[A pre-conception cohort study of fertility and its related factors among couples with the intention of conception].

Objective: To describe fertility and explore factors associated with it among pre-conception couples of childbearing age. Methods: Based on the pre-conceptional offspring trajectory study of the School of Public Health of Fudan University, couples of childbearing age who participated in the pre-conception physical examination in Shanghai Jiading District from 2016 to 2021 were recruited and followed up. Couples' time to pregnancy (TTP) was analyzed and Cox proportional hazards regression model was used to explore the factors associated with TTP. Kaplan-Meier was used to calculate each menstrual cycle's cumulative pregnancy rate. Results: A total of 1 095 preconception couples were included in the analysis, the M(Q1,Q3)of TTP was 4.33 (2.41, 9.78) menstrual cycles. Age of women (FR=0.90, 95%CI: 0.85-0.95, P<0.001), women who were overweight or obese before pregnancy (FR=0.36, 95%CI: 0.24-0.55, P<0.001), women who were exposed to second-hand smoking (FR=0.63, 95%CI: 0.44-0.92, P=0.016), women whose home or office had been renovated in the past 2 years and had a particular smell (FR=0.46, 95%CI: 0.26-0.81, P=0.008) were risk factors for impaired fertility. Regular menstrual cycles (FR=1.64, 95%CI: 1.16-2.31, P=0.005), females who often drank tea/coffee (FR=1.55, 95%CI: 1.11-2.17, P=0.011) and males who took folic acid before conception (FR=2.35, 95%CI: 1.38-4.23, P=0.002) were associated with better fertility. The cumulative pregnancy rate of 3, 6, and 12 menstrual cycles was 37.6%, 64.4%, and 78.4%, respectively. Conclusion: Older couples, overweight or obesity before pregnancy, irregular menstruation, exposure to secondhand smoke and decoration pollutants in females are associated with impaired fertility. Frequent tea/coffee drinking before pregnancy in females and taking folic acid before pregnancy in males are associated with shortened conception time.

Investigation of the motor skills assessments of typically developing preschool children in China.

BACKGROUND: The assessment of preschoolers' motor skills is essential to know young children's motor development and evaluate the intervention effects of promotion in children's sports activities. The purpose of this study was to review the motor skills assessment tools in Chinese pre-school-aged children, compare them in the international context, and provide guidelines to find appropriate motor skill assessment tools for developing children in China. METHODS: A comprehensive literature search was carried out using the WANFAGN, CNKI, VIP, ERIC, EMBASE, MEDLINE, and SPORT Discus databases. Relevant articles published between January 2000 and May 2020 were retrieved. Studies that described the discriminative and evaluative measures of motor skills among the population aged 3-6 years in China were included. RESULTS: A total of 17 studies were included in this study describing seven tools, including four self-developed tools and three international tools used in China. TGMD-2 appeared in a large proportion of the studies. The international tools used in China were incomplete in terms of translation, verification of reliability and validity, item selection, and implementation. Regarding the self-constructed tools, the CDCC was the most utilized self-developed tool, but it was mainly applied in intellectual development assessment. By comparing Chinese self-constructed and international tools, the construction of the CDCC and the Gross Motor Development Assessment Scale contained relatively complete development steps. However, the test content, validity and reliability, implementation instruction, and generalizability of self-constructed tools are still lacking. CONCLUSIONS: Both international and self-developed motor skills assessment tools have been rarely applied in China. Available tools lack enough validation and appropriate adjustments. Cultural differences in motor development between Chinese and Western populations should be considered when constructing a Chinese localized motor skill assessment tool.

[A pilot study of transcranial alternating current stimulation in the treatment of drug-naive adult patients with major depressive disorder].

Objective: To explore the efficacy, adverse reactions, feasibility, and acceptability of transcranial alternating current stimulation (tACS) treating drug-naive adult patients with major depressive disorder (MDD), and provide basis for further study with a large sample. Methods: The study was performed in the Neuromodulation laboratory, Department of Neurology of Xuanwu Hospital, Capital Medical University (Beijing, China) from July, 2017 to June, 2018. Thirty Eligible first-episode MDD outpatients were randomized 1∶1 to receive active tACS or sham intervention. The tACS was administered in a 40 minute, 77.5 Hz frequency, 15 mA session with one forehead (Fp1, Fpz, and Fp2, in the 10/20 international placement system, 4.45 cm×9.53 cm) and two mastoid (3.18 cm×3.81 cm) stimulation for 20 times in 4 consecutive weeks at fixed day time frame once daily from Monday through Friday, with weekends off (week 4), followed by 4 weeks with no tACS treatment (week 8). By utilizing the Hamilton rating scale for depression-17 item (HRSD-17) to assess the depressive severity of MDD patients, adverse events were administered by the treatment-emergent adverse events, the Young mania rating scale, and the self-made common questionnaire on cranial electrical stimulation. The primary efficacy outcome was the remission rate defined as HRSD-17 score ≤7 at week 8. Secondary outcomes included the rates of remission at week 4 and response at weeks 4 and 8. Safety was assessed by evaluation of adverse events. Also the proportions of participants accepting the intervention and this study procedure were evaluated at weeks 4 and 8. Results: Thirty MDD patients completed the study, and both groups had no statistical differences on their demographic characteristics (P>0.05). At week 8, the active group had a remission rate of 10/15, which was higher than 3/15 in the sham group (P<0.05). Also, the remission rate (14/15) in the active group was higher than 5/15 of the sham group at week 4 (P<0.05). For the response rates, significant differences were found between groups at week 8. For safety, both groups showed no severe adverse events and no mania/hypomania. One participant per group had 2 times of tinnitus cerebri during the intervention days. All patients accepted the intervention and the study procedure. Conclusions: The pilot study indicated that tACS with 77.5 Hz and 15 mA may have a therapeutic effect on depressive symptoms. It is well-tolerated and safe, as well as feasible and acceptable for adults with MDD.

Long non-coding RNA LINC00899 as a novel serum biomarker for diagnosis and prognosis prediction of acute myeloid leukemia.

OBJECTIVE: Recently, several long non-coding RNAs (lncRNAs) have been implicated in acute myeloid leukemia (AML). However, the clinical significance of lncRNAs in AML patients still remains unclear. We aimed to evaluate the expression level of lncRNA LINC00899 (LINC00899) and its potential for diagnosis and prognosis in AML. PATIENTS AND METHODS: Expression levels of LINC00899 in bone marrow and serum obtained from AML patients and healthy controls were assessed by quantitative real-time PCR. Receiver operating characteristic (ROC) curves were used to evaluate the sensitivity and specificity of serum LINC00899. The association between serum LINC00899 expression and clinicopathological factors as well as the overall survival were analyzed. RESULTS: We found that the levels of serum LINC00899 were frequently upregulated in the bone marrow and serum of AML patients. Higher expression of serum LINC00899 was positively associated with FAB classification (p = 0.002) and cytogenetics (p = 0.005). Moreover, ROC curve analyses showed that serum LINC00899 could discriminate AML patients from healthy controls with the area under the curve (AUC) of 0.807 (95% CI, 0.7262- 0.8752). In addition, the serum LINC00899 expression level was significantly reduced when the patients achieved complete remission. Kaplan-Meier analysis showed that patients with high serum LINC00899 expression had a shorter overall survival compared with the low serum LINC00899 expression group (p = 0.0013). Finally, Cox proportional hazards analysis showed that high serum LINC00899 expression was an independent prognostic marker of poor outcome. CONCLUSIONS: We firstly found that serum LINC00899 might be a potential and useful noninvasive biomarker for the early clinical detection and prognosis of AML.

Nucleotide variation in the Toxoplasma gondii micronemal protein 8 gene.

Toxoplasma gondii is a successful opportunistic protozoan distributed worldwide, which can infect all vertebrates, leading to serious infection, blindness, and abortion. Micronemal (MIC) proteins are critically important for T. gondii infection, as they participate in various stages of the Toxoplasma life cycle, including invasion and attachment to host cells. MIC8 secretion relies on the concentration of intracellular calcium, and can mediate the invasion of T. gondii by interacting with soluble MIC3. To investigate genetic diversity of the MIC8 gene, 16 T. gondii strains from different hosts and geographical locations, and two reference isolates (ToxoDB: TGME49_245490 and TGVEG_245490) were examined in this study. The results showed that all the examined MIC8 genes are 2055 bp, with an A+T content ranging from 50.2 to 50.6%. Conversely, lower levels of variation were detected within their nucleotide and amino acid sequences. Phylogenetic analyses indicated that three classical genotypes of T. gondii and the ToxoDB#9 genotype did not group exclusively via Bayesian inference, maximum parsimony, neighbor joining, and/or maximum likelihood assays based on the nucleotide and amino acid sequences of the MIC8 gene. In summary, the T. gondii MIC8 gene is not a suitable marker for population genetic studies of this parasite.

Characterization of Trichuris trichiura from humans and T. suis from pigs in China using internal transcribed spacers of nuclear ribosomal DNA.

Trichuris trichiura and Trichuris suis parasitize (at the adult stage) the caeca of humans and pigs, respectively, causing trichuriasis. Despite these parasites being of human and animal health significance, causing considerable socio-economic losses globally, little is known of the molecular characteristics of T. trichiura and T. suis from China. In the present study, the entire first and second internal transcribed spacer (ITS-1 and ITS-2) regions of nuclear ribosomal DNA (rDNA) of T. trichiura and T. suis from China were amplified by polymerase chain reaction (PCR), the representative amplicons were cloned and sequenced, and sequence variation in the ITS rDNA was examined. The ITS rDNA sequences for the T. trichiura and T. suis samples were 1222-1267 bp and 1339-1353 bp in length, respectively. Sequence analysis revealed that the ITS-1, 5.8S and ITS-2 rDNAs of both whipworms were 600-627 bp and 655-661 bp, 154 bp, and 468-486 bp and 530-538 bp in size, respectively. Sequence variation in ITS rDNA within and among T. trichiura and T. suis was examined. Excluding nucleotide variations in the simple sequence repeats, the intra-species sequence variation in the ITS-1 was 0.2-1.7% within T. trichiura, and 0-1.5% within T. suis. For ITS-2 rDNA, the intra-species sequence variation was 0-1.3% within T. trichiura and 0.2-1.7% within T. suis. The inter-species sequence differences between the two whipworms were 60.7-65.3% for ITS-1 and 59.3-61.5% for ITS-2. These results demonstrated that the ITS rDNA sequences provide additional genetic markers for the characterization and differentiation of the two whipworms. These data should be useful for studying the epidemiology and population genetics of T. trichiura and T. suis, as well as for the diagnosis of trichuriasis in humans and pigs.

Seroprevalence of Chlamydia psittaci infection in market-sold adult chickens, ducks and pigeons in north-western China.

Chlamydia psittaci, the agent of psittacosis in humans, infects a wide range of avian species. To assess the risk of psittacosis posed by domestic birds in the urban environment, the prevalence of C. psittaci antibodies in 413 chickens (Gallus domesticus; 305 caged and 108 free-range), 334 ducks (Anas spp.; 111 caged and 223 free-range) and 312 pigeons (Columba livia) in Lanzhou, north-western China, was detected using the indirect haemagglutination assay. The specific antibodies were found in sera of 55 (13.32 %) chickens, 130 (38.92 %) ducks and 97 (31.09 %) pigeons. Statistical analysis showed that the seroprevalence of C. psittaci infection in chickens was significantly lower than that in ducks and pigeons (P<0.05). The C. psittaci seroprevalence in caged and free-range chickens was 7.54 % and 29.63 %, respectively, and the difference was statistically significant (P<0.05). The C. psittaci seroprevalence in caged and free-range ducks was 26.13 % and 45.29 %, respectively (P<0.05). To our knowledge, this is the first study indicating the presence of C. psittaci infection in market-sold chickens, ducks and pigeons in north-western China. Close contact with these birds is associated with a risk of zoonotic transmission of C. psittaci. Public education should be implemented to reduce the risk of avian to human transmission of such a pathogenic agent.

Comparative characterization of microRNA profiles of different genotypes of Toxoplasma gondii.

The present study compared the miRNA expression profiles of five Toxoplasma gondii strains, namely RH (Type I, ToxoDB10), TgXD (Type I, ToxoDB10), PRU (Type II, ToxoDB1), QHO (Type II, ToxoDB1) and TgC7 (ToxoDB9), by Solexa deep sequencing, bioinformatics analysis and real-time quantitative PCR. A total of 7, 15, 10, 12 and 10 miRNAs were found from RH, TgXD, PRU, QHO and TgC7 strains, respectively. Thirteen miRNAs were shared by three genotypes, with only one miRNA shared by all of the 5 strains and others shared by 2 or more strains. A large number of targets ranging from 1 to 185 were identified for commonly shared miRNAs and strain-specific miRNAs with complete or nearly complete complementarity. Functional prediction showed that these targets were mostly focused on catalytic activity (191 targets) and binding activity (183 targets). Nonetheless, the majority of targets and most of the miRNAs are related to the virulence or invasion proteins of different strains of T. gondii, including ROP and MIC, as well as some other proteins, such as AMA1, GRA and RHO. The present study characterized comparatively the miRNA profiles of 3 different genotypes of T. gondii, identified genotype-shared miRNAs and strain-specific miRNAs.

Sequence variation in three mitochondrial DNA genes among isolates of Ascaridia galli originating from Guangdong, Hunan and Yunnan provinces, China.

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.

Seroprevalence of Toxoplasma gondii and Neospora caninum infection in dairy cows in subtropical southern China.

Toxoplasma gondii and Neospora caninum are closely related protozoan parasites which cause lowered production and increased abortion in dairy cows. The aim of the present study was to determine the seroprevalence of T. gondii and N. caninum infection in dairy cows in the Guangxi Zhuang Autonomous Region (GZAR), subtropical southern China. In total, 875 serum samples were collected from the tail veins of dairy cows in 6 main dairy cow-rearing districts of 4 administrative cities in GZAR. The samples were surveyed for T. gondii antibody using the Indirect Haemagglutination Test (IHA), and 365 of the serum samples were examined for N. caninum antibody by indirect Enzyme-Linked Immunosorbent Assay (ELISA). The overall seroprevalence of T. gondii in dairy cows was 13·71% (120/875), and the average seroprevalence of N. caninum was 15·07% (55/365). There were significant differences in the seroprevalence of N. caninum infection between different districts (P = 0·002, χ 2 = 9·261). The highest prevalences of T. gondii and N. caninum were found in cows older than 8 years and those that had completed 5-6 pregnancies. Five cows (1·37%) presented antibodies against both T. gondii and N. caninum, and dairy cows with both T. gondii and N. caninum antibodies had higher abortion rates. The present results indicate widespread exposure of dairy cows to T. gondii and N. caninum in GZAR, subtropical southern China.

The second transcribed spacer rDNA sequence: an effective genetic marker for inter-species phylogenetic analysis of trematodes in the order Strigeata.

In the present study, the second nuclear internal transcribed spacer (ITS-2) rDNA of Schistosoma japonicum isolates in mainland China was amplified, sequenced, and assessed for inferring the intra- and inter-species phylogenetic relationships of trematodes in the order Strigeata. The fragment containing ITS-2 rDNA was obtained from 24 S. japonicum isolates from eight epidemic provinces in mainland China. The length polymorphisms were observed among these ITS-2 rDNA sequences, ranging from 343 to 346 bp, and the intra- and inter-population variations in ITS-2 sequence were 0.0-2.1% among S. japonicum isolates in China. Phylogenetic analyses using the maximum parsimony and maximum likelihood methods revealed that the ITS-2 rDNA sequence is not a suitable marker for studying inter- and intra-population variation in S. japonicum. However, phylogenetic analysis of trematodes in the order Strigeata indicated that the ITS-2 rDNA sequence provides an effective molecular marker for studying inter-species phylogenetic relationships among trematodes in this order.

Genetic variation among Clonorchis sinensis isolates from different geographic regions in China revealed by sequence analyses of four mitochondrial genes.

The present study examined sequence variation in four mitochondrial (mt) genes, namely cytochrome c oxidase subunits 1 (cox1) and 2 (cox2), and NADH dehydrogenase subunits 1 and 2 (nad1 and nad2) among Clonorchis sinensis isolates from different endemic regions in China, and their phylogenetic relationships with other zoonotic trematodes were reconstructed. A portion of the cox1 and cox2 genes (pcox1 and pcox2), and nad1 and nad2 genes (pnad1 and pnad2) were amplified separately from individual liver flukes by polymerase chain reaction (PCR) and the amplicons were subjected to sequencing from both directions. The intra-specific sequence variations within C. sinensis were 0-1.6% for pcox1, 0-1.4% for pcox2, 0-0.9% for pnad1 and 0-1.0% for pnad2. Phylogenetic analyses based on the combined sequences of pcox1, pcox2, pnad1 and pnad2 revealed that all the C. sinensis isolates grouped together and were closely related to Opisthorchis felineus. These findings revealed the existence of intra-specific variation in mitochondrial DNA (mtDNA) sequences among C. sinensis isolates from different geographic regions, and demonstrated that mtDNA sequences provide reliable genetic markers for phylogenetic studies of zoonotic trematodes.

Sequence variability in two mitochondrial DNA regions and internal transcribed spacer among three cestodes infecting animals and humans from China.

Sequence variability in two mitochondrial DNA (mtDNA) regions, namely cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 4 (nad4), and internal transcribed spacer (ITS) of rDNA among and within three cestodes, Spirometra erinaceieuropaei, Taenia multiceps and Taenia hydatigena, from different geographical origins in China was examined. A portion of the cox1 (pcox1), nad4 genes (pnad4) and the ITS (ITS1+5.8S rDNA+ITS2) were amplified separately from individual cestodes by polymerase chain reaction (PCR). Representative amplicons were subjected to sequencing in order to estimate sequence variability. While the intra-specific sequence variations within each of the tapeworm species were 0-0.7% for pcox1, 0-1.7% for pnad4 and 0.1-3.6% for ITS, the inter-specific sequence differences were significantly higher, being 12.1-17.6%, 18.7-26.2% and 31-75.5% for pcox1, pnad4 and ITS, respectively. Phylogenetic analyses based on the pcox1 sequence data revealed that T. multiceps and T. hydatigena were more closely related to the other members of the Taenia genus, and S. erinaceieuropaei was more closely related to the other members of the Spirometra genus. These findings demonstrated clearly the usefulness of mtDNA and rDNA sequences for population genetic studies of these cestodes of socio-economic importance.

Sequences and gene organization of the mitochondrial genomes of the liver flukes Opisthorchis viverrini and Clonorchis sinensis (Trematoda).

Opisthorchis viverrini and Clonorchis sinensis are important trematodes infecting humans and animals, belonging to the family Opisthorchiidae. In the present study, we sequenced the nearly complete mitochondrial (mt) DNA (mtDNA) sequences of O. viverrini from Laos, obtained the complete mtDNA sequences of C. sinensis from China and Korea, and revealed their gene annotations and genome organizations. The mtDNA sequences of O. viverrini, C. sinensis (China isolate), C. sinensis (Korea isolate) were 13,510, 13,879, and 13,877 bp in size, respectively. Each of the three mt genomes comprises 36 genes, consisting of 12 genes coding for proteins, two genes for rRNA, and 20 genes (O. viverrini) or 22 genes (C. sinensis) for tRNA. The gene content and arrangement are identical to that of Fasciola hepatica, and Paragonimus westermani, but distinct from Schistosoma spp. All genes are transcribed in the same direction and have a nucleotide composition high in T. The contents of A + T of the mt genomes were 59.39% for O. viverrini, 60.03% for C. sinensis (China isolate), and 59.99% for C. sinensis (Korea isolate). Phylogenetic analyses using concatenated amino acid sequences of the 12 protein-coding genes, with three different computational algorithms [maximum parsimony, maximum likelihood, and Bayesian analysis], all revealed distinct groups with high statistical support, indicating that O. viverrini and C. sinensis represent sister taxa. These data provide additional novel mtDNA markers for studying the molecular epidemiology and population genetics of the two liver flukes and should have implications for the molecular diagnosis, prevention, and control of opisthorchiasis and clonorchiasis in humans and animals.

Modulation of mouse macrophage proteome induced by Toxoplasma gondii tachyzoites in vivo.

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.

Seroprevalence of Toxoplasma gondii infection in domestic pigeons (Columba livia) in Guangdong Province of southern China.

Little information on epidemiology of Toxoplasma gondii infection in pigeons was available in People's Republic of China. In the present study, sera from 275 pigeons raised in different commercial flocks in Guangdong Province of southern China were evaluated using modified agglutination test (MAT). Specific antibodies were found in sera of 8.7% of 275 pigeons (MAT titer ≥ 1:5), and the seropositivity of eight herds we surveyed varied ranging from 0 to 18.2%. The results demonstrated the circulation of T. gondii in the examined pigeon farms, which poses potential risk for human infection with T. gondii. To our knowledge, this is the first seroprevalence survey of pigeons infected by T. gondii in People's Republic of China.

Serological survey of Toxoplasma gondii infection in the domestic goose (Anser domestica) in southern China.

In the present study, the antibodies to Toxoplasma gondii in 191 farm-bred and 83 house-bred geese (Anser domestica) were assessed for the prevalence of T. gondii infection in southern China with the modified agglutination test. Antibodies to T. gondii (MAT ≥ 1 : 5) were found in 27 (14.14%) of farm-bred geese and 14 (16.87%) of house-bred geese. Geese infected with T. gondii may be a source of T. gondii infection for humans and cats.

Molecular and serological diagnosis of Toxoplasma gondii infection in experimentally infected chickens.

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.