Improved characterization of aminoglycoside penetration into human lung epithelial lining fluid via population pharmacokinetics.

Aminoglycosides are important treatment options for serious lung infections, but modeling analyses to quantify their human lung epithelial lining fluid (ELF) penetration are lacking. We estimated the extent and rate of penetration for five aminoglycosides via population pharmacokinetics from eight published studies. The area under the curve in ELF vs plasma ranged from 50% to 100% and equilibration half-lives from 0.61 to 5.80 h, indicating extensive system hysteresis. Aminoglycoside ELF peak concentrations were blunted, but overall exposures were moderately high.

Comprehensive stability analysis of 13 ß-lactams and ß-lactamase inhibitors in in vitro media, and novel supplement dosing strategy to mitigate thermal drug degradation.

Non-clinical antibiotic development relies on in vitro susceptibility and infection model studies. Validating the achievement of the targeted drug concentrations is essential to avoid under-estimation of drug effects and over-estimation of resistance emergence. While certain ß-lactams (e.g., imipenem) and ß-lactamase inhibitors (BLIs; clavulanic acid) are believed to be relatively unstable, limited tangible data on their stability in commonly used in vitro media are known. We aimed to determine the thermal stability of 10 ß-lactams and 3 BLIs via LC-MS/MS in cation-adjusted Mueller Hinton broth at 25 and 36°C as well as agar at 4 and 37°C, and in water at -20, 4, and 25°C. Supplement dosing algorithms were developed to achieve broth concentrations close to their target over 24 h. During incubation in broth (pH 7.25)/agar, degradation half-lives were 16.9/21.8 h for imipenem, 20.7/31.6 h for biapenem, 29.0 h for clavulanic acid (studied in broth only), 23.1/71.6 h for cefsulodin, 40.6/57.9 h for doripenem, 46.5/64.6 h for meropenem, 50.8/97.7 h for cefepime, 61.5/99.5 h for piperacillin, and >120 h for all other compounds. Broth stability decreased at higher pH. All drugs were ≥90% stable for 72 h in agar at 4°C. Degradation half-lives in water at 25°C were >200 h for all drugs except imipenem (14.7 h, at 1,000 mg/L) and doripenem (59.5 h). One imipenem supplement dose allowed concentrations to stay within ±31% of their target concentration. This study provides comprehensive stability data on ß-lactams and BLIs in relevant in vitro media using LC-MS/MS. Future studies are warranted applying these data to antimicrobial susceptibility testing and assessing the impact of ß-lactamase-related degradation.

Dietary Moringa oleifera mitigates Fluconazole-Induced immunological and spleen-histological alterations in Catfish (Clarias gariepinus).

Fluconazole (FCZ), an antifungal from the azole family, causes several detrimental effects in fish. In recent times, there has been a notable surge in interest regarding the utilization of Moringa oleifera (Mo) as a dietary antioxidant. This research aimed to evaluate the potential protective effects of dietary Moringa oleifera (MO) against the adverse impacts of fluconazole in the African catfish (Clarias gariepinus). The fish were allocated into four groups as follows: a control group fed a basal diet, an FCZ - exposed (200 ng/L) fed basal diet, 1% MO fed through basal diet, and an FCZ-exposed (200 ng/L) and 1% MO fed through basal diet fed group. The results showed that FCZ exposure decreased superoxide dismutase, total antioxidant capacity, and acetylcholine esterase levels. On the other hand, FCZ exposure increased malonaldehyde and cortisol levels as compared to control (P < 0.05). FCZ caused immunosuppressive effects in C. gariepinus as revealed by lower immunity indices (lysozyme and phagocytic activity and immunoglobulin level) and increased cytokine levels (IL-6 IL-1ß). Histological examination of the spleen from fish exposed to FCZ showed several splenic changes. We conclude that dietary MO supplementation has the potential to alleviate the oxidative stress, restore immune response balance, and mitigate histological damage induced by FCZ exposure, thus positioning MO as an immunostimulant in C. gariepinus when administered alongside FCZ.

Polystyrene nanoplastic and engine oil synergistically intensify toxicity in Nile tilapia, Oreochromis niloticus : Polystyrene nanoplastic and engine oil toxicity in Nile tilapia.

Polystyrene nanoplastic (PS-NPs) and Engine oil (EO) pose multiple ecotoxic effects with increasing threat to fish ecosystems. The current study investigated the toxicity of 15 days exposure to PS-NPs and / or EO to explore their combined synergistic effects on Nile tilapia, Oreochromis niloticus (O. niloticus). Hematobiochemical parameters, proinflammatory cytokines, and oxidative stress biomarkers as well as histological alterations were evaluated. The experimental design contained 120 acclimated Nile tilapia distributed into four groups, control, PS-NPs (5 mg/L), EO (1%) and their combination (PS-NPs + EO). After 15-days of exposure, blood and tissue samples were collected from all fish experimental groups. Results indicated that Nile tilapia exposed to PS-NPs and / or EO revealed a significant decrease in almost all the measured hematological parameters in comparison to the control, whereas WBCs and lymphocyte counts were significantly increased in the combined group only. Results clarified that the combined PS-NPs + EO group showed the maximum decrease in RBCs, Hb, MCH and MCHC, and showed the maximum significant rise in interleukin-1ß (IL-1ß), and interleukin-6 (IL-6) in comparison to all other exposed groups. Meanwhile, total antioxidant capacity (TAC) showed a significant (p < 0.05) decline only in the combination group, whereas reduced glutathione (GSH) showed a significant decline in all exposed groups in comparison to the control. Both malondialdehyde (MDA) and aspartate aminotransferase (AST) showed a significant elevation only in the combination group. Uric acid showed the maximum elevation in the combination group than all other groups, whereas creatinine showed significant elevation in the EO and combination group when compared to the control. Furthermore, the present experiment proved that exposure to these toxicants either individually or in combination is accompanied by pronounced histomorpholgical damage characterized by severe necrosis and hemorrhage of the vital organs of Nile tilapia, additionally extensively inflammatory conditions with leucocytes infiltration. We concluded that combination exposure to both PS-NPs and EO caused severe anemia, extreme inflammatory response, oxidative stress, and lipid peroxidation effects, thus they can synergize with each other to intensify toxicity in fish.

Hyaluronic acid impacts hematological endpoints and spleen histological features in African catfish (Clarias gariepinus).

Since its identification in the vitreous humour of the eye and laboratory biosynthesis, hyaluronic acid (HA) has been a vital component in several pharmaceutical, nutritional, medicinal, and cosmetic uses. However, little is known about its potential toxicological impacts on aquatic inhabitants. Herein, we investigated the hematological response of Clarias gariepinus to nominal doses of HA. To achieve this objective, 72 adult fish were randomly and evenly distributed into four groups: control, low-dose (0.5 mg/l HA), medium-dose (10 mg/l HA), and high-dose (100 mg/l HA) groups for two weeks each during both the exposure and recovery periods. The findings confirmed presence of anemia, neutrophilia, leucopoenia, lymphopenia, and eosinophilia at the end of exposure to HA. In addition, poikilocytosis and a variety of cytomorphological disturbances were observed. Dose-dependent histological alterations in spleen morphology were observed in the exposed groups. After HA removal from the aquarium for 2 weeks, the groups exposed to the two highest doses still exhibited a notable decline in red blood cell count, hemoglobin concentration, mean corpuscular hemoglobin concentration, and an increase in mean corpuscular volume. Additionally, there was a significant rise in neutrophils, eosinophils, cell alterations, and nuclear abnormalities percentages, along with a decrease in monocytes, coupled with a dose-dependent decrease in lymphocytes. Furthermore, only the highest dose of HA in the recovered groups continued to cause a significant increase in white blood cells. White blood cells remained lower, and the proportion of apoptotic RBCs remained higher in the high-dose group. The persistence of most of the haematological and histological disorders even after recovery period indicates a failure of physiological compensatory mechanisms to overcome the HA-associated problems or insufficient duration of recovery. Thus, these findings encourage the inclusion of this new hazardous agent in the biomonitoring program and provide a specific pattern of hematological profile in HA-challenged fish. Further experiments are highly warranted to explore other toxicological hazards of HA using dose/time window protocols.

Melatonin counteracts polyethylene microplastics induced adreno-cortical damage in male albino rats.

There are various substances that can disrupt the homeostatic mechanisms of the body, defined as endocrine-disrupting chemicals (EDCs). The persistent nature of microplastics (MPs) is a cause for concern due to their ability to accumulate in food chains and widespread use, making their toxic effects particularly alarming. The potential of MPs for disrupting the endocrine system was observed in multiple tissues. Moreover, the adrenal gland is known to be extremely sensitive to EDCs, while with the effect of MPs on the adrenal gland has not previously been studied. This study aimed to highlight the potential polyethylene microplastics (PE-MPs) induced adreno-toxic effects rather than exploring the implicated mechanisms and concluding if melatonin (Mel) can afford protection against PE-MPs induced adreno-toxicity. To fulfill the goal, six groups of rats were used; control, Mel, PE-MPs (3.75 mg/kg), PE-MPs (15 mg/kg), PE-MPs (3.75 mg/kg) +Mel, and PE-MPs (15 mg/kg) +Mel. PE-MPs induced toxic changes in the adrenal cortex, which was evident by increased adrenal weight, histopathological examination, and ultrastructural changes detected by electron microscope. A reduction in serum cortisol and an increase in serum adrenocorticotropic hormone resulted from the adreno-toxic effects of PE-MPs. Mechanisms may include the reduction of steroidogenesis-related genes, as PE-MPs drastically reduce mRNA levels of StAR, Nr0b1, Cyp11A1, as well as Cyp11B1. Also, oxidative stress that results from PE-MPs is associated with higher rates of lipid peroxidation and decreased superoxide dismutase and glutathione. PE-MPs inflammatory effect was illustrated by elevated expression of IL-1ß and NF-kB, detected by immunohistochemical staining, in addition to increased expression of caspase-3 and mRNA of Bax, markers of proapoptotic activity. The impacts of PE-MPs were relatively dose-related, with the higher dose showing more significant toxicity than the lower one. Mel treatment was associated with a substantial amelioration of PE-MPs-induced toxic changes. Collectively, this study fills the knowledge gap about the MPs-induced adrenal cortex and elucidates various related toxic mechanisms. It also supports Mel's potential protective activity through antioxidant, anti-inflammatory, anti-apoptotic, and gene transcription regulatory effects.

Oxidative stress, antioxidant defense responses, and histopathology: Biomarkers for monitoring exposure to pyrogallol in Clarias gariepinus.

Pyrogallol promotes free radicals leading to oxidative stress and toxicity. There are however a lack of studies on oxidative stress and the antioxidant system of fish following exposure to pyrogallol. This study measured oxidative stress markers, antioxidant responses, and histological changes in catfish exposed to pyrogallol. Fish were divided into one of four experimental groups: control only, or 1, 5 or 10 mg/L pyrogallol. After 15 days, glutathione-S-transferase in the serum was decreased in fish exposed to either 5 or 10 mg/L pyrogallol relative to controls while superoxide dismutase and total antioxidant capacity were decreased significantly in fish exposed to 1, 5, or 10 mg/L pyrogallol. Conversely, catalase was increased in serum of fish exposed to 1, 5, or 10 mg/L pyrogallol compared to controls. The liver of fish treated with 1, 5, or 10 mg/L pyrogallol had significantly higher levels of oxidative stress markers (malondialdehyde, lipid peroxidation, hydroperoxide content, oxidised protein content, and DNA fragmentation %) that varied with concentration. Catfish exposed to either 1, 5, or 10 mg/L pyrogallol presented with notable histological alterations in the intestine, kidney, and muscles with prominent fibrosis, as intense deposition of collagen fibre was observed by Masson's trichrome staining. Overall, endpoints related to oxidative stress and antioxidant defence enzymes in fish may be early biomarkers of pyrogallol exposure and contamination in aquatic ecosystems. Additional studies should characterize oxidative stress indicators for their utility as biomarkers of effect.

Combined effect of pantoprazole and mesenchymal stem cells on experimentally induced gastric ulcer: implication of oxidative stress, inflammation and apoptosis pathways.

Gastric ulcer (GU) is one of the most common diseases of the upper gastrointestinal tract that affects millions of people worldwide. This study aimed to investigate the possible alleviating effect of a combined treatment of pantoprazole (PANTO) and adipose tissue-derived mesenchymal stem cells (ADSCs) in comparison with each treatment alone on the healing process of the experimentally induced GU in rats, and to uncover the involved pathways. Rats were divided into five groups: (1) Control, (2) GU, (3) PANTO, (4) ADSCs and (5) ADSCs + PANTO. Markers of oxidative stress, inflammation and apoptosis were assessed. The current data indicated that PANTO-, ADSCs- and ADSCs + PANTO-treated groups showed significant drop (p < 0.05) in serum advanced oxidation protein products (AOPPs) and advanced glycation end products (AGEPs) along with significant elevation (p < 0.05) in serum TAC versus the untreated GU group. Moreover, the treated groups (PANTO, ADSCs and ADSCs + PANTO) displayed significant down-regulation (p < 0.05) in gastric nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), tumor necrosis factor alpha (TNF-α), cyclooxygenase-2 (COX-2), intercellular adhesion molecule-1 (ICAM-1), matrix metallopeptidase 9 (MMP-9) and caspase-3 along with significant up-regulation (p < 0.05) in vascular endothelial growth factor (VEGF) and peroxisome proliferator-activated receptor gamma (PPARγ) genes expression compared to the untreated GU group. Immunohistochemical examination of gastric tissue for transforming growth factor ß1 (TGF-ß1), epidermal growth factor (EGF) and proliferating cell nuclear antigen (PCNA) showed moderate to mild and weak immune reactions, respectively in the PANTO-, ADSCs- and ADSCs + PANTO-treated rat. Histopathological investigation of gastric tissue revealed moderate to slight histopathological alterations and almost normal histological features of the epithelial cells, gastric mucosal layer, muscularis mucosa and submucosa in PANTO-, ADSCs- and ADSCs + PANTO-treated rats, respectively. Conclusively, the co-treatment with ADSCs and PANTO evidenced sententious physiological protection against GU by suppressing oxidative stress, inhibiting inflammation and reducing apoptosis with consequent acceleration of gastric tissue healing process.

Penicillin-Binding Protein 5/6 Acting as a Decoy Target in Pseudomonas aeruginosa Identified by Whole-Cell Receptor Binding and Quantitative Systems Pharmacology.

The ß-lactam antibiotics have been successfully used for decades to combat susceptible Pseudomonas aeruginosa, which has a notoriously difficult to penetrate outer membrane (OM). However, there is a dearth of data on target site penetration and covalent binding of penicillin-binding proteins (PBP) for ß-lactams and ß-lactamase inhibitors in intact bacteria. We aimed to determine the time course of PBP binding in intact and lysed cells and estimate the target site penetration and PBP access for 15 compounds in P. aeruginosa PAO1. All ß-lactams (at 2 × MIC) considerably bound PBPs 1 to 4 in lysed bacteria. However, PBP binding in intact bacteria was substantially attenuated for slow but not for rapid penetrating ß-lactams. Imipenem yielded 1.5 ± 0.11 log10 killing at 1h compared to <0.5 log10 killing for all other drugs. Relative to imipenem, the rate of net influx and PBP access was ~ 2-fold slower for doripenem and meropenem, 7.6-fold for avibactam, 14-fold for ceftazidime, 45-fold for cefepime, 50-fold for sulbactam, 72-fold for ertapenem, ~ 249-fold for piperacillin and aztreonam, 358-fold for tazobactam, ~547-fold for carbenicillin and ticarcillin, and 1,019-fold for cefoxitin. At 2 × MIC, the extent of PBP5/6 binding was highly correlated (r2 = 0.96) with the rate of net influx and PBP access, suggesting that PBP5/6 acted as a decoy target that should be avoided by slowly penetrating, future ß-lactams. This first comprehensive assessment of the time course of PBP binding in intact and lysed P. aeruginosa explained why only imipenem killed rapidly. The developed novel covalent binding assay in intact bacteria accounts for all expressed resistance mechanisms.

Melatonin Alleviates Intestinal Barrier Damaging Effects Induced by Polyethylene Microplastics in Albino Rats.

There have been concerns about the potential health risks posed by microplastics (MP). The detection of MP in a variety of food products revealed that humans are ingesting MP. Nevertheless, there is a paucity of data about their impacts, as well as their uptake, on intestinal barrier integrity. This study examined the toxic effects of oral administration of two doses of polyethylene microplastics (PE-MP) (3.75 or 15 mg/kg/day for 5 weeks; mean particle size: 4.0-6.0 µm) on the intestinal barrier integrity in rats. Moreover, the effect of melatonin treatment with MP exposure was also assessed. The PE-MP particle uptake, histopathological changes, Alcian blue staining, Muc2 mRNA, proinflammatory cytokines (IL-1ß and TNF-α), and cleaved caspase-3, as well as tight junction proteins (claudin-1, myosin light-chain kinase (MLCK), occludin, and zonula occludens-1 (ZO-1)) were assessed. Oral administration of PE-MP resulted in apparent jejunal histopathological alterations; significantly decreased mucin secretion, occludin, ZO-1, and claudin-1 expression; and significantly upregulated MLCK mRNA, IL-1ß concentration, and cleaved caspase-3 expression. Melatonin reversed these altered parameters and improved the PE-MP-induced histopathological and ultrastructure changes. This study highlighted the PE-MP's toxic effect on intestinal barrier integrity and revealed the protective effect of melatonin.

Multi-biomarkers approach to assess the toxicity of novel insecticide (Voliam flexi®) on Clarias gariepinus: From behavior to immunotoxicity.

This study was conducted to determine for the first time the immunological, histopathological, histochemical, and ultrastructural changes; hematological and biochemical alterations; and poikilocytosis induced in Clarias gariepinus by Voliam flexi® 40% WG (thiamethoxam + chlorantraniliprole). Beside control fish, juvenile C. gariepinus were subjected to three sublethal concentrations of Voliam flexi® (43.5, 87.5, and 175 mg/L) for 15 days. Voliam flexi® induced immunotoxic impairments in C. gariepinus, such as a decrease in some immunity variables (lysozyme and phagocyte activity, immunoglobulin concentration, and nitro blue tetrazolium level). It also caused an extreme increase in the levels of primary cytokines (interleukin-1ß and IL-6), compared with the control. The toxic effects of Voliam flexi® increased gradually with the increasing concentrations tested. Histological examination of the liver demonstrated necrosis, vacuolated hepatocytes (fatty deposition), melanomacrophage centers, foci of inflammatory cells, congested and dilated blood sinusoids, hepatic degeneration, fibrosis increment (Sirius Red stain), and glycogen depletion, as well as cytopathological alterations. We conclude that the toxic effects of Voliam flexi® must be restricted or prevented by using control mechanisms in aquatic systems.

Implementation research on measuring quality in primary care: balancing national needs with learning from the Eastern Mediterranean Region.

BACKGROUND: Universal Health Coverage (UHC) is a core element of ensuring healthy lives, marking the third Sustainable Development Goal. It requires providing quality primary health-care (PHC) services. Assessment of quality of care considering a wide variety of contexts is a challenge. This study lists practical indicators to enhance the quality of PHC. OBJECTIVES: Demonstrating quality indicators for PHC that are feasible, comprehensive and adaptable to wide array of health systems and resource settings. METHODS: We applied the research framework: Exploration, Preparation, Implementation and Sustainment. Exploration included a scoping review to identify quality indicators. Preparation included an eDelphi to refine the primary indicators' list. A panel of 27 experts reviewed the list that was later pilot tested in PHC facilities. The outcomes were presented to two further expert consultations, to refine indicators and plan for broader testing. Implementation included testing the indicators through a five-step process in 40 facilities. A regional consultation in May 2016 discussed the testing outcomes. RESULTS: Initial efforts identified 83 quality indicators at the PHC level that were then refined to a 34-indicator list covering the six domains of quality. A toolkit was also developed to test the feasibility of each indicator measurement, data availability, challenges and gaps. Pilot testing provided insight into modifying and adding some indicators. Wide variability was encountered within and in between facilities, and timely initiation of antenatal care, for example, ranged 31-90% in Oman and 11-98% in Tunisia. Indicators were highly feasible, and 29 out of 34 were measured in 75% of facilities or more. While challenges included gaps in capacity for data collection, the tool showed high adaptability to the local context and was adopted by countries in the Eastern Mediterranean Region (EMR) including Libya, Oman, Iran, Pakistan, Sudan and Palestine. Stakeholders agreed on the high relevance and applicability of the proposed indicators that have been used to inform improvement. CONCLUSION: A cross-regional set of 34 quality indicators of PHC in the EMR was developed and adopted by a diverse group of countries. The toolkit showed high feasibility in pilot testing reflecting the practicality needed to encourage local uptake and sustainability. The core quality indicators are highly adaptable to different local and regional contexts regardless of current PHC strength or available resources. Continuous evaluation and sharing lessons of implementation and use are needed to ascertain the indicators' effectiveness in driving improvements in PHC and to refine and strengthen the evidence supporting the set of indicators for wider adoption.

First Penicillin-Binding Protein Occupancy Patterns for 15 ß-Lactams and ß-Lactamase Inhibitors in Mycobacterium abscessus.

Mycobacterium abscessus causes serious infections that often require over 18 months of antibiotic combination therapy. There is no standard regimen for the treatment of M. abscessus infections, and the multitude of combinations that have been used clinically have had low success rates and high rates of toxicities. With ß-lactam antibiotics being safe, double ß-lactam and ß-lactam/ß-lactamase inhibitor combinations are of interest for improving the treatment of M. abscessus infections and minimizing toxicity. However, a mechanistic approach for building these combinations is lacking since little is known about which penicillin-binding protein (PBP) target receptors are inactivated by different ß-lactams in M. abscessus We determined the preferred PBP targets of 13 ß-lactams and 2 ß-lactamase inhibitors in two M. abscessus strains and identified PBP sequences by proteomics. The Bocillin FL binding assay was used to determine the ß-lactam concentrations that half-maximally inhibited Bocillin binding (50% inhibitory concentrations [IC50s]). Principal component analysis identified four clusters of PBP occupancy patterns. Carbapenems inactivated all PBPs at low concentrations (0.016 to 0.5 mg/liter) (cluster 1). Cephalosporins (cluster 2) inactivated PonA2, PonA1, and PbpA at low (0.031 to 1 mg/liter) (ceftriaxone and cefotaxime) or intermediate (0.35 to 16 mg/liter) (ceftazidime and cefoxitin) concentrations. Sulbactam, aztreonam, carumonam, mecillinam, and avibactam (cluster 3) inactivated the same PBPs as cephalosporins but required higher concentrations. Other penicillins (cluster 4) specifically targeted PbpA at 2 to 16 mg/liter. Carbapenems, ceftriaxone, and cefotaxime were the most promising ß-lactams since they inactivated most or all PBPs at clinically relevant concentrations. These first PBP occupancy patterns in M. abscessus provide a mechanistic foundation for selecting and optimizing safe and effective combination therapies with ß-lactams.

Comparative histology of wild-type and p53-deficient medaka (Oryzias latipes): nephrotoxic effect of ultraviolet A radiation.

Ultraviolet radiation is an ecological factor that directly affects terrestrial organisms through suppression of immunity or damage to internal organs. The present study assessed the effects of ultraviolet A (UVA) radiation on the kidneys of both wild-type (WT) and p53-deficient medaka (Oryzias latipes) and evaluated which strain was more resistant to the effects of UVA. Fish were divided into four groups: control group 1 (Cwt and Cp53), kept for 3 days without UVA exposure; group 2 (1wt and 1p53), fish exposed daily to UVA for 1 h day-1 for 3 days; group 3 (2wt and 2p53), fish exposed daily to UVA for 2 h day-1 for 3 days; and group 4 (3wt and 3p53), fish exposed daily to UVA for 3 h day-1 for 3 days. Samples of tissues were obtained 24 h after UVA exposure. The most obvious histopathological changes induced by UVA radiation in kidney tissues of both strains of medaka (WT and p53-deficient) were high levels of vacuolation of tubular cells followed by necrosis. The tubular segments lost their normal shape which appeared like a network structure and their cells with clear cytoplasm. Necrosis of lymphoid tissues and spots of brown pigmentation (possibly melanomacrophages) were sporadically seen in interstitial lymphoid tissues, while shrinkage of glomeruli, diminution of periodic acid-Schiff staining, and increased amount of collagenous fibers were observed. Our results confirmed the harmful effects of UVA radiation on kidney tissues of both WT and p53-deficient medaka. However, WT medaka was affected more than p53-deficient medaka.

Biochemical and Molecular Characterization of Five Bacillus Isolates Displaying Remarkable Carboxymethyl Cellulase Activities.

Cellulases have many useful applications in industry and biotechnology. So, identification of new bacterial strains expressing cellulases with better properties is desired. Five soil bacterial strains screened for high carboxymethyl cellulase (CMCase) activities were characterized and identified by 16S rRNA analysis as Bacillus amyloliquefaciens (FAY088), B. velezensis (FAY0103), B. tequilensis (FAY0117), B. subtilis (FAY0136), and B. subtilis (FAY0182). Their CMCase activities were 1.49, 1.26, 1.21, 1.21, and 1.24 U/ml, respectively. The maximum CMCase production was attained by growth at 35 °C, pH 6, and 180 rpm for 5 days. Residual activities of CMCases from FAY088 and FAY0117 were 88% or more after growth at 40 °C, which is same as FAY0182 CMCase at 40 and 45 °C. Additionally, FAY0182 retained 73% residual activity at 50 °C. FAY088 and FAY0182 retained more than 85% at pH 7 and 8. Conversely, residual activities from FAY0103 and FAY0136 declined a lot by increasing growth temperature beyond 40 °C and pH beyond 7. The maximum CMCase stability in all isolates was observed at pH 7, 3-h incubation, and 40 °C except for FAY0103 CMCase showed optimum temperature at 30 °C. More than 70% CMCase stability was retained in case of FAY088 at 50 °C, FAY0117 at 50-70 °C, and FAY0136 at 50-60 °C. FAY088 CMCase seemed to be the lest sensitive to temperature variation as it displayed residual activities 67, 72, 78, 84, 77, 74, and 72% at pH 3, 4, 5, 6, 8, 9, and 10, respectively. Finally, the five CMCase-producing isolates are recommended further enzyme applications in biotechnology and industry.

Hepato-nephrotoxicity in late juvenile of Oreochromis niloticus exposed to gibberellic acid: Ameliorative effect of Spirulina platensis.

The current work intended to inspect the hepato-nephrotoxicity of gibberellic acid (GA3) in juvenile of Oreochromis niloticus as well as the possibility of restoration after dietary addition of different concentrations of Spirulina platensis (SP). Fishes were evenly assorted into five groups: Group I assigned as control, Group II fed on basal diet and exposed to 150 mg/L gibberellic acid (GA3). The 3rd, 4th, and 5th groups exposed to150 mg/L gibberellic acid (GA3) and previously fed for two months on SP supplemented diets at levels of 5, 20, and 100 g/kg, respectively. Fish serum were utilized to check glucose, total protein, hepatic and renal functions, enzymatic and non-enzymatic antioxidants activities (superoxide dismautase; SOD, catalase; CAT, and total antioxidant capacity; TAC) as well as histopathological alterations in liver and kidney. The results showed that creatinine, uric acid, liver enzymes, glucose, total protein, SOD, and CAT were significantly elevated in GA3-treated group. Liver of GA3-treated fish manifested some histopathological changes (hypertrophy, cytoplasmic vacuolization, and apoptotic cells with pyknotic nuclei, necrosis, dilated blood sinusoids, and lymphocytic aggregation around the central veins). Kidney of GA3-exposed fish revealed edema of the epithelium lining of some renal tubules and some showed vacuolar degeneration and dissociation. Hypertrophy in the glomerulus was observed with dilated blood capillaries. SP supplementation restored these biochemical, antioxidants, and histological changes near to control levels. This improvement was higher with 100 g/kg SP showing concentration dependency. According to this study we can conclude that SP supplementation can improve the hepato- and nephrotoxicity caused by GA3 exposure indicating its role as potent antioxidant food additive.

UVA-induced neurotoxicity in Japanese medaka (Oryzias latipes).

Ultraviolet radiation-induced neurodegeneration has been studied in the early stages of development in fish, but not extensively in the adult stage. The present study aimed at investigating the effects of ultraviolet radiation-A (UVA) in adult Japanese medaka (Oryzias latipes). The brain, spinal cord, and retina were examined histopathologically as nervous system target organs. Japanese medaka fish were exposed to 15, 30, and 60 min day-1 UVA for 3 days, and samples were obtained 24 h and 14 days after UVA exposure. Neurohistopathological alterations in brain tissue included vacuoles, blood congestion, degeneration of neuropils, and pyknotic nuclei in neurons. Alterations in the spinal cord included neuronal cell degeneration, reduction in the spinal cord area, and degeneration of Mauthner cells. Retinal tissue showed vacuolation in the nerve fiber layer (NFL), pyknotic nuclei in the ganglion cell layer (GCL), and decreased cell populations particularly in the inner nuclear layer (INL) and GCL. The degree of degeneration was dependent on the duration of UVA exposure. The signs of degeneration decreased gradually and disappeared completely after the 14-day recovery period. In addition, p53-deficient medaka fish were more tolerant than were wild-type (Hd-rR) Japanese medaka. In conclusion, UV radiation induced neurodegeneration in the brain, spinal cord, and retina of adult Japanese medaka (Oryzias latipes) but their normal histological architecture reappeared in these tissues after 14 days.

Cytotoxic and hemotoxic effects of silver nanoparticles on the African Catfish, Clarias gariepinus (Burchell, 1822).

The present study investigates the hemotoxic and cytotoxic impacts of two acute doses of silver nanoparticles (AgNPs) on the African catfish, Clarias garepinus in comparison to the impact of AgNO3 and the control fish. AgNPs-induced impacts were recorded on some biological and hematological indices of that species on the bases of their size (20 and 40 nm) and concentration (10 and 100 µg) but no significant interaction. AgNO3 had very low impact on these indices in comparison to AgNPs. Recovery period for 15 days was found to be valid to remove AgNPs and AgNO3 toxicity for most indices. The condition factor exhibited stability under stress whereas the hepatosomatic index was more sensitive to AgNPs. The AgNPs-induced hematological changes recorded were corresponding with different blood cell alterations which increased in frequency from the control and AgNO3 to 40 nm/100 µg; such blood cell alterations disappeared to great extent after recovery period of 15-days in a reverse order.

Silver nanoparticle-induced nephrotoxicity in Clarias gariepinus: physio-histological biomarkers.

The present study investigates the nephrotoxic effects of two acute doses of silver nanoparticles (AgNPs) and silver nitrate (AgNO3) on the African catfish, Clarias gariepinus, using biochemical, histochemical, and histopathological changes as biomarkers. AgNP-induced impacts were recorded in some of these characteristics on the bases of their size (20 and 40 nm) and concentration (10 and 100 µg/L) but no significant interaction between size and concentration. AgNO3 had low significant adverse effects on some parameters in comparison with those impacts of AgNPs. The concentrations of creatinine and uric acid exhibited different significant variations under stress in all exposed groups compared with those in the control group. On the tissue and cell levels, histopathological changes were observed. These changes include hypertrophies of glomeruli, proliferation in the haemopoietic tissue, dissociation in renal tubules, shrinkage of glomerulus, hydropic degeneration, dilatation of renal tubules, aggregation of melanomacrophages, rupture of Bowman's capsule, and the glomerular tuft and dilatation of Bowman's space. In more severe cases, the degenerative process leads to tissue necrosis in the kidney of AgNP-exposed fish as well as carbohydrate depletion; a faint coloration was also observed in the brush borders and basement membrane with a large amount of connective tissue fibers around the blood vessels and the renal tubules. Recovery period for 15 days led to improvement of most of the alterations in biochemical, histopathological, and histochemical parameters induced by AgNPs and AgNO3. In conclusion, one can postulate on the sensitivity of the kidney of C. gariepinus to AgNPs and recovery strategy is a must.

The protective role of Spirulina platensis to alleviate the Sodium dodecyl sulfate toxic effects in the catfish Clarias gariepinus (Burchell, 1822).

Sodium dodecyl sulfate (SDS) as anionic surfactant is common in household and personal care products and reach in the aquatic ecosystems from different applications. Present work aimed to study the effects of SDS and the potential ameliorative influence of Spirulina platensis (SP) in the African catfish Clarias gariepinus. Fish was exposed to SDS and SP, individually or in combination in four equal groups for two weeks. The 1st group (control), 2nd group (SDS-treated), 3rd group (SDS, 0.1 mg L-1 + SP, 100 mg L-1 water) and 4th group (SDS, 0.1 mg L-1 + SP, 200 mg L-1). Serum samples were used to analyze hepatic and renal functions, electrolytes, genetic, and antioxidant biomarkers. The results revealed that SDS exposure induced hepatic and renal dysfunction, electrolytes imbalance, as well as significant disruption in enzymatic and non-enzymatic antioxidants, and increase in alterations, micronuclei and apoptosis percentages in erythrocytes. SP addition restored these biochemical and genetic variations close to control levels. Thus, the present study suggests that SP could protect the catfish against SDS-induced injury by scavenging ROS, sustaining the antioxidant status and diminishing DNA oxidative damage.