transcriptome of E. pancreaticum should provide a useful resource for designing new strategies against pancreatic flukes and other trematodes of human and animal health significance.

Foot-and-mouth disease virus (FMDV) causes a highly contagious disease of cloven-hoofed animals, which leads to serious economical losses. FMDV is not adequately controlled by vaccination or biosecurity measures. To generate genetically modified FMDV-resistant animals, a combinatorial expression cassette producing three short hairpin (sh) RNAs was constructed using the lentivirus (LV) vector, LV-3shRNA. The three shRNAs were expressed under the regulation of DNA polymerase III promoters from a buffalo and a bovine source, with one targeted to the non-structural protein 3B, and the other two targeted to the viral polymerase protein 3D of FMDV, respectively. The role of LV-3shRNA in the inhibition of the replication of FMDV was determined in BHK-21 cells and in suckling mice. The results revealed that LV-3shRNA reduced viral growth 3-fold (24 h post-infection) when the cells were challenged with 107-times the tissue culture infective dose (TCID50)/ml of O serotype FMDV. The suckling mice pretreated with LV-3shRNA were completely protected on administration of 5-times the dose of FMDV otherwise sufficient to kill 50% of the experimental animals (LD50). These results demonstrated that the LV-mediated dual expression of three FMDV-specific shRNAs provided a novel strategy towards combating FMDV, which facilitates the permanent introduction of novel disease-resistance traits into the buffalo and bovine genomes in the future.

[Background] The parasitic nematodes Ascaris lumbricoides and A. suum are of great public health and economic significance, and the two taxa were proposed to represent a single species. miRNAs are known with functions of gene regulations at post-transcriptional level., [Results] We herein compared the miRNA profiles of A. lumbricoides and A. suum female adults by Solexa deep sequencing combined with bioinformatics analysis and stem-loop real-time PCR. Using the A. suum genome as the reference genome, we obtained 171 and 494 miRNA candidates from A. lumbricoides and A. suum, respectively. Among which, 74 miRNAs were shared between the two taxa, 97 and 420 miRNAs were A. lumbricoides and A. suum specific. Target and function prediction revealed a significant set of targets which are related to ovarian message protein, vitellogenin and chondroitin proteoglycan of the two nematodes. Enrichment analysis revealed that the percentages of most predicted functions of the miRNA targets were similar, with some taxon specific or taxon enhanced functions, such as different target numbers, specific functions (NADH dehydrogenase and electron carrier functions), etc., [Conclusions] This study characterized comparatively the miRNAs of adult A. lumbricoides and A. suum, and the findings provide additional evidence that A. lumbricoides and A. suum represent a single species. Due to the fast evolution nature of miRNAs and the different parasitic living conditions of humans and pigs, the phenomenon above might indicate a fast evolution of miRNAs of Ascaris in humans and pigs., This work was supported in part by the International Science & Technology Cooperation Program of China (Grant No. 2013DFA31840), the Science Fund for Creative Research Groups of Gansu Province (Grant No. 1210RJIA006) to XQZ, and the China Postdoctoral Science Foundation (Grant No. 201104363) to MJX.

Background Baylisascaris schroederi is one of the most significant threats to the giant panda’s survival, responsible for half of the deaths reported from 2001 to 2005. MicroRNA (miRNA) has been identified as one of the key factors for gene regulations at the post-transcriptional level, and also considered as a potential control and treatment target against infectious diseases. Methods The present study investigated the miRNA profile of B. schroederi via high throughput sequencing and real-time quantitative PCR. Results A total of 18.07 million raw reads were obtained and 18.01 million were identified with high quality. By analysis of standard stem-loop structures, 108 miRNA candidates were predicted, including 60 known miRNAs and 48 novel ones. Target prediction revealed that the “chitinase” was the most abundant target with 483 sequences, and 263 targets were related to ovarian and egg development. The ribosomal protein related sequences occupied 449 sequences. Conclusions Previous studies have shown that some parasites secrete chitinases for exsheathment and/or for penetrating the peritrophic matrix of the host. It therefore seems that B. schroederi may be effectively regulated by miRNAs for development, invasion, and reproduction. Given that chitinases have been identified as important biological control agents for pests, identification of microRNAs in B. schroederi of the giant panda would provide useful information for the development of biological control strategies and/or vaccines against B. schroederi infection in the giant panda.

Background The obligate intracellular parasite Toxoplasma gondii can interfere with host cell signaling pathways, alter host defense systems and cell cycle control, and establish a chronic infection in the central nervous system. T. gondii infection may alter the expression profile of host microRNAs (miRNAs) which have key regulatory functions at the post-transcriptional level. Methods Using high-throughput sequencing and real-time quantitative PCR technology, we compared the miRNA expression profiles of uninfected mouse brains with brains from mice at 14 days and 21 days after infection with cyst-forming T. gondii (Type II). Results A total of 51.30 million raw reads were obtained from all samples and 495 (14d infected mouse sample), 511 (14d sham-infected control), 504 (21d infected mouse sample) and 514 (21d sham-infected control) miRNA candidates identified. Among these, 414 miRNAs were consistent across all the studied groups, 17 were specific to the 14d infected group and 32 were specific to the 21d infected group. In addition, 9 miRNAs were common to both the 14d- and 21d-infected groups. Enrichment analysis for the targets of these miRNAs showed a high percentage of “protein tag” functions. Immune related targets including chemokines, cytokines, growth factors and interleukins were also found. Conclusions These results not only showed that the miRNA expression of the host can be changed by the invasion of cyst-forming T. gondii, but also indicated that the host attempts to respond using two tactics: marking proteins with “protein tags” and adaptation of immune related systems.

Chlamydia spp. are obligate intracellular gram-negative bacteria that cause a wide range of significant diseases in humans and animals worldwide, resulting in significant economic losses. Chlamydial infection in cattle has been reported in many countries including China. However, there has been no survey of chlamydial infection of dairy cattle in Guangzhou, southern China. The objective of the present investigation was to examine the chlamydial seroprevalence in dairy cattle in Guangzhou, subtropical southern China by using an indirect hemagglutination assay (IHA). The overall seroprevalence of chlamydial infection in dairy cattle was 7.25% (29/400). Greater than or equal to eight-yr-old dairy cattle had the highest seroprevalence (10.34%), followed by those that were ≥ 6 years old or P > 0.05). Dairy cattle with 5 pregnancies had the highest seroprevalence (10.81%). These results indicate that chlamydial infection was present in dairy cattle in Guangzhou, subtropical southern China, and integrated strategies and measures should be executed to control and prevent chlamydial infection and disease outbreak in the study region.

Toxoplasma gondii is a protozoan parasite infecting almost all warm-blooded animals and humans. There are three infective stages of T. gondii: the tachyzoites, the bradyzoites, and the oocysts. The tachyzoite is a rapidly multiplying stage and the main pathogenic factor. In North America and Europe, T. gondii is consisted of four major clonal lineages (namely Types I, II, III, and Type 12). In this study, we explored the proteomic profiles of different genotypes (Type I-RH strain, Type II-PRU strain, Type II-TgQHO strain, and ToxoDB 9-TgC7 strain) of T. gondii tachyzoites by using 2D DIGE combined with MALDI-TOF MS. Totally, 110 differentially abundant protein spots were selected. Of these, 98 spots corresponding to 56 proteins from T. gondii were successfully identified. These included surface antigen (SAG1), heat shock protein 70 (Hsp 70), disulfide isomerase, coronin, heat shock protein 60 (Hsp 60), pyruvate kinase, receptor for activated C kinase 1, and peroxiredoxin. Gene ontology enrichment analysis revealed that most of the differentially abundant proteins were involved in biological regulation, metabolic process, response to stress, binding, antioxidant activity, and transporter activity. According to the KEGG metabolic pathway maps of T. gondii, some identified proteins were involved in the glycolytic/gluconeogenesis pathway. The present study identified differentially abundant proteins among different genotypes of T. gondii and these findings have implications for the better understanding of the phenotypic differences among the examined T. gondii genotypes, which in turn may contribute to the better control of toxoplasmosis.

Background Eurytrema pancreaticum is one of the most common flukes, which mainly infects ruminants globally and infects human beings accidentally; causing eurytremiasis that has high veterinary and economic importance. MicroRNAs (miRNAs) are small non-coding RNAs and are now considered as a key mechanism of gene regulation at the post-transcription level. Methods We investigated the global miRNA expression profile of E. pancreaticum adults using next-generation sequencing technology combined with real-time quantitative PCR. Results By using the genome of the closely-related species Schistosoma japonicum as reference, we obtained 27 miRNA candidates out of 16.45 million raw sequencing reads, with 13 of them found as known miRNAs in S. japonicum and/or S. mansoni, and the remaining 14 miRNAs were considered as novel. Five out of the 13 known miRNAs coming from one family named as sja-miR-2, including family members from miR-2a to miR-2e. Targets of 19 miRNAs were successfully predicated out of the 17401 mRNA and EST non-redundant sequences of S. japonicum. It was found that a significant high number of targets were related to “chch domain-containing protein mitochondrial precursor” (n = 29), “small subunit ribosomal protein s30e” (n = 21), and “insulin-induced gene 1 protein” (n = 9). Besides, “egg protein cp3842” (n = 2), “fumarate hydratase” (n = 2), “ubiquitin-conjugating enzyme” (n = 2), and “sperm-associated antigen 6” (n = 1) were also found as targets of the miRNAs of E. pancreaticum. Conclusions The present study represents the first global characterization of E. pancreaticum miRNAs, which provides novel resources for a better understanding of the parasite, which, in turn, has implications for the effective control of the disease it causes.

Background Chlamydiaceae is a family of obligate intracellular pathogens with a worldwide distribution in many animal species, including humans. No information exists on the prevalence of Chlamydia felis infections in cats and dogs in Lanzhou, the geographical center of China. The aim of this study was to carry out a census of cats and dogs in Lanzhou and document the seroprevalence of C. felis exposure in these companion animals. Results In this study, blood samples were collected from 485 animals (221 cats and 264 pet dogs) in Lanzhou between November 2010 and July 2011 to identify antibodies against C. felis. Thirteen of 221 (5.9%) cats and 32 of 264 (12.1%) pet dogs were positive for C. felis infection using indirect hemagglutination at a cutoff of 1:16. The seroprevalence in household and stray cats was 3.9% and 14.3%, respectively, and the difference was statistically significant (P < 0.05). Among different age groups, the seroprevalence in cats varied from 1.9 to 7.9%, and that in dogs ranged from 9.6 to 20.4%; however, the differences were not statistically significant (P > 0.05). Conclusions This is the first report of the seroprevalence of C. felis exposure in cats and dogs in Lanzhou, northwestern China. Our results indicate that the presence of C. felis exposure in cats and dogs may pose a potential threat to human health.

Interleukin-15 (IL-15) is a pro-inflammatory cytokine and also a chemotactic factor for a variety of immune cells, which participates in and regulates the inflammatory response and immune response to organisms. Up to now, the molecular structure and receptor of IL-15 have been widely researched, with important advances in its signal transduction way. However, its application in parasitology is still a relatively new topic. This article summarizes the latest research development of IL-15 in anti-parasitic infection, prospects its mechanism and application.

Ascaris nematodes, which cause ascariasis in humans and pigs, are among the most important nematodes from both health and economic perspectives. microRNA (miRNA) is now recognized as key regulator of gene expression at posttranscription level. The public availability of the genome and transcripts of Ascaris suum provides powerful resources for the research of miRNA profiles of the parasite. Therefore, we investigated and compared the miRNA profiles of male and female adult A. suum using Solexa deep sequencing combined with bioinformatic analysis and stem-loop reverse transcription polymerase chain reaction. Deep sequencing of small RNAs yielded 11.71 and 11.72 million raw reads from male and female adults of A. suum, respectively. Analysis showed that the noncoding RNA of the two genders, including tRNA, rRNA, snRNA, and snoRNA, were similar. By mapping to the A. suum genome, we obtained 494 and 505 miRNA candidates from the female and male parasite, respectively, and 87 and 82 of miRNA candidates were consistent with A. suum miRNAs deposited in the miRBase database. Among the miRNA candidates, 154 were shared by the two genders, and 340 and 351 were female and male specific with their target numbers ranged from one to thousands, respectively. Functional prediction revealed a set of elongation factors, heat shock proteins, and growth factors from the targets of gender-specific miRNAs, which were essential for the development of the parasite. Moreover, major sperm protein and nematode sperm cell motility protein were found in targets of the male-specific miRNAs. Ovarian message protein was found in targets of the female-specific miRNAs. Enrichment analysis revealed significant differences among Gene Ontology terms of miRNA targets of the two genders, such as electron carrier and biological adhesion process. The regulating functions of gender-specific miRNAs was therefore not only related to the fundamental functions of cells but also were essential to the germ development of the parasite. The present study provides a framework for further research of Ascaris miRNAs, and consequently leads to the development of potential nucleotide vaccines against Ascaris of human and animal health significance.

Toxoplasma gondii infection is widely prevalent in humans and animals, including pigs throughout the world. In this study, the seroprevalence of T. gondii infection in Tibetan pigs in China was investigated for the first time. A total of 427 serum samples were collected from Tibetan pigs in Nyingchi prefecture, Tibet, between April and December 2010, and were assayed for antibodies to T. gondii using the modified agglutination test (MAT). Ninety-seven (22.72%) pigs were found to be positive with MAT titers of 1:25 or higher. Slaughter pigs had the highest seroprevalence, compared with seroprevalence in fattening pigs, growing pigs, or piglets, although the difference was not statistically significant (p≥0.05). The results of the present survey indicate that T. gondii is highly prevalent in Tibetan pigs in Tibet, which poses a significant public health concern in this unique region of the world.

Mycoplasma bovis is a major bacterial pathogen causing mastitis in dairy cattle, pneumonia and arthritis, and reduced weight gain in calves and reproductive problems in both dairy cattle and bulls, resulting in significant economic losses. The objective of the present investigation was to examine the M. bovis seroprevalence in dairy cattle in Guangzhou, subtropical Southern China by using an enzyme-linked immunosorbent assay (ELISA). A total of 370 serum samples of dairy cattle were collected between July, 2009 and March, 2010 from 5 different farms and examined for M. bovis antibodies using a commercially available ELISA kit. The overall seroprevalence of M. bovis infection in dairy cattle was 5.95% (22/370). One year-old dairy cattle had the highest seroprevalence (10.5%), followed by dairy cattle of 3 year-old (9.61%). Dairy cattle without pregnancy had the highest seroprevalence (9.26%), followed by dairy cattle with 2 pregnancies (8.62%). However, no statistically significant association was found between M. bovis infection and ages or numbers of pregnancies (P>0.05). These results indicate that M. bovis infection was present in dairy cattle in Guangzhou, subtropical Southern China, and integrated strategies and measures should be executed to control and prevent M. bovis infection and disease outbreak in the study region. Key words: Mycoplasma bovis, seroprevalence, dairy cattle, China, enzyme-linked immunosorbent assay (ELISA).

Chlamydophila is an obligate intracellular bacterium, found in a wide range of birds, which is frequently transmitted from infected birds to humans. In the present study, a total of 313 serum samples from free-living, clinically healthy house sparrows were examined using an indirect hemagglutination assay (IHA). Antibodies against Chlamydophila were found in 33 (10.54%) of 313 serum samples (IHA titer ≥ 1:16). The result indicated that clinical healthy status was not a reliable “standard” for the diagnosis of Chlamydophila infection and there was a potential risk for human infection with Chlamydophila in this region. The present study is the i¬rst seroprevalence survey of sparrows infected by Chlamydophila in China. Key words: Chlamydophila, Seroprevalence, house sparrows, Lanzhou, northwest China, indirect hemagglutination assay (IHA).

Canine and feline parasitic zoonoses have not been given high priority in China, although the role of companion animals as reservoirs for zoonotic parasitic diseases has been recognized worldwide. With an increasing number of dogs and cats under unregulated conditions in China, the canine and feline parasitic zoonoses are showing a trend towards being gradually uncontrolled. Currently, canine and feline parasitic zoonoses threaten human health, and cause death and serious diseases in China. This article comprehensively reviews the current status of major canine and feline parasitic zoonoses in mainland China, discusses the risks dogs and cats pose with regard to zoonotic transmission of canine and feline parasites, and proposes control strategies and measures.