Metabolic Profiling and In Vitro Assessment of the Biological Activities of the Ethyl Acetate Extract of Penicillium chrysogenum “Endozoic of Cliona sp. Marine Sponge” from the Red Sea (Egypt)

Marine sponge-derived endozoic fungi have been gaining increasing importance as promising sources of numerous and unique bioactive compounds. This study investigates the phytochemical profile and biological activities of the ethyl acetate extract of Penicillium chrysogenum derived from Cliona sp. sponge. Thirty-six compounds were tentatively identified from P. chrysogenum ethyl acetate extract along with the kojic acid (KA) isolation. The UPLC-ESI-MS/MS positive ionization mode was used to analyze and identify the extract constituents while 1D and 2D NMR spectroscopy were used for kojic acid (KA) structure confirmation. The antimicrobial, antioxidant, and cytotoxic activities were assessed in vitro. Both the extract and kojic acid showed potent antibacterial activity against Staphylococcus aureus and Pseudomonas aeruginosa with MIC 250 ± 0.82 µg/mL. Interestingly, the extract showed strong antifungal activity against Candida albicans and Cryptococcus neoformans with MIC 93.75 ± 0.55 and 19.53 ± 0.48 µg/mL, respectively. Furthermore, KA showed the same potency against Fusarium oxysporum and Cryptococcus neoformans with MIC 39.06 ± 0.85 and 39.06 ± 0.98 µg/mL, respectively. Ultimately, KA showed strong antioxidant activity with IC₅₀ 33.7 ± 0.8 µg/mL. Moreover, the extract and KA showed strong cytotoxic activity against colon carcinoma (with IC₅₀ 22.6 ± 0.8 and 23.4 ± 1.4 µg/mL, respectively) and human larynx carcinoma (with equal IC₅₀ 30.8 ± 1.3 and ± 2.1 µg/mL, respectively), respectively. The current study represents the first insights into the phytochemical profile and biological properties of P. chrysoenum ethyl acetate extract, which could be a promising source of valuable secondary metabolites with potent biological potentials.     

MicroRNA-21 expression,serum tumor markers,and immunohistochemistry in canine mammary tumors

Background

Canine mammary tumors (CMTs) are one of the most common malignancies in dogs and are associated with significant mortality. Serum tumor markers and non-coding microRNAs have gained widespread popularity in human oncology studies. The present study has two aims, first one is to investigate the miR-21 expression compared with changes in serum tumor markers (CEA and CA15-3) in CMT. The second aim is to detect the immunohistochemistry markers as vimentin, P63, and -SMA in CMT.

Methods

This study enrolled 17 female dogs: 10 with mammary tumors and seven controls without tumors. Blood samples were collected to measure miR-21, CEA, and CA 15-3, and histological samples were prepared for histological grading and immunohistochemistry.

Results

CA 15-3 was elevated in all animals, whereas CEA levels showed no change compared with controls. miR-21 was upregulated 12.84-fold in animals with CMT. The most frequently recorded CMT was the mixed type. Myoepithelial cells were identified by P63 immunoreactivity, but not SMA. High expression of miR-21 was observed with positive vimentin immunoreactivity, indicating the mesenchymal origin of the tumor cells.

Conclusion

The present study showed that miR-21 was elevated to a greater extent than CA 15-3 (12.84-fold vs. threefold). Tumors that was positive for vimentin immunoreactivity was also associated with an elevation in the levels of miR-21, showing that miR-21 is released from mesenchymal cells. These findings support the hypothesis that miR-21 may be a more sensitive, noninvasive indicator for CMT.

     

Effect of a single injection of gonadotropin-releasing hormone (GnRH) and human chorionic gonadotropin (hCG) on testicular blood flow measured by color doppler ultrasonography in male Shiba goats

Although color Doppler ultrasonography has been used to evaluate testicular blood flow in many species, very little has been done in goat. Eight male Shiba goats were exposed to a single intramuscular injection of either gonadotropin-releasing hormone (GnRH group; 1 µg/kg BW) or human chorionic gonadotropin (hCG group; 25 IU/kg BW). Plasma testosterone (T), estradiol (E2) and inhibin (INH) were measured just before (0 hr) and at different intervals post injection by radioimmunoassay. Testis volume (TV) and Doppler indices, such as resistive index (RI) and pulsatility index (PI) of the supratesticular artery, were measured by B-mode and color Doppler ultrasonography, respectively. The results indicated an increase in testicular blood flow in both groups, as RI and PI decreased significantly (P<0.05), but this increase was significant higher and earlier in hCG group (1 hr) than in the GnRH group (2 hr). A high correlation was found for RI and PI with both T (RI, r= −0.862; PI, r= −0.707) and INH in the GnRH group (RI, r=0.661; PI, r=0.701). However, a significant (P<0.05) correlation was found between E2 and both RI (r= −0.610) and PI (r= −0.763) in hCG group. In addition, TV significantly increased and was highly correlated with RI in both groups (GnRH, r= −0.718; hCG, r= −0.779). In conclusion, hCG and GnRH may improve testicular blood flow and TV in Shiba goats.     

Combining the use of molecular techniques and archival documentary evidence to trace the origin of <Emphasis Type="Italic">Populus alba</Emphasis> in a Central Mediterranean archipelago

The nativity of Populus alba in the Mediterranean has only been confirmed in the last decade, following the discovery of 8,000-year-old leaf imprints in Southern France. Recent evidence has even emerged from molecular studies suggesting that the species is native to some of the islands, and these populations may be relicts of a native flora that arrived there much earlier than previously thought. In view of this, samples obtained from the Central Mediterranean archipelago of Malta and other neighbouring regions were analysed to determine the native status of the Maltese populations and possibly trace their origins. All 38 samples were investigated in order to assess the genetic diversity and origin of Maltese trees. Nuclear microsatellite analysis revealed that all 28 trees sampled from the two islands of Malta belonged to one clone. Chloroplast data suggested relatedness of the Maltese clone to Italian P. alba samples. However, nuclear data suggested additional admixture through pollen from North Africa. Existing archival and palaeontological records were also examined for any supporting evidence. On considering the latter records in combination with molecular evidence, we arrived to the conclusion that arrival of this clone in Malta through human introduction in the sixteenth century is the most likely explanation, since alternative scenarios like autovegetative propagation or arrival by seed seem highly unlikely.     

Zingiber officinale and Glycyrrhiza glabra,individually or in combination,reduce heavy metal accumulation and improve growth performance and immune status in Nile tilapia,Oreochromis niloticus

This study evaluated the impact of dietary ginger and liquorice supplementation on growth performance, physiological and histopathological profiles and heavy metal accumulation in Nile tilapia fingerlings. Fish (n = 1,800, 17.5 ± 0.11 g BW) were randomly distributed into four treatment groups in triplicates and received no supplementation (control group), 5 ml aqueous ginger extract/kg feed (ginger group), 4 ml aqueous liquorice extract/kg feed (liquorice group) or 2.5 ml ginger plus 2 ml liquorice aqueous extracts/kg feed (mix group). The ginger‐liquorice mix supply improved the growth performance and feed efficiency (p < .05), increased the haematocrit and haemoglobin (p < .05), leucocytes (p = .108), neutrophils (p = .054), serum total protein (p < .05), albumin (p = .011) and globulin (p = .094) but decreased (p < .05) the blood urea nitrogen and creatinine than feeding liquorice or ginger lonely compared to the control. Heavy metal loads in pond water induced lamellar telangiectasis of gills and necrosis with sloughing of intestinal villi tips. These detrimental effects were alleviated, and the intestinal villus length (p = .041) and crypt depth (p = .069) were increased with liquorice supply. In all treatment groups, heavy metal contents in fish flesh were lower compared to the control. Thus, using ginger and/or liquorice aqueous extracts can decrease heavy metal accumulation in the fish flesh and exert positive effects on growth performance, metabolic profile and the intestinal and gill morphology of Nile tilapia.     

Exogenous xylanase improves growth,protein digestibility and digestive enzymes activities in Nile tilapia,Oreochromis niloticus,fed different ratios of fish meal to sunflower meal

A 3 × 2 factorial experiment was designed to evaluate the effect of different ratios of fish meal (FM): sunflower meal (SFM) with or without exogenous xylanase supplementation on growth, feed utilization, digestive enzymes activities, apparent digestibility, intestinal and liver morphology and chemical composition of Nile tilapia, Oreochromis niloticus. Three isonitrogenous (329.80 g/kg of crude protein) and isoenergetic (18.46 MJ/kg gross energy) experimental diets were formulated as SFM₁ (FM:SFM = 2:1), SFM₂ (FM:SFM = 1:1) and SFM₃ (FM:SFM = 1:2) based on protein content. Each diet was supplemented with 0 or 0.5 g/kg of exogenous xylanase and was fed to triplicate groups of twelve fish (with initial weight, 1.31 ± 0.02 g) for 84 days. After 84 days of feeding period, the highest weight gain, specific growth rate, protein efficiency, protein productive value and the best feed conversion ratio were recorded in fish fed either SFM₁ or SFM₂ supplemented with exogenous xylanase. Whereas lowest growth performance was recorded in fish fed SFM₂ and SFM₃ un‐supplemented with xylanase. The highest activities of chymotrypsin, trypsin, lipase, amylase, alkaline phosphatase and cholecystokinin were observed in fish fed SFM₁ and SFM₂ diets supplemented with xylanase. The highest ADCs of dry matter, protein, lipid and digestible energy were recorded in fish fed SFM₁ and SFM₂ diets supplemented with exogenous xylanase. Supplementation of exogenous xylanase improved muscularis mucosa thickness, height of mucosal folds and enterocytes of intestinal fish. Addition of exogenous xylanase increased the calcium and phosphorus retention. Results of this study indicated that the addition of exogenous xylanase to diet containing high inclusion level of sunflower meal improved growth, digestive enzymes, nutrient digestibility, histological morphometric of liver and intestine and nutrient retention.     

Influence of different microalgal diets on gonadal development of the carpet shell clam <Emphasis Type="Italic">Ruditapes decussatus</Emphasis> broodstock

Studies considering broodstock conditioning of Ruditapes decussatus mostly used flagellates and diatoms for feeding, mainly Isochrysis galbana and Chaetoceros calcitrans, respectively. The present study evaluated the effect of different microalgal diets on gonadic maturation of R. decussatus including three microalgae species (C. calcitrans, Tetraselmis suecica, and Nannochloropsis oculata) which were used to prepare six diets: three monospecific and three bispecific mixed diets at a proportion of 1:1. Clams were daily fed at a ratio of 1% dry weight of algae/live weight of clam and unfed treatment was used as control. Results showed that clams fed C. calcitrans alone or mixed with T. suecica attained earlier full maturity with the highest response to spawning induction (after 34 conditioning days) and largest proportion in the partial spawning phase for both, followed by T. suecica alone after 48 days. In contrast, N. oculata alone or mixed diets demonstrated poor performance for gonadal maturation. Unfed treatment did not show any sign of maturation. The results indicated that T. suecica has high conditioning potential like C. calcitrans and their mixture gave the highest benefit. Therefore, T. suecica can be recommended for R. decussatus broodstock conditioning.     

Response of European seabass (<Emphasis Type="Italic">Dicentrarchus labrax</Emphasis>) to canola oil diets: effect on growth performance,fish health and liver and intestine histomorphology

This study was undertaken to assess the effects of feeding European seabass (Dicentrarchus labrax) canola oil-added diets on growth, health status and liver and intestine histomorphology. Seabass (56.18 ± 0.16 g initial body weight) were fed one of three fish meal-based diets with ~48 % crude protein and ~16.0 % lipid, combining fish oil (FO) and canola oil (CO) for 12 weeks. The diets contained: zero (control, CTRL), 45 (CO50) or 63 (CO70) g CO kg^?1 assigned in triplicates to three dietary groups. The results indicated that neither dietary oil type (FO or CO) nor CO level adversely affected (P > 0.05) the growth, feed utilization or major blood constituents’ composition as an indicator of the overall health status of fish. Despite the CO diets influence on head kidney macrophage activity being unappreciable (P > 0.05), there was a reducing trend with an increase in CO level incorporation. The CO70 diet induced a minor fat infiltration in hepatocytes and leucocytes infiltration, hyperplasia of the basal nuclei and supranuclear vacuolization of the enterocytes of the distal intestine. The present observations suggest that it is possible to incorporate up to 63 g CO Kg^?1 in the feed for European seabass juveniles without major negative effects on growth, health status or liver and intestinal histomorphology.