Prevalence and genetic diversity of Entamoeba species infecting macaques in southwest China

Many colonies of macaques (Macaca fascicularis and Macaca mulatta) are maintained in China, especially in Guangxi and Guizhou. A total of 803 fresh stool samples infected with Entamoeba were obtained from three big colonies of macaques located in southwest China. The samples were examined for the presence of five Entamoeba species using PCR. Entamoeba nuttalli, Entamoeba dispar, Entamoeba coli, and Entamoeba chattoni infections were detected, but Entamoeba histolytica infection was not. This study is the first to report on the prevalence of E. nuttalli in wild macaques from China. Eighteen E. nuttalli isolates and five E. dispar isolates were obtained by culturing the samples in Tanabe–Chiba medium. The serine-rich protein (SRP), ribosomal RNA (rRNA), hexokinase (HXK), glucose-6-phosphate isomerase (GPI), and phosphoglucomutase (PGM) genes of E. nuttalli isolates were compared with other reported isolates. The results showed clear differences among the Chinese E. nuttalli isolates and other isolates based on the SRP gene sequences. However, HXK, GPI, and PGM genes of these strains were similar to those of other isolates. The rRNA genes of E. coli and E. chattoni were also amplified and analyzed from these samples. The results suggested that host species might be a more important factor than geographic location in amebic genetic diversity.     

Prevalence and genetic diversity of pneumococcal serogroup 6 in Australia

The prevalence of the newly discovered pneumococcal serotype 6C has increased in some countries since the introduction of seven-valent conjugate pneumococcal vaccine (PCV7). The distribution of invasive serogroup 6 serotypes, in Australia, including 6C and 6D, has not been reported previously. During the period 1999 to 2008, 6097 isolates were referred to the New South Wales Pneumococcal Reference Laboratory for serotyping. Of these, 847 were identified by Quellung reaction as belonging to serogroup 6 and 702 were available for further study. Serotypes were determined by serotype-specific PCR as follows: 6A, 197 (28.1%); 6B, 452 (64.4%); 6C, 52 (7.4%) and one 6D. The average numbers of invasive serogroup 6 isolates, per annum, fell from 62.2 before (2000– 2005) to 49.7 after (2006–2008) the introduction of PCV7. The proportions of invasive 6B fell (from 72.4% to 47.3%, p 0.03), those of 6C rose (from 3.3% to 17%, p 0.02) significantly and those of 6A remained fairly constant (24.3% vs 27%, p 0.69) between the two periods. All 6C and 6D and selected 6A and 6B isolates were further characterized by multilocus sequence typing and sequence analysis of cps genes cpsA-cpsB (wzg-wzh) and wchA-wciN^beta-wciO, wciP. Results showed considerable diversity within serotype 6C, apparently as a result of both mutation and recombination. Sequence typing indicates that, in Australia, 6C has been largely derived from 6A. The genetic diversity and rapid increase in incidence of serotype 6C causing invasive pneumococcal disease has potential implications for vaccine efficacy.     

Molecular characterization of a porcine kobuvirus strain in China

Porcine kobuvirus was first identified in 2007 in Hungary. The virus has been detected in several Asian countries. In our study, the complete genome of the recently identified porcine kobuvirus strain SH-W-CHN was amplified by RT-PCR and was sequenced. Dendrograms indicated that SH-W-CHN is closely related to other porcine kobuviruses. To identify sites of possible recombination within the genome of the SH-W-CHN strain, the SimPlot program was used to perform recombination analysis. The results showed that no significant recombination event between strain S-1-HUN and Y-1-CHI had occurred. However, certain possible recombination signals were identified, indicating that some early recombination events may have contributed to the genome of SH-W-CHN. This study further confirmed the existence of multiple lineages of porcine kobuvirus and indicated that homologous recombination may be a driving force in its evolution.     

Prevalence and genetic diversity of human caliciviruses (HuCVs) in Mexican children

Human caliciviruses (HuCVs) contain two genera: "Norwalk-like viruses" (NLVs) and "Sapporo-like viruses" (SLVs). The importance of the two genera as a cause of acute gastroenteritis of infants and children remains unknown. Beginning in 1989, a birth cohort of children in Mexico was enrolled and monitored for acute gastroenteritis. A subset of 115 diarrhea stool specimens from 76 children and 66 non-diarrhea stool specimens from 64 children was examined for HuCVs by RT-PCR by using a primer pair (p289/290) that detects both NLVs and SLVs. Twenty-two (19%) of the 115 diarrhea stool specimens and 5 (7%) of 66 non-diarrhea stool specimens produced RT-PCR products of expected size (319 bp for NLVs and 331 bp for SLVs). Twenty of the twenty-seven strains were cloned and sequenced. Pairwise sequence analysis showed that 9 (60%) and 6 (40%) of the 15 strains from the diarrhea stools were NLVs and SLVs, respectively. The same proportions of NLVs (60%) and SLVs (40%) were observed in the non-diarrhea stools. Strains in the NLV genus could be further divided into four clusters: Lordsdale, MxV, and HV and one potentially new cluster. Strains in the SLV genus could be divided into three clusters: Sapporo/82, Lon/92, and a potentially new cluster. Strains from the Lordsdale cluster were the most common among these children. The findings of both genera and multiple clusters of HuCVs co-circulating and the identification of new strains of HuCVs in the population justify the need for future studies of HuCVs in infants and children.     

Molecular characterization of a porcine kobuvirus variant strain in China

In this study, we report the complete nucleotide sequence of porcine kobuvirus CH/HZ/2011 (CH/HZ) isolated from infected piglets in China. Dendrograms indicated that CH/HZ is closely related to other porcine kobuviruses. The complete genome sequence of CH/HZ has an adenine insertion at position 124, an 18-nt deletion in the 5′UTR, and a 90-nt deletion in the 2B coding region. A neighbor-joining tree based on the VP1 gene indicated that 31 porcine kobuvirus strains are divided into four clusters (Thai-Japanese group, Hungarian group, Chinese group I and Chinese group II). The strain CH/HZ was included in Chinese group II.     

Prevalence and genetic diversity of human sapovirus associated with sporadic acute gastroenteritis in South China from 2013 to 2017

Human sapovirus (SaV) is an important viral agent for acute diarrhea worldwide, but timely prevalence data of human SaV in South China are still lacking. In this study, a 4-year surveillance was conducted to characterize the prevalence and genetic characteristics of the circulating SaV associated with sporadic diarrhea in South China. From November 2013 to October 2017, 569 fecal samples from patients with acute diarrhea were collected. SaV was detected in 11 samples with a positive rate of 1.93%. Three human genogroups of GI, GII, and GIV were identified, including five GI.1 strains, three GI.2 strains, one GI.3 strain, one GII.8 strain, and one GIV strain. Furthermore, multiple alignments of complete capsid protein VP1 genes of five local GI.1 strains and other available GI.1 strains in GenBank were performed. Average pairwise identities were calculated at 95.33% and 99.36% at nucleotide and amino acid levels, and only six variable amino acid sites were found during its 36-years’ evolution process. GI.1 strains could be further phylogenetically divided into four clusters with an approximate temporal evolution pattern, and local strains belonged to Cluster-d with other four strains from China and Japan. In summary, SaV was identified as an etiological agent responsible for sporadic gastroenteritis in Guangzhou with a low prevalence rate as in other Chinese cities, but its high genetic diversity suggested the necessity of continuous SaV surveillance in the future.     

Prevalence and genetic diversity of Staphylococcus aureus small-colony variants in cystic fibrosis patients

Staphylococcus aureus small-colony variants (SCVs) are being isolated more frequently in cystic fibrosis (CF) patients. We aimed to determine the prevalence of S. aureus SCVs and their phenotypic and genotypic properties in CF patients admitted to a university hospital. Specimens of 248 patients were examined during a period of 11 months. Colonies supposed to be SCVs were evaluated on Columbia blood agar, mannitol salt agar, and brain–heart infusion agar with 5% NaCl (BHIA 5% NaCl). Strains were confirmed by S. aureus nucA PCR. Antibiotic susceptibilities of SCVs and simultaneously isolated S. aureus strains were determined for oxacillin, gentamicin, trimethoprim–sulphamethoxazole, vancomycin, ciprofloxacin, linezolid, and tigecycline. Genetic relatedness between SCVs and normal S. aureus strains was determined with a pulsed-field gel electrophoresis (PFGE) method. S. aureus SCVs were detected in 20 of 248 patients (8.1%). The highest SCV isolation rate was obtained with MSA, followed by BHIA 5% NaCl. Auxotrophism for thymidine was demonstrated in six SCVs. The tigecycline susceptibilities of 48 SCV strains isolated in this study showed higher MIC values than those of 33 simultaneously isolated normal S. aureus strains. Whereas SCVs and normal S. aureus strains showed identical genotypes in 14 of the patients, five patients showed different genotypes. This first study from Turkey evaluating S. aureus SCVs in CF patients has indicated the importance of considering and reporting SCVs in chronic infections such as CF. The presence of SCVs will probably indicate persistent infection, and this might impact on antibiotic treatment decisions, as they are more resistant to antibiotics.     

Prevalence and genetic diversity of human astroviruses in Mexican children with symptomatic and asymptomatic infections

The prevalence and type diversity of human astroviruses (HAstV) in children with symptomatic and asymptomatic infections were determined in five localities of Mexico. HAstV were detected in 4.6 (24 of 522) and 2.6% (11 of 428) of children with and without diarrhea, respectively. Genotyping of the detected strains showed that at least seven (types 1 to 4 and 6 to 8) of the eight known HAstV types circulated in Mexico between October 1994 and March 1995. HAstV types 1 and 3 were the most prevalent in children with diarrhea, although they were not found in all localities studied. HAstV type 8 was found in Mexico City, Monterrey, and Mérida; in the last it was as prevalent (40%) as type 1 viruses, indicating that this astrovirus type is more common than previously recognized. A correlation between the HAstV infecting type and the presence or absence of diarrheic symptoms was not observed. Enteric adenoviruses were also studied, and they were found to be present in 2.3 (12 of 522) and 1.4% (6 of 428) of symptomatic and asymptomatic children, respectively.     

Studies on genetic diversity of bovine viral diarrhea viruses in Danish cattle herds

Scandinavian countries have successfully pursued bovine viral diarrhea virus (BVDV) eradication without the use of vaccines. In Denmark, control and eradication of BVDV were achieved during the last two decades, but occasionally new BVDV infections are detected in some Danish cattle herds. The aim of this study was to determine recent BVDV subtypes isolated from 4 Danish herds (A, B, C, and D) isolated in 2009–2012 and to analyze the genetic variation of these isolates within the same herd and its relation with those of other herds. The results showed that three herds (B, C, D) were BVDV 1-b and only one herd (herd A) was BVDV 1-d, no other subtypes were detected. The deduced E2 amino acids result showed a high identity percent (99–100 %) between isolates originating from the same herd, but with higher variation compared to isolates of the other herds. Some of these new Danish strains have closer relationship to BVDVs from outside Denmark than to older Danish strains indicating that these are new introductions to Denmark. In conclusion, BVDV-1 subtypes recently detected in Denmark were only subtypes 1b and 1d, and BVDV infections established in a herd is genetically stable over a long time period.     

Prevalence and genetic diversity of TT virus genotype 21 (YONBAN virus) in Brazil

Summary.  Isolates of the newly characterized, single-stranded DNA virus TTV, have been tentatively classified into four major phylogenetic groups and at least 28 genotypes. Four Japanese isolates, designated as YONBAN viruses, belong to the fourth group and to genotype 21. In this study, a genotype 21-specific PCR assay was standardized. With this assay, 48/184 (26%) serum samples and 76/167 (46%) saliva samples, collected from unselected ambulatory patients (aged 2 to 82) of a Brazilian public hospital, were positive. A total of 110 (66%) patients had TTV genotype 21 DNA in serum, saliva, or both fluids. Furthermore, 18/37 (49%) serum samples, collected from Indians belonging to three ethnic groups of the Western Brazilian Amazon, were also positive. Nucleotide sequences (253 bases at the 3′ end of the non-coding region of the genome) were determined, that derived from 25 individuals, i.e. 17 patients and eight Indians. Phylogenetic analysis showed that three isolates from Indians of a particular ethnic group formed a separate subgroup within genotype 21. Among non-Indians, a clustering of strains was observed according to their country of origin (Japan or Brazil), with all 17 sequences derived from Brazilian patients located in a unique subgroup. Received June 20, 2002; accepted September 17, 2002     

Prevalence of hepatitis E virus and sapovirus in post-weaning pigs and identification of their genetic diversity

Hepatitis E virus (HEV) and sapovirus (SaV) induce acute hepatitis and gastroenteritis, respectively, in humans. As pigs have been recognized as an important reservoir for these viruses, we evaluated the infection rates of both viruses using fecal samples collected from post-weaning pigs via RT-PCR methods. In the five swine farms assessed in this study, 3 farms were found to be HEV-positive and 4 farms were SaV-positive. The overall prevalence of HEV and SaV in the pigs was 17.0 and 23.1%, respectively. Phylogenetic tree analysis revealed that the isolated swine HEVs belonged to genotype 3 and the porcine SaVs belonged to genogroup III. This study proved that both HEV and SaV are prevailing in post-weaning pigs in Korea. J. N. Yu and M. Y. Kim are equally contributed to this work     

Molecular detection and genetic diversity of Theileria orientalis in cattle in China

Parasitology Research - The apicomplexan parasite Theileria orientalis is a tick-borne intracellular protozoan parasite that is widely distributed throughout China. It causes bovine theileriosis in...     

Prevalence and genetic diversity of Entamoeba histolytica in an institution for the mentally retarded in the Philippines

A total of 113 mentally retarded patients residing in a mental institution in Metropolitan Manila, Philippines, were screened for the presence of Entamoeba histolytica based on microscopy and polymerase chain reaction (PCR). Anti-E. histolytica antibodies were also screened in 97 serum samples collected using immunofluorescence antibody (IFA) test. Parasitological examination showed E. histolytica/Entamoeba dispar in 43 cases (38.05%), while PCR detected 74 cases (65.48%) positive for E. histolytica and 6 cases (5.30%) positive for E. dispar. Interestingly, these 6 samples were coinfected with E. histolytica. IFA test revealed that 80.41% (78/97) of the respondents possessed significant antibody titers for intestinal infection of E. histolytica. Of this number, there were 5 patients negative in IFA test but positive in PCR. The genetic diversity of E. histolytica isolates was also investigated by analyzing polymorphism in the serine-rich gene by nested PCR on DNA directly extracted from stool specimens. A combination of the nested PCR results and the AluI digestion of the PCR products examined yielded six distinct DNA banding patterns among the 74 stool isolates. An apparent clustering of E. histolytica strains was observed in patients living in different residential cottages of the institution. These results indicate the high prevalence of E. histolytica in an institution for the mentally retarded in the Philippines.     

Prevalence and mode of inheritance of major genetic eye diseases in China.

The prevalence and mode of inheritance of major genetic eye diseases have been investigated in China since the establishment of the Section of Ophthalmic Genetics of the Chinese Society of Genetics. Mass screening of genetic eye diseases has been undertaken in many districts in China, covering more than 700,000 people, and more than 5000 pedigrees of genetic eye diseases have been collected and analysed all over China. Based on these data, the prevalence and mode of inheritance of dyschromatopsia, degenerative myopia, retinitis pigmentosa, congenital ptosis, congenital microphthalmos, congenital cataract, congenital glaucoma, Leber's optic atrophy, corneal dystrophy, congenital nystagmus, coloboma of the eye, congenital aniridia, retinoblastoma, macular dystrophy, simple myopia, primary glaucoma, and strabismus have been investigated, and the results are presented.     

Complete sequencing and phylogenetic analysis of porcine kobuvirus in domestic pigs in Northwest China

Porcine kobuvirus, a member of the genus Kobuvirus that is associated with diarrhea, has been reported in many countries. We determined the complete genome sequence and investigated the genetic evolution of the kobuvirus strain swKoV CH441, which was detected in the highland of Gansu province in Northwest China. The viral genome is 8149 nucleotides (nt) long, including a 29-nt poly(A) tail of the 3′ end, and is 90 nt shorter in the 2B coding region than those of other kobuvirus strains whose sequences are available in the GenBank database. Phylogenetic analysis showed that swKoV CH441was most closely related to porcine kobuvirus CH/HNXX-4 but more distantly related to other strains, including the strains GS-1/2012/CH and GS-2/2012/CH, which were detected in Gansu province, indicating that porcine kobuvirus may have geographic and host differences in evolution.     

Prevalence and genetic diversity of norovirus infection in Tunisian children (2007–2010)

Viral gastroenteritis can be a life‐threatening disease in infants and young children, especially in developing countries. The aim of this study was to continue the epidemiological surveillance of norovirus (NoV) infections in Tunisian children suffering from acute gastroenteritis. Surveillance was initiated in January 2003, to monitor potential variations in strains over time, in terms of frequency and diversity of NoV genotypes, and more particularly the potential emergence of new GII.4 variants following the 2004 Hunter variant. From April 2007 to April 2010, a total of 407 stool specimens were collected from sporadic cases (238 inpatients and 169 outpatients). Furthermore, 28 stool samples were collected from children involved in 3 gastroenteritis outbreaks. Stool specimens were screened for NoV genogroup I (GI) and II (GII) by RT‐PCR. NoV strains were genotyped, and variants identified, based on sequence and phylogenetic analyses of the polymerase and capsid genes. NoVs were detected in 38 sporadic cases (9.3%) and 21 epidemic cases (75%). Great diversity was observed throughout the period, with seven distinct NoV genotypes characterized in sporadic cases, and three in outbreaks. GIIb/II.3 and GII.4 were predominant globally, with fluctuations of their prevalence over time. Interestingly, the Hunter variant, which was the unique GII.4 variant observed from 2003 to April 2007 in the region of Monastir, was replaced by the 2006b variant. NoV is an important enteropathogen responsible for viral gastroenteritis among infants and children in Tunisia, and the infecting strains between 2007 and 2010 were different from those in previous years. J. Med. Virol. 85: 1100–1110, 2013. © 2013 Wiley Periodicals, Inc.     

Canine kobuvirus infections in Korean dogs

To investigate canine kobuvirus (CaKoV) infection, fecal samples (n = 59) were collected from dogs with or without diarrhea (n = 21 and 38, respectively) in the Republic of Korea (ROK) in 2012. CaKoV infection was detected in four diarrheic samples (19.0 %) and five non-diarrheic samples (13.2 %). All CaKoV-positive dogs with diarrhea were found to be infected in mixed infections with canine distemper virus and canine parvovirus or canine adenovirus. There was no significant difference in the prevalence of CaKoV in dogs with and without diarrhea. By phylogenetic analysis based on partial 3D genes and complete genome sequences, the Korean isolates were found to be closely related to each other regardless of whether they were associated with diarrhea, and to the canine kobuviruses identified in the USA and UK. This study supports the conclusion that CaKoVs from different countries are not restricted geographically and belong to a single lineage.     

Viruses causing mosaic disease in sugarcane and their genetic diversity in southern China

A survey of cultivated hybrid sugarcane (Saccharum inter-specific hybrid) and noble sugarcane (Saccharum officinarum) in southern China for the presence of Sugarcane mosaic virus (SCMV), Sorghum mosaic virus (SrMV) and Sugarcane streak mosaic virus (SCSMV) was conducted by RT-PCR from the years 2003 to 2006. SCMV and SrMV, but not SCSMV, were found. A high incidence of SCMV and SrMV coinfection was revealed in both hybrid and noble sugarcanes. All coinfected plants showed mosaic symptom, whereas plants infected with a single virus were symptomatic or asymptomatic. It appears that virus mixtures are more virulent than single infections. The nucleotide sequences of the coat protein (CP) gene of 33 SCMV and 10 SrMV isolates from this study were compared to those of CP genes of SCMV and SrMV reported in GenBank. One hundred and seventy-three SCMV isolates, with the exception of MDB and Abaca strains, can be grouped into five groups, which include three previously known groups, the sugarcane (SCE), maize (MZ), and Thailand groups, and two newly identified groups, the noble sugarcane (NSCE) and Brazil groups. Twenty-two SrMV isolates were divided into two groups, HS (hybrid sugarcane) and NS (noble sugarcane) groups. Five out of eight SrMV hybrid isolates belonged to the HS group, and two SrMV noble isolates and three hybrid isolates were within the NS group. Interestingly, the three hybrid isolates within the NS group were isolated from hybrid sugarcane co-infected with SCMV. This indicates that SCMV helps the NS group SrMV to infect hybrid sugarcane.