Dietary peppermint (Mentha piperita) powder affects growth performance, hematological indices, skin mucosal immune parameters, and expression of growth and stress-related genes in Caspian roach (Rutilus caspicus).

Peppermint is a popular herbal medicine due to its several pharmaceutical applications. In this study, peppermint powder was used as a feed additive to evaluate growth performance, hematological parameters, protein profile of skin mucus, and immune parameters, as well as growth hormone (GH), insulin-like growth factor (IGF), and 70 kDa heat shock protein (HSP70) gene expression in Caspian roach (Rutilus caspicus). The fingerlings (average weight of 2.40 ± 0.12 g) were fed with diet containing 0 (control), 2, 3, and 4 g/kg peppermint for 8 weeks. The addition of peppermint significantly enhanced the growth parameters and decreased the food conversion ratio. Hematological indices of fish fed with peppermint-supplemented diets were significantly different from the control group (P < 0.05). Soluble protein, alkaline phosphatase, and lysozyme enzyme activity in mucus samples showed an incremental trend by increasing the peppermint levels in the diet. Evaluation of mucosal immunity indicated a remarkable difference between the protein profile in treatments fed with peppermint-supplemented diets and the control group. A new protein band (approximately 27 kDa) was also found in the skin mucus of fish fed with the diet containing 4 g/kg peppermint, and the highest band density was observed in this treatment. The highest IGF and GH gene expression were observed in 4 g/kg peppermint treatment. There was a significant difference in HSP70 expression between the fish fed with peppermint powder and the control group (P < 0.05). Overall, the results showed that dietary oral administration of peppermint at 4 g/kg of feed can act as a growth promoter and immunostimulant.

Effect of dietary supplementation with apple cider vinegar and propionic acid on hemolymph chemistry, intestinal microbiota and histological structure of hepatopancreas in white shrimp, Litopenaeus vannamei.

This experiment was conducted to evaluate the effects of dietary supplementation of Apple cider vinegar (ACV) and propionic acid (PA) on biochemical parameters of hemolymph, intestinal microbiota and histology of hepatopancreas in white shrimp (Litopenaeus vannamei). Five experimental diets were evaluated in this study including diets supplemented with 1.0, 2.0 and 4.0% of ACV, 0.5% propionic acid, and a control diet with no supplements. Shrimps (initial weight of 10.2 ±â€¯0.04 g) in triplicate groups with the density of 25 shrimps per tank were fed the diets for 60 days. At the end of the feeding trial, shrimps fed with ACV and PA supplemented diets had significantly higher total protein level than those fed the control diet (P < 0.05). The number of Vibrio spp., R-cells (lipid storage cells) of hepatopancreas and cholesterol level in shrimps fed the diets containing ACV and PA were lower compared to the control group (P < 0.05). However, there was no remarkable variations in glucose concentration, B-cell number and tubule diameter among the experimental diets (P > 0.05). In addition, shrimps fed the ACV diets had significantly lower total heterotrophic marine bacteria compared to the control or PA groups, and the lowest bacterial number was observed in shrimp fed 4% ACV supplemented diet (P < 0.05). Supplementation of 2 and 4% ACV as well as 0.5% PA in the diet led to a significantly higher calcium concentration than the control treatment (P < 0.05). The lowest triglyceride concentration was observed in the shrimps fed diets containing 2.0 and 4.0% ACV, which resulted in 15 and 20% reduction, respectively (P < 0.05). Overall, the findings indicates that ACV and PA possess antimicrobial activity and demonstrate beneficial effects on health status, so they can be potentially used as feed additive in the feeding of L. vannamei.

The effects of dietary supplement of galactooligosaccharide on innate immunity, immune related genes expression and growth performance in zebrafish (Danio rerio).

The present study investigates the effect of different levels of galactooligosaccharide (GOS) on innate immune parameters, immune related genes expression as well as growth performance in zebrafish (Danio rerio). Four hundred and twenty fish (mean weight 45 ±â€¯0.1 mg) were supplied, randomly stocked in twelve aquaria assigned to four treatments. Zebrafish were fed with either control diet or control diet enriched with different levels (0.5, 1 and 2%) of GOS for eight weeks. At the end of feeding trial innate immune parameters (total immunoglobulin, total protein and alkaline phosphatase activity), immune related genes expression (interleukin 1 beta [il1b], Lysozyme [lyz], tumor necrosis factor alpha [tnf-alpha]) as well as growth performance were measured. Evaluation of immune parameters revealed significant (P < .05) increase of total protein and total Ig in zebrafish fed 1 or 2% GOS compared other treatments. However, in case of lysozyme activity no significant (P > .05) differences were noticed between GOS fed fish and control group. Also, in case of ALP activity, significant increase (P < .05) was observed in 2% GOS treatment. Gene expression studies revealed significant upregulation (P < .05) of tnf-alpha and lyz genes in GOS fed fish. While no significant (P > .05) difference was observed in case of il1b gene expression. Evaluation of growth performance at the end of feeding trial revealed no significant (P < .05) improvement in GOS fed groups. The present results revealed positive effects of GOS on innate immune parameters and related gene expression in zebrafish.

Dietary Myrtle (Myrtus communis L.) improved non-specific immune parameters and bactericidal activity of skin mucus in rainbow trout (Oncorhynchus mykiss) fingerlings.

The present study examined the effects of dietary Myrtle (Myrtus communis L.) on non-specific immune parameters and bactericidal activity of skin mucus in rainbow trout (Oncorhynchus mykiss) fingerlings. Three hundred and sixty fingerlings (6.50 ± 0.55 g (were distributed in twelve cages (65 × 65 × 65 cm) with a metal framework. The study included four treatments repeated in triplicates. The treatments were feeding trouts with experimental diets containing different levels (0, 0.5, 1 and 1.5%) of Myrtle powder. The fingerlings were fed on experimental diet for sixty days and then skin mucus non-specific immune parameters as well as bactericidal activity were measured. At the end of the trial, the highest skin mucus soluble protein level was observed in group fed with 1.5% Myrtle (P < 0.05). The alkaline phosphatase (ALP) activity was significantly increased in fish groups fed 1 and 1.5% Myrtle compared with the control group (P < 0.05). However, evaluation of skin mucus lysozyme activity showed no significant difference between treatments and control group (P > 0.05). Also, no antibacterial activity was detected against Escherichia coli, Staphylococcus aureus and Salmonella enterica in all treatments and control group. Whereas skin mucus of rainbow trout showed antimicrobial activity against fish pathogens (Aeromonas hydrophila and Yersinia ruckeri) in 1 and 1.5% Myrtle treatments. These results indicated beneficial effects of dietary Myrtle on mucosal immune parameters of fingerling rainbow trout.

Dietary effect of apple cider vinegar and propionic acid on immune related transcriptional responses and growth performance in white shrimp, Litopenaeus vannamei.

This experiment was conducted to study the effect of various levels of ACV® and Propionic acid (PA) on expression of immune related genes and growth performance in white shrimp (Litopenaeus vannamei). Three hundred and seventy-five shrimps with an average initial weight of 10.2 ± 0.04 g were collected and acclimatized for two weeks. Five experimental diets including control diet, 0.5% PA diet and 1%, 2% and 4% ACV® diets were applied to feed the shrimps. They were fed 4 times a day with 2.5% of body weight. After 60 days of culture, shrimps fed with ACV® and PA diets showed no significant difference in growth performance. Expression of prophenoloxidase (proPo), lysozyme (Lys), penaeidin-3a (Pen-3a) and Crustin (Cru) genes were determined from hepatopancreas, using the real-time PCR after 15, 30 and 60 days. Expression of Lys and proPo genes was significantly up regulated in shrimps fed with ACV® and PA diets compared to the control group after 30 and 60 days of treatment. After 15 days, Pen-3a gene expression was significantly higher in PA group compared to the control group. Also, shrimps fed with 1% and 4% ACV® and PA diets showed significantly increased expression of Pen-3a after 30 days. In contrast, expression of Cru was significantly down regulated in response to ACV® diets, but, Cru expression in treated shrimps with PA diet was greater than the control group after 30 and 60 days. Overall, the results provided evidence that ACV® could be used as a natural immunostimulant for shrimps in order to adjust and enhance expression of the immune related genes.

Agaricus bisporus powder improved cutaneous mucosal and serum immune parameters and up-regulated intestinal cytokines gene expression in common carp (Cyprinus carpio) fingerlings.

The aim of the present study was to investigate immunomodulatory effects of Agaricus bisporus, white bottom mushroom powder (WBMP) on common carp (Cyprinus carpio) fingerlings. Carps were fed on different levels of WBMP (0, 0.5, 1 and 2%) for 8 weeks and at the end of feeding trial, skin mucus immune parameters (total Ig, lysozyme and protease activity), cytokines gene expression (TNF-alpha, IL1b, IL8) in intestine as well as serum non-specific immune parameters (total Ig, lysozyme and ACH50) were measured. The results showed significant dose dependent increase of skin mucus immune parameters in carps fed WBMP (P < 0.05). While, no significant difference was observed between 0.5% WBMP and control group (P > 0.05). In case of serum non-specific immune parameters, except lysozyme activity, other parameters (Ig total and ACH50) were significantly affected by dietary inclusion of WBMP (P < 0.05). Also, evaluation of cytokines gene expression in the intestine of carps revealed remarkable up-regulation of TNF-alpha in fish fed 2% WBMP supplemented diet compared other treatment (P < 0.05). Likewise, IL1b gene expression was significantly increased in 1 and 2% WBMP treatments compared to the 0.5% WBMP and control groups (P < 0.05). IL8 gene expression was not affected by inclusion of WBMP in carp diet (P > 0.05). Furthermore, feeding on WBMP supplemented diet significantly improved growth performance (P < 0.05). These results indicated that WBMP can be considered as a promising immunostimulants in early stage of common carp culture.

The effects of garlic-supplemented diets on skin mucosal immune responses, stress resistance and growth performance of the Caspian roach (Rutilus rutilus) fry.

This study was conducted to evaluate the effects of garlic supplementation on some skin mucus immune parameters, mucus antimicrobial activity and growth performance of the Caspian roach (Rutilus rutilus caspicus) fry. Fish (1 ± 0.07 g) were divided into four groups fed diets containing 0 (control), 5, 10 and 15 g kg(-1) garlic for 8 weeks. The results showed that there was a significant increase in weight gain and specific growth rate in those fish fed garlic diets compared with the control (P < 0.05). Condition factor was not significantly affected by garlic dosage. At the end of trial, the epidermal mucus protein level, alkaline phosphatase and antimicrobial activity against 2 g-negative bacteria (Escherichia coli and Serratia marcescens) and gram-positive bacteria (Streptococcus faecium and Micrococcus luteus) were measured. Skin mucus alkaline phosphatase, protein levels and antimicrobial activity were increased following garlic administration, and the bacterial growth inhibition zones were significantly elevated in garlic-fed fish (P < 0.05). In salinity stress experiment, no differences were observed for survival rate among the experimental diets. No mortality was recorded during the feeding trial. These results indicated that dietary garlic beneficially affects the skin mucus immune parameters and growth performance of the Caspian roach fry.

The effects of dietary vitamin C on mucosal immune responses and growth performance in Caspian roach (Rutilus rutilus caspicus) fry.

This study was conducted to investigate the effects of different levels of dietary vitamin C on some skin mucus immune parameters, mucus antimicrobial activity and growth performance of Caspian roach (Rutilus rutilus caspicus) fry. Six hundred sixty Caspian roach (1.4 ± 0.02 g) fry were allocated to 12 tanks (55 fish per tank), and triplicate groups were fed diets containing 0, 1,000, 1,500 and 2,000 mg kg(-1) vitamin C for 60 days. At the end of the trial, the epidermal mucus protein level, alkaline phosphatase and antimicrobial activity against two gram-positive bacteria (Streptococcus faecium and Micrococcus luteus) and gram-negative bacteria (Escherichia coli and Serratia marcescens) as well as growth performance were measured. The results demonstrated that feeding on vitamin C significantly elevated skin mucus alkaline phosphatase and protein levels compared to the control group (P < 0.05). However, lysozyme activity was undetectable in both the vitamin C-fed roach fry and the control group. Skin mucus antimicrobial activity was increased following vitamin C administration, and the bacterial growth inhibition zones were significantly elevated in vitamin C-fed roach (P < 0.05). Similar results were obtained in case of the minimum inhibitory concentration of skin mucus. Also fish fed the control diet had a significantly lower weight gain, specific growth rate and condition factor compared to the other treatments (P < 0.05). These results revealed that dietary vitamin C beneficially affects the skin mucus immune parameters and growth performance of Caspian roach fry.