In this study, we determine the curative effects of okra pods (Abelmoschus esculentus L.) extract against lead acetate toxicity in mice kidney. n-Hexane, ethyl acetate, and methanol solvent were used for extracting okra pods. The role of the extract as an antioxidant was tested by DPPH and FRAP methods. The methanol extract was used for experiments in animals. A total of 30 male BALB/c mice were randomly divided into six equal groups: normal control, negative control (lead-induced), and treatment groups (lead-induced for 28 days and administration of methanol extract at doses of 50, 100, 200, and 400 mg/kg BW for the 28 days). The following were analyzed in all groups: activity of the antioxidant enzymes, namely, superoxide dismutase (SOD) and catalase (CAT); oxidant level, namely, malondialdehyde (MDA) and nitric oxide (NO); and markers of kidney injury, namely, blood urea nitrogen (BUN) and creatinine (Cre). Kidney histopathology was also evaluated. This study showed that the methanol extract showed the highest antioxidant activity (IC50 is 35.21 µg/mL, and FRAP is 57.58 µM Fe2+/g). The CAT and SOD activities increased significantly in okra-treated groups (P<0.05). The okra administration groups experienced a significant decrease in MDA, NO, BUN, and Cre levels (P<0.05). Thickness of the epithelial proximal tubule, diameter of the proximal tubule, and percentage of necrotic cells in proximal tubule decreased, but the diameter ratio of glomerular Bowman’s capsule in mice treated with okra was optimally improved and repaired like normal control (P<0.05). The results of this study reveal that methanol extract has a very strong antioxidant effect and can reduce the influence of toxicity induced by lead acetate in mice kidney.

Enterocytozoon hepatopenaei (EHP), a microsporidia parasite known to hinder shrimp growth by infecting its hepatopancreas, is recently an emerging infection for Litopenaeus vannamei farms in Indonesia. The present study was conducted to investigate the relationship between nitrite and ammonia concentration with EHP infection in the super-intensive ponds. Shrimps and water samples were collected from six ponds in aquaculture super-intensive ponds with a stocking density of 5,000 shrimps in each pond which is located in Pasuruan, Lamongan, and Tuban Regency, Indonesia. The water sample was taken to the laboratory for the measurement of ammonia and nitrite. The samples of ammonia and nitrite were detected by spectrophotometer, and the PCR was used to detect 18S rRNA of EHP. The PCR analysis that showed two positive samples of L. vannamei, from the ponds with a high concentration of ammonia and nitrite, were infected by EHP. The statistical analysis showed a significant correlation between ammonia and nitrite with the prevalence of EHP infections, where the Pearson correlation (r) was 0.980 and 0.943, respectively. There was a high prevalence of EHP infection with the increase in nitrite concentration and ammonia in pond four and pond six. The concentration of over 1mg/l of ammonia and nitrite could influence EHP infection prevalence in the shrimp farms. Highlight ResearchAmmonia and nitrite concentration influence the prevalence of Enterocytozoon hepatopenaei infection in Super intensive ponds.The estimated prevalence of Enterocytozoon hepatopenaei was 16.7%.PCR amplification analysis was used to detect the 18S ss-rRNA of Enterocytozoon hepatopenaei and 510-bp was obtained.

Vegetables, drinking water, and preserved meats may contain sodium nitrite (NaNO2), which causes liver disease by inducing oxidative stress. Phytochemicals are highly recommended as an alternative to synthetic drugs and affordable medicines to treat liver disease because they have fewer or no side effects. Therefore, this study aims to determine the antioxidant and hepatoprotective potential of red okra fruit ethanol extract against NaNO2-induced liver damage. Thirty-six male mice were separated into six groups. The normal control group (WA) was given distilled water only, and the NaNO2 (SN) group was given only 50 mg/kg BW NaNO2. The other four groups (P1, P2, P3, and P4) were given NaNO2 and red okra ethanol extract at doses of 25, 50, 75, and 100 mg/kg BW, respectively. Gavage was administered orally for 21 consecutive days. Commercial kits define all biochemical parameters according to the manufacturer’s instructions. Liver tissue staining followed standard protocols using hematoxylin and eosin. The study revealed that NaNO2 induction causes oxidative stress and damages the liver. The activity of antioxidant enzymes (superoxide dismutase and catalase) significantly increased in the groups treated (P2–P4) with ethanol extract of red okra ( p < 0.05 ). Besides, the oxidants (malondialdehyde, F2-isoprostanes, and nitric oxide) in the liver homogenate significantly decreased in the P4 group, which were given red okra ethanol extract ( p < 0.05 ). Likewise, red okra pods decreased significantly for the serum biochemical parameters of liver damage (aspartate aminotransferase, alkaline phosphatase, and alanine aminotransferase) in the P3 and P4 groups ( p < 0.05 ). Then, it led to a restoration of the histological structure compared to exposed mice (SN), as the pathological scores decreased significantly in the P3 and P4 groups ( p < 0.05 ), as well as the number of the necrotic and swollen liver cells was reduced. Hepatocytes returned to normal. The results showed that the ethanol extract of red okra fruit could be helpful as an affordable medicine. It is an antioxidant and hepatoprotective agent to protect the liver from damage caused by NaNO2.

Red Okra (Abelmoschus esculentus Moench) is widely distributed in Indonesia and its antioxidant ability is unknown. This study was performed to evaluate the antioxidant activities of red okra pods extracts. The powder of red okra was extracted by reflux. The first extraction used n-hexane (three times). Ethyl acetate was used to extract the residue (three times). Then, the residue was extracted three times using ethanol. Antioxidant activities were tested using DPPH and FRAP assays. This study showed that ethanol extract showed the highest antioxidant activity, which had the lowest IC50 DPPH (39.8 µg/mL) and the highest FRAP (89.15 µmol Fe2+/g). The result of DPPH and FRAP methods indicates that ethanol extract of red okra pods is classified as a very strong antioxidant.

Polyester microplastics (PS) are toxic and hazardous chemicals in the ecosystem that can induce stress. Disposing PS articles into the environment can negatively impact health of aquatic biota, including fish. This study aimed to investigate the potential of probiotics or vitamin C supplementation in improving the histological structure of organs and cytokine secretion in tilapia fish exposed to PS. Thirty-six tilapia were divided into 12 groups consisting of treatment groups (four PS concentration variations: 0, 0.1, 1, and 10 mg/L). Each treatment was given three types of feed: Commercial feed alone, commercial feed containing probiotics (200 mL/kg), and commercial feed containing vitamin C (100 mg/kg). The study had a sample size of n=3. After treatment was completed, all parameters were measured. The result showed that the addition of probiotics and vitamin C could decrease TNF-α levels and increase IFN-γ levels. Probiotics and vitamin C prevent healthy cells to be damaged by pro-inflammatory cytokines. The percentage of normal hepatocytes increased significantly in all treatment groups with the addition of probiotics or vitamin C. Furthermore, the percentage of hepatocytes with swelling and necrosis decreased significantly in treatment groups (p < 0.05). Additionally, villi height, lamina propria width, submucosa height, and the number of goblet cells all increased significantly in all treatment groups with the administration of probiotics or vitamin C (p < 0.05). Overall, both probiotics and vitamin C supplements have the potential to maintain fish health. Vitamin C exhibits a greater potential than probiotics in regulating immune responses. Meanwhile, both probiotics and vitamin C supplements have potential to inhibit damage to the hepatic and intestine structures of fish exposed to PS.

Highlight Research The effect of immersion duration of shrimp to the immunostimulant from crude protein Zoothamnium penaei was determined. 20-minute immersion duration in crude protein from Zoothamnium penaei was found to increase the phagocytosis activities, phenoloxidase, and improve hepatopancreases of the shrimp. There is increase in phagocytosis activities, phenoloxidase and improved hepatopancreases cell within a three-week maintenance of shrimp with immersion after each seven days. There is interaction between immersion duration and maintenance time of shrimp into the crude protein from Zoothamnium penaei. Abstract Enterocytozoon hepatopenaei (EHP) is a spore-forming microsporidia intracellular parasite, which slough off the cells of the hepatopancreases of the shrimp. This study was intended to determine the effect of immersion duration and maintenance time to stimulate the immune system of shrimps by using a crude protein from Zoothamnium penaei to protect against EHP infections. The study used a completely randomized design where shrimps were divided into four groups each having five replicates. Groups P1, P2, P3 were immersed in immunostimulant for 10, 15, and 20 minutes respectively, except negative control. The phenoloxidase, phagocytosis activities, histopathology study were determined. Also, the molecular diagnosis of EHP was measured to confirm EHP infection in the shrimp samplesThe phenoloxidase activity significantly increased after immersion for 20 minutes (P3) from 0.64-, 0.72-, and 0.78-units min-1 mil -1 of protein on the first week, second week and third week, respectively. The phagocytosis activities of the sample immersed for 20 minutes were significantly increased to 68% on the second week. The sample also showed improved hepatopancreases epithelial cells with only 26% necrotic cell, less EHP spore and vacuolation. There was an interaction between immersion duration and maintenance time of the L. vannamei immersed in crude protein from Zoothamnium penaei. Twenty minutes immersion were significantly found to stimulate the immune system of the shrimp against EHP. The present work revealed that, the application of crude protein from Z. penaei is effective against EHP in shrimp culture.