Web architecture, behavior, and predatory potential of Larinia chloris from rice fields (Araneae: Araneidae).

Present study was carried out to investigate the variation in web architecture of Larinia chloris (Audouin 1826) in relation to seasonal differences and the biotic factors of the environment. In addition, relative abundance, behavior, and predatory potential of L. chloris were also recorded. For this purpose, 100 orb-webs of L. chloris were observed in rice fields (August-October, 2022) from 3 districts of Punjab (Lahore, Sheikhupura, and Kasur). Percent abundance of L. chloriswas found to be highest in rice fields from Barki road, Lahore (39.53%). All the webs of L. chloris were vertical at height equal to the height of vegetation (115.2 ±â€…9.7 cm). Time required to complete the web was 45 ±â€…5 min. There was positive correlation between web architecture and vegetation height. Web capture area and average mesh height of L. chloris also showed positive correlation with carapace length. There was a significant difference in various web parameters (number of spirals, number of radii, capture area, average mesh height, upper radii, lower radii, left radii, and right radii) among different trapping months. A total of 1,326 insects were recorded from the 100 webs of L. chloris. The prey abundance was found to be highest in the fields from Barki Road, Lahore. The majority of the prey collected from webs of L. chloris belonged to order Diptera, Hemiptera, Coleoptera, and Lepidoptera. However, prey items recorded during different growth stages (from vegetative to ripening) varied significantly. This is the first ever report describing the ecology of L. chloris in rice fields from Punjab, Pakistan.

Purification and Characterization of Keratinase from Bacillus licheniformis dcs1 for Poultry Waste Processing.

Feather wastes-byproduct of commercial poultry processing plant is produced in large amounts. Keratinolytic enzymes produced by feather degrading bacteria can easily degrade these waste products releasing pure keratin as a residue. The aim of present study was to isolate, and characterize feather degrading bacteria as well as assess the keratinolytic potential of purified enzyme. Three feather degrading bacteria (dps3, wps1 and dcs1) were isolated from feathers of domestic chickens. Preliminary characterization of isolated bacteria revealed these isolates belonging to genus Bacillus. 16S rRNA gene sequencing identified the isolates as B. subtilis dps3 (MW255302), B. cereus wps1 (MW255303) and B. licheniformis dcs1 (MW255304). Cell free supernatant of B. licheniformis dcs1 degraded feathers completely in 14 days indicating its keratinolytic ability. Purification of keratinase enzyme from B. licheniformis dcs1 was performed using column chromatography. SDS-PAGE indicated its molecular weight as 32 KDa. Kerotinolytice activity was maximum at optimum pH of 7 and 45℃ temperature. Enzyme showed the potential to degrade keratin material such as hairs and nails of humans. Findings of current study suggested that purified enzyme possess potential to upgrade nutritional quality of poultry waste containing keratin and might play as important biotechnological tool for keratin hydrolysis.

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.

Enzymatic and non-enzymatic detoxification in Lycosa terrestris and Pardosa birmanica exposed to single and binary mixture of copper and lead.

Organisms employ various enzymatic and non-enzymatic detoxification mechanisms to minimize the harmful effects of metal pollution in the terrestrial environment. We examined the effects of copper (Cu), lead (Pb) and their mixture (Cu + Pb) on glutathione (GSH), metallothionein (MTs), cytochrome P450 (CYP 450), carboxylesterase (CarbE), acetylcholinesterase (AchE) and glutathione S-transferase (GST) in Lycosa terrestris and Pardosa birmanica via two exposure routes, i.e., soil and food for 10, 20 and 40 days. The present results revealed that the accumulation of Cu and Pb in both spiders' species increase with exposure duration and depend on the route of exposure and type of metal. The activities of CarbE, GST, and MTs significantly increased with increasing metal body burden for all experimental treatments. The CYP 450 activity exhibited a significant time-dependent decrease with increasing Cu concentration in both species. The AchE activity was significantly inhibited on Pb exposure via soil and Cu + Pb exposure via both routes. The decrease in the level of GSH was measured on Cu + Pb exposure via both routes. Thus, all these enzymatic and non-enzymatic responses are sensitive to the metals tested and could serve as early warning indicators for assessing the effects of metal pollution in these species.

Assessment of bioaccumulation of cu and Pb in experimentally exposed spiders, Lycosa terrestris and Pardosa birmanica, using different exposure routes.

Major concerns exist regarding the environmental and human health risks caused by exposure to heavy metals. Spiders are often used as a model in ecotoxicological studies to assess soil pollution. Here, we measured the bioaccumulation of copper (Cu) and lead (Pb) in spiders, Lycosa terrestris and Pardosa birmanica, by inductively coupled plasma mass spectrometry (ICP-MS). We investigated whether Cu and Pb accumulation differs according to different spider species, single versus combined metal exposure, and routes of exposure. Spiders were exposed to 10 mM CuSO4 and 10 mM PbCl2 solutions separately or in combination (10 mM + 10 mM) through different exposure routes (spiked soil and food) for 6 weeks. The effect of metals on the survival and body mass of exposed and unexposed (control) spiders was determined. We found that in both spider species, accumulation of metals increased with exposure time. In single metal exposure, Cu accumulation from food was higher than soil exposure in both spider species, whereas the opposite was observed for Pb. The simultaneous uptake of Cu and Pb significantly decreased from food and soil, respectively. Soil exposure caused more accumulation of metals in L. terrestris than P. birmanica. Metal exposure via contaminated food caused higher mortality compared to soil exposure. Body mass of both spider species was significantly decreased and negatively correlated with metal's concentration. Overall, our results show that bioaccumulation efficiency of Cu and Pb differs significantly in spiders exposed to metal's mixture compared to single metal exposure and is dependent on the exposure route, the type of metal, and spider species. More understanding of the effects of exposure to metal mixture and exposure routes is essential for designing and supporting risk assessment and ecological monitoring programs.

Antioxidant enzymes as biomarkers of Cu and Pb exposure in the ground spiders Lycosa terrestris and Pardosa birmanica.

Heavy metal exposure induces oxidative stress in terrestrial organisms, which they counteract via activation of antioxidant biomarkers. The present study investigated the effects of copper (Cu) and lead (Pb) on the total antioxidant capacity (TAC) and antioxidant enzymes such as Catalase (CAT), Glutathione reductase (GR), Superoxide dismutase (SOD) and Glutathione peroxidase (GPX) in two spider species, namely Lycosa terrestris and Pardosa birmanica. The spiders were exposed to Cu and Pb separately (10 ppm) or in combination (10 ppm each) via two different exposure routes (i.e. food and soil) for 10, 20 and 40 days. The results showed that metal accumulation and antioxidant biomarker responses in spiders were metal- and species-dependent. Also, the levels of all antioxidant biomarkers increased significantly with increasing exposure time and metal load in the bodies of spiders via both exposure routes. The significant inhibition of TAC and antioxidant enzyme activities was only observed in single Pb treatment through soil exposure. In L. terrestris, the activities of detoxification enzymes and TAC were significantly enhanced on single Cu exposure than Pb via both exposure routes. However, in P. birmanica consistent variation among antioxidant parameters were observed depending on the metal load and exposure routes. The combined metal exposure caused more pronounced increase in the level of antioxidants compared to single metal exposure in both species, mainly via food exposure. These results suggest that the antioxidant enzymes and TAC are sensitive to single and combined metal exposure via both uptake routes. These data show that antioxidant parameters can be used potential biomarkers of oxidative stress associated with metal exposure and for monitoring environmental health using spiders as bioindicators.

Assembling a DNA barcode reference library for the spiders (Arachnida: Araneae) of Pakistan.

Morphological study of 1,795 spiders from sites across Pakistan placed these specimens in 27 families and 202 putative species. COI sequences >400 bp recovered from 1,782 specimens were analyzed using neighbor-joining trees, Bayesian inference, barcode gap, and Barcode Index Numbers (BINs). Specimens of 109 morphological species were assigned to 123 BINs with ten species showing BIN splits, while 93 interim species included representatives of 98 BINs. Maximum conspecific divergences ranged from 0-5.3% while congeneric distances varied from 2.8-23.2%. Excepting one species pair (Oxyopes azhari-Oxyopes oryzae), the maximum intraspecific distance was always less than the nearest-neighbor (NN) distance. Intraspecific divergence values were not significantly correlated with geographic distance. Most (75%) BINs detected in this study were new to science, while those shared with other nations mainly derived from India. The discovery of many new, potentially endemic species and the low level of BIN overlap with other nations highlight the importance of constructing regional DNA barcode reference libraries.

Bioaccumulation of cadmium, lead, and zinc in agriculture-based insect food chains.

Globally, the metal concentration in soil is increasing due to different anthropogenic and geogenic factors. These metals are taken up by plants and further transferred in the food chain through different routes. The present study was designed to assess the transfer and bioaccumulation of the heavy metals, cadmium (Cd), lead (Pb), and zinc (Zn), in food chains from soil to berseem plants (Triofolium alexandrinum), to insect herbivores (the grasshopper Ailopus thalassinus and the aphid Sitobion avenae) and to an insect carnivore (the ladybird beetle Coccinella septempunctata). The soil of studied berseem fields were slightly alkaline, silty loam in texture and moderate in organic matter. In soil, the concentration of Zn and Pb were under permissible level while Cd was above the permissible level. The accumulation of metals in T. alexandrinum were found in the order Zn > Cd > Pb. Grasshoppers showed higher accumulation of Pb than of Cd and Zn. In the soil-berseem-aphid-beetle food chain, metal enrichment was recorded. However, aphids did not show bioaccumulation for Cd. Metals accumulation in beetles showed that translocation of Zn, Cd, and Pb was taking place in the third trophic level. Our study highlights the mobility of metals in insect food chains and showed that insect feeding style greatly influenced the bioaccumulation. However, different metals showed variable bioaccumulation rates depending on their toxicity and retention.

New chrysilline and aelurilline jumping spiders from Pakistan (Araneae, Salticidae).

Epocillapakhtunkhwa Ali & Maddison, sp. n. and Stenaelurillusmardanicus Ali & Maddison, sp. n. are described from Khyber Pakhtunkhwa Province, Pakistan. Noted for the first time is the presence in Epocilla of a small bump just anterior to the fovea of the carapace, of unknown functional significance, otherwise known in the unrelated Opisthoncus L. Koch, 1880 and Cocalus Pocock, 1897. In addition, the female of Menemerusnigli Wesolowska & Freudenschuss, 2012 is described for the first time.

Genetic diversity in cytochrome c oxidase I gene of Anopheles mosquitoes.

Genetic diversity in cytochrome c oxidase I (coxI) among 7 species of Anopheles mosquitoes from Pakistan, and 37 species from different geographical regions of the world, was recorded. Automatic Barcode Gap Discovery (ABGD) analysis revealed a clear gap between intraspecific and interspecific distances of 7 species from Pakistan. However, genetic distances of 37 Anopheles species failed to adequately differentiate species in a global context. Intraspecific and interspecific divergences for 7 Anopheles species of Pakistan varied from 0.0% to 2.5% (mean = 0.49%) and 8% to 22.3% (mean = 12.77%), respectively. Similarly, intraspecific distances for 37 species from different parts of world ranged from 0.0% to 11.2% (mean = 0.65%) while values of interspecific divergences ranged from 3.4% to 35% (mean = 11.75%). Although phylogenetic tree revealed separate clades for 7 Anopheles species of Pakistan, it failed to produce separate clades for 37 species of the world. It is concluded that although standard barcode region is helpful for identifying Anopheles mosquitoes, combination of multi-locus approaches and morphology may be required to accurately identify species in this genus.