Integrated microbiome and metabolomic analysis of Spodoptera litura under Metarhizium flavoviride qc1401 stress.

Metarhizium spp. have emerged as an alternative to chemical pesticides for protecting crops from insect pest. Here, we investigated midgut microbial community and metabolites of Spodoptera litura at three different timepoints after infection with Metarhizium flavoviride. The innate immune system of S. litura was activated with levels of polyphenol oxidase, carboxylesterase, multifunctional oxidase, and glutathione S-transferase activity significantly increasing. Exposure to the fungal pathogen also altered bacterial abundance and diversity in host's midgut, and these changes varied depending on the time elapsed since exposure. We identified more operational taxonomic units in the treated samples as compared to the control samples at all tested time points. A total of 372 metabolites were identified, and 88, 149, and 142 differentially accumulated metabolites (DAMs) were identified between the treatment and control groups at 3 timepoints after treatment, respectively. Based on the changes of DAMs in response to M. flavoviride infection at different timepoints and significantly enriched KEGG pathways, we speculated that "tyrosine metabolism," "galactose metabolism," "ATP-binding cassette transporters," "neuroactive ligand-receptor interaction," "purine metabolism," "arginine and proline metabolism," "beta-alanine metabolism," "lysosome," and "carbon metabolism" may participate in the metabolic-level defense response. An integrated pathway-level analysis of the 16S-rDNA and metabolomic data illustrated the connections and interdependencies between the metabolic responses of S. litura and the midgut microorganisms to M. flavoviride infection. This work emphasizes the value of integrated analyses of insect-pathogen interactions, provides a framework for future studies of critical microorganisms and metabolic determinants of these interactions, establishes a theoretical basis for the sustainable use of M. flavoviride.

Screening and Identification of High-Yielding Strains of Conjugated Linoleic Acid and Optimization of Conditions for the Conversion of CLA.

Conjugated linoleic acid (CLA) is a class of naturally occurring octadecadienoic acid in humans and animals and is a general term for a group of conformational and positional isomers of linoleic acid. In order to obtain the development of excellent lactic acid strains with a high production of conjugated linoleic acid, 32 strains with a possible CLA conversion ability were obtained by initial screening using UV spectrophotometry, and then the strains were re-screened by gas chromatography, and finally, the strain with the highest CLA content was obtained. The strains were optimized for cultivation by changing the amount of substrate addition, inoculum amount, and fermentation time. The results showed that the yield of the experimentally optimized strain for the conversion of conjugated linoleic acid could reach 94.68 ± 3.57 µg/mL, which was 74.4% higher than the initial yield of 54.28 ± 2.12 µg/mL of the strain. The results of this study can provide some basis for the application of conjugated linoleic acid production by Lactobacillus paracasei in the fermentation of lactic acid bacteria.

Next step in Monachacantiana (Montagu, 1803) phylogeography: northern French and Dutch populations (Eupulmonata, Stylommatophora, Hygromiidae).

Features of shell and genitalia as well as nucleotide sequences of selected mitochondrial and nuclear genes of specimens of Monachacantiana from ten northern French and two Dutch populations were compared with the same features of British and Italian populations. They were found to be very similar to populations previously identified as belonging to the CAN-1 lineage of M.cantiana. This confirms previous suggestions that M.cantiana was introduced to western Europe (England, France and the Netherlands) in historical times.

Acinetobacter in fresh faecal samples is related to sperm motility: A case report in Piétrain boars housed on different deep-litter bedding materials.

The swine gastrointestinal tract contains a great variety of microbes, forming a complex and dynamic ecosystem. Various internal and external factors (e.g. age, breed and diet) may influence its composition. This study aimed to investigate the gut microbial diversity of German Piétrain boars housed on different deep-litter bedding materials (regional wood shavings, linen, hemp, spelt husks, and wood shavings) via 16S-rDNA sequencing. Additionally, short-chain fatty acids were analysed using gas chromatography. Fresh faecal samples (n = 80) from 40 Piétrain boars were collected twice during the trial. Although it can be assumed that boars ingest bedding orally, no differences in the microbiome composition could be found. The main phyla were Firmicutes and Bacteroides. Acinetobacter was identified as a biomarker for sperm quality differences (total sperm motility) in breeding boars.

Protective effect of Tao Hong Si Wu Decoction against inflammatory injury caused by intestinal flora disorders in an ischemic stroke mouse model.

BACKGROUND AND AIMS: Recent studies have shown that intestinal flora are involved in the pathological process of ischemic stroke (IS). The potential protective effect of the traditional Chinese prescription, Tao Hong Si Wu Decoction (THSWD), against inflammatory injury after IS and its underlying mechanisms of action were investigated in the current study. METHODS: Fifty SPF(Specefic pathogen Free) male C57 mice were randomly assigned to sham operation, model, THSWD low-dose (6.5 g/kg), medium-dose (13 g/kg) and high-dose (26 g/kg) groups (10 mice per group). Mouse models of transient middle cerebral artery occlusion were prepared via thread embolism. Neurological function score, hematoxylin-eosin (HE) staining, immunohistochemistry, enzyme-linked immunosorbent assay (ELISA), 16S ribosomal DNA (rDNA) sequencing, quantitative reverse transcription PCR (qRT-PCR) and other methods were employed to elucidate the underlying molecular mechanisms. RESULTS: Notably, THSWD induced a reduction in the neurological function score (P < 0.01) and neuronal injury in brain tissue, increase in protein expression of Claudin-5 and zonula occludens-1 (ZO-1) in brain tissue(P < 0.01), and decrease in serum lipopolysaccharide (LPS)(P < 0.01), diamine oxidase (DAO)(P < 0.01) and D-lactic acid(P < 0.01, P < 0.05) levels to a significant extent. THSWD also inhibited the levels of tumor necrosis factor-α (TNF-α)(P < 0.01) and interleukin - 1ß (IL-1ß)(P < 0.01) in brain tissue, and increased alpha and beta diversity in ischemic stroke mice, along with a certain reversal effect on different microflora. Finally, THSWD inhibited the polarization of microglia cells(P < 0.01) and decreased the protein and gene expression of toll-like receptor-4 (TLR-4)(P < 0.01, P < 0.05) and nuclear factor kappa B (NF-κB)(P < 0.01) in brain tissue. CONCLUSION: Our data indicate that THSWD may interfere with inflammatory response in ischemic stroke by regulating intestinal flora and promoting intestinal barrier repair.

Sequencing and multiple antimicrobial resistance of Pseudomonas fluorescens isolated from Nile tilapia fish in Egypt / Sequenciamento e resistência antimicrobiana múltipla de Pseudomonas fluorescens isoladas de tilápia-do-nilo no Egito

Pseudomonas fluorescens is one of the main causes of septicemic diseases among freshwater fish, causing severe economic losses and decreasing farm efficiency. Thus, this research was aimed to investigate the occurrence of P. fluorescens in Nile Tilapia (O. niloticus) fish in Egypt, gene sequencing of 16SrDNA gene, and antimicrobial susceptibility. P. fluorescens strains were detected in 32% (128/400) of apparently healthy (9%; 36/400) and diseased (23%; 92/400) Nile tilapia fish. The highest prevalence was observed in gills of fish, 31.3% followed by intestine 26.9%, liver 24.2%, and kidneys 17.6%. The PCR results for the 16SrDNA gene of P. fluorescens showed 16SrDNA gene in 30% of examined isolates. Moreover, Homogeny and a strong relationship between strains of P. fluorescens was confirmed using 16SrDNA sequences. Beside the responsibility of 16SrDNA gene on the virulence of P. fluorescens. The results of antimicrobial susceptibility tests revealed that all strains were resistant to piperacillin (100%), followed by ceftazidime (29.7%), and cefepime (25.8%). The strains of P. fluorescence were highly sensitive to cefotaxime (74.2%), followed by ceftriaxone and levofloxacin (70.3% each). Interestingly, 29.7% of strains of P. fluorescens were multiple antimicrobial-resistant (MAR).

Pseudomonas fluorescens é uma das principais causas de doenças septicêmicas em peixes de água doce, causando graves perdas econômicas e diminuindo a eficiência da fazenda. Assim, esta pesquisa teve como objetivo investigar a ocorrência de P. fluorescens em peixes de tilápia-do-nilo (O. niloticus) no Egito, sequenciamento do gene 16S rDNA e suscetibilidade antimicrobiana. Cepas de P. fluorescens foram detectadas em 32% (128/400) de peixes tilápia-do-nilo aparentemente saudáveis ​​(9%; 36/400) e doentes (23%; 92/400). A maior prevalência foi observada nas brânquias dos peixes, 31,3%, seguida pelo intestino 26,9%, fígado 24,2% e rins 17,6%. Os resultados da PCR para o gene 16SrDNA de P. fluorescens mostraram o gene 16SrDNA em 30% dos isolados examinados. Além disso, a homogeneidade e uma forte relação entre cepas de P. fluorescens foi confirmada usando sequências de 16SrDNA. Além da responsabilidade do gene 16SrDNA na virulência de P. fluorescens. Os resultados dos testes de suscetibilidade antimicrobiana revelaram que todas as cepas foram resistentes à piperacilina (100%), seguida pela ceftazidima (29,7%) e cefepima (25,8%). As cepas de P. fluorescens foram altamente sensíveis à cefotaxima (74,2%), seguida pela ceftriaxona e levofloxacina (70,3% cada). Curiosamente, 29,7% das cepas de P. fluorescens eram multirresistentes a antimicrobianos (MAR).

Sequencing and multiple antimicrobial resistance of Pseudomonas fluorescens isolated from Nile tilapia fish in Egypt

Abstract Pseudomonas fluorescens is one of the main causes of septicemic diseases among freshwater fish, causing severe economic losses and decreasing farm efficiency. Thus, this research was aimed to investigate the occurrence of P. fluorescens in Nile Tilapia (O. niloticus) fish in Egypt, gene sequencing of 16SrDNA gene, and antimicrobial susceptibility. P. fluorescens strains were detected in 32% (128/400) of apparently healthy (9%; 36/400) and diseased (23%; 92/400) Nile tilapia fish. The highest prevalence was observed in gills of fish, 31.3% followed by intestine 26.9%, liver 24.2%, and kidneys 17.6%. The PCR results for the 16SrDNA gene of P. fluorescens showed 16SrDNA gene in 30% of examined isolates. Moreover, Homogeny and a strong relationship between strains of P. fluorescens was confirmed using 16SrDNA sequences. Beside the responsibility of 16SrDNA gene on the virulence of P. fluorescens. The results of antimicrobial susceptibility tests revealed that all strains were resistant to piperacillin (100%), followed by ceftazidime (29.7%), and cefepime (25.8%). The strains of P. fluorescence were highly sensitive to cefotaxime (74.2%), followed by ceftriaxone and levofloxacin (70.3% each). Interestingly, 29.7% of strains of P. fluorescens were multiple antimicrobial-resistant (MAR).

Resumo Pseudomonas fluorescens é uma das principais causas de doenças septicêmicas em peixes de água doce, causando graves perdas econômicas e diminuindo a eficiência da fazenda. Assim, esta pesquisa teve como objetivo investigar a ocorrência de P. fluorescens em peixes de tilápia-do-nilo (O. niloticus) no Egito, sequenciamento do gene 16S rDNA e suscetibilidade antimicrobiana. Cepas de P. fluorescens foram detectadas em 32% (128/400) de peixes tilápia-do-nilo aparentemente saudáveis (9%; 36/400) e doentes (23%; 92/400). A maior prevalência foi observada nas brânquias dos peixes, 31,3%, seguida pelo intestino 26,9%, fígado 24,2% e rins 17,6%. Os resultados da PCR para o gene 16SrDNA de P. fluorescens mostraram o gene 16SrDNA em 30% dos isolados examinados. Além disso, a homogeneidade e uma forte relação entre cepas de P. fluorescens foi confirmada usando sequências de 16SrDNA. Além da responsabilidade do gene 16SrDNA na virulência de P. fluorescens. Os resultados dos testes de suscetibilidade antimicrobiana revelaram que todas as cepas foram resistentes à piperacilina (100%), seguida pela ceftazidima (29,7%) e cefepima (25,8%). As cepas de P. fluorescens foram altamente sensíveis à cefotaxima (74,2%), seguida pela ceftriaxona e levofloxacina (70,3% cada). Curiosamente, 29,7% das cepas de P. fluorescens eram multirresistentes a antimicrobianos (MAR).

Population genetics of the Ixodes affinis (Ixodida: Ixodidae) complex in America: new findings and a host-parasite review.

Ticks are hematophagous ectoparasites associated with a wide range of vertebrate hosts. Within this group, the Ixodidae family stands out, in which the Ixodes genus contains at least 245 species worldwide, from which 55 species are present in the Neotropical region. Ixodes affinis, a tick described in 1899, has a wide distribution from the Southern Cone of America to the United States. However, since its description, morphological variability has been reported among its populations. Furthermore, attempts have been made to clarify its status as a species complex using mitochondrial markers, but mainly in restricted populations of South and Central America. Thus, information related to populations of the transition region between the Neotropical and Nearctic zones is lacking. For these reasons, the objectives of the study were to evaluate the genetic diversity and structure of I. affinis across the Americas and to compile all the published records of I. affinis in America, to elucidate the host-parasite relationships and to identify their geographical distribution. For this, a phylogeny, and AMOVA analyses were performed to assess the genetic structure of samples obtained by field work in South Carolina, USA and Yucatán, Mexico. A total of 86 sequences were retrieved from a fragment of the 16S region. Phylogeny and genetic structure analysis showed four groups that were geographically and genetically related with high branch support and Fst values, all of them statistically significant. The results obtained support the hypothesis that I. affinis it corresponds to a complex of four species, which must be validated through future morphological comparisons.

Comparing sputum microbiota characteristics between severe and critically ill influenza patients.

Background: Currently, limited attention has been directed toward utilizing clinical cohorts as a starting point to elucidate alterations in the lower respiratory tract (LRT) microbiota following influenza A virus (IAV) infection. Objectives: Our objective was to undertake a comparative analysis of the diversity and composition of sputum microbiota in individuals afflicted by severe and critically ill influenza patients. Methods: Sputum specimens were procured from patients diagnosed with IAV infection for the purpose of profiling the microbiota using 16S-rDNA sequencing. To ascertain taxonomic differences between the severe and critically ill influenza cohorts, we leveraged Linear Discriminant Analysis Effect Size (LEfSe). Additionally, Spearman correlation analysis was employed to illuminate associations between sputum microbiota and influenza Ct values alongside laboratory indicators. Results: Our study encompassed a total cohort of 64 patients, comprising 48 within the severe group and 16 within the critically ill group. Intriguingly, Bacteroidetes exhibited significant depletion in the critically ill cohort (p=0.031). The sputum microbiomes of the severe influenza group were hallmarked by an overrepresentation of Neisseria, Porphyromonas, Actinobacillus, Alloprevotella, TM7x, and Clostridia_UCG-014, yielding ROC-plot AUC values of 0.71, 0.68, 0.60, 0.70, 0.70, and 0.68, respectively. Notably, Alloprevotella exhibited an inverse correlation with influenza Ct values. Moreover, C-reactive protein (CRP) manifested a positive correlation with Haemophilus and Porphyromonas. Conclusion: The outcomes of this investigation lay the groundwork for future studies delving into the connection between the LRT microbiome and respiratory disorders. Further exploration is warranted to elucidate the intricate mechanisms underlying the interaction between IAV and Alloprevotella, particularly in disease progression.

Characteristics of faecal bacterial flora and volatile fatty acids in Min pig, Landrace pig, and Yorkshire pig

BACKGROUND In order to study the influence of long-term growth process and evolution environment on intestinal bacteria of different breeds, the intestinal bacteria and volatile fatty acids among the faeces of Min, Landrace and Yorkshire pigs were analysed by Illumina high-throughput sequencing of the 16S-rDNA and gas chromatography. RESULTS The shared core microbiota of Landrace, Yorkshire and Min pig were 1273, accounting for 69.56% of total abundance of organisms. The proportion of Firmicutes in Min pig faeces (57.89%) was significantly higher than that in Landrace and Yorkshire pig faeces (47.01% and 46.40%, respectively) (P < 0.05), but that of Bacteroidetes was exactly opposite. Moreover, Min pig presented more highly efficient membrane transport, environmental adaptation, carbohydrate transport, and metabolism than Yorkshire pig (P < 0.05). The acetic acid/total volatile fatty acid ratio in Min pig was significantly higher than that in Landrace pig (P < 0.05), and the isobutyric acid/ total volatile fatty acid ratio in Min pig was significantly larger than that in Yorkshire pig (P < 0.05). Furthermore, the content of branched chain volatile fatty acids in Min pig was significantly higher than that in Yorkshire pig (P < 0.05). CONCLUSION This study demonstrated that Min pig, as an excellent breed in the cold area of China, possessed special intestinal floral structure compared to the imported pigs in order to satisfy their phys iological and metabolic demands, which may influence their characteristics such as resistance to cold, diseases, and crude feeding, and the ability to deposit intramuscular fat.