Bioprocessing in the Digital Age: The Role of Process Models

In this age of technology, the vision of manufacturing industries built of smart factories is not a farfetched future. As a prerequisite for Industry 4.0, industrial sectors are moving towards digitalization and automation. Despite its tremendous growth reaching a sales value of worth $188 billion in 2017, the biopharmaceutical sector distinctly lags in this transition. Currently, the challenges are innovative market disruptions such as personalized medicine as well as increasing commercial pressure for faster and cheaper product manufacturing. Improvements in digitalization and data analytics have been identified as key strategic activities for the next years to face these challenges. Alongside, there is an emphasis by the regulatory authorities on the use of advanced technologies, proclaimed through initiatives such as Quality by Design (QbD) and Process Analytical Technology (PAT). In the manufacturing sector, the biopharmaceutical domain features some of the most complex and least understood processes. Thereby, process models that can transform process data into more valuable information, guide decision‐making, and support the creation of digital and automated technologies are key enablers. This review summarizes the current state of model‐based methods in different bioprocess related applications and presents the corresponding future vision for the biopharmaceutical industry to achieve the goals of Industry 4.0 while meeting the regulatory requirements.     

Synopsis of the species of Myxidium Bütschli, 1882 (Myxozoa: Myxosporea: Bivalvulida)

A synopsis of the species of Myxidium Bütschli, 1882 (Myxozoa: Myxosporea: Bivalvulida) is presented. It includes a total of 232 nominal species, whose principal morphological and morphometric characteristics, site of infection within the host, and original hosts and locality are indicated in a tabulated format. A diagrammatic illustration of a spore of most of the species is also provided.     

First evidence of Panulirus argus Virus 1 (PaV1) in spiny lobster from Cuba and clinical estimation of its prevalence

The present study documents the first finding of Panulirus argus Virus 1 (PaV1) in spiny lobster Panulirus argus from Cuba. Samples originated from 2 nursery sites, Matias Keys and Bocas de Alonso Keys, and 2 fishing sites, La Grifa and El Ramajo. Lobsters from the nursery sites (artificial reefs) were collected by SCUBA diving, while those from the fishing sites were collected from artificial shelters known as 'casitas cubanas'. In these shelters it was observed that healthy lobsters tended to avoid infected lobsters. Prevalence of PaV1 in the sampling sites was assessed by using clinical signs such as lethargy, an opaque reddish shell coloration, and milky white hemolymph with loss of clotting activity. The presence of PaV1 was subsequently confirmed by histology and PCR of tissues and hemolymph samples from suspected individuals. Histological sections of the hepatopancreas, gills, gonads, and gut showed infected hemocytes with hypertrophied nuclei and eosinophilic intranuclear Cowdry type A inclusions. A 499 bp band was observed by PCR. The sequence of the amplified fragments was 96% similar to the PaV1 sequence in GenBank. The overall mean prevalence of PaV1 was 4.48% (range: 0 to 9.3%) after pooling the results of the 4 sampling sites.     

Pretreatment with the probiotic VSL#3 delays transition from inflammation to dysplasia in a rat model of colitis-associated cancer

Evidence supports involvement of microflora in the transition of chronic inflammation to neoplasia. We investigated the protective efficacy of the probiotic VSL#3 in a model of colitis-associated colorectal cancer. Chronic colitis was induced in Sprague-Dawley rats by administration of trinitrobenzene sulfonic acid (TNBS), followed 6 wk later by systemic reactivation. To induce colitis-associated dysplasia and cancer, the animals received TNBS (intravenously) twice a week for 10 wk. One group received VSL#3 in drinking water from 1 wk before colitis induction until death. The colons were examined for damage and presence of dysplasia or cancer. Samples were analyzed for cell proliferation and apoptosis, vitamin D receptor (VDR) expression, angiogenic factors, and presence of alkaline sphingomyelinase or phosphatase. Microbial community composition was evaluated by terminal restriction fragment-length polymorphism analysis of the bacterial 16S rRNA gene. None of the probiotic-treated animals developed carcinoma, and no high-grade dysplasia was found in either the proximal or mid colon. In contrast, 29% of the animals in the control group developed carcinoma in one or more regions of the colon. VSL#3-treated animals had significantly less damage than the vehicle treated-controls in all areas of the colon, and this correlated with decreased richness and diversity of the mucosally adherent microbiota. Treatment with the probiotic increased the antiangiogenic factor angiostatin, VDR expression, and alkaline sphingomyelinase. We concluded that pretreatment with the probiotic VSL#3 can attenuate various inflammatory-associated parameters, delaying transition to dysplasia and cancer, thus offering its potential therapeutic use in patients with long-standing colitis.     

Long-term preservation of anammox bacteria

Deposit of useful microorganisms in culture collections requires long-term preservation and successful reactivation techniques. The goal of this study was to develop a simple preservation protocol for the long-term storage and reactivation of the anammox biomass. To achieve this, anammox biomass was frozen or lyophilized at two different freezing temperatures (−60°C and in liquid nitrogen (−200°C)) in skim milk media (with and without glycerol), and the reactivation of anammox activity was monitored after a 4-month storage period. Of the different preservation treatments tested, only anammox biomass preserved via freezing in liquid nitrogen followed by lyophilization in skim milk media without glycerol achieved stoichiometric ratios for the anammox reaction similar to the biomass in both the parent bioreactor and in the freshly harvested control treatment. A freezing temperature of −60°C alone, or in conjunction with lyophilization, resulted in the partial recovery of the anammox bacteria, with an equal mixture of anammox and nitrifying bacteria in the reactivated biomass. To our knowledge, this is the first report of the successful reactivation of anammox biomass preserved via sub-zero freezing and/or lyophilization. The simple preservation protocol developed from this study could be beneficial to accelerate the integration of anammox-based processes into current treatment systems through a highly efficient starting anammox biomass.     

Effervescent tablets and ultrasonic devices against Candida and mutans streptococci in denture biofilm

doi: 10.1111/j.1741‐2358.2010.00378.x Effervescent tablets and ultrasonic devices against Candida and mutans streptococci in denture biofilm Objective: To evaluate the antimicrobial action of effervescent tablets and ultrasound on Candida spp. and mutans streptococci from denture biofilm. Background: It is not uncommon for edentulous patients to be elderly and find it difficult to brush their dentures. Hence, auxiliary methods are required for cleansing dentures as well as treating oral infections. Materials and methods: Seventy‐seven complete denture wearers were randomly assigned into four groups: (A) Brushing with water (control); (B) Effervescent tablets; (C) Ultrasonic device (Ultrasonic Cleaner, model 2840 D); (D) Effervescent tablets and ultrasonic device. All groups brushed their dentures with a specific brush and water, three times a day, before applying their treatments. Denture biofilm was collected at baseline and after 21 days. The samples were collected by brushing the dentures with saline and the detached microbial cells were quantified by plating. Counts [log (CFU+1) ml^?1] of total aerobes, Candida spp. and mutans streptococci were compared by one‐way anova or Kruskal–Wallis test (α = 0.05). Results: No significant difference was found among the methods from C. albicans (p = 0.76), C. tropicalis (p = 0.94) and C. glabrata (p = 0.80). Lower counts were found for methods B and D when compared with the other methods against mutans streptococci (p < 0.001). Method B showed lower total aerobic counts than A, whereas C and D showed intermediate results (p = 0.011). Conclusion: The effervescent tablets significantly reduced mutans streptococci and total aerobes from denture biofilm. However, they was not as effective against C. albicans. Ultrasonic cleansing presented a discrete antimicrobial effect and was less effective than the tablets for complete denture disinfection.     

Phylogeny and reclassification of the species of two neotropical grunt genera,Anisotremus and Genyatremus (Perciformes: Haemulidae), based on morphological evidence

The haemulid genera Anisotremus and Genyatremus contain eleven valid New World species. We examined 52 morphological characters of all Anisotremus and Genyatremus species and five relatives. Based on phylogenetic analysis, we propose that two Anisotremus species (Pristipoma dovii and Conodon pacifici) are members of the genus Genyatremus. The primary synapomorphies leading to the definition of these clades come from the oral and pharyngeal jaws. Reclassification of both genera, as well as an identification key, is included. A review of early literature indicated that the type species of Genyatremus is Diagramma cavifrons, as stated by the original describer, and is not Lutjanus luteus, which is not a haemulid.     

Optimisation of entrapped activated carbon conditions to remove coloured compounds from winery wastewaters

The objective of this work was to study the entrapped conditions of activated carbon in calcium-alginate beads for the clarification of winery wastewaters. An incomplete 3³ factorial design was carried out to study the efficiency of activated carbon (0.5-2%); sodium alginate (1 - 5%); and calcium chloride (0.050 - 0.900 M), on the following dependent variables: colour reduction at 280, 465, 530 and 665 nm. The activated carbon and calcium chloride were the most influential variables in the colour reduction. Nearly 100% colour reductions were found for the wavelengths assayed when employing 2% of activated carbon, 5% of sodium alginate and intermediate concentrations of calcium chloride (0.475 M). Instead, other conditions like, 2% of activated carbon, 4% of sodium alginate and 0.580 M of calcium chloride can also give absorbance reductions close to 100%.