scholarly journals Isolation and primary culture of Galleria mellonella hemocytes for infection studies

F1000Research ◽  
2021 ◽  
Vol 9 ◽  
pp. 1392
Author(s):  
Nicola J. Senior ◽  
Richard W. Titball
Keyword(s):  
Primary Culture ◽  
Alternative Model ◽  
Galleria Mellonella ◽  
Microbial Pathogens ◽  
Needle Stick Injury ◽  
Antimicrobial Compounds ◽  
Phagocytic Cells ◽  
Needle Stick ◽  
Research Institutes ◽  
Pre Treatment

Galleria mellonella larvae are increasingly used to study the mechanisms of virulence of microbial pathogens and to assess the efficacy of antimicrobials.  The G. mellonella model can faithfully reproduce many aspects of microbial disease which are seen in mammals, and therefore allows a reduction in the use of mammals. The model is now being widely used by researchers in universities, research institutes and industry. An attraction of the model is the interaction between pathogen and host. Hemocytes are specialised phagocytic cells which resemble neutrophils in mammals and play a major role in the response of the larvae to infection. However, the detailed interactions of hemocytes with pathogens is poorly understood, and is complicated by the presence of different sub-populations of cells. We report here a method for the isolation of hemocytes from Galleria mellonella.  A needle-stick injury of larvae, before harvesting, markedly increased the recovery of hemocytes in the hemolymph. The majority of the hemocytes recovered were granulocyte-like cells. The hemocytes survived for at least 7 days in culture at either 28°C or 37°C. Pre-treatment of larvae with antibiotics did not enhance the survival of the cultured hemocytes. Our studies highlight the importance of including sham injected, rather than un-injected, controls when the G. mellonella model is used to test antimicrobial compounds. Our method will now allow investigations of the interactions of microbial pathogens with insect hemocytes enhancing the value of G. mellonella as an alternative model to replace the use of mammals, and for studies on hemocyte biology.

scholarly journals Isolation and primary culture of Galleria mellonella hemocytes for infection studies

F1000Research ◽  
2020 ◽  
Vol 9 ◽  
pp. 1392
Author(s):  
Nicola J. Senior ◽  
Richard W. Titball
Keyword(s):  
Primary Culture ◽  
Alternative Model ◽  
Galleria Mellonella ◽  
Microbial Pathogens ◽  
Needle Stick Injury ◽  
Antimicrobial Compounds ◽  
Phagocytic Cells ◽  
Needle Stick ◽  
Research Institutes ◽  
Pre Treatment

Galleria mellonella larvae are increasingly used to study the mechanisms of virulence of microbial pathogens and to assess the efficacy of antimicrobials.  The G. mellonella model can faithfully reproduce many aspects of microbial disease which are seen in mammals, and therefore allows a reduction in the use of mammals. The model is now being widely used by researchers in universities, research institutes and industry. An attraction of the model is the interaction between pathogen and host. Hemocytes are specialised phagocytic cells which resemble neutrophils in mammals and play a major role in the response of the larvae to infection. However, the detailed interactions of hemocytes with pathogens is poorly understood, and is complicated by the presence of different sub-populations of cells. We report here a method for the isolation of hemocytes from Galleria mellonella.  A needle-stick injury of larvae, before harvesting, markedly increased the recovery of hemocytes in the hemolymph. The majority of the hemocytes recovered were granulocyte-like cells. The hemocytes survived for at least 7 days in culture at either 28°C or 37°C. Pre-treatment of larvae with antibiotics did not enhance the survival of the cultured hemocytes. Our studies highlight the importance of including sham injected, rather than un-injected, controls when the G. mellonella model is used to test antimicrobial compounds. Our method will now allow investigations of the interactions of microbial pathogens with insect hemocytes enhancing the value of G. mellonella as an alternative model to replace the use of mammals, and for studies on hemocyte biology.

scholarly journals A novel restraint device for injection of Galleria mellonella larvae that minimises the risk of accidental operator needle stick injury

2016 ◽  
Author(s):  
James P Dalton ◽  
Benedict Uy ◽  
Simon Swift ◽  
Siouxsie Wiles
Keyword(s):  
Antimicrobial Agents ◽  
Galleria Mellonella ◽  
Cost Effective ◽  
Manual Dexterity ◽  
Needle Stick Injury ◽  
Drug Resistant ◽  
Highly Pathogenic ◽  
Needle Stick ◽  
Risk Of Injury ◽  
Pathogenic Microbes

Larvae of the insect Galleria mellonella are increasingly being used for studying pathogenic microbes and their virulence mechanisms, and as a rapid model for screening novel antimicrobial agents. The larvae (waxworms) are most frequently infected by injection of pathogenic organisms into the haemocoel through the insect's prolegs. The mostly widely used method for restraining the waxworms for injection is by grasping them between the operator's fingers, which puts the operator at risk of needle stick injury, an important consideration when working with highly pathogenic and/or drug-resistant microorganisms. While use of a stab proof glove can reduce this risk of injury, it does so at the loss of manual dexterity and speed, resulting in a more labour-intensive and cumbersome assay. We describe a simple cost effective device (the so-called 'Galleria Grabber') for restraining waxworms for injection that keeps the operator's fingers clear of the needle thus reducing the risk of injury.

scholarly journals The novel Dbl homology/BAR domain protein, MsgA, of Talaromyces marneffei regulates yeast morphogenesis during growth inside host cells

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Harshini Weerasinghe ◽  
Hayley E. Bugeja ◽  
Alex Andrianopoulos
Keyword(s):  
Pathogenic Fungi ◽  
Host Cells ◽  
Microbial Pathogens ◽  
Mammalian Host ◽  
Host Immune System ◽  
Nucleotide Exchange ◽  
Phagocytic Cells ◽  
Macrophage Infection ◽  
Bar Domain ◽  
Talaromyces Marneffei

AbstractMicrobial pathogens have evolved many strategies to evade recognition by the host immune system, including the use of phagocytic cells as a niche within which to proliferate. Dimorphic pathogenic fungi employ an induced morphogenetic transition, switching from multicellular hyphae to unicellular yeast that are more compatible with intracellular growth. A switch to mammalian host body temperature (37 °C) is a key trigger for the dimorphic switch. This study describes a novel gene, msgA, from the dimorphic fungal pathogen Talaromyces marneffei that controls cell morphology in response to host cues rather than temperature. The msgA gene is upregulated during murine macrophage infection, and deletion results in aberrant yeast morphology solely during growth inside macrophages. MsgA contains a Dbl homology domain, and a Bin, Amphiphysin, Rvs (BAR) domain instead of a Plekstrin homology domain typically associated with guanine nucleotide exchange factors (GEFs). The BAR domain is crucial in maintaining yeast morphology and cellular localisation during infection. The data suggests that MsgA does not act as a canonical GEF during macrophage infection and identifies a temperature independent pathway in T. marneffei that controls intracellular yeast morphogenesis.

Transmission of hepatitis C virus by needle-stick injury in community settings

2007 ◽  
Vol 22 (11) ◽  
pp. 1882-1885 ◽  
Author(s):  
Paul S Haber ◽  
Margaret M Young ◽  
Lloyd Dorrington ◽  
Andrew Jones ◽  
John Kaldor ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Needle Stick Injury ◽  
Community Settings ◽  
Needle Stick

scholarly journals Economic Burden of Needle-Stick Injury Among Healthcare Workers in China

2018 ◽  
Vol 21 ◽  
pp. S86 ◽  
Author(s):  
L Zhang ◽  
Y Ai ◽  
J Liu ◽  
N Yue ◽  
J Xuan ◽  
...  
Keyword(s):  
Economic Burden ◽  
Healthcare Workers ◽  
Needle Stick Injury ◽  
Needle Stick

scholarly journals Prevalence of Needle Stick Injury among Surgeons

2021 ◽  
pp. 89-94
Author(s):  
Nasima Iqbal ◽  
Faiza Quraishi ◽  
Muhammad Aslam Bhatti ◽  
Faizah Mughal ◽  
Tayyaba Mumtaz ◽  
...  
Keyword(s):  
Reporting System ◽  
Medical Center ◽  
Working Hours ◽  
Work Load ◽  
Needle Stick Injury ◽  
Post Exposure Prophylaxis ◽  
Cross Sectional ◽  
Needle Stick ◽  
Post Exposure ◽  
Exposure Prophylaxis

Aim: To find out the prevalence of needle stick injury, its reporting system and the reasons behind it. Study design: Descriptive cross-sectional Place and duration of study: Study was conducted at Jinnah post-graduate medical center (JPMC) Karachi during the period of March to September 2019 Methodology: A self-designed, self-explanatory questionnaire was used, consisting of two parts, the first part was about demographic information while second part is for information related to needle stick injury like probable cause, frequency, response after injury, post-exposure prophylaxis and about reporting of the incident. Questionnaire was validated by calculating the Cronbach’s alpha which was 0.78. data was analyzed by using the Statistical Package for the Social Sciences (SPSS) version 20. Results: Majority of the study participants were female (67.2%) and about 50% were postgraduate students. Out of total 134 doctors about 64.2% of the doctors had needle stick injury during their career. Finding out the most probable cause of needle stick injury during the survey it was found out that increased work load and prolonged working hours were the main reasons. Majority of the cases occurred in emergency department (41.9%). About 95.5% of the doctors didn’t get any post-exposure prophylaxis. Majority of the participants (96.3%) did not report to any authority because of the lack of knowledge about the reporting policy, it was noted that about 38.8% were confused either the reporting system exist or not. Most of the injuries occur during the procedure of suturing followed by recapping syringes. Conclusion: It has been concluded that majority of the doctors had faced needle stick injury during their career and a very negligible number of them got any post-exposure prophylaxis. Majority of them did not report to any authority. So there is a need of implication of safety measures and reporting policies for early detection and treatment of infections after needle stick injury.

scholarly journals Manajemen Resiko Kesehatan dan Keselamatan Kerja Asuhan Keperawatan di IGD

2019 ◽  
Author(s):  
Dewi Kurniati
Keyword(s):  
Needle Stick Injury ◽  
Needle Stick ◽  
National Safety Council

Undang-undang No.44 Tahun 2009 tentang Rumah Sakit pasal 7 ayat 1, bahwa salah satu persyaratan Rumah Sakit adalah harus memenuhi unsur keselamatan dan kesehatan kerja. Laporan National Safety Council (NSC) tahun 1988 menunjukkan terjadinya kecelakaan di RS 41% lebih besar dari pekerja di industri lain. Kasus yang sering terjadi di antaranya tertusuk jarum atau needle stick injury (NSI), terkilir, sakit pinggang, tergores/terpotong, luka bakar, penyakit dan infeksi. Salah satu upaya pencegahan terjadinya kecelakaan kerja adalah dengan melakukan analisis risiko. membuat job hazard analisis, kemudian dilakukan analisis risiko dengan pendekatan AS/NZS 4360: 2004 dan menilai dengan tabel W.T.Fine. Hasil studi menunjukkan bahwa faktor bahaya di instalasi gawat darurat terdiri dari bahaya fisik, biologi, ergonomi, perilaku, dan psikologis. Faktor bahaya fisik merupakan yang dominan yaitu jarum suntik (benda tajam) yang berdampak luka tusuk dan tertular penyakit menular dari pasien. Nilai risiko tertinggi bahaya fisik dan biologi pada proses pekerjaan pemasangan infus pada pasien sebesar 150 (tinggi) mengharuskan adanya perbaikan secara teknis. Nilai risiko ini didapatkan apabila telah melakukan rekomendasi pengendalian dari peneliti.

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