Dr. Chen Chin Chang
Adjunct Professor, Department of Food and Beverage Management, Hebei Normal University, China.

ISBN 978-81-960551-0-3 (Print)
ISBN 978-81-960551-5-8 (eBook)
DOI: 10.9734/bpi/ramb/v1

This book covers key areas of Microbiology and Biotechnology. The contributions by the authors include body fluids, beta-lactamase, infection control, liver disease, gastrointestinal helminthes, nanoparticle, bacterial pathogenicity, infectious diseases, prokaryotic and eukaryotic organisms, immuno-electron tomography, integrin clustering, limiting membrane rupturing, echovirus host-entry, 3D multivesicular compartments, endosomal uncoating, intraluminal vesicles, caveolar domains, retinoblastoma like protein 2, human papillomavirus infection, oncoprotein, cervical cancer, histopathology, pathogenic microorganisms, T cell immunity, mucosal immunity, respiratory viral vaccine, human filariasis, microfilaria, knott technique, dirofilaria, gene regulatory network inference, and gene expression. This book contains various materials suitable for students, researchers and academicians in the field of Microbiology and Biotechnology.


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Pathogenic Bacteria Isolated from Presumably Sterile Sites and their Drug Susceptibility at a University Hospital in North India

Mitra Kar, Chinmoy Sahu, Radhika Choudhary, Akanksha Dubey

Research Advances in Microbiology and Biotechnology Vol. 1, 28 December 2022, Page 1-15

The objective of this study was to identify commonly isolated bacteria from the body fluid samples and access their antibiotic sensitivity pattern.

All body fluid samples (except blood, cerebrospinal fluid, and contaminated samples) received in the Bacteriology section of the Department of Microbiology from November 2019 to May 2020 were included in our study.

Out of 363 samples of body fluids, 113 (31.12%) were found to be positive for bacterial culture and sensitivity. Male predominance among liver disease patients was deemed statistically significant when compared to those without liver disease. Comorbidities such as hypertension and encephalopathy were statistically significant in patients with liver disease compared to those without liver disease. Escherichia coli was the most commonly isolated bacteria (20.35%) which was followed by Pseudomonas spp (15.92%) and Klebsiella pneumoniae (11.50%). Fifty-one (45.13%) isolates in our study were Multidrug-resistant. In our investigation, cancer patients had statistically significant MDR microbe isolation from bodily fluid samples. With the Extended-spectrum beta-lactamase (ESBL) character and total resistance to Carbapenems, Klebsiella pneumoniae was considered the most resistant MDR bacterium. However, all isolates were vulnerable to Colistin. This study aids in the empirical treatment of patients based on antibiotic susceptibility patterns by displaying the range of bacterial isolates identified from bodily fluid samples collected in our laboratory. It also emphasises the importance of adhering to antibiotic sensitivity results and infection control practises in the hospital setting to prevent the spread of multidrug-resistant infections.

This chapter aims to evaluate the incidence of Spirometra species and other gastrointestinal helminthes in wild lions (Panthera leo) in Tarangire National Park, Northern Tanzania. Faecal samples from seven lions were examined by using floatation and sedimentation methods. There are three different taxa of intestinal helminthes. The parasite genera identified were Spirometra, Toxocara, and Taennidae, with 100%, 71.4%, and 42.8% prevalence rates, respectively. The outcomes demonstrated that the sedimentation method had a 100% success rate in detecting Spirometra (7 infected samples).Results were better for single infections (100%) than for mixed infections. Toxocara and Taennidae had high prevalences according to the flotation method (salt solution SG 1.210) and sugar solution (SG 1.459), respectively. As one of the most often found gastrointestinal helminths, Spirometra sp. is a significant source of zoonotic helminthes and provides a starting point for future research.  

Nanoparticles against Viruses

Liudmila B. Boldyreva

Research Advances in Microbiology and Biotechnology Vol. 1, 28 December 2022, Page 24-35

The possibility of using nanoparticles against viruses is proved by the following. At the beginning of the chapter (Section 2) the properties of unique process of transfer of angular momentum (spin supercurrent) are considered. Then it is shown in Section 3 that the effect of metal nanoparticles on biological systems may be performed by spin supercurrent. Further, it is shown in Section 4 that spin supercurrent may influence viruses. Thus, from data presented in Sections 3 and 4 it follows: metal nanoparticles may influence viruses.

Biological Concept of Bacterial Pathogenicity (Theoretical Review)

Yurii V. Ezepchuk

Research Advances in Microbiology and Biotechnology Vol. 1, 28 December 2022, Page 36-47

Biological nature of the bacterial pathogenicity phenomenon is based on the interaction of prokaryotic and eukaryotic organisms. The phenomenon is the poly-functional biological potency of germs that are realized by factors (determinants) of pathogenicity. Some fundamental biological functions are responsible for bacterial pathogenicity in a multi-cellular host organism: the adhesive function, the function of invasion and penetration into the cell, the function of evasion of host defense, and the damage function. The action of adhesion, invasion and evasionis directed to towards establishing an ecological niche in multi-cellular host while the aim of the damaging function is destruction of the environment.

3D Multivesicular Structures: Triggered Permeability by Integrin-mediated Picornavirus Entry

Pan Soonsawad, Lassi Paavolainen, Varpu Marjoma, R. Holland Cheng

Research Advances in Microbiology and Biotechnology Vol. 1, 28 December 2022, Page 48-65

We elucidated the structural features of vMVBs (virus-induced multivesicular body) compared to antibody-induced control as mock infection. High-pressure cryo fixation of cells is employed to conduct the study with immunoelectron tomography during early virus entry. ILV (intraluminal vesicle) grew concurrently with the dramatic alterations in vMVB size and membrane integrity brought on by EV1 infection. ILV breakages started to appear at 2 hours post-injury, followed by vMVB limiting membrane rupture. These breakages likely enable effective replication and are connected to EV1 genome egress from vMVBs. The increased internalization of EV1 appears to be caused by a2b1-integrin clustering, where the cellular uptake further develops membrane compartments forming MVBs. Three-dimensional tomograms revealed a marked increase in the size and complexity of these vMVBs and ILVs at 2 and 3.5 hours post-infection (p.i.), in contrast to the control MVBs without the virus. Breakages in the membranes of vMVBs were detected from tomograms after 2 and especially after 3.5 h, suggesting that these breakages could facilitate the genome release to the cytoplasm. The in situ neutral-red labeling of the viral genome showed that virus uncoating starts as early as 30 min p.i., while an increase of permeability was detected in the vMVBs between 1 and 3 hours p.i., based on a confocal microscopy assay. Here, the rupture of endosomes upon the entry of a non-enveloped enterovirus unveils how the viral genome is transferred to the cytoplasm for subsequent replications.

High Risk and Cutaneous Types of HPV Suppress p130 to Induce Host Cell Cycle

Nurshamimi Nor Rashid, Zi Ling Yong, Rohana Yusof, Roger J. Watson

Research Advances in Microbiology and Biotechnology Vol. 1, 28 December 2022, Page 66-81

p130, also known as retinoblastoma like protein 2 (RBL2), is a member of the pocket protein family that is rarely mutated in human tumours. Its expression is largely G0 restricted and posttranscriptionally regulated. We previously demonstrated that E6/E7 oncoproteins encoded by HPV type 16, a high-risk type for cervical cancer progression, must target p130 in order to promote the host cell to exit the quiescence (G0) state and enter the S phase of the cell cycle. P130 is also associated with the DREAM (DP, RB-like, E2F and MuvB) complex in G0/G1, which prevents S phase development by repressing transcription of E2F-regulated genes. In this chapter,  we have used p130 mutants deficient in binding the E7 LXCXE domain (p130mE7), unphosphorylatable by CDK2 (p130PM22) or a combination of both (p130PM22/mE7) to investigate these mechanisms used by E7 proteins to disrupt the p130-DREAM complex and promote cell cycle progression. As HPV16 E7 was unable to suppress p130mE7 but could do so with p130PM22, we discovered that binding of HPV16 E7 to p130 through its LXCXE domain was definitely necessary to disrupt p130-DREAM and promote S phase of the cell cycle. In comparison, the E7 protein encoded by HPV 48 E7, a cutaneous HPV type lacking a functional LXCXE domain, was also capable of disrupting p130-DREAM to promote cell cycling, but via a different mechanism. Thus, HPV48 E7 could suppress a cell cycle block induced by p130mE7 but not p130PM22. Overall, these findings suggest that p130 suppression is needed for HPV-induced cell cycling, and that different HPV E7 proteins can achieve this through various mechanisms.

Biochemical and Histopathological Changes in the Liver of Oreochromis niloticus Experimentaly Infected with Escherichia coli

Amal M. Yacoub, Sherifa Mostafa M. Sabra, Mona Khaled D. Al-Kourashi

Research Advances in Microbiology and Biotechnology Vol. 1, 28 December 2022, Page 82-101

Fish of Oreochromis niloticus were treated with different concentrations of Enterotoxigenic Escherichia coli (ETEC) as (103-105, 106-107 109-1010/ml water). The study was carried out to detect the impact of E. coli toxins on Oreochromis niloticus fish throughout physiological liver function and histopathological changes in the liver. The values of GPT, GOT and alkaline Phosphatase ranged between (905.90 IU/L -75.40 IU/L), (4827.7 IU/L - 385.50 IU/L) and (106- 107 CFU /ml - 103 - 105 CFU / ml) for the three liver enzymes respectively. When Oreochromis niloticus liver was exposed to Escherichia coli, it showed signs of fatty degeneration, ballooning degeneration, pyknotic hepatocyte nuclei, necrosis of the hepatocytes and pancreatic acini, haemorrhage, inflammatory WBC infiltration, and hemosiderin pigment buildup. The experiment's findings showed that the liver of O. niloticus was extremely vulnerable to E. coli infection, which caused higher liver enzyme levels, reversible and irreversible liver damage, and ultimately the highest concentration of mortality.

The Development of Long-term Vaccine against Viral Respiratory Infection is Difficult, What is the Enduring Solution?

Rudra Prasad Roy, Laxmi Devi

Research Advances in Microbiology and Biotechnology Vol. 1, 28 December 2022, Page 102-114

Safe and effective vaccine formation is very important for viral respiratory infection which is also an ambiguous process. There are many difficulties in the development of effective vaccine such as regular mutations in viruses and lacking of long term immunity. To avoid the difficulties in vaccine development against viral respiratory infection evolution of mucosal vaccine plays an important role in protection against these viral infections. In mucosal immunity, memory CD8+ T cell populations play a crucial role in producing high levels of gamma interferon (IFN-\(\gamma\)) and tumor necrosis factor (TNF) may be essential for protection as well as memory CD8+ T cell being able to elicit long-lived immunity. Hence, the hypothesis related to the present study novel form of combination in between viroporin as an antigenic site or vaccine candidate used with immunotherapeutic adjuvant (OX40 or OX40 and OX40L complex or IL12) may become to resolve the development of a long-term immunity containing vaccine.

Human Filariasis Diagnosed by Modified Knott Technique Randomly in Iraq: A Recent Study

Afkar M. Hadi, Farah A. Al-Zahawi

Research Advances in Microbiology and Biotechnology Vol. 1, 28 December 2022, Page 115-126

The current study aimed to the detection of Microfilariae in humans for clinically unsuspected cases random samples and for different ages; Applied modified Knott technique for the first time in Iraq; used three types of stains for blood samples and compression the results. A total of 103 blood samples healthy and patient people, the current study revealed to 37 (35.92%) were infected with Filariasis. The current study recorded positive rates for four groups of ages: children (5-15 year), young (16-27 year), Adult (28-49 year) and old (50 year & above) that the higher rate 55.55% in children group. The statistic test revealed non-significant differences between ages of positive samples for Filariasis. The current study revealed out of 56 serum samples of male 19 were positive (33.92%) and out of 47 serum sample of female 18 were positive (38.29) for Filariasis. The statistic test revealed non-significant differences between male and female. The current study applied three types of stains Methylene blue, Giemsa and Lieshmen for compression; all them appeared positive rates 38.15%, 35.71% and 23.07% respectively This study recorded three genera of filariae according to the morphological characteristics (Brugia, Mansonella and Wucheraria).

Study on Gene Regulatory Network Inference Using PLS-based Methods

Hailing Xu , Zenan Huang, Shun Guo, Donghui Guo

Research Advances in Microbiology and Biotechnology Vol. 1, 28 December 2022, Page 127-151

We introduce an ensemble gene regulatory network inference method PLSNET, which decomposes the GRN inference problem with p genes into p subproblems and solves each of the subproblems by using Partial least squares (PLS) based feature selection algorithm. Numerous potential uses for inferring the topology of gene regulatory networks (GRNs) from microarray gene expression data include identifying potential drug targets and offering important insights into biological processes. The data is noisy, high dimensional, and there are many potential interactions, so it continues to be a challenge. Then, a statistical technique is used to refine the predictions in our method. The proposed method was evaluated on the DREAM4 and DREAM5 benchmark datasets and achieved higher accuracy than the winners of those competitions and other state-of-the-art GRN inference methods.

Superior accuracy achieved on different benchmark datasets, including both in silico and in vivo networks, shows that PLSNET reaches state-of-the-art performance.