Molecular detection of piroplasms in domestic donkeys in Xinjiang, China.

BACKGROUND: Piroplasmosis is a common and prevalent tick-borne disease that affects equids. OBJECTIVES: To determine the infection and molecular characteristics of the piroplasms in donkeys from Xinjiang, northwestern China, we undertook a cross sectional study by collecting representative samples across several counties within the region. METHODS: A total of 344 blood samples were collected from adult domestic donkeys from 13 counties in Xinjiang. PCR was conducted to test for T. equi and B. caballi in the blood samples based on the equine merozoite antigen-1 (Ema-1) gene and the 48 kDa rhoptry protein (BC48) gene, respectively. RESULTS: Sixteen blood samples tested positive for piroplasms and the overall infection rate was 4.7% (16/344). Seven of the 13 counties were positive for piroplasms. Among the 16 piroplasm-positive samples, 15 were singly infected with T. equi with an infection rate of 4.4% (15/344), and coinfection with T. equi and B. caballi was detected in one sample (0.3%, 1/344) from Wushi. Four T. equi sequence genotypes were identified and grouped into different branches of the evolutionary trees. CONCLUSION: These findings suggest that the infection rate of piroplasms is low in domestic donkeys in southern Xinjiang and that T. equi genotypes have a regional distribution.

Preparation and Characterization of Prickly Ash Peel Oleoresin Microcapsules and Flavor Retention Analysis.

Prickly ash peel oleoresin (PPO) is a highly concentrated oil of Prickly ash essential oil and has a stronger aroma. However, its low water solubility, high volatility, difficulty in transport and storage, and decomposition by light, heat, and oxygen limit its wider application. To solve this problem, this study used freeze-drying or spray-drying, with soybean protein isolate (SPI) or gum Arabic (GA), combined with aqueous maltodextrin (MD) as the encapsulating agents to prepare four types of PPO microcapsules (POMs). Spray-dried microcapsules with GA as the encapsulating agent achieved a high encapsulation efficiency (EE) of 92.31 ± 0.31%, improved the thermal stability of the PPO, and had spherical morphology. (Headspace solid-phase microextraction/gas chromatography-mass spectrometry) HS-SPME/GC-MS detected 41 volatile compounds in PPO; of these, linalool, ß-myrcene, sabinene, and D-limonene were identified as key flavor components. Principal component analysis (PCA) effectively distinguished the significant differences in flavor between PPO, spray-dried SPI/MD microcapsules (SS), and spray-dried GA/MD microcapsules (SG). During 15 days of air-exposure, the loss of flavor from SG (54.62 ± 0.54%) was significantly lower than PPO (79.45 ± 1.45%) and SS (57.55 ± 0.36%). During the air-exposure period, SG consistently had the highest antioxidant capacity, making it desirable for PPO packaging, and expanding its potential applications within the food industry.

Development and application of one-step multiplex Real-Time PCR for detection of three main pathogens associated with bovine neonatal diarrhea.

Introduction: Neonatal calf diarrhea (NCD) is one of the most common diseases in calves, causing huge economic and productivity losses to the bovine industry worldwide. The main pathogens include bovine rotavirus (BRV), bovine coronavirus (BCoV), and Enterotoxigenic Escherichia coli (ETEC) K99. Since multiple infectious agents can be involved in calf diarrhea, detecting each causative agent by traditional methods is laborious and expensive. Methods: In this study, we developed a one-step multiplex Real-Time PCR assay to simultaneously detect BRV, BCoV, and E. coli K99+. The assay performance on field samples was evaluated on 1100 rectal swabs of diseased cattle with diarrhea symptoms and compared with the conventional gel-based RT-PCR assay detect BRV, BCoV, and E. coli K99+. Results: The established assay could specifically detect the target pathogens without cross-reactivity with other pathogens. A single real-time PCR can detect ~1 copy/µL for each pathogen, and multiplex real-time PCR has a detection limit of 10 copies/µL. Reproducibility as measured by standard deviation and coefficient of variation were desirable. The triple real-time PCR method established in this study was compared with gel-based PT-PCR. Both methods are reasonably consistent, while the real-time PCR assay was more sensitive and could rapidly distinguish these three pathogens in one tube. Analysis of surveillance data showed that BRV and BCoV are major enteric viral pathogens accounting for calves' diarrhea in China. Discussion: The established assay has excellent specificity and sensitivity and was suitable for clinical application. The robustness and high-throughput performance of the developed assay make it a powerful tool in diagnostic applications and calf diarrhea research. ​.

Molecular characterization of Enterocytozoon bieneusi genotypes in wild Altai marmot (Marmota baibacina) in Xinjiang, China: host specificity and adaptation.

Enterocytozoon bieneusi is responsible for opportunistic infections leading to gastrointestinal diseases in humans and animals worldwide. A total of 334 fresh fecal samples were collected from wild Altai marmots (Marmota baibacina) in Xinjiang, China, and E. bieneusi was screened via PCR amplification of the internal transcribed spacer (ITS) region of the small submit ribosomal RNA (SSU rRNA). The results indicated that 22.8% (76/334) of the wild Altai marmot fecal samples were positive for E. bieneusi, and the highest positive rate was detected in Akqi (51.9%, 27/52), with a significant difference from other sampling sites (p < 0.01). Four known genotypes (BEB6, CHG3, GX2, and YAK1) and three novel genotypes (XJHT2 to XJHT4) were identified in the present study. Genotype XJHT3 was dominant and detected in 48 fecal samples. In the phylogenetic analysis, the novel genotypes XJHT2 and XJHT3 were clustered in Group 1 together with the known genotype YAK1, while genotypes CHG3 and BEB6 were clustered in Group 2. The novel genotype XJHT4 was clustered together with other rodent-derived genotypes and generated a novel Group 14. These data confirmed the host specificity and adaptation of E. bieneusi in rodents. These findings enrich our understanding of the prevalence and genetic diversity of E. bieneusi in wild Altai marmots in Xinjiang, China.

Molecular evaluation of Cryptosporidium spp. in sheep in southern Xinjiang, China.

Cryptosporidium spp. are diarrheagenic intestinal parasites with multiple hosts worldwide. A total of 1252 fresh fecal samples of sheep were collected from 10 large-scale farms in southern Xinjiang. Based on the small subunit ribosomal (SSU rRNA) gene of Cryptosporidium, 100 Cryptosporidium-positive samples (8.0%, 100/1252) were detected by PCR. Nine out of 10 farms were positive for Cryptosporidium, with the highest infection rate being 18.4% (23/125) on farm 9 in Qira. The infection rates of Cryptosporidium in pre-weaned lambs, weaned lambs, fattening sheep, and adult sheep were 20.3% (61/301), 10.3% (34/329), 0.9% (3/327), and 0.7% (2/295), respectively. Three Cryptosporidium species were identified, namely, C. xiaoi (n = 61), C. parvum (n = 22), and C. ubiquitum (n = 17). Of them, C. xiaoi was detected on all positive farms and in different age groups of sheep. The subtypes of C. parvum and C. ubiquitum were identified by PCR at the 60 kDa glycoprotein (gp60) gene. Two C. parvum subtypes were identified: IIdA19G1 (n = 21) and IIdA15G1 (n = 1). One C. ubiquitum subtype was identified with XIIa (n = 17). These results indicated the common transmission and genetic diversity of Cryptosporidium in sheep in southern Xinjiang, and further investigations are needed on the zoonotic potential of C. parvum and C. ubiquitum in this region.

Molecular detection and genetic characteristics of Cryptosporidium spp. in Chinese racehorses.

BACKGROUND: Cryptosporidium is a protozoan parasite causing diarrhoea in humans and animals. Although Cryptosporidium has been found in domestic horses (farmed or kept at pasture), there has been only one published study of Cryptosporidium infections in Chinese racehorses, which was restricted to a very small geographical area. OBJECTIVES: To investigate the presence of Cryptosporidium spp. in the faeces of racehorses in China and to perform molecular characterisation of the parasite. STUDY DESIGN: Cross-sectional. METHODS: A total of 621 fresh faecal samples were collected for DNA extraction from racehorses at 17 equestrian clubs from 12 provinces of China from December 2016 to May 2018. All the DNA were analysed for the presence of Cryptosporidium species/genotypes and subtypes by PCR amplification of the small subunit ribosomal RNA and 60 kDa glycoprotein genes respectively. RESULTS: PCR analysis revealed that 11 samples (1.8%) were positive for Cryptosporidium spp. Among them seven samples were identified as C. parvum and four were C. hominis. The C. parvum isolates were identified as subtype IIdA14G1 (n = 4) and IIdA15G1 (n = 3), while all C. hominis isolates were identified as subtype IkA18G1 (n = 4). MAIN LIMITATIONS: A single faecal sample from each horse was used instead of multiple samples that could improve the detection rates of the parasite. CONCLUSIONS: Although Cryptosporidium infection rate was relatively low in the investigated racehorses, the presence of zoonotic subtypes IIdA14G and 1IIdA15G1 of C. parvum and IkA18G1 of C. hominis, suggesting that these animals are a potential source of Cryptosporidium in humans.

Prevalence and genetic characterization of Theileria equi and Babesia caballi in grazing horses in Xinjiang, northwestern China.

Piroplasmosis is a disease that negatively affects equine health worldwide. Hence, 324 blood samples were collected from grazing horses in ten sites in Xinjiang and testing them for the presence of Theileria equi and Babesia caballi by PCR of the EMA-1 gene and BC48 gene, respectively. Of the 324 blood samples, 161 (49.7%) were positive for equine piroplasms. The prevalence of T. equi was 38.9% (126/324), while that of B. caballi was 30.2% (98/324). The T. equi and B. caballi co-infection rate was 19.4% (63/324). From the 126 EMA-1 gene sequences and 98 BC48 gene sequences we obtained, 21 and 27 genotypes were identified, respectively. The EMA-1 sequences together with the GenBank reference sequences grouped into four clusters, with those from the present study forming two distinct clusters. In contrast, the BC48 sequences formed eight clusters with the GenBank reference sequences, while those obtained in the present study formed five distinct clusters. Our results highlight the widespread distribution and abundant gene polymorphism of T. equi and B. caballi in grazing horses from Xinjiang.

Molecular detection and characterization of Enterocytozoon bieneusi in captive alpine musk deer (Moschus chrysogaster) in Central and Eastern of Gansu Province, China.

The most ubiquitously diagnosed microsporidian species in animals and humans is Enterocytozoon bieneusi (E. bieneusi). In this case, this work aimed to probe the occurrence and genotypes of this pathogen in captive alpine musk deer (Moschus chrysogaster) in Gansu Province, China. After fecal sample collection (n = 201) from three farms in Gansu, the internal transcribed spacer (ITS) region of the rRNA gene was probed by PCR for detection and genotyping of E. bieneusi. The infection rate of E. bieneusi in alpine musk deer was 6.0% (12/201), with 0% (0/36), 7.7% (5/65) and 7.0% (7/100) in farms 1 to 3, respectively. The infection rate of E. bieneusi in young alpine musk deer (3.2%; 1/31) is lower than that of adults (6.5%; 11/170), with no evident significant differences between age groups (P > 0.05). Three known genotypes D (n = 8), EbpA (n = 3) and BEB6 (n = 1) were identified by sequence analysis. This is the first such scrutiny of E. bieneusi infection in alpine musk deer in China as per our knowledge. Genotypes D and EbpA were common in humans and animals that is suggestive of the plausible zoonotic role in E. bieneusi transmission by alpine musk deer.

Genetic diversity of Blastocystis subtypes in the Alpine musk deer (Moschus chrysogaster) in Gansu province, northwestern China.

Blastocystis sp. is a gastrointestinal pathogen that is frequently found in humans and animals worldwide. In this study, 201 fecal samples were collected from captive Alpine musk deer (Moschus chrysogaster) at three farms in Gansu province. Blastocystis was detected and subtyped by amplifying and sequencing the small subunit ribosomal DNA gene. The overall prevalence of Blastocystis was 39.8% (80/201). Five known Blastocystis subtypes (STs), including ST1 (n = 1), ST4 (n = 12), ST10 (n = 50), ST14 (n = 6), and ST24 (n = 11) were identified using subtyping and evolutionary analysis. ST10 was the most common ST observed in each farm. This study showed the infection status and genetic characteristics of Blastocystis in M. chrysogaster. Based on the surveyed data, because various potentially zoonotic STs, such as ST1, ST4, ST10, ST14, and ST24, were detected, it is believed that the zoonotic risk of Blastocystis from the Alpine musk deer in this area cannot be ignored.

Molecular detection and genotypes of Enterocytozoon bieneusi in farmed mink (Neovison vison), blue foxes (Alopex lagopus), and raccoon dogs (Nyctereutes procyonoides) in Xinjiang, China.

Enterocytozoon bieneusi is a zoonotic pathogen that infects a variety of hosts including humans, livestock, wildlife, companion animals, and birds, as well as being abundant in the environment. Humans and nonhuman animals could be infected with E. bieneusi via consumption of food or water that contains zoonotic and host-adapted genotypes. In this study, 288 fecal specimens were collected from farmed minks, blue foxes, and raccoon dogs, in Xinjiang, China. Enterocytozoon bieneusi was examined by PCR amplification based on sequence analysis of the internal transcribed spacer (ITS) region. The overall infection rate of E. bieneusi was 4.9% (14/288), with mink samples showing the highest infection rate (5.6%, 12/214), followed by blue foxes (2.9%, 1/35), and then raccoon dogs (2.6%, 1/39). Six E. bieneusi genotypes were identified, including D (n = 5), PigEBITS7 (n = 4), EbpA (n = 2), CAM5 (n = 1), WildBoar3 (n = 1), and a novel genotype XJMI-1 (n = 1). Phylogenetic analysis showed that all E. bieneusi genotypes belonged to group 1, which composed of over 300 genotypes and most of them have been identified in human and variety of animals, suggesting a risk of zoonotic transmission from farmed wildlife to humans.

Host-adaptation of the rare Enterocytozoon bieneusi genotype CHN4 in Myocastor coypus (Rodentia: Echimyidae) in China.

BACKGROUND: Enterocytozoon bieneusi is a zoonotic gastrointestinal pathogen and can infect both humans and animals. The coypu (Myocastor coypus) is a semi-aquatic rodent, in which few E. bieneusi infections have been reported and the distribution of genotypes and zoonotic potential remains unknown. METHODS: A total of 308 fresh fecal samples were collected from seven coypu farms in China to determine the infection rate and the distribution of genotypes of E. bieneusi from coypus using nested-PCR amplification of the internal transcribed spacer (ITS) region of the ribosomal RNA (rRNA) gene. RESULTS: Enterocytozoon bieneusi was detected with an infection rate of 41.2% (n = 127). Four genotypes were identified, including three known genotypes (CHN4 (n = 111), EbpC (n = 8) and EbpA (n = 7)) and a novel genotype named CNCP1 (n = 1). CONCLUSIONS: The rare genotype CHN4 was the most common genotype in the present study, and the transmission dynamics of E. bieneusi in coypus were different from other rodents. To the best of our knowledge, this is the first report of E. bieneusi infections in coypus in China. Our study reveals that E. bieneusi in coypus may be a potential infection source to humans.

Longitudinal identification of Enterocytozoon bieneusi in dairy calves on a farm in Southern Xinjiang, China.

Enterocytozoon bieneusi is the most common species responsible for human and animals microsporidiasis. A total of 250 samples were collected weekly from 25 newborn dairy calves of a farm in Southern Xinjiang, China at one to ten weeks of age. Enterocytozoon bieneusi was identified and genotyped by nested PCR amplification and sequencing of internal transcribed spacer (ITS) region.The cumulative prevalence of E. bieneusi infection was 100% (25/25), and the average infection was 52.0% (130/250). The highest infection rate was recorded at six weeks of age (92.0%, 23/25), and no infection was observed at one and two weeks of age. Sequencing analysis showed nine E. bieneusi genotypes (J, EbpC, PigEBITS5, CHV4, CHC3, CS-9, KIN-1, CH5, and CAM5) were identified. The highest genetic polymorphism was observed at ten weeks of age. Genotype J was the predominant E. bieneusi genotype. Phylogenetic analysis clustered genotype J into Group 2 and other eight genotypes (EbpC, PigEBITS5, CHV4, CHC3, CS-9, KIN-1, CH5, and CAM5), detected in 22 (16.9%, 22/130) samples, into Group 1. Among the genotypes, EbpC, KIN-1, and J have been identified in humans. The highest E. bieneusi infection rate (57.9%, 124/214) was observed in fecal samples with formed feces with no diarrhea (p < 0.01), and high genetic polymorphism was observed in class I fecal samples. The presence of zoonotic E. bieneusi genotypes in dairy calves suggests the possibility of transmitting zoonotic infections to humans. It provides the basic data on dynamic change of E. bieneusi in calves.

Molecular characterization of Cryptosporidium spp. in minks (Neovison vison), blue foxes (Vulpes lagopus), and raccoon dogs (Nyctereutes procyonoides) in farms from Xinjiang, Northwest China.

The objective of this study was to determine the infection rate and genetic diversity of Cryptosporidium spp. in minks, foxes, and raccoon dogs, farmed in the Xinjiang Uygur Autonomous Region, Northwest China. Fresh fecal specimens were collected from individual cages of farmed minks (n = 214), blue foxes (n = 35), and raccoon dogs (n = 39) and examined using nested PCR based on the Cryptosporidium spp. small subunit rRNA gene. Cryptosporidium spp. was detected in 35 cages (12.2%, 35/288), with a higher infection rate detected in raccoon dogs (20.5%) compared with minks (12.1%) and blue foxes (2.9%). Sequence analysis showed that Cryptosporidium canis was the only species identified in blue foxes and raccoon dogs, while in the 26 Cryptosporidium-positive mink specimens, Cryptosporidium mink genotype (n = 17), C. canis (n = 7), and Cryptosporidium parvum (n = 2) were identified. Further analysis based on the 60-kDa glycoprotein (gp60) gene determined that both C. parvum isolates belonged to the subtype IIdA15G1, while eight of the 17 Cryptosporidium mink genotype isolates were a novel subtype that we have named XeA5G1. To the best of our knowledge, this is the first report of C. parvum subtype IIdA15G1 infection in minks. Since all the Cryptosporidium species/genotypes identified in minks, foxes, and raccoon dogs from Xinjiang have been previously found in humans, our results suggest that these fur animals may play a role in the transmission of zoonotic Cryptosporidium.

Enterocytozoon bieneusi in donkeys from Xinjiang, China: prevalence, molecular characterization and the assessment of zoonotic risk.

BACKGROUND: Enterocytozoon bieneusi, a zoonotic pathogen, has the potential to infect both immunocompromised and immunocompetent humans. It is found in large number of animals; however, not much is known regarding its prevalence in equine animals, particularly donkeys. This is the first molecular epidemiological evaluation of E. bieneusi in 178 free-ranging donkeys from five countrysides; and 502 farmed donkeys from 18 farms in 12 cities of Xinjiang, China by Nested PCR. RESULTS: E. bieneusi was detected in 2.5% (17/680) donkeys, with 2.6% (13/502) in farmed and 2.2% (4/178) in free-ranging ones. Sequence analysis identified eight ITS genotypes, all belonging to zoonotic Groups 1 or 2, including six known genotypes: horse1 (n = 5), D (n = 3), NCD-2 (n = 3), BEB6 (n = 2), BEB4 (n = 1), and NIAI (n = 1); and two new genotypes: XJD1 (n = 1) and XJD2 (n = 1). CONCLUSIONS: This is the first report confirming the presence of E. bieneusi in donkeys in Xinjiang, China, and indicates the possibility of zoonotic transmission of this pathogenic parasite.

Prevalence and genotypic identification of Cryptosporidium in free-ranging and farm-raised donkeys (Equus asinus asinus) in Xinjiang, China.

The prevalence and zoonotic potential of Cryptosporidium in donkeys is poorly understood. Here, 680 fecal specimens were collected from 178 free-ranging and 502 farmed donkeys in Xinjiang, China. Cryptosporidium was identified using PCR amplification of the small subunit of ribosomal DNA. Cryptosporidium-positive isolates were subtyped using PCR analysis of the 60 kDa glycoprotein gene (gp60). The overall prevalence of Cryptosporidium was 2.4% (16/680), with 3.2% (16/502) in farmed donkeys and 0% (0/178) in free-ranging donkeys. Cryptosporidium hominis (n = 13), C. parvum (n = 1) and Cryptosporidium horse genotype (n = 2) were identified. The C. hominis isolates belonged to two subtypes, IkA16 (n = 9) and IkA16G1 (n = 4). The subtype of C. parvum was IIdA15G1, whereas the two Cryptosporidium horse genotype isolates were of subtype VIaA15G4. The predominance of C. hominis in donkeys suggests that these animals are infected through human contact.

TITLE: Prévalence et identification génotypique des Cryptosporidium chez les ânes (Equus asinus asinus) en liberté et en élevage au Xinjiang, Chine. ABSTRACT: La prévalence et le potentiel zoonotique de Cryptosporidium chez les ânes sont mal connus. Dans cet article, 680 spécimens fécaux ont été prélevés de 178 ânes en liberté et 502 ânes en élevage dans le Xinjiang, en Chine. Les Cryptosporidium ont été identifiés en utilisant une amplification par PCR de la petite sous-unité de l'ADN ribosomique. Les isolats positifs pour Cryptosporidium ont été sous-typés en utilisant une analyse PCR du gène de la glycoprotéine 60 kDa (gp60). La prévalence globale de Cryptosporidium était de 2,4 % (16/680), avec 3,2 % (16/502) chez les ânes d'élevage et 0 % (0/178) chez les ânes en liberté. Cryptosporidium hominis (n = 13), C. parvum (n = 1) et Cryptosporidium génotype chevalin (n = 2) ont été identifiés. Les isolats de C. hominis appartenaient à deux sous-types, IkA16 (n = 9) et IkA16G1 (n = 4). Le sous-type de C. parvum était IIdA15G1, tandis que les deux isolats de Cryptosporidium génotype chevalin étaient du sous-type VIaA15G4. La prédominance de C. hominis chez les ânes suggère que ces animaux sont infectés par contact humain.

Unusual dominant genotype NIA1 of Enterocytozoon bieneusi in children in Southern Xinjiang, China.

Enterocytozoon bieneusi is the mainly pathologies or intestinal disorders that causes approximately 90% of reported cases of human microsporidiosis. To understand the prevalence and genotype distribution of E. bieneusi in the Xinjiang Uygur Autonomous Region, China, 609 fecal samples were collected from children in kindergarten in Southern Xinjiang and screened for this pathogen by PCR and sequencing of the internal transcribed spacer (ITS). Thirty-six fecal samples (5.9%, 36/609) were positive for E. bieneusi, with the highest prevalence observed in children from Yopurga (17.5%, 11/63). Nine genotypes were identified, of which six were known (A, CHN6, D, EbpA, KB-1, and NIA1) and three were novel (CXJH1, CXJH2 and CXJH3). Genotype NIA1 was most prevalent (52.8%, 19/36), followed by genotypes D (16.7%, 6/36), A (8.3%, 3/36), and EbpA (8.3%, 3/36). The remaining five genotypes were detected in one sample each. Phylogenetic analysis revealed that the E. bieneusi isolates clustered into two groups, one consisting of six genotypes (Group 1: A, CXJH1, D, EbpA, KB-1, and NIA1) and another consisting of three genotypes (Group 2: CHN6, CXJH2, and CXJH3). Our results confirmed that infection of E. bieneusi unusual dominant genotype NIA1 occurs in children in Xinjiang, China. Further epidemiological studies must be conducted to clarify potential sources of E. bieneusi infection in this area.

Cloning, expression profile of the complement component C9 gene and influence of the recombinant C9 protein on peripheral mononuclear leukocytes transcriptome in half-smooth tongue sole (Cynoglossus semilaevis).

The ninth complement component (C9) is a terminal complement component (TCC) that is involved in creating the membrane attack complex (MAC) on the target cell surface. In this study, the CsC9 (C9 of Cynoglossus semilaevis) cDNA sequence was cloned and characterized. The full-length CsC9 cDNA measured 2,150 bp, containing an open reading frame (ORF) of 1,803 bp, a 5'-untranslated region (UTR) of 24 bp and a 3'-UTR of 323 bp. A domain search revealed that the CsC9 protein contains five domains, including two TSP1s, an LDLRA, an EGF, and a MACPF. Quantitative real-time PCR analysis showed that CsC9 at the mRNA level was expressed in all the tested tissues, with the highest expression being observed in the liver. CsC9 expression is significantly upregulated in the tested tissues after challenge with Vibrio anguillarum. To further characterize the role of CsC9, peripheral blood mononuclear cells of C. semilaevis were used for transcriptome analysis after incubation with recombinant CsC9 (rCsC9) protein. A total of 3,775 significant differentially expressed genes (DEGs) were identified between the control and the rCsC9-treated group, including 2,063 upregulated genes and 1,712 downregulated genes. KEGG analyses revealed that the DEGs were enriched in cell adhesion molecules, cytokine-cytokine receptor interactions, T cell receptor signaling pathways, B cell receptor signaling pathways and Toll-like receptor signaling pathways. The results of this study indicate that in addition to participating in MAC formation, CsC9 might play multiple roles in the innate and adaptive immunity of C. semilaevis.

Molecular detection of Anaplasma spp. in dairy cattle in southern Xinjiang, China.

Bovine anaplasmosis is caused by a group of obligate intracellular bacteria belonging to the genus Anaplasma, which are transmitted by ticks. This study was conducted to determine the prevalences and molecular characterization of Anaplasma spp. in dairy cattle in the upper reaches of the Tarim River in Xinjiang, China. Using polymerase chain reaction (PCR) and sequencing approaches, DNA of Anaplasma spp. was detected in 16 of 493 (3.2%) blood samples from dairy cattle. Positive rates were 0.2% (1/493), 0.4% (2/493), 0.2% (1/493), 2.4% (12/493) and 2.4% (12/493) for A. bovis, A. ovis, A. phagocytophilum like strain, A. phagocytophilum and A. platys like strain, respectively. Anaplasma phagocytophilum and A. platys like strain co-infection was detected in 12 samples. To our knowledge, this is the first report of A. ovis infection in dairy cattle in Xinjiang. This study provides new data on the prevalences of Anaplasma spp. in cattle in Xinjiang, which will help to formulate appropriate control strategies for these pathogens in this area.

Genotypes and zoonotic potential of Enterocytozoon bieneusi in edible bullfrogs (Lithobates catesbeiana) in China.

Enterocytozoon bieneusi, an obligate intracellular pathogen of the intestinal epithelium, is commonly identified in humans and many other animals and is ubiquitous in water sources and the environment generally. To determine the molecular prevalence of E. bieneusi in edible bullfrogs (Lithobates catesbeiana) and evaluate the possibility of its potential zoonotic transmission to humans via food or water, the intestinal contents of 295 bullfrogs were intermittently collected from two open markets in Aksu, China. The samples were screened for the internal transcribed spacer by polymerase chain reaction amplifications, revealing that 20.7% (61/295) of them were infected with E. bieneusi, with no significant differences found between the two sampling locations (p > 0.05). Twenty-two different E. bieneusi genotypes were identified, including one known genotype (EbpC) and 19 novel ones (named BLC1 to BLC19). The zoonotic genotype EbpC was identified in most of the E. bieneusi-positive samples (65.6%, 40/61). The remaining genotypes were identified in either one or three samples each. Our phylogenetic analysis showed that 20 of the E. bieneusi genotypes belonged to Group 1. As far as we are aware, this is the first report of E. bieneusi infections in edible bullfrogs. Our findings suggest that E. bieneusi can be maintained in edible bullfrogs and potentially transmitted via food or water. It is possible that these amphibians are unsuspected zoonotic reservoirs of E. bieneusi.

Molecular characterization of Cryptosporidium and Enterocytozoon bieneusi in Père David's deer (Elaphurus davidianus) from Shishou, China.

Cryptosporidium and Enterocytozoon bieneusi are important intestinal pathogens that infect humans and various animals. Few reports are available regarding the infections of the two pathogens in Père David's deer. In this study, polymerase chain reaction (PCR) confirmed Cryptosporidium infection in two (1.6%) and E. bieneusi in 45 (35.2%) of 128 fecal samples collected from Père David's deer in the National Nature Reserve of Shishou, Hubei Province, China. C. parvum (n = 1) and Cryptosporidium deer genotype (n = 1) were identified using the small subunit rRNA (SSU rRNA) gene. The C. parvum was further subtyped as IIdA20G1 by sequencing analysis of the 60-kDa glycoprotein (gp60) gene. The identity of E. bieneusi was confirmed by an internal transcribed spacer (ITS) gene; the HLJD-V (n = 42) and MWC_d1 (n = 3) genotypes were identified, with the former clustering in group 2 and the latter in group 1. These data suggest that the Père David's deer were infected with host-specific and/or zoonotic genotypes of these pathogens, implicating Père David's deer could be a potential source of human Cryptosporidium infection.