Role of Human Body Composition Analysis and Malnutrition Risk Questionnaire in the Assessment of Nutritional Status of Patients With Initially Diagnosed Crohn's Disease.

Objective: This study was carried out to investigate the role and necessity of human body composition analysis in assessing the nutritional status of initially diagnosed Crohn's disease (CD) patients. Methods: A total of 47 initially diagnosed CD patients were recruited. The skeletal muscle mass index (SMI), fat-free mass index (FFMI), body fat mass, body fat percent, visceral fat area (VFA), and body cell mass were determined with the Biospace Inbody S10 composition analyzer. Results: In 47 patients with initially diagnosed CD, SMI could determine the muscular mass reduction that could not be determined by the body mass index (BMI) (35.3%), albumin (ALB) (65.6%), nutrition risk screening (NRS)2002 (25.0%), and Patient-Generated Subjective Global Assessment (PG-SGA) (55.6%). FFMI could determine the malnutrition that could not be determined by the BMI (58.8%), albumin (90.6%), NRS2002 (50.0%), and PG-SGA (55.6%). VFA in the fistulizing CD patients was significantly higher than in the stricturing and non-fistulizing, non-stricturing patients (P < 0.05). SMI and BMI had the same performance (P = 1.000) and general consistence (Kappa = 0.487, P = 0.001) in the assessment of malnutrition; SMI and ALB had different performance (P < 0.001) and inconsistence was noted (Kappa = 0.069, P = 0.489) in the assessment of malnutrition; the results of the nutrition assessment were different between SMI and NRS2002 (P = 0.002), and inconsistence was observed (Kappa = 0.190, P = 0.071). SMI and PG-SGA had the same performance in the assessment of nutrition (P = 0.143), but there was inconsistence (Kappa = 0.099, P = 0.464). FFMI and BMI had general consistence in the assessment of malnutrition (Kappa = 0.472, P < 0.001), but the positive rate determined by FFMI (85.1%) was markedly higher than that by BMI (63.8%) (P = 0.002). FFMI and ALB had different performance in the assessment of malnutrition (P < 0.001) and there was inconsistence (Kappa = -0.008, P = 0.877). FFMI and NRS2002 had the same performance in the assessment of malnutrition (P = 0.453), but the consistence was poor (Kappa = 0.286, P = 0.039). The results determined by SMI and PG-SGA were consistent (P = 0.727), but the consistence was poor (Kappa = 0.399, P = 0.006). Conclusion: Human body composition analysis can identify the patients with muscular mass reduction that cannot be identified by commonly used nutrition assessment scales/parameters. Thus, it is helpful for the assessment of disease severity and also important for the nutrition assessment in CD patients.

Identification of the first case of SFTSV infection in the Hunan Province of China and epidemiological surveillance in the locality.

This study reports the etiological identification, clinical diagnosis, and the results of the local epidemiological surveillance of the first case of severe fever with thrombocytopenia syndrome virus (SFTSV) infection in 2014 in Hunan Province, China. The infected patient was isolated and closely monitored. The virus is a member of the Bunyaviridae sandfly family and is characterized by real-time PCR, electron microscopy, immunofluorescence, and whole-genome sequencing. We also detected IgG and IgM antibodies against SFTSV among the local human population and domestic animals in a serological surveillance. Prevalence of SFTSV-specific antibodies was monitored in the local population for two years after the identification of the first SFTS case. Approximately 5% (4/77) of the people who had direct contact with the patient were seropositive, which is significantly higher than the seropositivity of the general local population [1.57% (44/2800), P < 0.05]. Furthermore, the percentage of the general population who were seropositive was higher in 2015 than in 2014 (χ2 = 7.481, P = 0.006). The epidemiological investigation found that the SFTSV is epidemic in goats, cattle, and chickens in Hunan Province. The risk of infection of domestic animals can be minimized by feeding in pens rather than allowing foraging.

Cloning of the gene and characterization of the enzymatic properties of the monomeric alkaline phosphatase (PhoX) from Pasteurella multocida strain X-73.

We have identified a new phoX gene encoding the monomeric alkaline phosphatase from Pasteurella multocida X-73. This gene was not found in the published genome sequence of Pasteurella multocida pm70. Characterization of the recombinant PhoX of Pasteurella multocida X-73 showed that it is a monomeric enzyme, activated by Ca(2+) and possibly secreted by the Tat pathway. These features distinguish phosphatases of the PhoX family from those of the PhoA family. All proteins of the PhoX family were found to contain a conserved motif that shares significant sequence homology with the calcium-binding site of a phosphotriesterase known as diisopropylfluorophosphatase. Site-directed mutagenesis revealed that D527 of PhoX might be the ligand bound to the catalytic calcium. This is the first report on identification of homologous sequences between PhoX and the phosphotriesterase and on the potential calcium-binding site of PhoX.

Molecular characterization of plasmids with antimicrobial resistant genes in avian isolates of Pasteurella multocida.

The complete nucleotide sequences of two plasmids from avian isolates of Pasteurella multocida that caused outbreaks of fowl cholera in Taiwan were determined. The entire sequences of the two plasmids, designated as pJR1 and pJR2, were 6792 bp and 5252 bp. Sequence analysis showed that the plasmid pJR1 contained six major genes: the first gene (sulII) encoded a type II sulfonamide resistant dihydropteroate synthase, the second gene (tetG) encoded a tetracycline resistance protein, the third gene (catB2) encoded a chloramphenicol acetyltransferase, the fourth gene (rep) encoded a replication protein, and the fifth and sixth genes (mbeCy and deltambeAy) encoded proteins involved in the mobilization of plasmid. The plasmid pJR2 contained five major genes: the first gene (deltaintI1) encoded a truncated form of a type I integrase, the second gene (aadA1) encoded an aminoglycoside adenylyltransferase that confers resistance to streptomycin and spectinomycin, the third gene (blaP1) encoded a beta-lactamase that confers resistance to ampicillin and carbenicillin, and the fourth and fifth genes might encode proteins involved in the plasmid replication or segregation. Sequence comparisons showed that the antibiotic resistance genes found in pJR1 and pJR2 exhibited a high degree of sequence homology to the corresponding genes found in a great variety of gram-negative bacteria, including Escherichia coli, Salmonella enterica Typhimurium DT104, Psedomonas spp., P. multocida, Mannheimia spp., and Actinobacills pleuropneumoniae, which suggests that these resistance genes were disseminated in these bacteria. Although sulII and tetG genes were found previously in P. multocida or Mannheimia spp., this is the first report on the presence of catB2, aadA1, and blaP1 genes in bacteria of the family Pasturellaceae. Moreover, the aadA1 and blaP1 genes found in pJR2 were organized into an integron structure, which is a site-specific recombination system capable of capturing and mobilizing antibiotic resistance genes. This is also the first report on the presence of an integron in bacteria of the family Pasteurellaceae. The presence of a P. multocida integron might facilitate the spreading of antibiotic resistance genes between P. multocida and other gram-negative bacteria.

Recombinant proteins containing the hypervariable region of the haemagglutinin protect chickens against challenge with Avibacterium paragallinarum.

The haemagglutinin (HA) protein plays a key role in the immunogenicity and pathogenicity of Avibacterium paragallinarum, but the domain organization and antigenicity exhibited by different domains of this protein remain unknown. This study reports the presence of a hypervariable region in the HA proteins of strains of serovars A and C of A. paragallinarum. This hypervariable region is located approximately at residues 1100-1600 of the HA protein. The sequence identity found in this hypervariable region was only 18.1%, whereas those upstream and downstream of this region were 83.8 and 97.8%, respectively. Western blot analyses using antisera against the whole-cell antigens of A. paragallinarum showed that the hypervariable region was more antigenic than other regions of the HA protein. Moreover, the antigenicity of the hypervariable region was serovar-specific. Chickens immunized with recombinant proteins that contained the hypervariable region were protected (83-100% protection rate) against challenge infection with A. paragallinarum of the homologous serovar. These results suggest that recombinant proteins containing the hypervariable region may be useful antigens for use in the development of a vaccine against A. paragallinarum.

Polymerase chain reaction-restriction fragment length polymorphism analysis of the genes involved in the biosynthesis of the lipopolysaccharide of Pasteurella multocida.

The lipopolysaccharide, also known as the somatic antigen or O-antigen, is an important virulence factor of Pasteurella multocida. In the current study, the genes involved in the biosynthesis of the outer core region of the lipopolysaccharide, which were obtained from somatic type reference strains and field strains of P. multocida, were subjected to polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. The PCR-RFLP analysis classified 11 out of the 16 serotypes into 5 PCR-RFLP types (I-V). Types I and V contain strains belong to serotypes 1 and 13, respectively. The rest of the PCR-RFLP types contain strains belong to certain groups of serotypes. Typing of 38 field strains from poultry using PCR-RFLP analysis and the gel diffusion precipitation test showed consistent results. These results indicate that the PCR-RFLP analysis can be a useful tool for rapid somatic typing of some strains of P. multocida.

Analysis of the biosynthesis genes and chemical components of the capsule of Avibacterium paragallinarum.

The aim of this study was to investigate biosynthesis genes and chemical components of the capsule of Avibacterium paragallinarum. The sequence of a 10-kb region containing the capsule biosynthetic locus of Av. paragallinarum was determined. Two reference strains, i.e., 221 (serovar A) and H18 (serovar C), together with four Taiwanese field strains (all serovar C) were sequenced. The results showed that there are two genotypes (I and II) of the capsule biosynthetic locus in Av. paragallinarum, and the capsule genotype is independent of the serovar. The capsule biosynthetic loci of genotypes I and II consisted of six and five genes, respectively. The genotype I genes encoded proteins that are most similar to proteins from Pasteurella multocida capsule types A and F while the genotype II genes encoded proteins most similar to proteins from P. multocida capsule type D and Escherichia coli K5. The results suggested that genotype I strains contain hyaluronan or chondroitin in the capsule wall while genotype II contain heparosan. Enzymatic digestion of the capsule materials extracted from Av. paragallinarum showed that genotype I strains contained chondroitin while genotype II strains contained heparosan in the capsule. This is the first report on the existence of different genotypes of capsule biosynthesis genes in Av. paragallinarum and the presence of chondroitin and heparosan as chemical components of the capsule of Av. paragallinarum.

Development of blocking ELISA for detection of antibodies against avian influenza virus of the H7 subtype.

BACKGROUND AND PURPOSE: The conventional method used for subtyping of antibodies against avian influenza viruses is hemagglutination inhibition (HI) test. However, the HI test is laborious and requires preparation of antigen from viable viruses that might be hazardous. The aim of this study was to develop a blocking enzyme-linked immunosorbent assay (B-ELISA) for detection of antibody of avian influenza of the H7 subtype. The B-ELISA is fast and avoids the need to culture whole viruses. METHODS: The B-ELISA was based on the reaction between a monoclonal antibody and a recombinant hemagglutinin protein purified from Escherichia coli. The specificity of the B-ELISA was determined by testing H7-negative field sera and the sensitivity of the B-ELISA was determined by testing sera collected from experimentally immunized chickens. RESULTS: The specificity of the B-ELISA was found to be 97.7% when compared with the HI test. The sensitivity was found to vary with the HI titer of sera. A sensitivity of 100% was achieved when test sera had HI titers >or=2(7). The sensitivity dropped to 33% and 20% when test sera had HI titers of 2(6) and 2(5), respectively. Nearly all test sera with HI titers

Protective immunity conferred by recombinant Pasteurella multocida lipoprotein E (PlpE).

The genes encoding Pasteurella multocida lipoprotein E (PlpE) and lipoprotein B (PlpB) were cloned from P. multocida strain X-73 (serotype A:1) and expressed in Escherichia coli. The protective immunity conferred by recombinant PlpE (r-PlpE) and PlpB (r-PlpB) on mice and chickens was evaluated. The results showed that mice immunized with 10microg of purified r-PlpE were protected (80-100% survival rate) against challenge infection with 10 or 20 LD(50) of P. multocida strains X-73 (serotype A:1), P-1059 (serotype A:3) and P-1662 (serotype A:4). In contrast, mice immunized with r-PlpB were not protected. Chickens immunized with 100microg of purified r-PlpE were protected (63-100% survival rate) against lethal challenge infection with strains X-73 and P-1662, whereas those immunized with r-PlpB were not. Sequence analyses showed that PlpE from different strains of P. multocida exhibited 90.8-100% sequence identity to each other, suggesting that PlpE might serve as a cross-protective antigen. This is the first report of a recombinant P. multocida antigen that confers cross protection on animals.