Implications of Heavy Metal Accumulation in Fish Feed, Water, Sediment, and Different Fish Species in a Polyculture System.

Heavy metal bioaccumulation in organisms is primarily a result of dietary uptake. The current study examines the concentrations of heavy metals (Pb, Cd, Cr, and Cu) in fish feed, water, sediment, and three fish species (Catla catla, Labeo rohita, and Cyprinus carpio) from different feeding zones in a polyculture pond system. Furthermore, associated human health risks were also evaluated. The fish samples (n = 25 for each species) were collected from 10 different fish ponds in the Kohat district, Pakistan. Heavy metals were determined using an atomic absorption spectrometer. Results revealed higher concentrations of heavy metals in sediment, followed by water. However, the concentration of heavy metals in fish feed was lower than the standard limits. In the case of fish, the bottom feeder (C. carpio) notably exhibited higher (P < 0.05) levels of heavy metals than the column feeder (L. rohita) and surface feeder (C. catla) fish. Moreover, in the liver of all fish species, the bioaccumulation of heavy metals was higher, followed by the gills. Principal component analysis (PCA) demonstrated a strong correlation of heavy metals in C. carpio gills, flesh, feed, and pond water, while the heavy metals in the liver correlated with the detected metals in sediment. The human health risk analysis shows that bottom feeder fish had higher estimated daily intake (EDI), target hazard quotient (THQ), and hazard index (HI) values (> 1). Consequently, the exposed population may experience adverse health effects. The findings of this study suggest that the bottom feeder (C. carpio) bioaccumulates a higher concentration of heavy metals than column (L. rohita) and surface feeder (C. catla) in the polyculture system.

Insights into sequence characteristics and evolutionary history of DGATs in arthropods.

Triacylglycerol (TAG) is crucial in animal energy storage and membrane biogenesis. The conversion of diacylglycerol (DAG) to triacylglycerol (TAG) is catalyzed by diacylglycerol acyltransferase enzymes (DGATs), which are encoded by genes belonging to two distinct gene families. Although arthropods are known to possess DGATs activities and utilize the glycerol-3-phosphate pathway and MAG pathway for TAG biosynthesis, the sequence characterization and evolutionary history of DGATs in arthropods remains unclear. This study aimed to comparatively evaluate genomic analyses of DGATs in 13 arthropod species and 14 outgroup species. We found that arthropods lack SOAT2 genes within the DGAT1 family, while DGAT2, MOGAT3, AWAT1, and AWAT2 were absent from in DGAT2 family. Gene structure and phylogenetic analyses revealed that DGAT1 and DGAT2 genes come from different gene families. The expression patterns of these genes were further analyzed in crustaceans, demonstrating the importance of DGAT1 in TAG biosynthesis. Additionally, we identified the DGAT1 gene in Swimming crab (P. trituberculatus) undergoes a mutually exclusive alternative splicing event in the molt stages. Our newly determined DGAT inventory data provide a more complete scenario and insights into the evolutionary dynamics and functional diversification of DGATs in arthropods.

Assessment and Bioaccumulation of Heavy Metals in Water, Fish (wild and Farmed) and Associated Human Health Risk.

Worldwide, anthropogenic activities are causing the natural environment and aquaculture systems to become heavily contaminated with heavy metals, which can lead to consumer's health problems. In the current study, wild and farmed fish (n = 30) and water samples (n = 6) have been collected from the Chashma barrage and fish farm to assess the heavy metals concentration, i.e., Cu, Cd, Pb, Zn and Cr, in the water and some important organs (gills, liver, muscle, brain and bones) of wild and farmed fish (Labeo rohita) using Graphite furnace Atomic absorption spectrometry. Bioaccumulation factor and human health risk assessment were calculated to measure the health status of both fish and humans. Results show that in wild and farm fish's gills, muscles and bones, the trend of the heavy metals was Zn > Pb > Cu > Cd > Cr. On the other hand, the brain and liver show Zn > Cu > Pb > Cd > Cr trend. Comparatively, the heavy metals concentration was mostly higher (P < 0.05) in wild fish. Further; in both fish habitats (water) the heavy metals (Cd and Pb) concentration was higher than the WHO standard level, while in the body, Cd was higher (P < 0.05) in all studied organs except the muscle, Cr was only lower (P > 0.05) in muscle and brain. Pb was higher (P < 0.05) in all studied organs of both fish. Bioaccumulation of heavy metals was mostly higher (P < 0.05) in wild fish than in farmed. EDI and THQ were higher in wild fish, but the HI value was lower than 1 for both fish. Moreover, the PCA analysis suggests a positive correlation between heavy metals concentration in fish organs and the water of both fish (wild and farmed). Results indicated that farmed fish showed less potential risk to humans than wild fish.

Evaluation of the antibacterial activity and protein profiling of Nile tilapia (Oreochromis niloticus) epidermal mucus under different feeds and culture systems (biofloc technology and earthen pond).

The mucus layers of fish serve as the main interface between the organism and the environment. They play an important biological and ecological role. The current study focuses on Nile tilapia epidermal mucus reared under different commercial feeds (coded A and B) and environments (biofloc technology and earthen pond systems). Crude protein levels in feed A and B were 30% and 28%, respectively. Water parameters in all culturing systems were suitable for tilapia throughout the study period. The antimicrobial potency of tilapia (n = 5 from each) epidermal mucus was tested in vitro against human and fish pathogenic strains viz. Staphylococcus epidermidis, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Francisella noatunensis, and Aeromonas hydrophila. To determine the antimicrobial activity, zones of inhibition (ZOI) were measured in millimetres and compared with two antibiotics (chloramphenicol and ciprofloxacin). SDS-PAGE analysis was performed on skin mucus samples of tilapia to determine protein quantity and size (molecular weight). Results of tilapia skin mucus (crude and aqueous) revealed a strong antibacterial effect against all the selected pathogenic strains. However, variation has been observed in the mucus potency and ZOI values between the biofloc and pond tilapia mucus. The crude mucus of tilapia fed on feed A and cultured in the pond exhibited strong antibacterial effects and high ZOI values compared to the mucus of biofloc tilapia, aqueous mucus extracts and positive control chloramphenicol (antibiotic). The SDS-PAGE results showed that the high molecular weight proteins were found in the collected epidermal mucus of BFT-B (240 kDa) and EP-B (230 kDa). Several peptides in fish skin mucus may play a crucial role in the protection of fish against disease-causing pathogens. Thus, it can be utilized in the human and veterinary sectors as an 'antimicrobial' for treating various bacterial infections.

Investigation of the incidence of heavy metals contamination in commonly used fertilizers applied to vegetables, fish ponds, and human health risk assessments.

Overuse of fertilizers on agricultural lands and fish ponds may result in serious pollution problems, such as heavy metals that can enter the food chain and pose serious health problems. Due to this, the present study investigates the incidence of heavy metals in commonly used fertilizers and its association with heavy metals in vegetables, soil, fish species, and pond water. Samples were collected from different sites (fields and ponds) in district Kohat, where the application of fertilizers was common and control groups (no fertilizers used). Heavy metal analysis was carried out through a spectrophotometer. Results showed higher Cd and Cr concentrations in triple superphosphate (TSP), Cu and Pb in nitrogen, phosphorus, and potassium (NPK), while lower concentrations were found in gypsum. In vegetables (onion, tomato, brinjal, and potato) and associated soil, most of the heavy metals concentrations were significantly higher (P < 0.05) in fertilizer-applied sites than in the control. Also, the Cd concentration in potatoes and Pb level in all vegetables obtained from sites were greater than the WHO/FAO standard limit. In the case of fish species (Hypophthalmichthys molitrix and Cyprinus carpio) muscles and their habitat (water), all the understudy heavy metals were notably higher (P < 0.05) in fertilizer-applied sites (ponds) than the control group. Collectively, in all vegetables and muscles of fish species, the bioaccumulation factor was higher in sites compared to the control. The estimated daily intake (EDI) and target hazard quotient (THQ) values were also higher in fertilizer-applied sites (fields and ponds) than control. The health index (HI) value was > 1 in vegetables (onion, tomato, and potato) and fish muscles collected from different sites compared to the control. Thus, there is the possibility of severe health risks. The use of fertilizers must be carefully monitored in order to ensure that humans and animals are safe from exposure to heavy metals.

Evaluation and Association of Heavy Metals in Commonly Used Fish Feed with Metals Concentration in Some Tissues of O. niloticus Cultured in Biofloc Technology and Earthen Pond System.

For successful aquaculture, the primary need is the quality of fish feed, which determines fish flesh quality. The current study was conducted to evaluate the number and concentration of heavy metals in commonly used fish feeds and fish gills, liver, and muscle of biofloc technology and earthen pond systems. Besides this, the correlation between heavy metals in fish feeds with detected metals in the gills, liver, and muscle of fish was also determined. Results revealed that heavy metals concentration, including Cu and Cd, in feed B was significantly greater than in feed A, but the Zn level in feed A was significantly higher than in feed B. Furthermore, the concentration of heavy metals in fish of both aquaculture systems was significantly higher in the liver than in the gills and muscle. The metal concentration in fish feeds and fish edible parts (muscle) was lower than the WHO standard level; however, the amount of Pb was higher in the fish muscle, which is harmful for human consumption. Though the correlation test revealed that all of the metals from the feeds were positively correlated to the metals detected in the fish, but most of the estimated correlation was insignificant. From the current study, it can be concluded that the fish feed producers need to measure feed quality adequately to avoid hazardous contamination by heavy metals in the feed. The ultimate consumer, fish and humans, may, otherwise, be predisposed to assimilate and accumulate these heavy metals.

Withania coagulans fruit extract: a possible useful additive in ameliorating growth and immunity of Labeo rohita (Hamilton, 1822).

This study aimed to assess the Withania coagulans fruit extract's effects on growth and haematological parameters of Labeo rohita. Healthy fish (n = 120) were divided in four groups. Experimental diet was prepared with different extraction of W. coagulans (0%, 1%, 1.5% and 2%). Diet was provided for eight weeks to all groups except the control group which fed on basal diet. The 1.5% and 2% W. coagulans extracts lead improvement in feed conversion ratio and specific fish growth rate; haematological parameters in fish group fed with 2% of W. coagulans showed increased in WBC, RBC, Hb, MCHC, PCV. The values of plasma proteins and lysozyme activity were higher in the experimental group compared control group. Fish fed with W. coagulans extracts showed increased of immunity and less mortality respect to others. It suggested that W. coagulans could be considered valuable support for the improve growth and immunity in Labeo rohita.

Assessment and Bioaccumulation of Heavy Metals in Fish Feeds, Water, and Some Tissues of Cyprinus carpio Cultured in Different Environments (Biofloc Technology and Earthen Pond System).

Fish feed quality is the main determinant of fish flesh quality, so it is important for successful aquaculture. The current study determines the concentration of heavy metals in fish feeds (A and B), water, and their bioaccumulation in gills, liver, and muscle of C. carpio cultured in different environments (biofloc technology and earthen pond systems). In addition, the correlation between heavy metals in fish feeds with bioaccumulated metals in fish tissues was also determined. Results revealed that most heavy metal concentration was significantly greater (P < 0.05) in feed B than in feed A but in permissible range, while all the heavy metal concentration was notably higher in earthen ponds than in biofloc technology. Result from the bioaccumulation factor and concentration of the metals showed that heavy metals were highly accumulated in the fish liver followed by gills. The metal concentration in fish feeds and fish edible parts (muscle) was lower than the WHO standard level; however, the amount of Pb was higher in the fish muscle, liver, and gills, which is harmful for human consumption and also for fish health. Though the correlation test revealed that all of the metals from the feeds were positively correlated to the metals in fish tissues, but most of the estimated correlation was significant and linearly correlated. It can be concluded that producers must measure feed quality correctly to avoid heavy metal contamination because it may assimilate and accumulate in the food chain.

Cadmium Sub-Lethal Concentration Effect on Growth, Haematological and Biochemical Parameters of Mystus seenghala (Sykes, 1839).

The current study aims to assess the cadmium sub-lethal concentration influence on growth and haematological and biochemical parameters of Mystus seenghala. A total of 60 fish of three different length groups (20 each) were collected from Head Qadirabad, Pakistan. The fish were treated to the sub-lethal concentration viz. one-third of LC50, for 16 weeks except for the control groups. Water quality parameters were kept constant during the entire course of the research, and the major parameters were measured as temperature (28.03 ± 0.03 °C), DO (5.82 ± 0.14 mg L-1), pH (8.00 ± 0.01) and total hardness (249.98 ± 0.01 mg L-1). Findings revealed that the growth of three treated variant length groups was affected negatively by cadmium exposure and showed significantly (P < 0.05) lower average wet weight, body length and condition factor as compared to control groups, while the feed conversion ratio (FCR) increases by increasing the exposure duration. The haematological parameters including values of Hct, Hb and MCHC were significantly (P < 0.05) reduced in all Cd-treated groups than control groups, whereas the level of MCH and MCV were significantly higher, but no significant difference was found in the value of RBCs in all treated groups. Biochemical parameters such as ALT, AST, total lipid and glucose level in Cd exposure groups were significantly higher, while the total protein level was significantly (P < 0.05) reduced in all treated groups as compared to control groups. From the current study, it has been concluded that the growth, haematology and biochemical parameters are important indicators of ecotoxicology particularly contamination of the cadmium and health of the fish.

Effect of fortified feed with olive leaves extract on the haematological and biochemical parameters of Oreochromis niloticus (Nile tilapia).

Olive leaves have been used traditionally to improve the immunity against several infections. Current study aimed to examine the effect of fortified feed with olive leaves extract on haematological and biochemical parameters of Oreochromis niloticus. Total 400 healthy fish were divided randomly into four groups and were cultured for two months in glass tanks of 250 L water capacity in triplicate. Fortified feed was prepared with various extract, 1%, 1.5% and 2% concentration in diet, while, control-group diet was without extraction. Results showed that all the haematological parameters including WBC, RBC, Hb, PCV and plasma protein were significantly increased while the biochemical parameters such as globulin albumen ratio increased in fish which fed on 1% of extract, but the level of cholesterol and triglycerides was decreasing by increasing the concentration of extract. It was concluded that olive leaves extract can be used to enhance the immunity, growth and health of fish.

Effect of Fortified Feed with Phyto-Extract on the First Physical Barrier (Mucus) of Labeo rohita.

The aim of the current study was to assess the effect of two different fortified feeds with different concentrations of two important medicinal plants (Withania coagulans and Zingiber officinale) on the mucosal immunity of Labeo rohita. After a dietary intervention, mucus was tested against five pathogenic bacteria (in-vitro), while experimental fish were tested against the ectoparasite (Lernaea) (in-vivo). Our results revealed that all fish groups fed with different concentrations (1, 1.5, and 2%) of Z. officinale had low molecular weight proteins and did not develop any significant signs of parasitic infection, with low mortality rate; whereas the groups that were fed with W. coagulans (particularly with 1% and 2%), including a control group, developed rapid signs of infection with high mortality rate. The highest hemagglutination titer value was recorded for the fish fed with 1% and 1.5% of Z. officinale. The lowest value was found for the fish fed with 2% of W. coagulans. The mucus of all fish of fortified groups was active and inhibited the growth of tested bacterial pathogens as compared to the control group. Further, Z. officinale groups showed greater efficacy against bacteria as compared to the W. coagulans groups. In conclusion, Z. officinale can be considered as a potential and functional ingredient in aquaculture feed. Furthermore, future studies should be conducted to investigate more details on the subject.

Accuracy of Specimen Radiography in Assessing Complete Local Excision with Breast-Conservation Surgery

Objective: The aim of this study was to evaluate the accuracy of "X- ray examination of surgically resected specimen'' in assessing complete local excision (CLE). Materials and Methods: In this retrospective cross sectional study, data were collected for all female breast cancer cases who underwent breast-conserving surgery after needle localization of mammographically visible disease. Males, patients with mammographically invisible disease and cases with benign or inconclusive histopathology, those undergoing modified radical mastectomy and individuals with dense breast parenchyma were excluded. We evaluated radiography of resected specimens to assess margin spiculation, distance of mass/microcalcification from the excised margin, presence of a mass, and presence of any adjacent microcalcification, Other features including mass size, nuclear grade and patient's age were also recorded and all were analyzed for any association with CLE. Results: Absence of adjacent microcalcification and the presence of a mass on radiographs showed significant associations with CLE, but no links were evident with other features. Specimen radiography was found to be a sufficient tool to predict CLE with a positive predictive value of 83.3%, a sensitivity of 80.7% and a specificity of 81%. Conclusion: Specimen radiography is an important and sensitive tool to predict CLE.

Negative Predictive Value of Ultrasound in Predicting Tumor-Free Margins in Specimen Sonography.

OBJECTIVE: To evaluate the success of ultrasound in post-excision specimen visualization, and negative predictive value of ultrasound for estimation of tumor-free margins using histopathology as the gold standard. STUDY DESIGN: Cross-sectional analytical study. PLACE AND DURATION OF STUDY: The Aga Khan University Hospital, Karachi, Pakistan, from May 2010 till January 2013. METHODOLOGY: Sonography of all breast nodules was done before and after exicision by two female radiologists with at least five years clinical experience. All surgeries were performed by the same referring breast surgeons. All nodules were non-palpable and had histopathology as well as specimen sonography performed at AKUH. Subjects were excluded, if histopathology was not available, post-procedure sonogram not done or done in another hospital and nodules that were not seen on ultrasound. After needle localization in 47 patients using ultrasound and in 7 patients using mammogram was done, sonogram was conducted in all 54 lesions. These were then assessed by ultrasound for detection of lesion and tumor-free margins in malignant lesion. Post-excision ultrasound was performed for the evaluation of lesion whether visualized or absent with localizing needle in situ, lesion dimensions, depth measurement between the superior margin of the lesion and its edge. RESULTS: All 54 lesions were present on post-exicison scan, out of which 28 were documented as malignant and 26 as benign. Ultrasound declared all specimens as tumor-free. On histopathology, two lesions were documented as having tumor-positive margins and were proven to be invasive lobular carcinoma. Therefore, the negative predictive value of the specimen sonography for margin detection was 26/28 (92.8%). CONCLUSION: Ultrasound of the excised breast tumor specimen is a simple and reliable technique for confirmation of the tumor-free margins in non-palpable breast lesions.

Transverse Vaginal Septum: Report of Two Patients with MRI Findings.

The transverse vaginal septum is a rare form of Mullerian duct anomaly, and is a disorder of vertical fusion of Mullerian system with the sinovaginal bulb. Magnetic resonance imaging (MRI)is very useful in the detection, classification and management planning of this rare anomaly. We present two cases of transverse vaginal septum in two patients of different age groups and with different clinical presentations. The findings on MRI, related to presence of vaginal septum and the management work-up, are also discussed.