One year of surgical mask testing at the University of Bologna labs:Lessons learned from data analysis

The outbreak of SARS-CoV-2 pandemic highlighted the worldwide lack of surgical masks and personal protective equipment, which represent the main defense available against respiratory diseases as COVID-19. At the time, masks shortage was dramatic in Italy, the first European country seriously hit by the pandemic: aiming to address the emergency and to support the Italian industrial reconversion to the production of surgical masks, a multidisciplinary team of the University of Bologna organized a laboratory to test surgical masks according to European regulations. The group, driven by the expertise of chemical engineers, microbiologists, and occupational physicians, set-up the test lines to perform all the functional tests required. The laboratory started its activity on late March 2020, and as of the end of December of the same year 435 surgical mask prototypes were tested, with only 42 masks compliant to the European standard. From the analysis of the materials used, as well as of the production methods, it was found that a compliant surgical mask is most likely composed of three layers, a central meltblown filtration layer and two external spunbond comfort layers. An increase in the material thickness (grammage), or in the number of layers, does not improve the filtration efficiency, but leads to poor breathability, indicating that filtration depends not only on pure size exclusion, but other mechanisms are taking place (driven by electrostatic charge). The study critically reviewed the European standard procedures, identifying the weak aspects; among the others, the control of aerosol droplet size during the bacterial filtration test results to be crucial, since it can change the classification of a mask when its performance lies near to the limiting values of 95 or 98%

Fungal contamination assessment in healthcare environments: a bibliographic review

FCT_UIDB/05608/2020. FCT_UIDP/05608/2020.In Healthcare environments fungal presence depends on the medical activities performed, number and types of patients, cleaning frequency and procedures, and the type of ventilation systems. The aim of this review article was to identify different methodologies applied to assess fungal contamination in Healthcare environments, as well as to describe the most reported fungi in these environments. This study was based on a systematic search for information and data that have been published in free access sources during the period of 1st January 2000 to 31st December 2020. PRISMA methodology was applied to identify and select studies referring to Healthcare environments where the fungal assessment was performed. The most common Healthcare environments assessed were hospitals (26 out of 56) and the most used sampling methods were active (27 articles). Passive methods were exclusively used in 8 papers, and the combined use of both methods was verified in 21 papers. Concerning analytical procedures, the exclusive use of morphological identification was the most frequent approach (40 out of 56). Aspergillus sp., Cladosporium sp., and Penicillium sp. were the predominant genera found indoors (24 out of 56). There is scientific evidence of fungal contamination present in Healthcare environments. Thus, in order to have an accurate and reliable risk characterization, the combined use of active and passive sampling methods and the use of culture based-methods and molecular tools are of upmost importance.info:eu-repo/semantics/publishedVersio

An investigation and examination of the levels and types of bacterial contamination on the surface of clean room operators' garments.

The contamination of sterile pharmaceutical products is a serious event which has in the worst case scenario led to patient death. Operators are the primary source of clean room contamination, with the majority of their detritus being identified as skin squames and their related microorganisms. The ability of operator associated bacterial contamination to disseminate through specialist garments worn in the clean room environment is apparent in the literature. However, despite the fibres of such garments being identified as a suitable substrate for bacteria to adhere to and grow upon, the bacterial bioburden of the surface of clean room operators garments is an area which severely lacks in published research. Reported here is the recovery, enumeration and comparison of the levels of bacteria on the surface of reusable antistatic carbon filament polyester clean room garments, using the direct agar contact method, following their laundering with and without terminal gamma sterilisation, immediately following their donning with operators dressing wearing either no gloves, non “ sterile gloves or sterile clean room gloves, and following their wear within the clean room environment, with respect to gender. The aforementioned method, with its recovery efficiency shown to be unaffected by agar composition (NA or TSA), recovered bacteria from the surface of garments laundered with and without gamma sterilisation. Such terminal decontamination was shown to reduce the surface bacterial bioburden of the garments, especially at the chest and umbilicus regions, which were shown to harbour higher levels of bacteria than the other sites tested. The direct agar contact method, showing an increase in recovery efficiency following a 48 hour agar incubation period as opposed to a 24 hour period, also recovered bacteria from the surface of clean room garments donned by operators dressing wearing either no gloves, non “ sterile gloves and sterile clean room gloves. Bacteria were transferred onto the surface of these garments via the hand borne route, with the chest and oral cavity regions being found to harbour more bacteria than the other sites tested. Overall, glove type was shown to have no effect upon the resultant bacterial bioburden of the surface of the garments, suggesting expensive clean room gloves could be substituted for their cheaper non “ sterile equivalents or no gloves during the donning process without subsequently increasing the surface bacterial bioburden of the garment. The direct agar contact method also recovered bacteria from the surface of clean room garments worn by male and female operators, following their working period within a clean room environment. Gender was found to significantly affect the surface bacterial bioburden of the garments, with the surface of those garments worn by male operators being more contaminated than the surface of those worn by their female counterparts. In addition, the donning of a clean room hood was shown to reduce the levels of bacteria at the chest and posterior cervicis regions of suits worn by both genders. Overall, the direct agar contact method was identified as a successful tool to recover, enumerate and estimate the surface bacterial bioburden of reusable antistatic polyester carbon filament clean room garments. Finally, using 16S rRNA gene sequencing, found to be more reliable and accurate at identifying unknown isolates than traditional phenotypic first - stage tests, which were subsequently found to misidentify > 85 % of the isolates tested, a self - selected representative number of isolates recovered from the surface of garments during the laundering and gender comparison studies were predominantly identified as skin commensal species of Staphylococcus and Micrococcus, as well as environmental species of Bacillus. The knowledge contained within this thesis, with respect to clean room operators and their specialist garments, contributes towards improving contamination control standards within clean room facilities

Proceedings of the NASA Microbiology Workshop

Long-term spaceflight is characterized by extraordinary challenges to maintain the life-supporting instrumentation free from microbial contamination and the crew healthy. The methodology currently employed for microbial monitoring in space stations or short spaceflights within the orbit of Earth have been instrumental in safeguarding the success of the missions, but suffers certain shortcomings that are critical for long spaceflights. This workshop addressed current practices and methodologies for microbial monitoring in space systems, and identified and discussed promising alternative methodologies and cutting-edge technologies for pursuit in the microbial monitoring that hold promise for supporting future NASA long-duration space missions

Critical parameters in manufacturing process validation of different forms of pharmaceutical injectable products to assess products´ risk framework

Tese de mestrado, Engenharia Farmacêutica, Universidade de Lisboa, Faculdade de Farmácia, 2014The main goal of this work was to create a valuable Risk Management Approach that enables a Process Validation over products lifecycle. Quality risk management (QRM) has been described in regulatory guidance for several aspects of process validation, such as product lifecycle, extent of validation, determination of critical quality attributes (CQAs) and critical process parameters (CPPs), process design space (DS), and sampling plans and statistical confidence levels. Verification of the process in every single produced batch, over the product life time is now an expectation from regulatory authorities. Based on this Hikma is required to implement a Process Validation as collection and evaluation of data, from the process design phase and continuing through commercial production phase, establishing scientific evidence that a process is in state of control and therefore capable of consistently and effectively assure product quality. Since pharmaceutical products and processes are complex and multivariate by nature, a scientific understanding of relevant multi-factorial relationships requires risk-based approach. In this context, a risk management approach to assess risk of injectable products manufacturing was created. The aim is to reduce or even eliminate potential failures and make more resourceful and efficient, qualitatively and quantitatively, processes over lifecycle. The goal was successfully achieved, and a systematic process for the assessment, control, communication and review of risks, targeting the highest quality of an injectable product is now available to be applied - Hikma Process Validation Program of Injectable Products Lifecycle.O principal objetivo deste trabalho foi criar uma ferramenta valiosa para uma abordagem de Gestão de Risco que permita Validação de Processos em todo o ciclo de vida do produto. A Gestão da qualidade e do Risco tem sido descrita nos guias regulatórios por diversos aspetos da validação de processo, como o ciclo de vida do produto, a extensão da validação, a determinação dos atributos críticos de qualidade (CQAs) e dos parâmetros críticos do processo (CPPs), espaço de desenho do processo (DS), planos de amostragem e intervalos estatíscos de confiança. A verificação do processo em cada lote produzido ao longo do tempo de vida do produto é agora uma expectativa das autoridades reguladoras. Com base nisto é necessário que a Hikma implemente a Validação de Processo como uma coleção e avaliação de dados, desde a fase de desenho do processo e continuamente durante a fase de produção comercial, estabelecendo evidências científicas que o processo está em estado controlado e que por isso é capaz de consistentemente e eficientemente assegurar um produto de qualidade. Sendo que os produtos e processos farmacêuticos são complexos e multivariados por natureza, um entendimento científico das relações multi-factoriais relevantes pede uma abordagem baseada no risco. Neste contexto, foi criada uma abordagem de gestão de risco para avaliar o risco da produção de produtos injetáveis. A finalidade é reduzir ou até mesmo eliminar potenciais falhas e facultar ao processo mais recursos e tona-lo mais eficiente, qualitativamente e quantitativamente, durante o seu ciclo de vida. O objetivo foi alcançado com sucesso, e um processo sistemático de avaliação, controlo, comunicação e revisão dos riscos com o alvo da máxima qualidade do produto injetável está agora disponível para ser aplicado – Programa de Validação de Processo Hikma do Ciclo de Vida dos Produtos Injetáveis

Hypromelloosin aiheuttamat ongelmat silmätippavalmisteiden steriilisuodatuksessa lääketeollisuudessa

Tutkimusprojektissa tutkittiin hypromelloosin aiheuttamia ongelmia silmävalmisteiden steriilisuodatuksessa lääketeollisuudessa. Tutkimusprojekti suoritettiin NextPharma Oy:n silmälääketehtaalla Tampereella syksyllä 2020. Hypromelloosi on silmävalmisteissa yleisesti käytetty apuaine, jonka tarkoitus on nostaa liuoksen viskositeettia ja täten pidentää valmisteen kontaktiaikaa silmän pinnalla. Valmistusprosessissa hypromelloosi ensin dispergoidaan ripottelemalla hitaasti kuumaan liuokseen, jonka jälkeen liuos sekoitetaan huolellisesti ja lopuksi jäähdytetään huoneenlämpöön. Jäähdytyksen aikana hypromelloosi liukenee ja geeliytyy, jolloin liuoksen viskositeetti nousee. Hypromelloosin epätäydellinen dispersio tai liuotus valmistusprosessin aikana voi aiheuttaa suodatusnopeuden hidastumista tai jopa tukkia suodattimen kokonaan. Hypromelloosi on teollisesti valmistettu selluloosajohdannainen, joka usein sisältää epäpuhtautena reagoimatonta selluloosaa ja muita niukkaliukoisia polymeeripartikkeleita, mitkä voivat myös aiheuttaa ongelmia suodatusprosesseissa. Silmävalmisteissa yleisesti käytetty sterilointikeino on steriilisuodatus, jossa valmistettu liuos suodatetaan 0,1 – 0,2 µm huokoskoon suodatinmembraanin läpi steriiliin vastaanottoastiaan. Johtuen erittäin pienestä huokoskoosta, steriilisuodattimet tukkeutuvat herkästi, mikäli liuos sisältää liukenematonta ainesta. Työn tarkoituksena oli tutkia hypromelloosin aiheuttaman suodattimien tukkeutumisen mahdollisia syitä ja selvittää, voidaanko hypromelloosia sisältävän valmisteen suodattuvuutta parantaa optimoimalla valmistuksen prosessiparametreja. Koesuunnittelu luotiin neljälle eri prosessiparametrille (sekoitusaika, sekoitusnopeus, dispersiolämpötila ja jäähdytyslämpötila) kaksitasoinena full factorial -koematriisina ilman toistoja, kolmella keskipisteellä. Prosessiparametreille haettiin alkutestauksissa sopivat minimi- ja maksimiarvot, jonka jälkeen koeliuokset valmistettiin ja suodatettiin matriisin mukaisessa satunnaistetussa järjestyksessä. Seulonnan perusteella pyrittiin selvittämään suodattuvuuteen vaikuttavat parametrit ja niiden optimaalinen kombinaatio, jolla suodatusnopeus ja suodattuneen liuoksen määrä saataisiin maksimoitua. Lopuksi optimoiduilla parametreilla testattiin eri hypromelloosieriä verraten tuloksia aikaisempiin suodattuvuustestituloksiin, sekä testattiin vaihtoehtoista hypromelloosin dispersiomenetelmää, jolla pyrittiin minimoimaan dispersio- ja jäähdytysvaiheessa valmistusastiaan/liuokseen jäävä liukenematon aines. Testatuista parametreista sekoitusnopeudella oli pienin vaikutus suodattuvuuteen, ja jäähdytyslämpötilalla suurin. Kylmemmäksi jäähdytetyt liuokset suodattuivat paremmin, mikä voi johtua polymeeriketjujen lisääntyneestä hydraatiosta ja siitä aiheutuvasta hypromelloosin vähentyneestä aggregoitumisesta. Hypromelloosiliuosten lämpötilakäyttäytyminen voisikin olla hyödyllinen aihe tarkemmille jatkotutkimuksille. Pidempi sekoitusaika ja korkeampi dispersiolämpötila tuottivat keskimäärin hieman parempia suodatustuloksia, mutta erot eivät olleet tilastollisesti merkittäviä. Tutkimuksessa eniten haasteita aiheutti lämpötilan ja sekoituksen kontrollointi, sekä liukenemattoman hypromelloosiaineksen jääminen valmistusastian reunoille. Vaihtoehtoisella dispersiomenetelmällä saatiin alustavasti lupaavia tuloksia, mutta menetelmä vaatii vielä lisätutkimuksia. Tärkeää olisi myös löytää juurisyy suodattimen tukkeutumiselle, esim. analysoimalla tukkeutunutta suodatinmembraania tarkemmin. Tutkimuksesta saatiin hyödyllistä lisätietoa hypromelloosiliuosten käyttäytymisestä liuosvalmistuksessa sekä steriilisuodatuksessa, mistä on ollut apua tuotannon ongelmien ratkaisemisessa.The problems caused by hypromellose in sterile filtration of ophthalmic products in the pharmaceutical industry were investigated. The research project was performed at NextPharma Oy's ophthalmics manufacturing facility in Tampere during the autumn of 2020. Hypromellose is an excipient commonly used in ophthalmic products as a viscosity enhancer to prolong the contact time of the preparation on the eye surface. In the ophthalmics compounding process, hypromellose is first dispersed by slowly sprinkling it into a hot solution and thoroughly mixing, after which the solution is cooled to room temperature. During cooling, the hypromellose dissolves and gels, increasing the viscosity of the solution. Incomplete dispersion or dissolution of hypromellose during the manufacturing process can slow down the filtration rate or even clog the filter completely due to undissolved hypromellose polymer material. Hypromellose is an industrially produced cellulose derivative that often contains some amounts of unreacted cellulose and other sparingly soluble polymer particles as impurities, which can also cause problems in filtration processes. Sterile filtration is a commonly used sterilization method for ophthalmic products, in which the prepared bulk solution is filtered through a 0.1 to 0.2 µm pore size filter membrane into a sterile receiving vessel. Due to the very small pore size, sterile filters are easily clogged if the solution contains poorly dissolved material. The purpose of this work was to collect additional information on the possible causes of clogging caused by hypromellose and to determine whether the filterability of a solution containing hypromellose can be improved by optimizing the manufacturing process parameters. The design of experiments was prepared, creating a two-level full-factorial test matrix without replicates and with three centre points. Four different process parameters were used (mixing time, mixing speed, dispersion temperature, and cooling temperature). Minimum and maximum levels for the parameters were obtained in the initial tests, after which the test solutions were prepared and filtered in a randomized order according to the test matrix. The aim of the screening was to find out which parameters were affecting the filterability and what would be their optimal combination that would maximize the filtration rate and the yield of filtration. Finally, the optimized parameters were used to test different batches of hypromellose, comparing the results to previous filtration tests. Additionally, an alternative hypromellose dispersion method was tested to minimize the amount of insoluble material remained during the dispersion and cooling steps. Of the parameters tested, mixing speed was the least significant, while cooling temperature had the most effect on the filtration results. The solutions with lower cooling temperature had better filtration results, which may be due to reduced aggregation of hypromellose due to increased hydration of the polymer chains. The temperature behaviour of hypromellose solutions could be an interesting subject for further investigation. Longer mixing times and higher dispersion temperatures produced slightly better filtration results on average, but the differences were not statistically significant. Most challenging in the study was controlling the temperature and mixing of the solutions, and the retention of insoluble hypromellose material at the walls of the compounding vessel. The alternative dispersion method gave promising preliminary results, but the method still requires further testing. It would be important to also find the root cause of the filter clogging mechanism e.g., by further analysing the clogged filter membrane. The study provided additional useful information of the behaviour of hypromellose solutions in solution preparations and during sterile filtration, which has been helpful in solving production problems

Guideline for disinfection and sterilization in healthcare facilities, 2008. Update: May 2019

The Guideline for Disinfection and Sterilization in Healthcare Facilities, 2008, presents evidencebased recommendations on the preferred methods for cleaning, disinfection and sterilization of patientcare medical devices and for cleaning and disinfecting the healthcare environment. This document supercedes the relevant sections contained in the 1985 Centers for Disease Control (CDC) Guideline for Handwashing and Environmental Control. Because maximum effectiveness from disinfection and sterilization results from first cleaning and removing organic and inorganic materials, this document also reviews cleaning methods. The chemical disinfectants discussed for patient-care equipment include alcohols, glutaraldehyde, formaldehyde, hydrogen peroxide, iodophors, ortho-phthalaldehyde, peracetic acid, phenolics, quaternary ammonium compounds, and chlorine. The choice of disinfectant, concentration, and exposure time is based on the risk for infection associated with use of the equipment and other factors discussed in this guideline. The sterilization methods discussed include steam sterilization, ethylene oxide (ETO), hydrogen peroxide gas plasma, and liquid peracetic acid. When properly used, these cleaning, disinfection, and sterilization processes can reduce the risk for infection associated with use of invasive and noninvasive medical and surgical devices. However, for these processes to be effective, health-care workers should adhere strictly to the cleaning, disinfection, and sterilization recommendations in this document and to instructions on product labels.In addition to updated recommendations, new topics addressed in this guideline include1. inactivation of antibiotic-resistant bacteria, bioterrorist agents, emerging pathogens, and bloodborne pathogens;2. toxicologic, environmental, and occupational concerns associated with disinfection andsterilization practices;3. disinfection of patient-care equipment used in ambulatory settings and home care;4. new sterilization processes, such as hydrogen peroxide gas plasma and liquid peracetic acid; and5. disinfection of complex medical instruments (e.g., endoscopes).This guideline discusses use of products by healthcare personnel in healthcare settings such as hospitals, ambulatory care and home care; the recommendations are not intended for consumer use of the products discussed.disinfection-guidelines-H.pd

Guideline for disinfection and sterilization in healthcare facilities, 2008

2008, last update: Feburary 15, 2017"The Guideline for Disinfection and Sterilization in Healthcare Facilities, 2008, presents evidence-based recommendations on the preferred methods for cleaning, disinfection and sterilization of patient-care medical devices and for cleaning and disinfecting the healthcare environment. This document supercedes the relevant sections contained in the 1985 Centers for Disease Control (CDC) Guideline for Handwashing and Environmental Control. Because maximum effectiveness from disinfection and sterilization results from first cleaning and removing organic and inorganic materials, this document also reviews cleaning methods. The chemical disinfectants discussed for patient-care equipment include alcohols, glutaraldehyde, formaldehyde, hydrogen peroxide, iodophors, ortho-phthalaldehyde, peracetic acid, phenolics, quaternary ammonium compounds, and chlorine. The choice of disinfectant, concentration, and exposure time is based on the risk for infection associated with use of the equipment and other factors discussed in this guideline. The sterilization methods discussed include steam sterilization, ethylene oxide (ETO), hydrogen peroxide gas plasma, and liquid peracetic acid. When properly used, these cleaning, disinfection, and sterilization processes can reduce the risk for infection associated with use of invasive and noninvasive medical and surgical devices. However, for these processes to be effective, health-care workers should adhere strictly to the cleaning, disinfection, and sterilization recommendations in this document and to instructions on product labels. In addition to updated recommendations, new topics addressed in this guideline include 1) inactivation of antibiotic-resistant bacteria, bioterrorist agents, emerging pathogens, and bloodborne pathogens; 2) toxicologic, environmental, and occupational concerns associated with disinfection and sterilization practices; 3) disinfection of patient-care equipment used in ambulatory settings and home care; 4) new sterilization processes, such as hydrogen peroxide gas plasma and liquid peracetic acid; and 5) disinfection of complex medical instruments (e.g., endoscopes)." - p. 7Environmental Fogging [December 2009]Clarification Statement: CDC and HICPAC have recommendations in both 2003 Guidelines for Environmental Infection Control in Health-Care Facilities and the 2008 Guideline for Disinfection and Sterilization in Healthcare Facilities that state that the CDC does not support disinfectant fogging. Specifically, the 2003 and 2008 Guidelines state:\ue2\u20ac\ua2 2003: \ue2\u20ac\u153Do not perform disinfectant fogging for routine purposes in patient-care areas. Category IB\ue2\u20ac?\ue2\u20ac\ua2 2008: \ue2\u20ac\u153Do not perform disinfectant fogging in patient-care areas. Category II\ue2\u20ac?These recommendations refer to the spraying or fogging of chemicals (e.g., formaldehyde, phenol-based agents, or quaternary ammonium compounds) as a way to decontaminate environmental surfaces or disinfect the air in patient rooms. The recommendation against fogging was based on studies in the 1970\ue2\u20ac\u2122s that reported a lack of microbicidal efficacy (e.g., use of quaternary ammonium compounds in mist applications) but also adverse effects on healthcare workers and others in facilities where these methods were utilized. Furthermore, some of these chemicals are not EPA-registered for use in fogging-type applications.These recommendations do not apply to newer technologies involving fogging for room decontamination (e.g., ozone mists, vaporized hydrogen peroxide) that have become available since the 2003 and 2008 recommendations were made. These newer technologies were assessed by CDC and HICPAC in the 2011 Guideline for the Prevention and Control of Norovirus Gastroenteritis Outbreaks in Healthcare Settings, which makes the recommendation:\ue2\u20ac\u153More research is required to clarify the effectiveness and reliability of fogging, UV irradiation, and ozone mists to reduce norovirus environmental contamination. (No recommendation/unresolved issue)\ue2\u20ac?The 2003 and 2008 recommendations still apply; however, CDC does not yet make a recommendation regarding these newer technologies. This issue will be revisited as additional evidence becomes available.CurrentHICPACPrevention and ControlInfectious Diseas

Microbiological contamination in Portuguese firefighters headquarters

Tese de mestrado, Biologia Humana e Ambiente, 2021, Universidade de Lisboa, Faculdade de CiênciasIn occupational environments, employees might be exposed to a diversity of microbiological agents. However, less is known about firefighter’s exposure in the work environment. Thus, this study aims to characterize the microbiological contamination in firefighter’s headquarters from Lisbon, Portugal. SARS-CoV-2 and fungal resistance profile assessment were also performed. Eleven firefighters headquarters (FFH) were assessed and the sampling campaign comprised active and passive sampling methods. The microbiological assessment covered culture-dependent and culture-independent methods. Additionally, to perform the azole resistance screening the samples extracts were inoculated in Sabouraud dextrose agar (SDA) azole supplemented media (Itraconazole (ITR), Voriconazole (VOR) and Posaconazole (POS)). In general, from all the matrices the predominant genera were Cladosporium and Penicillium. Regarding microbiological load indoors, in accordance with the scientific criteria for occupational exposure assessment (I /O ≤1), from the 11 FFH sampled, 5 (45.45%) surpassed the stipulated value in what concerns fungal levels. The same trend was reported in 7 FFH (63.63%) for bacteria. Moreover, when using the World Health Organization (WHO) suggested indoor guideline (maximum value of 150 CFU.m-3 ), the same FFH were above the stipulated limits for fungal and bacterial load respectively. Overall, the use of a multi-approach protocol for sampling and analyses allowed a broader microbial characterization. Also, fungal growth in azole supplemented media suggests the presence of multidrug resistance. Thus, further antifungal susceptibility tests should be preconized. The molecular detection of sections Fumigati and Nidulantes in additional matrices suggests molecular tools as a suitable approach overcoming culture-dependent methods limitations. Regarding SARS-CoV-2 assessment, no detection was obtained which suggest the effectiveness of cleaning in viral control. In short, these facilities are an occupational environment to have into consideration regarding the microbiological contamination. In fact, more research is needed in order to implement proper measures to reduce workers risk and minimize the exposure