The clear relationship between the microbiome and the general well-being of metazoans has motivated the characterization of bacterial communities in cultured species for future microbiome monitoring ...and modulation to improve productivity. To study the bacterial diversity associated with farmed adult P. monodon at harvest, guts, gastric mills (GM), pond water and sediments were investigated using next-generation sequencing technology in two control ponds (CP1 and CP2) where animals showed a healthy appearance, and two low productive ponds (LP1 and LP2) where animals showed signs of disease. Animals from CP2 were on average at least 4 g heavier than animals from other ponds, and the final yield of both CP was more than double that of LP. LP2 showed the lowest diversity indices, which can potentially be attributed to high abundance of Vibrio spp. and warm water conditions (31.6 ± 0.3 °C on average). The structure of the bacterial community in the gut, GM and water samples was different between CP and LP. Vibrio spp. and anaerobic bacteria (e.g. Fusibacter) were enriched in the gastrointestinal tract of animals from LP (LDA ≥4). While Vibrio can potentially act against the host and other members of the bacterial community, Fusibacter might be an indicator of sulfur compounds accumulation in the animal as the hepatopancreas becomes affected by a PirA toxin. Enrichment of Cyanobacteria in the water (LDA = 4.38) and members of Desulfobacteraceae in sediments of LP (5% on average) could be environmental stress indicators in summer conditions. Characterizing bacterial microbiomes in aquaculture is important to detect indicators of health and productivity that can be used as monitoring tools in the animal's gastrointestinal tract and the environment, ultimately enabling a targeted modulation of the bacterial community for improved production.
•Control and low productive P. monodon ponds at harvest exhibit different bacterial communities.•Bacterial assemblages in P. monodon gut from control ponds differ from those in low productivity ponds.•Bacterial communities in guts and gastric mills show indicators of decay in productivity.•Bacterial community in the environment is more diverse than in Penaeus monodon's gut.•Indicators of low productivity include enrichment of Vibrio, Propionigenium and Fusibacter in the gut.
Under what circumstances do paramilitary groups place limits on their use of political violence? This article considers why one of the most prominent paramilitary groups active during the "Troubles" ...in Northern Ireland, the Provisional IRA (PIRA), exercised restraint and limited their use of political violence, both in the lead up to, and in the wake of the Good Friday Agreement. It applies Busher, Holbrook, and Macklin's "internal brakes" typology to understand why despite the PIRA's proven capacity to sustain a campaign of armed violence for more than thirty years, they reined in on paramilitarism during the 1990s peace process. Through the lens of various brakes, the analysis identifies seven developments that were particularly salient in limiting the PIRA's use of armed violence. These developments included: a weakened capacity to continue their armed campaign, waning popular support, an opening of political opportunities, the need to maintain ties with strategically important non-violent allies, the gradual softening of out-group boundaries, the decommissioning process, as well as the creation of spaces for non-violent activism within the wider movement. This study's findings contribute to the scholarly debate on understanding how militant groups manage the parameters of their violence, highlighting several factors that may mitigate the risk of extremist views escalating to political violence.
A 4-bp deletion in the ATP-binding cassette subfamily B member 1 (ABCB1) gene, also referred to as the multidrug resistance gene (MDR1), produces stop codons that cause premature termination of ...P-glycoprotein 1 (P-gp) synthesis. Dogs with the homozygous mutation do not express functional P-gp, which increases their sensitivity markedly to many common veterinary drugs. We detected the nt230 (del4) ABCB1 mutation in Border Collie dogs in western Mexico with a simple and affordable primer-introduced restriction analysis PCR (PIRA-PCR). PIRA-PCR clearly identified all genotypes in our sample of 104 dogs. Genotype frequencies were 0.952 (wild/wild), 0.029 (wild/mut) and 0.019 (mut/mut). Allele frequencies were 0.033 (mutant alleles) and 0.966 (wild-type alleles). In this small subset of the Mexican dog population, we found a higher prevalence of the nt230 (del4) MDR1/ABCB1 gene mutation than reported in other countries.
Acute hepatopancreatic necrosis disease (AHPND), caused by emerging strains of Vibrio Parahaemolyticus, is of concern in shrimp aquaculture. Secreted proteins PirA and PirB, encoded by a plasmid ...harbored in V. parahaemolyticus, were determined to be the major virulence factors that induce AHPND. To better understand pathogenesis associated with PirA and PirB, recombinant proteins rPirA and rPirB were produced to evaluate their relative toxicities in shrimp. By challenging shrimp at concentration of 3 μM with reverse gavage method, rPirA and rPirB (approximately 0.4 and 1.5 μg per g of body weight, respectively) caused 27.8 ± 7.8% and 33.3 ± 13.6% mortality, respectively; combination of 3 μM rPirA and rPirB resulted in 88.9 ± 7.9% mortality. Analysis of protein mobility in native gel revealed that rPirB was apparently in the form of monomer while rPirA was oligomerized as an octamer-like macromolecule, suggesting that inter- and intra-molecular interactions between rPirA and rPirB enhanced the toxic effect. An attempt to block or reduce rPirA activity with a putative receptor, N-acetyl-galactosamine, was unsuccessful, implying that remodeling analysis of PirA molecule, such as the octamer observed in this study, is necessary. Results of this study provided new insight into toxic mechanism of PirA and PirB and shall help design strategic antitoxin methods against AHPND in shrimp.
•Recombinant proteins rPirA and rPirB were produced to evaluate their relative toxicities in shrimp.•Both rPirA and rPirB were potent in causing acute death but combination of the two was more deadly.•Formation of an octamer rPirA was observed, suggesting intra- and inter-molecule interaction.
Toxins PirvpA and PirvpB were first described in 2014 from Vibrio parahaemolyticus (VP) isolates discovered in 2013 as the cause of acute hepatopancreatic necrosis disease (AHPND) that emerged as a ...new disease of Chinese cultivated shrimp in 2009. It was subsequently reported from Thailand in 2012. By using PirvpA and PirvpB specific monoclonal antibodies (MAbs) together with reference VPAHPND isolates to screen archived bacterial cultures, we discovered 3 isolates of Vibrio (previously identified as V. harveyi) including VH 639, VH 1526 and VH Surat that were positive for both PirA and PirB toxins, even though the isolates were collected in Thailand over 10 years prior to reported Thai AHPND outbreaks. Western blot analysis revealed that the PirA/B toxins from these Vibrio isolates matched the molecular masses of the PirvpA/B toxins of previously described VPAHPND isolates. PCR analysis using VPAHPND detection methods that target the PirvpA and PirvpB toxin genes (AP4 and TUMSAT-Vp3) gave amplicons of the expected size from the 3 Vibrio isolates, and sequence analysis of the amplicons revealed 99% identity with the sequences of the PirvpA and PirvpB genes of previously described VPAHPND isolates. Pathogenicity tests performed by reverse gavage and immersion challenge tests demonstrated the pattern of AHPND pathology. Species confirmation of these 3 Vibrio isolates by multilocus sequence analysis (MLSA) of 16S rRNA, rpoD, rctB, and toxR genes revealed that they belong to V. campbellii clade. The results revealed that V. campbellii isolates capable of producing PirA and PirB toxins were present in Thailand up to 7 years before AHPND was reported from China and up to a decade before it was reported from Thailand. The plasmid or chromosomal location of the PirA and PirB toxin genes in these archived V. campbellii isolates and their phylogenetic relationship to current VPAHPND isolates are being determined.
•Three archived isolates of V. campbellii were positive for PirvpA and PirvpB toxins by using specific monoclonal antibodies.•Those three virulent isolates were collected in Thailand 5–10 years prior to reported Thai AHPND outbreaks.•The PirvpA and PirvpB toxin genes from V. campbellii shared 99% identity with that of previously described VPAHPND isolates.
A quantitative polymerase chain reaction (qPCR) assay was developed, based on a TaqMan probe, to detect and quantify a virulence plasmid harbored by the bacterium Vibrio parahaemolyticus which can ...cause acute hepatopancreatic necrosis disease (AHPND). The assay uses a pair of PCR primers, which amplify a 135-bp DNA fragment, and a TaqMan probe selected from the plasmid pirA-like gene. This qPCR assay reacted with AHPND-pathogenic isolates of V. parahaemolyticus collected from Vietnam and Mexico, but not with non-pathogenic strains of Vibrio spp. For quantification, a plasmid (pVpPirA-1) containing the target pirA-like gene was constructed, purified and serially diluted to be used as a standard. With this standard, the qPCR assay was then used to quantify the virulence plasmid in shrimp samples collected from different farms. Up to 5.8×105 copy per mg tissue were detected in AHPND-affected shrimp collected from Vietnam. Lower quantities, up to 1.5×104 copies per mg of tissues were detected in affected shrimp collected from a Chinese farm. In the laboratory bioassays, similar plasmid quantities, 1.8×103 to 4.7×106 copies of plasmid per mg of tissues were found in the moribund/dead shrimp, 3.5×102 to 2.2×106 copies of plasmid per mL were detected in the water samples. This assay is specific with high sensitivity (10 copies of virulence plasmid) and can be used to detect AHPND-pathogenic V. parahaemolyticus in shrimp and water samples.
•We developed a qPCR method to detect and quantify AHPND (EMS).•This assay is specific with high sensitivity (10 copies of virulence plasmid).•This method can be used for quantification in water and shrimp samples from fields.
Background:
Odour discrimination and identification (DI) are markers associated with disability worsening and neuroaxonal damage in multiple sclerosis (MS).
Objective:
The main objective of this ...research is to investigate whether longitudinal change of DI predicts long-term MS disease course.
Methods:
This is a 6-year prospective longitudinal study on MS patients at the MS Clinic Innsbruck. Clinical, bi-annual visits assessed patients’ history and Expanded Disability Status Scale (EDSS) score. DI and cognitive function were assessed at baseline (BL), Year 1 (Y1), Year 2 (Y2) and Year 6 (Y6) by the ‘Sniffin’ Sticks’/Symbol Digit Modalities Test.
Results:
Around 92 of 139 patients were available for Y6 follow-up. Mean DI scores significantly decreased over time (BL = 27.8, Y1 = 27.5, Y2 = 26.3 and Y6 = 26.3; p < 0.001) and negatively correlated with patients’ age (rs = −0.120, p = 0.032) and disease duration (rs = −0.103, p = 0.041). Multivariable regression analyses revealed that lower absolute DI scores and larger DI score loss over time were associated with higher probability of EDSS worsening (per −1 point: hazard ratio (HR) = 1.40 (1.16–1.68) and 2.34 (1.27–4.21)), progression independent of relapse activity (PIRA) (HR = 1.49 (1.20–1.85) and 2.22 (1.33–3.31)) and cognitive deterioration (HR = 1.75 (1.35–2.27) and 4.29 (1.26–2.84)) at Y6, but not with time to first relapse.
Conclusion:
Odour DI is an irreversible marker of neuroaxonal damage, associated with PIRA, cognitive deterioration and EDSS worsening.
This article examines the murder of two British Army corporals by the Provisional Irish Republican Army (PIRA) in March 1988. In doing so it reveals much about the micro dynamics of the political ...violence which occurred during the Northern Ireland conflict. The central theoretical contention of the article is that most human beings are not particularly good at violence, and for the most part, irrespective of motive, can only participate in such actions when they have what is defined as attacker advantage. The attacker advantage principle stipulates that on most occasions when most humans feel it necessary or choose to resort to offensive violence, they do so on the premise of having the advantage, or a perceived advantage, over their intended target.
While magnetic resonance imaging (MRI) is established in diagnosing and monitoring disease activity in multiple sclerosis (MS), its utility in predicting and monitoring disease progression is less ...clear.
The authors consider changing concepts in the phenotypic classification of MS, including progression independent of relapses; pathological processes underpinning progression; advances in MRI measures to assess them; how well MRI features explain and predict clinical outcomes, including models that assess disease effects on neural networks, and the potential role for machine learning.
Relapsing-remitting and progressive MS have evolved from being viewed as mutually exclusive to having considerable overlap. Progression is likely the consequence of several pathological elements, each important in building more holistic prognostic models beyond conventional phenotypes. MRI is well placed to assess pathogenic processes underpinning progression, but we need to bridge the gap between MRI measures and clinical outcomes. Mapping pathological effects on specific neural networks may help and machine learning methods may be able to optimize predictive markers while identifying new, or previously overlooked, clinically relevant features. The ever-increasing ability to measure features on MRI raises the dilemma of what to measure and when, and the challenge of translating research methods into clinically useable tools.
