Molecular identification and genotyping of Blastocystis sp. in sheep and goats from some areas in Inner Mongolia, Northern China.

Blastocystis sp. is a kind of unicellular intestinal commensal which is widely distributed in humans and animals, and frequently found in the people who are in close contact with animals. To investigate the prevalence and evaluate the zoonotic potential of Blastocystis sp. in sheep and goats from Inner Mongolia, China, a total of 1037 samples were collected from them, and subjected to nested PCR amplification based on the small subunit ribosomal RNA (SSU rRNA) gene of Blastocystis sp. The sanger sequencing was used for Blastocystis sp. subtype identification. The results indicated that the average infection rate of Blastocystis sp. was 10.70% [95CI: 8.82%-12.58%] (111/1037), including 11.30% [95CI: 7.96%-14.64%] for sheep (39/345) and 10.40% [95CI: 8.13%-12.67%] for goats (72/692). Five Blastocystis subtypes (ST5, ST10, ST14, ST21 and ST26) were identified in the present study. Among them, ST10 was the most dominant subtype in sheep and goats, accounting for 70.27% (78/111) of the total identified positive samples. This is the first report regarding Blastocystis sp. subtypes ST21 and ST26 in goats in China. This study has provided a detail epidemiological data on the prevalence and subtypes distribution of Blastocystis sp. in sheep and goats in Inner Mongolia, China. Our results indicated that sheep and goats could be reservoir host for multiple Bastocystis subtypes, including the zoonotic subtypes. Further studies among humans, livestock and wild animals are needed to better understand their role in the spread of Blastocystis sp.

Molecular characterization and prevalence of Cryptosporidium spp. in sheep and goats in western Inner Mongolia, China.

Cryptosporidium spp. are zoonotic intestinal parasites that infect fish, birds, reptiles and mammals. Cryptosporidium spp. are common cause of diarrhea. In this study, a total of 1032 fecal samples were collected from the rectums of sheep and goats. The samples were analyzed using nested polymerase chain reaction (nested PCR) based on the small subunit ribosomal RNA (SSU rRNA) gene of Cryptosporidium spp. The average infection rate of Cryptosporidium spp. was 2.23% (n = 23), and three Cryptosporidium species were identified, namely Cryptosporidium ubiquitum (8/23), Cryptosporidium andersoni (5/23) and Cryptosporidium xiaoi (10/23). Subtyping of C. ubiquitum and C. xiaoi was carried out by DNA sequence analysis of the 60-kDa glycoprotein (gp60) gene. Eight C. ubiquitum isolates were identified as zoonotic subtype XIIa. Nine C. xiaoi isolates were identified as subtypes XXIIIc (n = 1), XXIIIf (n = 3) and XXIIIg (n = 5). Subtype XXIIIg was first found in Chinese sheep. C. ubiquitum subtype XIIa was found in both sheep and goats, suggesting that sheep and goats are important sources of C. ubiquitum infections.

Molecular characterizations of Giardia duodenalis based on multilocus genotyping in sheep, goats, and beef cattle in Southwest Inner Mongolia, China.

Giardia duodenalis is an important zoonotic parasite that causes economic losses to animal husbandry and threatens public health. In the present study, a total of 1466 fresh fecal samples were collected from sheep (n = 797), goats (n = 561) and beef cattle (n = 108) in Southwest Inner Mongolia, China. Giardia duodenalis was initially screened via nested polymerase chain reaction (PCR) targeting the ß-giardin (bg) gene, and bg-positive samples were subjected to PCR amplification targeting the glutamate dehydrogenase (gdh) and triose phosphate isomerase (tpi) genes. A total of 4.0% of samples (58/1466) were positive for G. duodenalis, with a prevalence of 3.4% in sheep, 3.7% in goats and 5.2% in beef cattle. Three G. duodenalis assemblages (A, B, and E) were identified, with E as the prevalent assemblage. Four and one novel assemblage E sequences were obtained for the gdh and tpi loci, respectively and four assemblage E multilocus genotypes (MLG) were obtained. This study demonstrates high genetic variations in G. duodenalis assemblage E, and provides baseline data for preventing and controlling G. duodenalis infection in livestock in Inner Mongolia.

Title: Caractérisation moléculaire de Giardia duodenalis basée sur le génotypage multilocus chez les ovins, les caprins et les bovins dans le sud-ouest de la Mongolie intérieure, en Chine. Abstract: Giardia duodenalis est un parasite zoonotique important, qui cause des pertes économiques à l'élevage et menace la santé publique. Dans la présente étude, un total de 1466 échantillons fécaux frais ont été prélevés sur des moutons (n = 797), des chèvres (n = 561) et des bovins de boucherie (n = 108) dans le sud-ouest de la Mongolie intérieure, en Chine. Giardia duodenalis a été initialement criblé via une réaction en chaîne par polymérase imbriquée ciblant le gène de la ß-giardine (bg), et les échantillons bg-positifs ont été soumis à une amplification par PCR ciblant les gènes de la glutamate déshydrogénase (gdh) et de la triose phosphate isomérase (tpi). Au total, 4,0 % (58/1466) des échantillons étaient positifs pour G. duodenalis, avec une prévalence de 3,4 % chez les ovins, 3,7 % chez les caprins et 5,2 % chez les bovins. Trois assemblages de G. duodenalis (A, B et E) ont été identifiés, E étant l'assemblage prédominant. Respectivement quatre et une nouvelle séquences de l'assemblage E ont été obtenues dans les loci gdh et tpi, et quatre génotypes multilocus (MLG) de l'assemblage E ont été mis en évidence. Cette étude montre des variations génétiques élevées dans l'assemblage E de G. duodenalis et fournit des données de base pour prévenir et contrôler l'infection à G. duodenalis chez le bétail en Mongolie intérieure.

Molecular characterization and zoonotic potential of Enterocytozoon bieneusi in ruminants in northwest China.

This study was conducted to examine the molecular characteristics and assess the zoonotic potential of Enterocytozoon bieneusi in ruminants in northwest China. A total of 1581 fresh fecal samples were collected from eight categories of ruminants. The E. bieneusi was screened and genotyped via nested polymerase chain reaction (PCR) amplification targeting the internal transcribed spacer (ITS) of the small subunit rRNA (ssu rRNA) gene. The result indicated that the average infection rate of E. bieneusi in ruminants was 16.0% (253/1581), with infection rates of E. bieneusi in Mongolian sheep, Mongolian goats, Chifeng cattle, red deer, alpine musk deer, and blue sheep at 21.8% (169/777), 8.2% (46/561), 25.9% (28/108), 13.2% (5/38), 20.0% (4/20), and 6.3% (1/16), respectively. The infections of E. bieneusi varied by different categories. For the different age groups, the infection rates in lambs (29.3%, 108/369) and calves (57.1%, 8/14) were significantly higher than that in ewes (21.1%, 215/1020) and cows (21.3%, 20/94). For the molecular characterization, diverse E. bieneusi ITS genotypes were identified, with a total of 13 genotypes were observed, including 10 known genotypes (BEB6, COS-I, J, CHC8, I, CHG1, BEB4, CHG3, CHS7, and NCF2) and 3 novel genotypes (CNR1 to CNR3). Genotype BEB6 was predominant (59.7%, 151/253). Phylogenetic analysis revealed that most E. bieneusi ITS genotypes clustered into group 2 and only one (NCF2) genotype belonged to group 1. The zoonotic genotypes identified in ruminants in the present study indicated the zoonotic potential of E. bieneusi. In addition, simultaneous identification of genotypes, such as BEB6, COS-I, and BEB4, in the same eco-geographical system indicated some host multiplicity transmission potential of E. bieneusi.

Molecular characterization of Balantioides coli in pigs from Shaanxi province, northwestern China.

Balantioides coli (syn. Balantidium coli) is an important zoonotic but usually neglected protozoa infecting human and a great number of animals, and the pig was considered to be the most important natural host and reservoir. However, no information about the infection of B. coli in pigs in northwestern China was available. In the present study, the prevalence and genetic diversity of B. coli in pigs in Shaanxi province were investigated. A total of 560 fecal samples were collected from pigs of four age groups in five different geographical regions and analyzed by using PCR targeting the ITS1-5.8S rRNA-ITS2 gene fragment. The infection of B. coli was detected in all age groups and regions, with the total prevalence of 16.8% (94/560). Significant differences (P < 0.01) in prevalence were found among four investigated age groups, with the highest in fatteners (38.8%) and the lowest in adults (5.7%). The prevalence was also significantly (P < 0.01) different among pigs from five sampling regions. Sequence analysis revealed two genetic variants, namely, A and B, in these investigated pigs, and both of them were detected in all age groups and regions, with the latter as the predominant one. Further, sixty-eight different haplotypes were found, with 19 and 49 belonged to genetic variants A and B, respectively. The findings in the present study indicated wide distribution and high diversity of B. coli in pigs in Shaanxi province and provided fundamental data for implementing control strategies on B. coli infection in pigs as well as other hosts in this province.

A canine model of experimental infection with Cryptosporidium canis.

Cryptosporidium is a genus of protozoal parasites that affects the gastrointestinal epithelium of a variety of hosts. Several models of experimental infection have been described to study the susceptibility, infectivity and pathogenicity among different Cryptosporidium species and isolates. This study aimed to establish an experimental infection of Cryptodporidium canis in canids. Infectivity and pathogenicity have been measured by evaluating the clinical status, pattern of oocyst excretion and histological examination. Results showed that C. canis was not infective for immunocompetent dogs or mice with severe combined immunodeficiency syndrome (SCID). Oocysts were first detected in the feces of immunosuppressed dogs on day 3 post-infection (p.i.), with levels peaking twice on days 10 and 17 p.i. during the patent period. cryptosporidial developmental stages were found in the duodenum and jejunum of dogs in histological sections stained with hematoxylin and eosin (H & E) and using scanning electron microscopy (SEM). Histopathological changes in the intestinal tract of infected dogs were characterized by epithelial metaplasia and dilatation; the integrity of intestinal mucosal epithelial cells was distinctly damaged with whole sheets of cilia sloughed away. Ultrastructural observation data were consistent with histological observations. Based on these findings, the canine model described in this work will be useful to evaluate clinical, parasitological and histological aspects of C. canis infection and will be useful for the further understanding of cryptosporidiosis, drug development, and vaccine development.

Presence of aliphatic and polycyclic aromatic hydrocarbons in near-surface sediments of an oil spill area in Bohai Sea.

In order to determine the source of organic matter and the fingerprint of the oil components, 50 samples collected from the near-surface sediments of the oil spill area in Bohai Sea, China, were analyzed for grain size, total organic carbon, aliphatic hydrocarbons (AHs), and polycyclic aromatic hydrocarbons (PAHs). The concentrations of C15-35 n-alkanes and 16 United States Environmental Protection Agency (US EPA) priority pollutant PAHs were found in the ranges of 0.88-3.48µg g(-1) and 9.97-490.13ng/g, respectively. The terrestrial organic matters characterized by C27-C35 n-alkanes and PAHs, resulting from the combustion of higher plants, are dominantly contributed from the transportation of these plants by rivers. Marine organic matters produced from plankton and aquatic plants were represented by C17-C26 n-alkanes in AHs. Crude oil, characterized by C17-C21 n-alkanes, unresolved complex mixture (UCM) with a mean response factor of C19 n-alkanes, low levels of perylene, and a high InP/(InP+BghiP) ratio, seeped into the oceans from deep hydrocarbon reservoirs, as a result of geological faults.

An analysis of organic matter sources for surface sediments in the central South Yellow Sea, China: evidence based on macroelements and n-alkanes.

By analyzing the composition of n-alkane and macroelements in the surface sediments of the central South Yellow Sea of China, we evaluated the influencing factors on the distribution of organic matter. The analysis indicates that the distribution of total organic carbon (TOC) was low in the west and high in the east, and TOC was more related to Al2O3 content than medium diameter (MD). The composition of n-alkanes indicated the organic matter was mainly derived from terrestrial higher plants. Contributions from herbaceous plants and woody plants were comparable. The comprehensive analysis of the parameters of macroelements and n-alkanes showed the terrestrial organic matter in the central South Yellow Sea was mainly from the input of the modern Yellow River and old Yellow River. However, some samples exhibited evident input characteristics from petroleum sources, which changed the original n-alkanes of organic matter in sediments.

Occurrence and molecular characterization of Cryptosporidium in dogs in Henan Province, China.

BACKGROUND: Cryptosporidiosis in dogs has been reported worldwide, involving both asymptomatic and diarrheic dogs. Large-scale surveys of Cryptosporidium infection in dogs have been performed in some countries using different diagnostic methods. But, few data are available on the infection rate and molecular characteristics of Cryptosporidium spp. in dogs in China. RESULT: In this study, 770 fecal samples from 66 locations in Henan Province were examined. The average Cryptosporidium infection rate was 3.8%, with dogs in kennels having the highest rate of 7.0% (χ² = 14.82, P < 0.01). The infection rate was 8.0% in dogs younger than 90 days, which was significantly higher than that in the other age groups (1.1-3.8%;χ² = 18.82, P < 0.01). No association was noted between the infection rate and the sex of the dogs. Twenty-nine Cryptosporidium-positive samples were amplified at the small subunit rRNA (SSU rRNA), 70-kDa heat shock protein (HSP70), and actin loci using PCR. Sequence analysis of these amplicons identified only Cryptosporidium canis, which showed 100% identity with the published sequences of the SSU rRNA, HSP70, and actin genes. CONCLUSIONS: Our results confirm that C. canis is popular in the dog population in China, considering the large number of dogs in China and the close contact between dogs and humans, the role of C. canis in the transmission of human cryptosporidiosis warrants attention.

Fabrication of biomimetic high performance antireflective and antifogging film by spin-coating.

In this paper, we report a facile way to fabricate biomimetic high performance optical hybrid films with excellent antireflective and antifogging properties by one-step spin-coating the mixture of mesoporous SiO(2) particles and SiO(2) sol. The production process of the films is easy, low-cost, and time-efficient. Mesoporous SiO(2) particles containing surfactants disperse in SiO(2) sol stably without any chemical modification, which decrease the effective refractive index and increase the transmittance of the films. In addition, such films possess superhydrophilic properties and exhibit high performance antifogging properties. Due to the good film forming performance of SiO(2) sol, mesoporous SiO(2) particles are embedded in the films and impart the films high mechanical stability and durability. The surface morphology of the films can maintain well after repeated friction, and the performances of antireflective and antifogging also do not change as well.

Bioinspired silica surfaces with near-infrared improved transmittance and superhydrophobicity by colloidal lithography.

In this paper, we report a kind of bioinspired high performance near-infrared improved transmittance silica surfaces with superhydrophobic properties by colloidal lithography, with transmittance about 99% from 1300 to 2000 nm. Meanwhile, the optical properties of such surfaces can be controlled by the antireflective structure morphologies resulting from the different reactive ion etching conditions. Using proper microspheres as mask, the high-performance near-infrared telecommunication optics can be achieved. Besides, the antireflective surfaces possess superhydrophobic properties after modified by fluorosilane. Such antireflective surfaces are promising for fabrication of highly light transmissive, antireflective, and superhydrophobic near-infrared optical materials to be used in many important fields.

Multilocus phylogenetic analysis of Cryptosporidium andersoni (Apicomplexa) isolated from a bactrian camel (Camelus bactrianus) in China.

This is the first report of cryptosporidiosis in a bactrian camel (Camelus bactrianus) in China. Two Cryptosporidium isolates derived from the same bactrian camel (3-year-old) in November 2005 and April 2006 were characterized using sequence and phylogenetic analysis of the small-subunit rRNA (18S rRNA), 70-kDa heat shock protein (HSP70), actin and Cryptosporidium oocyst wall protein (COWP) genes. The sequences of the 18S rRNA and COWP were identical to all other Cryptosporidium andersoni isolates although minor differences were noticed between the isolates and the USA isolate at the actin locus (99.2% of similarity). The sequence of the HSP70 was identical to the Japanese C. andersoni isolate, with a minor difference from the Australian C. andersoni isolate (99.7% of similarity). Cross-transmission studies demonstrated that the C. andersoni isolates did not infect immunosuppressed or immunocompetent Kun-ming mice, severe combined immunodeficiency mice, and immunosuppressed or immunocompetent calves. Among the C. andersoni isolates reported so far, only isolates from Japan could infect SCID mice. Thus, the C. andersoni isolates from the bactrian camel were biologically similar to most bovine C. andersoni isolates characterized so far, but are different from bovine isolates from Japan.