Volumen 22 (2022): Heft 3 (July 2022)

The use of antibiotics in aquatic feed reduces the incidence of disease and enhances growth performance, although it presents harmful effects, such as development of resistant bacteria and accumulation in the natural environment. A variety of immune stimulants including probiotics, prebiotics, synbiotics, phytobiotics, organic acids, nucleotides, antioxidants, microalgae, yeast and enzymes have been used in the aquaculture industry. In recent decades, much attention has been paid to finding a variety of immunostimulants with lower cost which also affect specific and non-specific immunity and improve fish resistance against a wide range of pathogens. These stimulants strengthen the fish’s immune system by increasing the number of phagocytes, lysozyme activity and level of immunoglobulin. The use of immune stimulants as an effective tool to overcome diseases and strengthen the immune system of farmed species, leads to the promotion of cellular and humoral defense mechanisms and increases resistance to infectious diseases. Among these immunostimulants used in aquaculture, β-glucans are of particular importance. Glucans are complex polysaccharide compounds extracted from the cell wall of yeasts and fungi. These compounds can stimulate fish growth, survival, and immune function. Therefore, this review discusses the role and importance of β-glucan as a food additive in aquaculture and examines the impact of these compounds on the growth performance, immunity and biochemical parameters of farmed species.

The recent pandemic stress and the impacts of climatic changes on humans’ and animals’ health status and well-being resulted in severe drawbacks. Initially, stress-induced oxidation resulting from the generation of free radicals led to the impairment of cellular function and a high possibility of attack with infection. Astaxanthin is a bioactive material derived from fish, crustaceans, and algae with high antioxidative potential. Astaxanthin is a lipid-soluble carotenoid that can easily cross through the cellular membrane layers to catch the reactive oxygen metabolites. Astaxanthin also has pigmentation properties making it suitable for pharmaceutical, cosmetic, nutraceutical, agriculture, and aquaculture sectors. Recently, astaxanthin is suggested as a natural scavenger for free radicals induced by COVID-19. Besides, using astaxanthin as antioxidative and immunostimulant agents is well-reported in several clinical studies. The output of these investigations should be simplified and presented to the scientific community to utilize the available information and fill the gap of knowledge. Also, it is necessary to update the researchers with the recent recommendations of applying astaxanthin in vivo and in vitro to help in proposing new horizons for engaging natural antioxidative agents to protect human and animal health. Herein, this review article tackled the nature, sources, potential roles, applicable sides, and availability of astaxanthin to fortify the scientific community with the required knowledge for further research efforts.

The association of microorganisms with livestock as endosymbionts, opportunists, and pathogens has been a matter of debate for a long time. Several livestock-associated bacterial and other microbial species have been identified and characterized through traditional culture-dependent genomic approaches. However, it is imperative to understand the comprehensive microbial network of domestic animals for their wellness, disease management, and disease transmission control. Since it is strenuous to provide a niche replica to any microorganisms while culturing them, thus a substantial number of microbial communities remain obscure. Metagenomics has laid out a powerful lens for gaining insight into the hidden microbial diversity by allowing the direct sequencing of the DNA isolated from any livestock sample like the gastrointestinal tract, udder, or genital system. Through metatranscriptomics and metabolomics, understanding gene expression profiles of the microorganisms and their molecular phenotype has become unchallenging. With large data sets emerging out of the genomic, metagenomic, and other meta-omics methods, several computational tools have also been developed for curation, assembly, gene prediction, and taxonomic profiling of the microorganisms. This review provides a detailed account of the beneficial and pathogenic organisms that dwell within or on farm animals. Besides, it highlights the role of meta-omics and computational tools in a comprehensive analysis of livestock-associated microorganisms.

In recent decades, the number of birds reared per unit area has dramatically spiked to increase profitability in egg and meat production. However, nowadays, the increase in sensitivity to animal welfare and consumer demands brings along with it a raised interest in stocking density. Stocking density is defined either as the number of animals or body weight per unit area or as the area per animal. High stocking density, which is a stress factor, can be defined as an increase in the number of animals per unit area or a decrease in the area per animal. Stress caused by high stocking density negatively affects the bird’s physiology and performance as well as the quality of the product obtained. The ideal stocking density should be 9 laying hens, 35 kilograms for broilers, and 45 quails per square metre. Otherwise, one will observe stress indicators in birds reared in more than the recommended stocking density per unit area and, consequently, a decrease in bird growth, egg production, feed efficiency, and egg or meat quality. Apart from increasing the concentrations of amino acids such as lysine, methionine, tryptophan and arginine, minerals such as selenium and chromium, and vitamins such as C and E in the diet, the addition of additives such as probiotics, humates, phytophenol compounds, and propolis is also effective in reducing or eliminating these negative effects caused by high stocking density. As a result, regulations in the nutrition of animals are effective in reducing/preventing such negative effects, thus improving animal welfare and ensuring the maintenance of optimum yield.

Sudden death syndrome (SDS) is a condition in which apparently healthy broiler chickens die suddenly. There are short convulsions and frantic wing-beating prior to death, and the weight of internal organs is the same as in healthy chickens. The exact etiology of SDS is unknown. Heart problems have been implicated as a potential cause of the disease in broiler chickens. Despite considerable research, effective methods of prevention are still being sought. Furthermore, the prophylactic measures often make production less profitable (reduced body weight gain resulting from restricted feeding) and prolong the growth period. It is necessary to continue research on SDS, in particular on stimulation of the cardiovascular system to reduce the susceptibility of broilers to sudden death syndrome under intensive production systems.

The human population is expected to reach 9.7 billion by 2050. This in turn will put more pressure on the limited available resources such as land and freshwater. Combined with the high food demand, highly virulent pathogens, and worsening effects of climate change, cases of chronic hunger and malnutrition are expected to escalate in the future. Therefore, the implementation of sustainable food production systems is crucial in safeguarding food security. Recirculating aquaculture systems (RAS) have gained much attention today for the intensive production of certain aquatic species in controlled conditions. In these systems, wastewater is purified via several water purification steps and recycled back into the system. As such, water quality parameters such as water temperature, dissolved oxygen, dissolved carbon dioxide, pH, total ammonia-nitrogen, nitrites, nitrates, and total soluble solutes are maintained within the desirable range required for proper growth and survival of the reared species. However, maintenance of good water quality largely depends on certain factors, most noticeably, the stocking density. Stocking densities below and above the recommended optimal levels negatively impact the behavior, growth performance, and immunity of reared animals. As a consequence, huge production losses are incurred. This review, therefore, aims to discuss the effect of stocking density on behavior, growth performance, feed utilization, and immunity of reared species in RAS. Moreover, optimum stocking densities of several aquatic species reared in RAS under certain culturing conditions are highlighted for sustainable production of food.

Canine DNA is widely used in forensic investigations, particularly in cases of dog attacks on humans. Nowadays, STR markers are employed worldwide in forensic laboratories to test human and animal genotypes. In the study we analysed the effectiveness of 18 STR panel as previously recommended by ISAG and the same panel with three additional markers – 21 STR, which has been recommended by ISAG as the core panel for dog identification since 2016. We calculated the PD, P_ID for these sets of panels and estimated RMP based on the DNA profile obtained during an investigation of a woman bitten by a dog. The high combined CPD value for 18 and 21 STRs showed values close to 1.0. The CP_ID value for theses panels was 5.2 × 10⁻¹⁰ to 6.4 × 10⁻¹⁴. Statistical analysis estimated the random DNA match, in the case of the woman bitten by a dog, with a probability of 4.3×10¹⁹ and 2.8×10²², using 18 and 21 STR panels respectively, and that the canine DNA profile from the crime scene originated from the suspected dog and not from another random dog. Our results show that both STR panels can be used effectively for individual identification and forensic casework.

The aim of the study was a search for new polymorphisms in the genes related to lactogenesis, including lactose synthesis in colostrum and milk, and determine their relationship with the content of colostrum and milk components and body weight of the piglets. Colostrum and milk were sampled during the second lactation on d 1, 7, 14 and 21 from 112 sows of Polish Large White (PLW) and Polish Landrace (PL). The piglets’ rearing performance was determined based on the number and body weight at 1, 7, 14 and 21 d of age. In the study six polymorphisms located in different genes were analysed. Among the six polymorphisms under analysis, those identified in the B4GALT1 genes had the most significant effect on colostrum and milk composition and the piglets’ body weight changes. Sows of the B4GALT1^GG genotype, when compared to those of B4GALT1^AA, produced colostrum and milk with a significantly lower lactose content and reared piglets with a substantially higher body weight (by 0.30 kg) at 21 d of age.

The aim of the study was to identify differences in the expression of splice variants of the PRMT2, LTF and C4A genes in the mammary glands of healthy dairy cows and those infected with staphylococci. An expression study was conducted on 38 Polish Holstein-Friesian dairy cows who were removed from the herd owing to subclinical and chronic mastitic or reproductive issues. Two days before slaughter, milk samples were taken for microbiological analysis and examined for the presence of bacteria. The mammary gland parenchyma samples with a predominance of secretory tissue were taken; these were divided into three groups according to the health status of the mammary gland: H (without pathogenic bacteria in milk), CoNS (with coagulase-negative staphylococci in milk), and CoPS (with coagulase-positive staphylococci in milk). Two of the investigated genes, LTF and C4A, demonstrated variants unequivocally expressed in infected tissue. Two LTF gene variants were found to be associated with cow health status, and with the type of bacteria causing mastitis (CoPS or CoNS). In addition, the expression of C4A isoforms differed with regard to mastitis etiology groups. The comprehensive evaluation of PRMT2 transcript suggested that the gene may also be involved in course of mastitis: two of four PRMT2 transcripts showed increased expression in the mammary gland of the CoPS group compared to controls. The obtained results are important for the knowledge on the etiology of bovine mastitis. The effects of the identified mastitis-relevant splice variants need to be further explored on the protein level to verify the suitability of splice variants and recognize their contribution towards the disease phenotypes and course.

The aetiology and pathogenesis of many canine tumours are likely to be similar to cancers found in humans. This study aimed to evaluate a plausible link between changes in the tRNA-Leu (UUR) gene and the carcinogenesis process in dogs with mammary gland tumours. The whole mitochondrial DNA (mtDNA) isolated from blood and tumour tissues of 13 dogs with malignant mammary gland tumours was sequenced. The present work is the first report showing that some polymorphisms might occur at the corresponding positions in the human and canine mtDNA genome, which in turn may provoke similar deleterious effects. The homology between the human MT-TL1 and canine tRNA-Leu (UUR) genes was 84%. After resequencing of the whole mitochondrial DNA genome with the use of the NGS technology, two polymorphisms in two haplotypes were identified: m.2683G>A (observed in 18 out of 27 samples) and m.2678_2679insG (27 out of 27 samples). The m.2683G>A polymorphism corresponded to a deleterious change at m.3243A>G, which is linked with MELAS (Mitochondrial Encephalomyopathy, Lactic Acidosis, Stroke-like episodes) syndrome and with different types of cancers in humans as well. The comparative analysis of MT-TL1 and tRNA-Leu (UUR) led us to hypothesise that the m.2678_2679insG and m.2683G>A polymorphisms might influence the dog’s condition and might be linked with tumourigenesis, as observed in humans.

The role of reactive oxygen species (ROS) during oocyte in vitro maturation (IVM) is still controversial. Although an increase in ROS production may cause deleterious effects in cells, these reactive species may also act as signaling molecules influencing different cell functions. The aim of this study was to examine the effect of varying endogenous ROS levels during IVM on the process of bovine oocyte maturation. To do so, different enzymatic antioxidant (catalase, or superoxide dismutase + catalase, or diphenyl iodonium) or pro-oxidant systems (xanthine + xanthine oxidase, or xanthine + xanthine oxidase + catalase) were added to the culture medium. ROS levels were determined by 2′,7′-dichlorodihydrofluorescein diacetate stain, nuclear maturation was evaluated by the presence of the metaphase II chromosome configuration at 22h of IVM and cleavage rate was recorded 48h post- in vitro fertilization. ROS levels were only significantly increased (P<0.05) by the O₂^.- generating system (xanthine + xanthine oxidase + catalase), but meiotic maturation rates were significantly lower (P<0.05) in all the evaluated systems compared with the control, except for the diphenyl iodonium group. However, this last group presented a significantly lower (P<0.05) cleavage rate in comparison to the control group. These results indicate that ROS would play an essential role during oocyte maturation, since its increase or decrease beyond a physiological level significantly reduced nuclear or cytoplasmic maturation rates in bovine oocytes.

The aim of this study was to evaluate the effect of high hydrostatic pressure (HHP) on the in vitro developmental abilities of nano-transfected rabbit zygotes, their transfection efficiency, and the molecular quality of the blastocysts generated. This quality was assessed by estimating the quantitative profiles of Oct4, Casp7, and Bcl2 mRNA transcripts. The nano-transfection efficiencies of zygotes that had been pre-treated with either 20 MPa or 40 MPa of HHP (13.5% and 13.7%, respectively) were insignificantly lower than those found in zygotes not exposed to HHP prior to their nano-transfection (20.1%; P≥0.05). Moreover, applying HHP treatment with the parameters of 20 MPa and 40 MPa followed by the nano-transfection of zygotes brought about an insignificant decrease in the rates of embryos at the blastocyst stage (30.4% and 23.0%, respectively) as compared to the control group of nano-transfected zygotes (40.4%; P≥0.05). Furthermore, analyzing the transcriptional activity of Oct4, Bcl2, and Casp7 genes revealed that HHP enhances the relative abundance (RA) of all mRNA transcripts in blastocysts derived from non-transfected rabbit zygotes. In turn, the augmented RAs found in the pro-apoptotic Casp7 and anti-apoptotic Bcl-2 transcripts confirmed the onset and progression of programmed cell death in blastocysts developed from nano-transfected zygotes that had undergone HHP pre-treatment. The conceptualization based not only on a novel nano-transfection approach used to genetically modify in vivo-fertilized rabbit zygotes but also on their HHP pre-treatment is elaborated here for the first time, with an emphasis on further investigations aimed at producing transgenic rabbit and other mammalian species embryos by somatic cell cloning.

It has been suggested that nanowater (NW-water declustered in the cold plasma generator and characterized by a low freezing point and high diffusivity) could improve ram semen quality after freezing in glycerol-containing extenders. Eighteen ejaculates from six Olkuska rams were divided into six equal portions each, and then diluted (800×10⁶ spermatozoa/ml) and frozen in the fructose-skimmed milk-egg yolk Kareta extenders containing 3% or 7% of glycerol. The extenders were prepared with deionized water (DW-3% and DW-7%) or NW declustered for 15 min (NW15’) or 30 min (NW30’). Post-thaw sperm motility, proportions of sperm defects and percentages of apoptotic, necrotic, and live spermatozoa were determined. The proportion of spermatozoa with midpiece defects was lower (P<0.05) in NW15’-3% compared with DW-3%. Sperm progressive motility was greater (P<0.05) for spermatozoa cryopreserved in both NW30’ (NW30’-3%/7%) extenders compared with their respective controls (DW30’-3%/7%). The proportion of necrotic spermatozoa 1 h after thawing was lower (P<0.05) in NW30’-7% compared with DW-7%, whereas the proportion of live cells detected immediately and 1 h after thawing was greater (P<0.05) in NW30’-7% than in DW-7%. In summary, NW enhanced cryoprotective effects of glycerol-containing extenders with an increase in sperm viability being greater with 7% than 3% of glycerol. Different declustering times appear to alter NW properties. These observations merit future studies of the utility of NW for semen cryopreservation in rams and other mammalian species. The specific mechanisms whereby NW ameliorates the quality of frozen-thawed ram spermatozoa remain to be elucidated.

Adding antioxidants to semen extender has become one of the major strategies to reduce the damage caused by freezing to stallion sperm. Isoespintanol (Iso) is a natural antioxidant that is known as a good free radical scavenger. The aim of this research was to evaluate the effect of supplementation of freezing media with Iso on functional integrity of equine semen. Semen from five Colombian Creole horses was frozen with 0, 10, 20, 30, 40 and 50 µM Iso. Post-thawing motility and kinetics, structural membrane integrity (SMI), normal morphology (NM) and functional membrane integrity (FMI) of sperm were evaluated. Seminal quality (SQi) and seminal velocity (SVi) indexes were calculated. Through a flow cytometric analysis, mitochondrial membrane potential (ΔΨM), viability and DNA integrity of frozen-thawed semen were assessed. General linear models were adjusted, and means comparisons were performed using Tukey’s test. The significance level was P<0.05. No differences for post-thaw sperm quality parameters were found, except for total motility, which showed a reduction by adding 40 µM Iso (P<0.05). However, the estimation of indexes, showed that supplementation with 20 or 30 µM Iso before semen freezing, can increase post-thaw SQi and SVi values of some horses. A higher proportion of high-∆ΨM sperm for 20 µM Iso was found. It is concluded that Iso can improve the quality and mitochondrial activity of frozen-thawed equine semen, however its effect can vary according to the horse and in high concentrations it could have detrimental effects.

Sperm functions may be influenced by seminal plasma (SP) proteins with affinity to zinc ions. The aim of the study was to isolate and characterise zinc-binding proteins (ZnBPs) from stallion SP using proteomic tools. Zinc-binding proteins were isolated from the SP of eight stallions by zinc-affinity chromatography. They were analysed in the SDS-PAGE system, and peptide extracts were prepared. Samples of ZnBPs isolated from stallion SP were injected onto the nLC-1000 nanoflow HPLC system coupled via a nano-electrospray ion source to the Orbitrap Elite FTMS mass spectrometer (Thermo Fisher Scientific). Raw MS data were analysed using MaxQuant software with label-free quantification (LFQ). Protein sequences were obtained from the UniProt database. Forty-seven proteins in ZnBPs were annotated in a gene ontology (GO) analysis. The LFQ intensity analysis of individual proteins revealed that ZnBPs comprised mainly clusterin (CLU, 27%), serin protease inhibitor F1 (SERPINF1, 13%), actin, cytoplasmic 1 (ACTB, 9%), nucleobindin 2 (NUCB2, 8%) and polymeric immunoglobulin receptor (PIGR, 6%). This is the first proteomic study to analyse ZnBPs in stallion SP. The present findings show that ZnBPs in stallion SP could play an important role in the regulation of sperm function.

The aim of this study was to evaluate the effect of prostaglandins E₂ (PGE₂) and F_2α (PGF_2α) on orexins (OXA and OXB) secretion (ELISA), prepro-orexin (PPO) gene expression and the content of orexin receptors (OX1R, OX2R) mRNA (qPCR) and proteins (Western blot) in porcine endometrial and myometrial tissue slices during early pregnancy (days 10–28) and on days 10–11 of the oestrous cycle. On days 10–11 of pregnancy, prostaglandins (PGs) decreased the expression of the PPO gene and OXR gene and protein in the endometrium. On days 12– 13, PGs increased OXB secretion, PGE₂ enhanced OXA secretion, and PGF_2α suppressed PPO expression. On days 15–16, both PGs inhibited PPO expression and OXB secretion, and PGF_2α increased OXA release. On days 27–28, PGs decreased PPO mRNA and OX1R protein levels, PGE₂ decreased OX2R protein content, and PGF_2α decreased OXB secretion. On days 10-11 of the cycle PGs increased the expression of PPO mRNA and OX2R protein, whereas PGE₂ increased OX1R protein content. The effect of PGs on orexin system expression in the porcine myometrium was dependent on the animal physiological status and the concentrations of specific PGs. The study demonstrated that PGs exert regulatory effects on orexin system expression in the porcine uterus, which suggests that a local regulatory mechanism could be responsible for modulating uterine metabolism. The observed differences in PGs’ influence on orexin system expression could be attributed to changes in the sensitivity of the studied tissues, associated with the phase of the oestrous cycle, the stage of early pregnancy or PGs concentration.

Neem (Azadirachta indica) belongs to Meliaceae family, represented mainly by trees, and widely cultivated and adapted in many tropical regions. The objective of this study was to evaluate the effect of increasing doses of neem seed oil (NSO) on ruminal methane (CH₄) formation, diet degradability, and fermentation characteristics after 24 h of in vitro incubation. Treatments were randomly designed to four doses of NSO supplemented to the basal diet (0, 20, 40, or 60 ml/kg DM). Increasing NSO dose resulted in a quadratic decrease (P < 0.05) in net gas (expressed as ml/g DM and ml/g TDOM) and CH₄ (expressed as ml/g TDNDF) production, while CH₄ (expressed as ml/g TDOM), acetate and propionate proportions decreased linearly confirming a dose-related effect. A quadratic increase in TDOM and linear increase (P = 0.023) in DNDF, NH₃-N concentrations, and total protozoal counts were observed. However, a linear increase (P = 0.009) was found in the ruminal butyrate proportion and partitioning factor as dietary NSO supplementation increased. In conclusion, dietary NSO supplementation mediated some desirable fermentation patterns, reducing ruminal NH₃-N concentration and CH₄ production with some adverse effects on fiber degradability. However, practical research under long-term conditions is required for further investigation.

The objective of the present study was to investigate dose-response effects of the essential oil (EO) and dry extract (EX) of Satureja khuzistanica (SK) on in vitro gas production kinetics, rumen fermentation, ruminal methanogenesis and microbial protein synthesis. So, EO and EX were tested at 0 (as control); 150 (low dose); 300, 450 (intermediate doses) and 600 mg/L (high dose). The gas produced over 24 h of incubation (GP₂₄) decreased linearly with both EO and EX dosages (P<0.01). In vitro methane production was reduced by both EO (14–69%, depending on the included dose) and EX (7–58%). Microbial protein (MP) as well as the efficiency of microbial protein synthesis (EMPS) were improved by EO (18.8–49.8% and 20.4–61.5% for MP and EMPS, respectively) and to a lesser extent by EX (8.3–25.7% and 4.6–24.2% for MP and EMPS, respectively). Ammonia concentration was dropped in linear and quadratic manners with EO (P<0.05), and linearly with EX dosages (P<0.01). EO and EX exhibited depressive effects (in linear and quadratic (P<0.05), and linear manners (P<0.01), respectively) on total protozoa count. A mixed linear and quadratic effect was observed from both EO and EX on total VFA concentration (P<0.01). Total VFA concentration increased at 300 mg/L of EX, but decreased at high dose of both EO and EX. The acetate proportion increased with EO intermediate and high dosages, but it decreased at the expense of propionate at low and intermediate doses of EX. In total, these findings confirmed previous research on the great capacity of plant-based feed additives in positively modulating rumen fermentation that their effects may vary depending on the doses used. Specifically, these results suggest that EO and EX have high potentials to improve rumen functions at intermediate doses, which needs to be confirmed by in vivo experiments.

Modifying the chemical composition of a diet can be a good strategy for reducing methane emission in the rumen. However, this strategy can have adverse effects on the ruminal microbial flora. The aim of our study was to reduce methane without disturbing ruminal function by stimulating the growth and propagation of methanotrophs. In this study, we randomly divided twenty multiparous Holstein dairy cows into 4 groups in a 2×2 factorial design with two forage levels (40% and 60%) and two nitrate supplementation levels (3.5% and zero). We examined the effect of experimental diets on cow performance, ruminal fermentation, blood metabolites and changes of ruminal microbial flora throughout the experimental period (45-day). Additionally, in vitro methane emission was evaluated. Animals fed diet with 60% forage had greater dry matter intake (DMI) and milk fat content, but lower lactose and milk urea content compared with those fed 40% forage diet. Moreover, nitrate supplementation had no significant effect on DMI and milk yield. Furthermore, the interactions showed that nitrate reduces DMI and milk fat independently of forage levels. Our findings showed that nitrate can increase ammonia concentration, pH, nitrite, and acetate while reducing the total volatile fatty acids concentration, propionate, and butyrate in the rumen. With increasing nitrate, methane emission was considerably decreased possibly due to the stimulated growth of Fibrobacteria, Proteobacteria, type II Methanotrophs, and Methanoperedense nitroreducens, especially with high forage level. Overall, nitrate supplementation could potentially increase methane oxidizing microorganisms without adversely affecting cattle performance.

This preliminary study was conducted to evaluate the effects of β-mannanase in diets of broilers on performance, pododermatitis, intestinal morphology, and digesta Clostridium perfringens counts. A total of 264 broilers (Ross 308) were randomly allotted to four treatments. A wheat-soybean based diet (CO) was supplemented with either 0.15 g xylanase/kg (XY), 0.15 g xylanase/kg + 110 mL β-mannanase/ton (MA1) or 0.15 g xylanase/kg + 220 mL β-mannanase/ton (MA2). At the end of the experiment, only significant differences in body weight were noted between groups fed MA1 and MA2 diets (2501 g vs 2641 g, respectively). Birds fed MA1 had significantly the lowest pododermatitis scores (2.26) compared to other groups. Ileal villus height measurements revealed that birds fed CO and MA2 diets had significantly higher values (1614 and 1651 µm, respectively) than those fed the MA1 diet (1489 µm). The positive percent of Clostridium perfringens in intestinal digesta at d 36 was significantly the highest in the XY group. Overall, although not improving performance, supplementing the diet with β-mannanase (110 mL/ton) contributed positively to foot pad health.

The aim of the study was to determine the effect of two proportions of arginine (95% and 105%) relative to lysine (Lys), where Lys content in the diet is in accordance with NRC (1994) recommendations or 10% higher, on the metabolism, antioxidant status, and growth performance of turkeys. The experiment had a 2x2 factorial design with two levels of Lys and Arg. The diets with a low level of Lys were according to the NRC (1994) requirements. In the diets with a high level of Lys, the content of Lys was increased by 10% relative to the low level Lys. The two Arg levels in the experimental diets were determined so as to provide 95% and 105% Arg relative to the content of dietary Lys. An increase in the amount of Lys in the diet of turkeys by 10% relative to NRC nutritional recommendations (1994) was not shown to improve growth performance, but had beneficial effects on the metabolism and antioxidant status of the birds, as evidenced by the improvement of hepatic indices (reduction of AST and ALT activity at 9^th week of life) and renal indices (reduction of UREA at 9th week of life and reduction of TP and increase level of ALB levels at 16th week of life), as well as an increase in the level of glutathione with strong antioxidant properties at 16th week of life. In comparison to the lower level of Arg in the diet, an increase in the amount of this amino acid to 105% Lys did not improve growth performance, metabolism, or antioxidant status. An Arg level of 95% Lys can be used in a diet for turkeys containing 10% more Lys than the level recommended by the NRC (1994).

Red seaweeds have several biomedical derivatives making them healthy additives for the aquaculture industry. Previously we reported enhanced growth performance, feed utilization, and immunity of Persian sturgeon treated with Gracilaria gracilis. Herein, we investigated the effects of G. gracilis on the intestinal microflora, thyroid hormones, and resistance against Aeromonas hydrophila in Persian sturgeon. Fish fed G. gracilis at 0, 2.5, 5, and 10 g/kg for eight weeks, then challenged with A. hydrophila for ten days. The results showed that the total bacterial count in the fish intestines had no meaningful differences among the groups of fish fed varying levels of G. persica (P˃0.05). Fish fed 10 g/kg of G. persica had a higher lactic acid bacteria (LAB) count than fish fed 0, 2.5, and 5 g/kg (P<0.05). Serum thyroid-stimulating hormone (TSH) showed higher levels in fish treated with 2.5 and 5 g/kg of G. persica than the other groups (P<0.05). Besides, fish fed 2.5 g/kg G. persica had higher thyroxine (T4) and triiodothyronine (T3) levels than the other groups (P<0.05). At the end of the challenge test, the highest mortality was seen in the fish fed the G. persica free diet. In summary, Persian sturgeon treated G. gracilis had improved intestinal microflora, thyroid hormones, and resistance against A. hydrophila.

The inclusion of exogenous digestive enzymes and probiotics is well established in the aquafeed industry. The mixture of multi-enzymes and probiotics improves the feed utilization and wellbeing of aquatic animals compared to the individual supplementation. Herein, we evaluated the exogenous multi-enzyme mixture (beta-glucanase, cellulase, alpha-amylase, protease, xylanase, and phytase) at 250 mg/kg and multi-species probiotic (Bacillus subtilis, Lactobacillus acidophilus, L. delbrueckii, L. rhamnosus, L. plantarum, and Pediococcus acidilactici; 1 × 10¹⁰ CFU/g for each bacterial strain) at 2 g/kg on the performances of Siberian sturgeon. The final weight, weight gain, SGR, and PER were markedly enhanced while the FCR was reduced in fish fed multi-enzyme and probiotics premix (P<0.05). Multi enzymes and probiotic mixture significantly increased the total body protein content (P˃0.05). Multi enzymes and probiotic mixture also improved the digestibility of crude protein, dry matter, and crude lipids nutrients (P<0.05). The count of goblet cells, microvilli diameter, microvilli length, outer muscle wall diameter, and enterocyte total absorptive surface were markedly increased (P<0.05) by dietary multienzymes and probiotics mixture. The WBCs and neutrophils showed marked improvements (P<0.05). The levels of glucose, triglycerides, blood urea nitrogen, and total bilirubin were markedly higher in fish fed the control than fish fed the multi-enzymes and probiotics mixture (P<0.05). Significantly, Siberian sturgeon-fed dietary multi-enzymes and probiotics had improved lysozyme activity, total immunoglobulin, and total protein in the skin mucus and serum samples (P<0.05). Further, the serum complement C3 and C4 was higher in fish-delivered multi-enzymes and probiotic mixture than in control (P<0.05). In conclusion, dietary probiotics synergistically enhanced the activity of multi-enzymes and resulted in increased feed utilization, nutrient digestibility, and health status of Siberian sturgeon.

Most probiotic yeast supplement in fish exhibit beneficial effect at ≤1% of the dietary proportion. This study aimed at evaluating the specific effects of Lypomyces starkeyi CB1807 yeast strain supplemented at ≤1% of dietary proportion on the performance of juvenile red sea bream (Pagrus major, 1.9 ± 0.04 g). Five diets were supplemented with yeast at graded levels of 0% (control diet ‘CD1’), 0.05% (D2), 0.1% (D3), 0.5% (D4), and 1.0% (D5). After 45 days of feeding trial, significant (P<0.05) improvement was detected on final body weight (FBW) and body weight gain (BWG) in fish fed D3 and D5 compared to control. Low values of total cholesterol (T-Cho) and aspartate aminotransferase (AST) were recorded in fish groups fed on D2, D4, and D5, respectively. Fish fed on D3, D4 and D5 diets showed high (P<0.05) values of serum, mucus and liver lysozyme compared to control. Fish fed on D5 showed high values of total immunoglobulin (Ig) compared to control. Fish fed on D2 showed strong correlation with biological antioxidant activity (BAP), superoxide dismutase (SOD) and catalase activity (CAT). The biological antioxidant potential (BAP) activity in fish fed on D2 was significantly higher compared to control (P<0.05). The reactive oxygen metabolites (d-ROM) were significantly lower in fish fed on D2 and D3 compared to CD1 (P<0.05). Peroxidase activity was improved significantly (P<0.05) in fish fed on D3, D4 and D5 compared to control. The tolerance ability (LT₅₀) of fish fed on D5 against low salinity stress was significantly higher compared to control (P<0.05). It was concluded that dietary benefits of spent L. starkeyi yeast at ≤1% showed considerable improvement in antioxidant capacity in red sea bream, P. major.

The present study was aimed to evaluate the moderating properties of lavender (Lavandula officinalis) essential oil (LEO) against immunotoxic effects of the organophosphate pesticide, malathion in rainbow trout, Oncorhynchus mykiss. For this purpose, fish were supplemented with LEO at dietary concentrations of 0.5, 1, 2 and 4 ml/kg diet LEO for 56 days. A non-LEO supplemented group was also considered as control. After 57 days feeding trial, biochemicals were assayed in the blood and kidney tissue and then fish exposed to a sub-lethal concentration of malathion [0.24 mg/l equal to 30% of LC₅₀ (0.8 mg/L)]. After 57 days feeding trial, the serum total immunoglobulin, respiratory burst activity, lysozyme activity and complement activity significantly elevated in fish treated with 1 and 2 ml LEO/kg diet compared to non-LEO supplemented individuals (P<0.01). Such changes were not observed in non-LEO supplemented fish (P>0.01). Significant elevations were observed in the expression of the immune genes (iNOS and C₃ genes) in fish treated with 0.2–2 mg LEO/kg diet compared to non-supplemented ones (P<0.01). The lysozyme and complement activity significantly decreased in fish fed 4 ml/kg diet (P<0.01). After exposure to malathion, all immune components significantly declined in control and those treated with 0.5, 1 and 4 ml LEO/kg diet (P<0.01). In contrast, the immunity components exhibited no significant changes in fish treated with 2 ml LEO/kg diet after exposure (P>0.01). The expression of iNOS and C₃ genes significantly reduced in control and fish fed 0.5, 1 and 4 ml LEO/kg diet in response to malathion (P<0.05). Furthermore, the expression of these genes showed no significant changes in fish fed with 2 ml LEO/kg diet after exposure (P>0.01). The findings of the present study suggested an immunoprotective role for dietary LEO at optimized dietary concentrations of 1 and 2 ml LEO/kg diet against oxidative stress and toxicity induced by malathion. Nevertheless, LEO at high dietary concentration (4 ml/kg diet) had reducing effects on the fish immunity.

An 8-week feeding trial was conducted to examine the efficacy of stachyose as a prebiotic on immune parameters, antioxidant-/immune-related genes’ expression, and lipid metabolism of zebrafish. Three hundred zebrafish (0.45 ± 0.08 g) were fed four diets containing different stachyose levels at 0, 1, 2 and 4 g kg⁻¹, respectively. After eight weeks of the feeding trial, immunity, antioxidant defence and lipid metabolism were tested. It was observed that the addition of stachyose to the diet induced no significant influence (P>0.05) in SOD, GPX, and CAT, gene’s expression, compared to the control diet. The inclusion of stachyose resulted in no significant changes in immune gene expression (Lyz, IL-1, IL-6, and TNF) in zebrafish (P>0.05) compared to the control diet. Total cholesterol, triglyceride and LDL (low-density lipoprotein) significantly (P<0.05) decreased with the addition of 2 and 4 g kg⁻¹ stachyose, while fish fed the control diet and 1 g.kg⁻¹ recorded the highest significant value of LDL (P<0.05). Fish fed diet, either control diet or diet supplemented with 0.5 g kg⁻¹ stachyose, recorded the lowest HDL value (P<0.05) compared to other treatments. In conclusion, stachyose can be potentially used as a feed additive to modulate lipid metabolism. However, this prebiotic did not benefit immune parameters and antioxidant defence.

India remains as a top supplier of value-added seafoods to the global markets. Cochin is one of the leading fishing ports in India contributing to country’s major seafood export. As part of risk assessment, we analyzed the prevalence of multi-drug resistant Vibrio parahaemolyticus; seafood borne gastroenteritis-causing pathogen, in shellfishes collected from major fish markets in Cochin. This bacterial species was initially isolated on thiosulfate-citrate-bile salts-sucrose agar and HiCrome™ Vibrio agar, further confirmed by detection of V. parahaemolyticus species-specific toxR gene and by 16S r RNA sequencing. A total of 113 confirmed V. parahaemolyticus were recovered. Almost all the strains exhibited resistance towards three or more antibiotics (multiple-drug resistant) and harbored virulence related exoenzymes especially hemolysin (Kanagawa phenomenon); which is indeed a matter of concern. The multiple antibiotic resistance (MAR) index of the isolates ranged from 0.21 to 0.57. Furthermore, our results also indicate a substantial increase in the incidence of the pathogen in seafood from the area compared to previous years. This may pose a risk to the local and international consumers of the seafood. Practicing complete hygiene and adequate cooking is strongly recommended. Regular monitoring of aquaculture areas and fish markets is urged to reduce dissemination of pathogenic V. parahaemolyticus in seafood and to prevent fatal cases of human infection induced by this species in the country.

In organic pig husbandry, the use of roughage is mandatory as dietary supplement. This study investigated the effects of oat silage on the fatty acid composition, in entire males and gilts, as well as indole and skatole levels in perirenal adipose tissue of entire males. Entire males and gilts (forty-five to forty-eight pigs/sex) were assigned to two dietary roughage feeds (control with straw vs. oat silage). There was no significant effect of silage or sex on total SFA and MUFA in pork fat. However, the oat silage increased the total PUFA n-3 and decreased the PUFA n-6/n-3 ratio. The content of boar taint compounds (skatole and indole) in the entire male pigs did not differ between diets, although human nose scoring rejected in a greater extent more pork fat from entire males supplemented with oat silage, compared with those only supplied with straw. Approximately 50% of the entire males (90 to 97 kg of carcass) had low skatole values (≤0.1 μg/g), that were below the range of boar taint detection, regardless of the feeding regime. This finding indicates that more studies should be performed to avoid the problem of taint detection in entire males under organic production.