Diversity and Abundance of Delineated Earthworm (Annelida: Clitellata) in Pakistan: A Review.

Earthworm, a ubiquitous (but neglected) macro-invertebrate, is found in terrestrial vicinity of Pakistan. Moreover, the occurrence of earthworms is often diverse with fluctuating quantity depending upon abiotic factors and land usage patterns. The aim of this study was to summarize all the reported information related to earthworm diversity in different areas of Pakistan. Almost all the data published from year 2001 to 2021 were collected. Following data organization, total 42 earthworm's species including five families (Acanthodrilidae, Lumbricidae, Moniligastridae, Octochaetidae and Megascolecidae) were reported from various researchers. Among five families, family Acanthodrilidae was found to have only one specie (Ramiella bishambari), Lumbricidae consist of 10 species (Apporactodea rosea, Allolobophora trapezoids, Allolobophora chloroticaa, Aporrectodea longa, A. caliginosa, Bimastus parvus, Eisenia fetida, Helodrilus foetidus, Lumbricus terrestris and L. rubillus), Moniligastridae has two species (Drawida nepalensis and D. pellucida) while Octochaetidae possess only one specie (Eutyphoeus incommodus). The most abundant and diverse family Megascolecidae consist of 28 earthworm species in all habitats of different regions of Pakistan. Among geographical areas, Faisalabad was found as the richest territory with most reported earthworm species (i.e. 28). The current study suggests further in depth research to explore the unidentified and/ missing species of earthworms in Pakistan.

Medicinal Perspective of Antibacterial Bioactive Agents in Earthworms (Clitellata, Annelida): A Comprehensive Review.

Humoral practice is a fundamental natural biological phenomenon in earthworm defensive system which protects them from infectious bacteria and irritating agents by different mechanisms. The defensive system of earthworms is highly complicated because they lack antibodies in their blood circulatory system but their body extracts and coelomic fluid comprise of different bioactive agents (i.e. peptides and proteins) that defend these worms. There are various groups of bioactive agents such as proteases (name depends on proteins/peptide function or formal earthworm species name), metabolites (total 59 metabolites found in Eisenia fetida), metal binding proteins (2 proteins such as Ca2+ binding calmodulin and metallothionein), active proteins (include lysozyme, lysenin and eiseniapore etc.), antimicrobial peptides (antibacterial vermi-peptides family (AVPF), antimicrobial peptide I (PP-I), coelomic cytolytic factor (CCF, CCF-I and CCF like protein), fetidin, lysenin, lumbricin (lumbricin I, lumbricin PG, and lumbricusin), organic acids (fatty acids, succinic acids, and lauric acid) and other organic compounds (such as purine and vitamin D). The presence of above mentioned molecules confer therapeutic potential that affect energy intake and involve in decreasing oxidative stress, metabolic disturbances and pro-inflammatory conditions. The future perspectives of earthworm bioactive compounds are concerned with the development of provisional standards, purification and classification for utilizations in pharma industry.

Impact of Interplay between Obese Gut Microbiota and Diet in Developing Obesity in Synthetic Community Mice.

This study aims at investigating the effects of cultured gut microbiota (GM) of obese human coupled high fat diet (HFD) or chow diet (CD) in development of obesity in mice. 20 stool samples were collected from obese patients and isolated bacteria were identified morphologically and biochemically. Identified isolates were mixed in equal proportions to synthesize obese GM. In vivo study was performed using obese GM combined with HFD/CD using mouse model for three months. Albino mice were treated with ampicillin from one week prior to birth until weaning of the pups at seven weeks of age and then inoculated with obese GM. Sixteen mice were divided into four groups: i.e. group 1 (G1) mice fed with CD, group 2 (G2) mice with HFD, group 3 (G3) mice with GM + HFD and group 4 (G4) mice with GM + CD. Mice from groups 3-4 were considered synthetic community (SC) mice due to transfer of synthesize human GM. 16S rRNA sequencing identified five abundant bacteria as Pseudomonas aeruginosa, Staphylococcus sp., Escherichia coli, Morganella morganii, and Klebsiella oxytoca (accession numbers: MZ150742-MZ150746). In vivo study indicated that GM combination with either HFD/CD caused significantly increased body weight in SC mice (BMI; Kg/m2) compared to HFD or CD fed mice groups. One way ANOVA revealed highly significant increase (p ≤ 0.001) in levels of total cholesterol (TC), triglycerides and low density lipoprotein (LDL) in GM coupled diet groups (G3-G4; SC mice) compared to significant increase in HFD group (G2) versus CD group (G1). Our study is first of its kind to report significant effects of using purified strains as obese GM plus diet (HFD/CD) in inducing obesity in SC mice and elevated serum liver parameters as metabolic indicators, hence providing strong evidence about significance of modified GM combination with HFD in developing obesity in SC mice.

Role of modified diet and gut microbiota in metabolic endotoxemia in mice.

This study was aimed at investigating the effect of cultured gut microbiota (GM) from obese humans coupled HFD in inducing metabolic endotoxemia in humanized mice. In total, 30 strains were isolated from 10 stool samples of obese patients. Following morphological and biochemical characterization, 16S rRNA gene sequencing of six abundant isolates identified these Klebsiella aerogenes, Levilactobacillus brevis, Escherichia coli, Staphylococcus aureus, Bacillus cereus and Bacillus subtilis (MZ052089-MZ052094). In vivo trial using above isolates, known as human gut microbiota (HGM), was performed for six months. Sixteen mice were distributed into four groups, i.e., G1 (control) mice fed with chow diet, group 2 (G2) with HFD, group 3 (G3) with HFD + HGM and group 4 (G4) with chow diet + HGM. Body mass index (BMI) and plasma endotoxins were measured pre- and post-experiment. In vivo study revealed that HFD + HGM caused significant increase (3.9 g/cm at 20 weeks) in the body weight and BMI (0.4 g/cm post-experiment) of G3 mice compared to the other groups. One-way ANOVA showed significantly higher level of endotoxins (2.41, 4.08 and 3.7 mmol/L) in mice groups G2, G3 and G4, respectively, indicating onset of metabolic endotoxemia. Cecal contents of experimental mice groups showed a shift in microbial diversity as observed by all isolates belonging to either Firmicutes or Bacteroidetes phyla, respectively. In conclusion, current study reported that minor alteration in GM composition through HFD feeding and cultured GM transfer has significant impact in development of metabolic endotoxemia, possibly via modified intestinal permeability.