Genetic Variation Analysis between Wild and Cultured Pangasianodon hypopthalmus Using COI and Cytochrome b Among Asian Countries

One of the top species in the aquaculture sector, known as striped catfish or Pangasianodon hypophthalmus, is an important and valuable freshwater fish in many countries. Due to the high demand for this species, their number has declined to "threatened" levels. Hence, the purpose of this study is to analyse the genetic variation of wild and cultured striped catfish collected from five producers in Asian countries; Thailand, Vietnam, Indonesia, India, and Philippines, by using mitochondrial DNA partial region data sequence; CO1 and cytochrome b gene. Population analyses using 395 base pairs length for CO1 and 275 base pairs length of cytochrome b partial region nucleotide sequence have shown no significance difference between wild and cultured striped catfish. Vietnam species had shown a wide range of genetic distance of the intrapopulation compared with other countries in the range of 0.000-0.040 for CO1 gene and 0.003-0.008 for cytochrome b gene. The Neighbour-joining method has also been used to construct phylogenetic trees using CO1 gene; the tree formed few subclades with mixed populations, and the tree using cytochrome b showed only Vietnam species divided into a few sub-populations. For the other four countries, Thailand, Indonesia, India, and Philippines were in the same group. Hence, this study's findings may provide a reference for inter and intra-relationships of P. hypophthalmus that may help in the aquaculture activity of this striped catfish.

In Silico Study of the Effectiveness of 16S rRNA Gene as a Universal Genetic Marker to Identify Closely Related Burkholderia Spp. in Panicle Blight of Rice

The 16S rRNA gene is a housekeeping genetic marker that is available in almost all bacterial species and it is used in bacterial phylogeny and taxonomy studies. In many studies, the 16S rRNA gene is used in identification of certain bacterial species. Being a less conserved genetic marker, certain studies found it is a useful tool to infer the genome-wide similarity levels among the closely related prokaryotic organisms. Thus, this study aimed to compare the variation in the 16S rRNA partial region of Burkholderia spp. that infect the panicle of rice from eight different geographical areas. 58 sequences with total of 688 base pairs (bp) of 16S rRNA gene in B. glumae and B. gladioli were retrieved from public database based on several countries namely United State, Panama, Ecuador, Thailand, China, India, Korea and Malaysia. Then, the data sequences were analysed and validated using MEGAX and ABGD software respectively. The result of phylogenetic tree confirmed that B. glumae and B. gladioli were species that present in the panicle blight of rice. However, Data Analysis in Molecular Biology and Evolution (DAMBE) and Automatic Barcode Gap Discovery (ABGD) software were not able to detect substitution saturation and divergence between B. glumae and B. gladioli respectively based on the 58 sequences of the 16S rRNA partial region. Hence, it proves that 16S rRNA gene is an ineffective genetic marker to be used to differentiate the closely related species of bacteria from similar genus.

The Full Region of N-Terminal in Polymerase of IBDV Plays an Important Role in Viral Replication and Pathogenicity: Either Partial Region or Single Amino Acid V4I Substitution Does Not Completely Lead to the Virus Attenuation to Three-Yellow Chickens

Infectious Bursal Disease Virus (IBDV) has haunted the poultry industry with severe, prolonged immunosuppression of chickens when infected at an early age and can easily lead to other secondary infections. Understanding the pathogenic mechanisms could lead to effective prevention and control of Infectious Bursal Disease (IBD). Evidence suggests that the N-terminal domain of polymerase in segment B plays an important role, but it is not clear which part or residual is crucial for the pathogenicity. Using a reverse genetics technique, a molecular clone (rNN1172) of the parental vvIBDV strain NN1172 was generated, and its pathogenicity was found to be the same as the parental virus. Then, three recombinant chimeric viruses were rescued based on the rNN1172 and substituted with the counterparts in the N-terminal domain of the attenuated vaccine strain B87: the rNN1172-B87VP1a (substituting the full region of the 1–167 aa residuals), the rNN1172-B87VP1a∆4 (substituting the region of the 5–167 aa residuals), and the rNN1172-VP1∆4 (one single aa residual substitution V4I), to better explore the role of the N-terminal domain of polymerase on the viral pathogenicity. Interestingly, all these substitutions played different roles in the viral pathogenicity: the mortality of the rNN1172-B87VP1a-challenged chickens was significantly reduced from 30% to 0%. No obvious lesion was found in the histopathological examination, and the lowest viral genome copy number was also detected in the bursa when compared to the parental and two other recombinant viruses. The mortalities caused by rNN1172-B87VP1a∆4 and rNN1172-B87VP1∆4, respectively, were all reduced to 10% and had a delayed onset of death. Our results also revealed that the pathogenicity of the IBDV was consistent with the viral replication efficiency in vivo (bursae). This study demonstrated that the full region of the N-terminal of polymerase plays an important role in viral replication and pathogenicity, but the substitutions of its partial region or a single residual do not completely lead to the virus attenuation to Three-Yellow chickens, although that significantly reduces its pathogenicity.

Molecular Detection and Genetic Analysis of Theileria equi Detected in Apparently Healthy Horses in Nigeria

Equine theileriosis, an apicomplexan debilitating tick-borne parasitic disease of horses has caused considerable havoc to equine production all over the world. There is a dearth of information on the molecular characteristic of the parasites, Theileria equi Laveran, 1901, in Nigeria. Thus, in this study microscopy techniques and PCR were used to detect the T. equi of horses in Ogun, Oyo and Lagos States of Nigeria. We also characterized the partial region of 18S ribosomal RNA gene by sequencing and sequences analysis. One hundred and two horses consisting of Argentine 34 (33.3 %), Sudanese 21 (20.6 %) and local breeds 47 (46.1 %) including 2 females and 100 males were randomly sampled from the Polo Clubs in Ibadan, Lagos and from privately owned horse stables in Abeokuta. Blood samples were collected from the jugular vein, thin smears were prepared and stained with a field stain. The DNA was extracted from the blood and a partial region of the 18S ribosomal RNA gene was amplified. The amplified products were sequenced unidirectionally and subjected to phylogenetic analysis with those sequences obtained from the Gen-Bank. Of the 102 horses tested, 12 (11.7 %) were positive for T. equi by microscopy which included 9 (19.1 %) local breeds, 2 (5.8 %) Argentine breed and 1 (4.8 %) Sudanese breed. In contrast, 7 (6.8 %) were positive by the PCR method; out of which 5 (10.6 %) of these samples were from the local breed of horses while the remaining 2 (5.8 %) were from the Argentine breed. The Packed Cell Volume (PCV) of the infected and non-infected horses did not show any significant (P < 0.05) difference. The sequences lengths obtained were 311 bp and they had 97.43—98.07 % homologies with available sequences in the GenBank. The phylogenetic analysis of the sequences suggested that the strain of T. equi detected in the study area formed a new genotype different from the established genotypes around the world. In conclusion, the prevalence of T. equi was very low in the study area and one strain of the parasite may be in circulation among the studied horses.

Characteristics of GII.4 Norovirus Versus Other Genotypes in Sporadic Pediatric Infections in Davidson County, Tennessee, USA

Background Norovirus is a leading cause of epidemic acute gastroenteritis (AGE), with most outbreaks occurring during winter. The majority of outbreaks are caused by GII.4 noroviruses; however, data to support whether this is true for sporadic medically attended AGE are limited. Therefore, we sought to compare the clinical characteristics and seasonality of GII.4 vs non-GII.4 viruses. Methods Children aged 15 days -17 years with AGE symptoms were recruited from the outpatient, emergency department, and inpatient settings at Vanderbilt Children’s Hospital, Davidson County, Nashville, Tennessee, from December 2012 -November 2015. Stool specimens were tested using qRT-PCR for GI and GII noroviruses and subsequently genotyped by sequencing a partial region of the capsid gene. Results A total of 3705 patients were enrolled, and stool specimens were collected and tested from 2885 (78%) enrollees. Overall, 636 (22%) samples were norovirus-positive, of which 567 (89%) were GII. Of the 460 (81%) genotyped GII-positive samples, 233 (51%) were typed as GII.4 and 227 (49%) as non-GII.4. Compared with children with non-GII.4 infections, children with GII.4 infections were younger, more likely to have diarrhea, and more likely to receive oral rehydration fluids. Norovirus was detected year-round and peaked during winter. Conclusions Approximately 40% of sporadic pediatric norovirus AGE cases were caused by GII.4 norovirus. Children infected with GII.4 had more severe symptoms that required more medical care. Seasonal variations were noticed among different genotypes. These data highlight the importance of continuous norovirus surveillance and provide important information on which strains pediatric norovirus vaccines should protect against.

High-Resolution Crystal Structure of RpoS Fragment Including a Partial Region 1.2 and Region 2 from the Intracellular Pathogen Legionella pneumophila

Legionella pneumophila RpoS (LpRpoS) is an alternative sigma factor of RNA polymerase (RNAP) essential for virulence and stress resistance. To investigate the mechanism of RpoS in the intracellular pathogen L. pneumophila, we determined the high-resolution crystal structure of the LpRpoS (residues 95-194) containing a partial region 1.2 and region 2. The structure of LpRpoS (residues 95-194) reveals that the conserved residues are critical for promoter melting, DNA and core RNAP binding. The differences in regulatory factor binding site between Escherichia coli RpoS and LpRpoS suggest that LpRpoS may employ a distinct mechanism to recruit alternative regulatory factors controlling transcription initiation.