Co-infections of Klebsiella pneumoniae and Elizabethkingia miricola in black-spotted frogs (Pelophylax nigromaculatus).

Pelophylax nigromaculatus is a common commercial specie of frogs that generally cultured throughout China. With the application of high-density culture, P. nigromaculatus can be co-infected by two or more pathogens, which thereby induce synergistic influence on the virulence of the infection. In this study, two bacterial strains were simultaneously isolated from diseased frogs by incubating on Luria-Bertani (LB) agar. Isolates were identified as Klebsiella pneumoniae and Elizabethkingia miricola by morphological, physiological and biochemical features, as well as 16S rRNA sequencing and phylogenetic analysis. The whole genome of K. pneumoniae and E. miricola isolates consist single circular chromosome of 5,419,557 bp and 4,215,349 bp, respectively. The genomic sequence analysis further indicated that K. pneumoniae isolate conserved 172 virulent and 349 antibiotic-resistance genes, whereas E. miricola contained 24 virulent and 168 antibiotic resistance genes. In LB broth, both isolates could grow well at 0%-1% NaCl concentration and pH 5-7. Antibiotic susceptibility testing revealed that both K. pneumoniae and E. miricola were resistant to kanamycin, neomycin, ampicillin, piperacillin, carbenicillin, enrofloxacin, norfloxacin and sulfisoxazole. Histopathological studies showed that co-infection caused considerable lesions in the tissues of brain, eye, muscle, spleen, kidney and liver, including cell degeneration, necrosis, hemorrhage and inflammatory cell infiltration. The LD50 of K. pneumoniae and E. miricola isolates were 6.31 × 105 CFU/g and 3.98 × 105 CFU/g frog weight, respectively. Moreover, experimentally infected frogs exhibited quick and higher mortality under coinfection with K. pneumoniae and E. miricola than those single challenge of each bacterium. To date, no natural co-infection by these two bacteria has been reported from frogs and even amphibians. The results will not only shed light on the feature and pathogenesis of K. pneumoniae and E. miricola, but also highlight that co-infection of these two pathogen is a potential threat to black-spotted frog farming.

Characterization of the complete mitochondrial genome of Khawia sinensis belongs among platyhelminths, cestodes.

Khawia sinensis is an important species in freshwater fish causing considerable economic losses to the breeding industry. This is the first mt genome of a caryophyllidean cestode characterised. The entire mt genome of K. sinensis is 13,759 bp in length. This mt genome contains 12 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes and two non-coding regions. The arrangement of the K. sinensis mt genome is the same as other tapeworms, however, the incomplete stop codon (A) is more frequent that other species. Phylogenetic analyses based on concatenated amino-acid sequences of the 12 protein-coding genes of 17 tapeworms including K. sinensis were conducted to assess the relationship of K. sinensis with other species, the result indicated K. sinensis was closely related with cestode species. This complete mt genome of K. sinensis will enrich the mitochondrial genome databases of tapeworms and provide important molecular markers for ecology, diagnostics, population variation and evolution of K. sinensis and other species.

Research of System Design and Automatic Detection Method for Excretion Nursing Equipment.

(1) Background: The nursing of the elderly has received more and more attention, especially the nursing of urination and defecation for the elderly. (2) Purpose: Design an excretion nursing equipment that can accurately identify and deal with urine and stool. (3) Methods: In this paper, based on the analysis of the requirements of excretion nursing equipment, a split mechanical design method and a modular control method are used to design the equipment. The Dempster-Shafer (D-S) evidence theory is used in the identification of urine and stool. (4) Results: The excretion nursing equipment designed in this paper works well according to functional test, and the success rate of stool and urine identification method using D-S evidence theory is 20% higher than that of traditional methods, reaching 90%. (5) Conclusions: The urine and stool recognition and detection algorithm based on the D-S evidence theory used in this paper can improve the recognition accuracy of traditional detection methods, and the designed excretion nursing equipment can realize the function of excretion care for patients.

Probiotic Supplementation During Human Pregnancy Affects the Gut Microbiota and Immune Status.

The consumption of probiotics and fermented foods has been very popular in recent decades. The primary aim of our study was to evaluate the effect of probiotics on the gut microbiota and the changes in inflammatory cytokines after an average of 6.7 weeks of probiotic administration among normal pregnant women. Thirty-two healthy pregnant women at 32 weeks of gestation were recruited and divided into two groups. The probiotic group ingested combined probiotics until after birth. The base characteristics of the probiotics and control groups showed no significant differences. The structure of the fecal microbiota at the genus level varied during the third trimester, and administration of probiotics had no influence on the composition of the fecal microbiota however, many highly abundant taxa and core microbiota at the genus level changed in the probiotic group when compared to the control group. The analysis of cytokines showed that IL-5, IL-6, TNF-α, and GM-CSF had equal levels between the baseline and control groups but were significantly increased after probiotic administration (baseline = control < probiotics). Additionally, levels of IL-1ß, IL-2, IL-12, and IFN-γ significantly increased among the three groups (baseline < control < probiotics). This result demonstrated that probiotics helped to shift the anti-inflammatory state to a pro-inflammatory state. The correlation analysis outcome suggested that the relationship between the microbiota and the cytokines was not strain-dependent. The gut microbiota varied during the third trimester. The probiotics demonstrated immunomodulation effects that helped to switch over to a pro-inflammatory immune state in the third trimester, which was important for labor.

Complete mitochondrial genome of Lernaea cyprinacea (Copepoda: Cyclopoida).

The complete mitochondrial genome of Lernaea cyprinacea is 14,656 bp in size, containing 13 protein-coding genes (PCGs), 22 transfer RNA genes, two rRNA genes (12S and 16S), as well as two non-coding regions (NCRs, the control regions). The genome organization, nucleotide composition and codon usage do not differ significantly from other Copepodas. The complete mitogenome sequence information of L. cyprinacea provides useful data for further studies on phylogenetics, stock evaluation and conservation genetics.

Complete mitochondrial genome of Sinergasilus polycolpus (Copepoda:Poecilostomatoida).

The full mitochondrial (mt) genome of Sinergasilus polycolpus is 14 000 bp in length, and it contains 13 protein-coding genes, 22 transfer RNA genes, 2 rRNA genes and 1 non-coding region. The genome organization, nucleotide composition and codon usage are different compared to other Copepodas, such as Lernaea cyprinacea. The full mt genome of S. polycolpus provides useful information for further studies on epidemiology and phylogenetics.

Complete mitochondrial genome of Philometra carassii (Nematoda: Philometridae).

The complete mitochondrial genome of Philometra carassii is 14,378 bp in size, containing 12 protein-coding genes (PCGs), 22 transfer RNA genes, 2 rRNA genes (12S and 16S), as well as one non-coding region (NCR, the control region), but lacks an atp8 gene. The genome organization, nucleotide composition and codon usage do not differ significantly from other nematodes. The complete mitogenome sequence information of P. carassii can provides useful data for further studies on phylogenetics, stock evaluation and conservation genetics.