The potential use of daphnia meal as substitute for fishmeal in diets of hybrid red tilapia affects growth performance, activities of digestive enzymes, antioxidant, immune status and intestinal histological parameters.

The current study aimed to evaluate growth performance, digestive enzyme activities, antioxidant status, nonspecific immune response and intestinal histological status of red tilapia fed Daphnia meal (DM) as a substitute for fishmeal (FM). Hybrid red tilapia (Oreochromis mossambicus × Oreochromis aureus) fry (0.54 ± 0.05 g fish-1) was allocated in nylon haba cages (100 fry m-3) for 2 weeks as an acclimation period. The fish were divided into five groups (three replicates each). The experimental diets were prepared by replacing FM with DM at concentrations of 25%, 50%, 75% and 100% respectively. The results indicated that fish fed increasing levels of DM (50%-75%) experienced high growth performance, feed utilisation and protein content. The activities of digestive enzymes were significantly increased in all groups fed DM diets compared to the control. The antioxidant balance was improved by decreasing the level of malondialdehyde and increased the total antioxidant capacity, catalase, superoxide dismutase and glutathione reductase activities in the liver of fish fed DM. The nonspecific immune response, including lysozyme, alkaline phosphatase activities and total protein level improved significantly with increasing FM substitution levels by DM in a dose-dependent manner. Histometric analysis of the intestinal wall revealed an increase in the villus length, crypts depth and goblet cells number in groups fed DM meal up to 50% substitution level compared to other treatments. It may be concluded from results of this feeding trial that in the aquaculture of hybrid tilapia, FM may be substituted with up to 50% DM without compromising intestinal health, growth performance and immune status of the fish.

Diversity and genetic structure of freshwater shark Wallago attu: an emerging species of commercial interest.

Pakistan has natural freshwater resources acting as a hotspot for diverse fish fauna. However, this aquatic fauna is declining at an alarming rate due to over-exploitation, habitat degradation, water pollution, climate change, and certain anthropogenic activities. The freshwater shark, Wallago attu, is a popular edible catfish inhabiting these freshwater ecosystems. Habitat degradation, overfishing, and human activities are heavily impacting the natural population of this species. So, sound knowledge about its population structure is necessary for its proper management in natural waters. The current study involves utilizing two mtDNA markers (COI, Cytb) to assess the genetic structure and differentiation among W. attu populations of Pakistani Rivers. Genetic variability analysis indicated a high haplotype (0.343 ± 0.046-0.870 ± 0.023) and low nucleotide diversity (0.0024 ± 0.012-0.0038 ± 0.018) among single and combined gene sequences, respectively. Overall, River Indus was populated with more diverse fauna of Wallago attu as compared to River Chenab and River Ravi. Population pairwise, Fst values (0.40-0.61) were found to be significantly different (p < 0.01) among three Riverine populations based upon combined gene sequences. The gene flow for the combined gene (COI + Cytb) dataset among three populations was less than 1.0. The transition/transversion bias value R (0.58) was calculated for testing of neutral evolution, and it declared low genetic polymorphism among natural riverine populations of Wallago attu. The current study's findings would be meaningful in planning the management and conservation of this economically important catfish in future.

Antifungal activity of copper oxide nanoparticles derived from Zizyphus spina leaf extract against Fusarium root rot disease in tomato plants.

Incorporating green chemistry concepts into nanotechnology is an important focus area in nanoscience. The demand for green metal oxide nanoparticle production has grown in recent years. The beneficial effects of using nanoparticles in agriculture have already been established. Here, we highlight some potential antifungal properties of Zizyphus spina leaf extract-derived copper oxide nanoparticles (CuO-Zs-NPs), produced with a spherical shape and defined a 13-30 nm particle size. Three different dosages of CuO-Zs-NPs were utilized and showed promising antifungal efficacy in vitro and in vivo against the selected fungal strain of F. solani causes tomato root rot disease, which was molecularly identified with accession number (OP824846). In vivo  results indicated that, for all CuO-Zs-NPs concentrations, a significant reduction in Fusarium root rot disease occurred between 72.0 to 88.6% compared to 80.5% disease severity in the infected control. Although treatments with either the chemical fungicide (Kocide 2000) showed a better disease reduction and incidence with (18.33% and 6.67%) values, respectively, than CuO-Zs-NPs at conc. 50 mg/l, however CuO-Zs-NPs at 250 mg/l conc. showed the highest disease reduction (9.17 ± 2.89%) and lowest disease incidence (4.17 ± 3.80%). On the other hand, CuO-Zs-NPs at varied values elevated the beneficial effects of tomato seedling vigor at the initial stages and plant growth development compared to either treatment with the commercial fungicide or Trichoderma Biocide. Additionally, CuO-Zs-NPs treatments introduced beneficial results for tomato seedling development, with a significant increase in chlorophyll pigments and enzymatic activity for CuO-Zs-NPs treatments. Additionally, treatment with low concentrations of CuO-Zs-NPs led to a rise in the number of mature pollen grains compared to the immature ones.  however the data showed that CuO-Zs-NPs have a unique antifungal mechanism against F. solani, they  subsequently imply that CuO-Zs-NPs might be a useful environmentally friendly controlling agent for the Fusarium root rot disease that affects tomato plants.

Isolation, molecular characterization, and genetic diversity of recently isolated foot-and-mouth disease virus serotype A in Egypt.

Foot-and-mouth Disease (FMD) is a highly contagious viral disease affecting all hoof-cloven animals. Serotypes A, O and SAT 2 of the foot-and-mouth disease virus (FMDV) are circulating in Egypt. The present study aimed to identify and molecularly characterize the FMDV strains circulating in Northern Egypt during an epidemic that struck the nation in 2022. RNA was extracted from the epithelial specimens, vesicular fluid from affected cattle. The samples were screened using real-time reverse-transcription polymerase chain reaction (RT-PCR) targeting the RNA-dependent RNA polymerase (RdRp) gene. Positive samples underwent individual serotype-specific amplification using primers designed for VP1 of O, A, and SAT 2 serotypes. Subsequently, direct sequencing was performed on the positive samples. The real-time RT-PCR detected positive samples from epithelial and vesicular fluid samples, but not in the blood of infected animals. Out of the 16 samples, seven tested positive for FMDV serotype A. Of these seven positive samples, six were categorized as serotype A-African topotype-G-IV, and these positive samples were isolated in BHK-21 cells, yielding an overt cytopathic effect caused by the virus. In conclusion, it is necessary to sustain continuous surveillance of the evolution of circulating FMDV strains to facilitate the assessment and aid in the selection of vaccine strains for the effective control of FMDV in Egypt.

UPLC/ESI/MS profiling of red algae Galaxaura rugosa extracts and its activity against malaria mosquito vector, Anopheles pharoensis, with reference to Danio rerio and Daphnia magna as bioindicators.

BACKGROUND: Anopheles pharoensis has a major role in transmitting several human diseases, especially malaria, in Egypt?. Controlling Anopheles is considered as an effective strategy to eliminate the spread of malaria worldwide. Galaxaura rugosa is a species of red algae found in tropical to subtropical marine environments. The presence of G. rugosa is indicative of the ecosystem's overall health. The current work aims to investigate UPLC/ESI/MS profile of G. rugosa methanol and petroleum ether extracts and its activity against An. pharoensis and non-target organisms, Danio rerio and Daphnia magna. METHODS: Galaxaura rugosa specimens have been identified using DNA barcoding for the COI gene and verified as G. rugosa. The UPLC/ESI/MS profiling of G. rugosa collected from Egypt was described. The larvicidal and repellent activities of G. rugosa methanol and petroleum ether extracts against An. pharoensis were evaluated, as well as the toxicity of tested extracts on non-target organisms, Dan. rerio and Dap. magna. RESULTS: The UPLC/ESI/MS analysis of methanol and petroleum ether extracts led to the tentative identification of 57 compounds belonging to different phytochemical classes, including flavonoids, tannins, phenolic acids, phenyl propanoids. Larval mortality was recorded at 93.33% and 90.67% at 80 and 35 ppm of methanol and petroleum ether extracts, respectively, while pupal mortality recorded 44.44 and 22.48% at 35 and 30 ppm, respectively. Larval duration was recorded at 5.31 and 5.64 days by methanol and petroleum ether extracts at 80 and 35 ppm, respectively. A decrease in acetylcholinesterase (AChE) level and a promotion in Glutathione-S-transferase (GST) level of An. pharoensis 3rd instar larvae were recorded by tested extracts. The petroleum ether extract was more effective against An. pharoensis starved females than methanol extract. Also, tested extracts recorded LC50 of 1988.8, 1365.1, and 11.65, 14.36 µg/mL against Dan. rerio, and Dap. magna, respectively. CONCLUSIONS: Using red algae derivatives in An. pharoensis control could reduce costs and environmental impact and be harmless to humans and other non-target organisms.

Relative expression levels of growth hormone gene and growth rate in Indian major carp species.

The phenomenon of growth is a leading factor for aquaculture success. The uneven growth of major Indian carps (Labeo rohita, Catla catla, and Cirrhinus mrigala) is a serious issue in fish culture from an economic point of view. The growth hormone (GH) gene is crucial for selection in commercially cultivated fish species for better growth and production. Indian major carp (L. rohita, C. catla, and C. mrigala) are commonly cultured in Pakistan. The GH expression was examined using qPCR to understand growth in fish species better. Muscle tissue samples (n=480) from 160 individuals of the same age were collected from three species (L. rohita, C. catla, and C. mrigala). Individuals were divided into two groups (high-weight and low-weight groups), cultured under normal conditions. The housekeeping gene ß-actin validated GH expression in fast and slow-growing fishes from the same species. Results showed that GH expression varies across species and fish specimens that overweight their counterpart feature have higher GH expression. A selection for overweight fish in the aquaculture breeding systems is preferable as those fish could inherit their genomics to the future cohort, enhancing production, and commercial profit for farmers. Comprehensive research about different growth genes and the environmental aspects that influence fish growth is mandatory. No work has been reported regarding the growth gene analysis of fish from Pakistan. This report was Pakistan's first and baseline study regarding growth analysis of main culturable fish species at the molecular level.

Environmental impacts of industrial activities on floral coverage with special emphasis on detoxification enzyme activities in Cataglyphis savignyi as pollution biomarker.

The present study investigates the environmental impact of industrial activities on floral coverage within the major industrial district of Borg El-Arab City, Egypt. Additionally, it aims to evaluate the detoxification enzymatic activity of Cataglyphis savignyi as a pollution biomarker. To achieve this objective, seasonal soil samples were collected from the studied sites to determine soil properties and heavy metal concentrations. Furthermore, a seasonal specimen of C. savignyi was collected to study the enzymatic activity of acetylcholinesterase (AChE) and glutathione-S-transferase (GST). Heavy metal contamination pollution indices were calculated, and fourteen plant species were identified at the investigated sites for four successive seasons from 2020 to 2021. The soil physicochemical parameters significantly varied in the industrial sites compared to the control site. The accumulation of heavy metal contamination in soil for investigated sites followed the order Ni > As > Pb > Hg. Calculated Cdeg and PLI for industrial 3 revealed a very high degree of contamination, attributed to increased industrial activity from the chemical and silicate factories that characterize this region. The current results highlight the inhibition of GST levels in C. savignyi at the industrial site compared to the control site. In contrast, AChE increases, which might be due to heavy metals enhancing acetylcholine activity at synapses. Consequently, the antioxidant enzymatic activities are useful as biomarkers for assessing and monitoring environmental contamination. In conclusion, this study underscores insects as potent biomarkers for heavy metal contamination, marking a significant advancement in environmental monitoring. These bioindicators offer crucial insights into the impacts of climate change and industrial pollution. The research reveals distinct plant diversity variations and higher heavy metal content in industrial sites, indicating pronounced contamination. Additionally, the study highlights altered enzyme activities in insects, emphasizing their utility as biomarkers for assessing environmental contamination. This work represents a substantial leap forward in comprehending the complex dynamics between contamination and ecological balance.

Molecular docking and antibacterial activity of Sargassum fusiforme extracts against major coral pathogens.

The present study evaluates the antibacterial properties of alkaloids and the crude extracts (ethanol, n-hexane and ethyl acetate) from seaweed Sargassum fusiforme against coral pathogens (Photobacterium galatheae, Vibrio harveyi, Bordetella trematum, and Ochrobactrum pseudogrignonese) isolated from coral Porites lutea. To our knowledge, this is the first in vitro assay for such extracts on Porites lutea coral pathogens. Bacterial pathogens have been identified using 16S RNA and BankIt into gene bank and given the accession numbers (OR401000; OR401001; OR401336, and OR400998 respectively). GC-Mass profiling conducted for n-hexane compounds confirmed the presence of thirty-eight molecules, twelve of which have been previously reported for their bioactivity. The results revealed that alkaloids and n-hexane extract demonstrated eminent antibacterial activity compared to the other extracts against the tested coral pathogenic bacteria. Molecular docking was conducted to evaluate the twelve previously mentioned bioactive molecules to get a full understanding of the interaction of those bioactive molecules on vital bacterial proteins (Hemolysin protein (PDB ID: 1XEZ) and Cytoplasmic proteins (PDB ID: 3TZC)). Docked twelve molecules against hemolysin protein (PDB ID: 1XEZ) came exactly in line with the docked result of the same molecules with cytoplasmic proteins (PDB ID: 3TZC), proving the bioactivity of 6-O-Palmitoyl-L-ascorbic acid, 3TMS derivative; Glycerol monostearate, 2TMS derivative and Eicosanoic acid complexes in antibacterial activity action and score higher than reference ligand. Those three compounds will be investigated separately in future in vitro assay soon. Our conclusions align with the study's antibacterial in vitro assay results. The present study reports the novelty of different extracts of S. fusiforme as an antibacterial agent against coral pathogenic bacteria that trigger diseases in Porites lutea.

Bioaccumulation, Biosedimentation, and Health Hazards of Elements in Crayfish, Procambarus clarkii from El-Rahawi Drain and El-Qanatir in the River Nile, Egypt.

Elements accumulation in crayfish is proportional to the increase in bioavailability (direct contact) with the surrounding water, sediment, and feeding. Five heavy metals (Cu, Cr, Mn, Ni, and Ag) and lithium (Li) were analyzed in the sediment, water, and crayfish tissues. Elements (heavy metals and lithium) concentrations in sediment, water, and crayfish tissues showed significant differences between the two sampling stations (El-Qanatir and El-Rahawi drain). However, the levels of elements in crayfish tissues were arranged in declining order as hepatopancreas > gills > exoskeleton > muscles for Cu and Cr; hepatopancreas > exoskeleton > gills > muscles for Ni and Ag; and exoskeleton > gills > hepatopancreas > muscles for Li and Mn. The human health hazard evaluation of heavy metals and lithium exposure via edible tissue consumption was assessed for both children and adult consumers. The target hazard quotient THQ values of crayfish edible tissues (less than 1) will not impose any health implications for consumers who ingest edible tissues in sufficient quantities. Furthermore, the hazard index (HI) values reported for children and adult consumers were lower than one, indicating non-carcinogenic and carcinogenic hazards, suggesting that crayfish edible tissues are safe for human ingestion. This evidence also found that Procambarus clarkii could be a good bio-indicator organism for monitoring potentially metals in aquatic systems.

Daphnia magna and Gammarus pulex, novel promising agents for biomedical and agricultural applications.

Various studies have shown the importance of using different types of Zooplankton biomasses as an additional substance in the diet of fish. In addition, the drainage water of the fish cultures could be used in plant irrigation. In this study, biomasses of water flea Daphnia magna and Gammarus pulex collected and tested, for the first time, their effect against pathogenic microorganisms and on plant germination. The results showed significant antibacterial activity of D. magna and G. pulex against Staphylococcus aureus and Pseudomonas aeruginosa bacteria, as well as antifungal activity against Alternaria solani and Penicillium expansum, which gives the possibility to be used as biocontrol against these bacteria and plant pathogenic fungi. Furthermore, both animals showed positive activity in the germination rate of Vicia faba seed, reaching 83.0 ± 3.5 and 86.0 ± 3.8%, respectively. In conclusion, the biomasses of D. magna and G. pulex are promising and effective agents for their use in the medical field against some pathogenic microbes and as stimulators of plant growth.

Effect of replacing fish meal in fish diet with shrimp by-product meal on growth performance, feed utilization, length-weight relationship and condition factors of Nile tilapia, Oreochromis niloticus (Linnaeus, 1758) / Efecto de reemplazar la harina de pescado en la dieta de pescado por harina de subproducto de camarón sobre el rendimiento del crecimiento, la utilización del alimento, la relación longitud-peso y los factores de condición de la tilapia del Nilo, Oreochromis niloticus (Linnaeus, 1758)

SUMMARY: The present work aimed to study the effect of replacing fish meal (FM) in the fish diet with shrimp by-product meal (SBM) on the growth performance parameters of the Nile Tilapia, Oreochromis niloticus. A total of 200 specimens of monosex O. niloticus fries were obtained from a private fish farm at Kafr El-Sheikh Governorate. They were transported to the fish laboratory at the Animal House of Zoology Department, Faculty of Science, Al-Azhar University. After two weeks for acclimation, the fish were divided into 5 groups, 2 replicates for each group (20 fish in each replicate). The five experimental diets were: C: control group with20 % fish meal (FM) and 0 % shrimp by-product meal (SM); T1, T2, T3, and T4 FM was replaced with SBM as 25 %, 50 %, 75 % and 100 %, respectively. Results indicated that the highest final length, growth in length, length gain, daily length gain and other growth performance parameters including the relationship between length and weight and condition factors of O. niloticus were recorded in T4 group (fed diet in which fish meal was completely replaced with shrimp by-product meal); while, their lowest values were recorded in control group. On the other hand, feed utilization parameters (feed intake, food conversion ratio, the maximum values of feed efficiency ratio and protein efficiency ratio) were recorded in T4 group and the minimum values were recorded in T3 group (fed diet in which 75 % of fish meal was replaced with shrimp by-product meal).

RESUMEN: El presente trabajo tuvo como objetivo estudiar el efecto de sustituir la harina de pescado (HP) en la dieta del pescado por harina de subproductos de camarón (HSC) sobre los parámetros de rendimiento de crecimiento de la Tilapia del Nilo, Oreochromis niloticus. Se obtuvieron un total de 200 especímenes de monosexo de O. niloticus de una piscifactoría privada en la gobernación de Kafr El-Sheikh. Fueron transportados al laboratorio de peces en el Departamento de Zoología de la Facultad de Ciencias, de la Universidad Al-Azhar. Después de dos semanas de aclimatación, los peces se dividieron en 5 grupos: Se realizaron dos repeticiones para cada grupo (20 peces en cada repetición). Las cinco dietas experimentales fueron: C: grupo control con 20 % de harina de pescado (HP) y 0 % de harina de subproductos de camarón (HSC); T1, T2, T3 y T4 FM se reemplazó con HSC con 25 %, 50 %, 75 % y 100 %, respectivamente. Los resultados indicaron que la longitud final más alta, el crecimiento en longitud, la ganancia de longitud, la ganancia de longitud diaria y otros parámetros de rendimiento del crecimiento, como además la relación entre la longitud y el peso, y los factores de condición de O. niloticus, se registraron en el grupo T4 (con una dieta reemplazada con harina de subproducto de camarón); mientras que, sus valores más bajos se registraron en el grupo control. Por otro lado, los parámetros de utilización del alimento (ingesta de alimento, índice de conversión de alimento, los valores máximos de índice de eficiencia alimenticia e índice de eficiencia proteica) se registraron en el grupo T4 y los valores mínimos se registraron en el grupo T3 (alimentación con dieta en la que el 75 % de la harina de pescado fue reemplazada por harina de subproductos de camarón).