Enhancing Early Life Performance of Pangasianodon hypophthalmus with Probiotic Feed Additives for Sustainable Aquaculture

The shortage of fish meal and fish oil provokes the search for alternative sources of these feed components. Insects that are part of the diet of animals, birds and fish, rich in protein and fat, can serve as such an alternative. The article presents the results of testing of compound feed for fish containing the fat of the larvae of the black soldier fly (Hermetia illucens) and a feed additive with probiotic activity. The use of the fat of the larva of the black soldier fly (Hermetia illucens) and a probiotic feed additive allows increasing the survival rate of fish from 90 to 95%, increase the conversion of feed and the average daily increase. The calculation of the economic efficiency from the use of compound feed, which includes feed additives of animal origin and additives with probiotic activity, showed an increase in profit when replacing compound feed made according to a standard recipe with compound feed with new feed additives. The economic effect of using compound feed with new feed additives amounted to more than 650 thousand rubles per year on small and medium-sized trout farms up to 20 tons per year.

In order to normalize metabolic processes in the body of cattle and adsorb mold toxins coming with feed, the use of probiotic feed additives is promising. Until now, animal science does not have sufficient scientific and practically substantiated information on the effect of probiotics Provitol and Vetosporin Active on the reproductive capacity of cows and their use as sorbents in mycotoxicosis. The purpose of the work was to analyze the effectiveness of using probiotic feed additives when feeding cows of Black-and-White breed. Comprehensive research has been carried out to study the effect of using probiotics Provitol and Vetosporin Active on the productivity and reproductive trait of cows in the Middle Volga region, the optimal rate of introducing these feed additives into the ration of cows during the dry period and the period of increasing of milk has been determined. Five groups were formed such as the 1st experimental group received the probiotic Provitol together with the main ration at a dose of 1.5 kg/t of concentrated feed (15 g/head/day). For cows of the 2nd experimental group the dose of the probiotic Provitol was increased to 2.5 kg/t of concentrated feed (25 g/head/day). For cows of the 3rd experimental group the probiotic feed additive Vetosporin Active was introduced at a dose of 1.0 kg/t (10 g/head/day), and for cows of the 4th experimental group the dose of Vetosporin Active was increased to 2 kg/t of concentrated feed (20 g/head/day). The control group of cows did not receive the feed additive to the main ration. The highest level of milk production profitability was 78.6 % in the 4th experimental group of cows that received the feed probiotic Vetosporin Active at a dosage of 20 g/head/day. Thus, to increase milk productivity and improve the reproductive capacity of cows, it is necessary to introduce the probiotic feed additive Vetosporin Active into the ration of highly productive cows at a dosage of 20 g/head/day or 2.0 kg/t of concentrates when preparing complete mixed ration.

In the development of effective methods of prevention and treatment of poultry today, importance is given to probiotics based on microbial cultures, in particular, lactobacilli, bifidobacteria, yeast, etc. In contrast to antibiotics, the use of probiotics stimulates the immune response of animals, restores the microflora of the gastrointestinal tract, and ensures its optimal ratio. At the same time, livestock products remain safe for the consumer. The purpose of our work was to conduct comparative clinical trials of the efficiency of the probiotic feed additive Probion-forte in terms of productivity, blood indices, and histological structure of immune organs of chicken broilers during fattening. The clinical studies were carried out with broiler chickens of “Cobb-500” cross at the age of 2 days, which were formed into four groups (300 units in each one). In order to determine the efficiency of application, probiotic feed additives were given to chickens in different concentrations during the growing period, together with the main diet. The first group (1st) was given probiotic Probion-forte in the dose of 1 g/kg of feed, the second group (2nd) – Probion-forte in a dose of 0.5 g/kg, the third (3rd) one – a probiotic-analog “Bio plus 2B” in a dose of 0.4 g/kg; the forth (4th) group was a control group – chickens received an essential diet, without adding any feed additives. The compound feed was provided according to norms recommended for the cross “Cobb-500”, considering age. On the 15th, 30th, and 43rd day of the test, 20 units were selected for hematological, pathomorphological and microbiological tests. The productivity of birds of all studied groups was evaluated by the average daily gains, safety, feed conversion, and slaughter output. The stabilized with EDTA chicken blood was used for morphological studies, and blood serum – for biochemical and immunological studies. Comparative clinical studies have shown that adding to the main diet of broiler chickens probiotic feed additive Probion-forte at a dose of 0.5 and 1.0 g/kg throughout the growing period increases the safety and improves feed digestibility, and increases the bird’s European Efficiency Index. Based on the obtained hematological, biochemical, and immunological parameters of blood, it can be argued about the activating effect on the body of broiler chickens probiotic feed additives, namely the intensification of reanimation of free amino acids, which increases the content of the studied metabolites of lipid metabolism, which chickens use as energy and plastic material. Stimulating cellular and humoral protection, nonspecific resistance is established, confirmed by a high level of lysozyme activity in blood serum and significantly higher content of T- and B-lymphocytes, NK-cells, and γ-globulins. The macro- and microscopic structure of the studied immune organs is preserved in all groups of broiler chickens. Compared with the control group, morphometric examination in the immune organs of broiler chickens of the first and second groups revealed an increase in the area and density of lymphoid elements, an increase in the number of plasmablasts and plasma cells, much higher structural and functional capacity of cells, which was reflected at the ultrastructural level. The most significant difference was found in the 1st group of chickens fed with the feed additive Probion-forte at a dose of 1.0 g/kg of feed.

The application of local probiotics in commercial feed not only contributes to aquaculture efficiency but also opens up opportunities for utilizing inexpensive and easily accessible local materials as alternative environmentally friendly feed additives. Biologically, probiotics work by enhancing the balance of intestinal microflora, improving nutrient absorption, and strengthening the fish's immune system against environmental stress and pathogenic infections. Through this mechanism, the inclusion of local probiotics in feed can be an effective approach to naturally and sustainably enhance the physiological performance of fish fry. This study aimed to determine the effect of adding locally-sourced probiotics to commercial feed on the growth and survival of Siamese Pangasius (Pangasianodon hypophthalmus) fry. The probiotics were formulated using local ingredients such as banana fronds, turmeric, fine rice bran, yeast, molasses, and Yakult, and were applied to commercial feed at various concentrations. The research was conducted experimentally using a Completely Randomized Design (CRD) with four treatments and three replications: P0 (control, no probiotics), P1 (10 mL probiotics/kg feed), P2 (20 mL/kg feed), and P3 (30 mL/kg feed). Observed parameters included growth, feed conversion ratio (FCR), and survival rate (SR). The results showed that treatment P2 yielded the best outcomes, with the highest growth rate, lowest FCR, and significantly higher survival rate compared to the control. In conclusion, enrichment of commercial feed with local probiotics at a dosage of 20 mL/kg significantly improved the growth performance and survival of Siamese Pangasius fry (p<0.05).

In vivo and in vitro assessment of commercial probiotic and organic acid feed additives in broilers challenged with Campylobacter coli

Simple SummaryThe aquaculture industry is facing several challenges, including water quality, stocking density and disease outbreaks due to bacterial pathogens. Pathogens and challenging conditions in aquaculture are common problems that cause mortality, reduce growth performance and consequently lead to high economic losses. To prevent those losses, antibiotics are often used for treatment or prophylaxis. With the increasing concern for antibiotic resistance and the demand to reduce the application of antibiotics, alternative solutions are needed. Incorporating probiotics in the diet can be one of the solutions to support feed efficiency as well as the resilience of the animals to pathogen pressure. The results of two experiments demonstrate that probiotic feed additives are promising strategies to improve shrimp production and provide increased protection against bacterial infection, independent of the diet formulation.Probiotic feed additives can support the gut health of shrimp and thereby improve performance, production efficiency and disease resistance. Two experiments in white leg shrimp aimed to investigate the effects of a multi-species probiotic feed supplement (AquaStar®, 3 g/kg feed, Biomin GmbH, Getzersdorf, Austria) in feed formulations with different marine meal levels (32% and 15%) on growth performance and resistance against Vibrio parahaemolyticus. Juvenile shrimp were stocked in a recirculating aquaculture tank system at a density of 20 shrimp/46.8 L and were fed diets with and without the probiotic supplementation for 8 weeks. Afterwards, a bath immersion with V. parahaemolyticus was performed and mortality was observed over a period of 14 days. Independent of the diet formulation, probiotic supplementation significantly improved the survival rate of the shrimp and the specific growth rate while decreasing feed consumption and feed conversion ratio when compared to the control (p ≤ 0.042). After the Vibrio immersion challenge, mortality was significantly decreased by 13.33% with probiotic supplementation in the high marine meal diet experiment (p = 0.042) and numerically decreased by 11.67% in the low marine meal diet experiment (p = 0.133). Overall, the results suggest that the beneficial effects of the probiotic can occur independently of the diet formulation.

The spread of resistance to antibiotics is a major health concern worldwide due to the increasing rate of isolation of multidrug resistant pathogens hampering the treatment of infections. The food chain has been recognized as one of the key routes of antibiotic resistant bacteria transmission between animals and humans. Considering that lactic acid bacteria (LAB) could act as a reservoir of transferable antibiotic resistance genes, LAB strains intended to be used as feed additives should be monitored for their safety. Sixty-five LAB strains which might be potentially used as probiotic feed additives or silage inoculants, were assessed for susceptibility to eight clinically relevant antimicrobials by a minimum inhibitory concentration determination. Among antimicrobial resistant strains, a prevalence of selected genes associated with the acquired resistance was investigated. Nineteen LAB strains displayed phenotypic resistance to one antibiotic, and 15 strains were resistant to more than one of the tested antibiotics. The resistance to aminoglycosides and tetracyclines were the most prevalent and were found in 37 and 26% of the studied strains, respectively. Phenotypic resistance to other antimicrobials was found in single strains. Determinants related to resistance phenotypes were detected in 15 strains as follows, the aph(3″)-IIIa gene in 9 strains, the lnu(A) gene in three strains, the str(A)-str(B), erm(B), msr(C), and tet(M) genes in two strains and the tet(K) gene in one strain. The nucleotide sequences of the detected genes revealed homology to the sequences of the transmissible resistance genes found in lactic acid bacteria as well as pathogenic bacteria. Our study highlights that LAB may be a reservoir of antimicrobial resistance determinants, thus, the first and key step in considering the usefulness of LAB strains as feed additives should be an assessment of their antibiotic resistance. This safety criterion should always precede more complex studies, such as an assessment of adaptability of a strain or its beneficial effect on a host. These results would help in the selection of the best LAB strains for use as feed additives. Importantly, presented data can be useful for revising the current microbiological cut-off values within the genus Lactobacillus and Pediococcus.

The normal functioning of a gastrointestinal microflora in poultry and livestock is of significant importance, since its imbalance negatively influences an organism’s functions. In this study, the UV mutagenesis and selection were used to obtain two Bacillus strains possessing antagonistic activity towards Escherichia coli and Staphylococcus aureus, and their potential as a probiotic feed additive was evaluated. Compared to the parental strains, the ability of B. subtilis RBT-7/32 and B. licheniformis RBT-11/17 strains to suppress E. coli increased by 77 and 63%, respectively; the corresponding ability of these strains to suppress S. aureus increased by 80 and 79%, respectively. RBT-11/17 could not utilize microcrystalline cellulose and carboxymethyl cellulose, whereas cellulolytic activity of RBT-7/32 was doubled compared to the initial strain. The amylolytic activity of new strains was increased by 40%. Cultivation of strains on media containing soybean, pea, and corn meal did not provide any difference in the biomass production compared to the control. The heating of a water suspension of a dried biomass of the strains for 10–20 min at 80 and 100 °C or incubation in water solutions of citric, ascorbic, acetic, and formic acids (pH 3.0) for 3 and 24 h at 40 °C did not provide any negative influence on the spore survivability. Both strains were evaluated for their resistance to a number of veterinary antibiotics. Thus, RBT-7/32 and RBT-11/17 strains have good prospects for use in feed additives.

Ensuring a well-organised and sustainable feed base is the main condition for the development of animal husbandry, increasing its productivity and product quality. To ensure active growth and high productivity, probiotic feed additives are added to the feed, which increase the immune response of the host organism to pathogenic microorganisms, increases the conversion of feed and live weight gain. One of the main processes of feed production is extrusion, which can be cold, warm and hot. The hot extrusion process takes place at a temperature above 130℃ and cannot be used in the production of compound feeds with probiotics that withstand temperatures up to 85℃. During cold extrusion, the temperature reaches no higher than 70℃, which allows the extrusion of mixed feed mass, which contains probiotic feed additives.

To improve the adaptation of the pancreatic microflora to changes in the diets of calves and for the rational use of feed, in intensive cultivation, it is desirable to change the composition of feed probiotic supplements according to the age of the animals. Each age group of calves responds differently to live weight gain on probiotic feed additives. Feed supplement Immunobacterin-D containing Bacillus subtilis, Bacillus licheniformis has a positive effect on natural resistance, increases the average daily gain and has a preventive effect on diseases of the digestive tract of calves. But for 2 months of continuous feeding can cause suppression of natural resistance of some animals. When baker's yeast is added to the feed additive (B. subtilis, B. licheniformis 1 × 1010 CFU / g, Sascharomyces cerevisiae 1 × 109 CFU / g), the dry form of the product makes it possible to feed it with compound feed. The use of immunobacterin-D with yeast in 3-month-old calves in experimental animals in the clinical examination for fatness and coat condition looked better. Calves of the experimental group during this period added in increments of 276 g in the previous month, the arithmetic mean of the average daily gain of live weight was 820 ± 180 g, and in the current 1096 ± 90 g, which indicates a positive effect of yeast (especially live) on the productivity of calves at this age. For studies on heifers in a farm with lower average daily gains in live weight, we received a smaller percentage increase in gains with the use of feed probiotic supplement Immunobacterin-D with baker's yeast. When adding to the feed additives at a dose of 40 g and 20 g to heifers 350–400 kg live weight of the animal had an average daily gain of live weight in the first group of 500 ± 17 g, in the second 440 ± 30 g, and in the control 414 ± 12 g, which is 20.8 and 6.3 % more than in the control group. The use of yeast in feed additives confirms the pattern of increasing the percentage of additional average daily gain. That is, with higher average daily gains, we get a higher percentage of additional growth from feeding a probiotic feed additive containing baker's yeast.

Probiotics and phytogenics for poultry: Myth or reality?

In order to improve the digestibility and absorption of feed, metabolic processes, growth and development of animals, increase the resistance of the body immunomodulatory agents, probiotics, prebiotics, combined enzyme-probiotic feed additives are widely used. The purpose of our work was to conduct comparative clinical trials of the effectiveness of the probiotic feed additive Probion-forte in terms of productivity, blood indices and histological structure of individual internal pigs organs during fattening. The researches were carried out on 120 piglets of big white breed at the age of 28 days, which were divided into 4 groups with 30 units in each one. The probiotics were added to feed in different concentrations in order to determine their efficiency and examine influence on piglets organism: the first group was given probiotic Probion-forte in dose of 1.0 g/kg (10 weeks); the second one – Probion in dose of 1.0 g/kg (6 weeks) and 0.5 g/kg (4 weeks); the third one was given probiotic Bio Plus 2B in dose of 0.4 g/kg for 10 weeks; and the fourth one was a control group. The compound feed was provided according to norms recommended for big white breed taking into account age. On the 42nd day (6 weeks) and on the 70th day of test 10 units were selected for haematological, pathomorphological and microbiological tests. The productivity of animals of all studied groups was evaluated by the average daily gains, safety, feed conversion and slaughter output. The stabilized with EDTA piglets’ blood was used for morphological studies, and blood serum – for biochemical studies. The clinical trials have shown that the application of Probion-forte, as a feed additive for fattening of piglets within 10 weeks after weaning, did not cause adverse reactions, was well tolerated by animals and contributed to the improvement of the processes of erythropoiesis and leucopoiesis, increased of serum total protein content. The increasing activity of serum transaminases indicated more intense metabolic processes in experimental animals’ organism, which was confirmed by increase in average daily weight gains and slaughter output compared to control. In the microscopic examination of the thymus, spleen, lymph nodes, intestines, liver, the characteristic histological structure of the organs was preserved and indicated their active morphofunctional state throughout the study period. Morphometrically the increase in the height of the villi in the duodenum and the size of the thymus lobes in piglets, which were fed with probiotic feed additives was established. The efficiency and appropriateness of the application of these products was confirmed in the first and second experimental groups. However, the most significant difference was observed in piglets fed with Probion-forte for 10 weeks at a dose of 1.0 g/kg of feed.

The study was aimed to evaluate the effect of various categories of feed additives on rumen fermentation. Fermentation kinetics and substrate utilization by rumen microbes was studied by fortifying the substrate having roughage to concentrate of 60:40 with glucogenic viz., glycerol and propylene glycol (PG), rumen fermentation modifiers viz., para aminobenzoic acid (PABA), exogenous fibrolytic enzymes (EFE) and probiotic feed additives by applying first order kinetics model to in vitro gas production (IVGP) technique. Glycerol was more effective than PG for substrate fermentation kinetics and its utilization, particularly IVTVFA production. Buffer addition was more effective with PG but not with glycerol. PABA had non-significant impact on either substrate fermentation kinetics or utilization. EFE fortification significantly improved the extent of fermentation, IVNDFD and IVTVFA; however, probiotics showed only significant increase in IVDMD (P<0.01) and IVTVFA (P<0.05) by 11%. Study concluded that PG and glycerol being glucogenic compounds improved the extent of fermentation and TVFA production but glycerol was more effective than PG. Probiotics supplementation also had positive effect on IVTVFA production. EFE fortification improved fibre digestion. Buffer addition along with feed additives may enhance substrate kinetic efficiency of fermentation but it was selective.

The paper represents the impact of a probiotic feed supplement on the clinical and biochemical parameters of laying hens in an amateur private farm setting for organic product production. The experiment was conducted on laying hens aged 160 days (n=200) of the Leghorn breed of egg production, floor-housed on deep litter, weighing 1500-1600 g. Before the experiment, the birds were categorised into groups of 100 based on the principle of analogues. Following a 14-day acclimatisation period, a probiotic feed additive (L. casei, L. delbrueckii, B. licheniformis, B. subtilis at a concentration of 10^6 CFU/mL each) was incorporated into the diet of the experimental group in addition to the complete diet at a dosage of 1.0 cm³ per kg of feed for 30 days. The control group of poultry got a full meal according to the standards for laying hens. Before administering the medications, and on 15th and 30th days post-initiation of probiotic feeding, the birds were slaughted under preparatory inhalation chloroform anaesthesia (n=10). Blood samples, together with tissue and organ specimens, were collected for microbiological and biochemical analyses. The results suggest that the use of probiotic feed additives did not significantly alter the quantitative composition of indicator microflora. However, there was a tendency (on the 30th day of the experiment) to increase the content of Lactobacillus spp. and Bacillus spp. by 14.6% and 8.7%, respectively; a tendency to decrease the content of potential pathogenic microorganisms was also detected - Staphylococcus spp. 19.5%, Candida spp. 15.2%, Enterococcus spp. 1.47%, E. coli 13.0%. Administration of probiotics to laying hens resulted in a significant increase in serum immunoglobulin G levels throughout the experiment, with the peak IgG concentration in the experimental group observed on the 30th day, demonstrating an increase of 11.3% (p˂0.05). On the 30th day of the trial, immunoglobulin M and A levels rose by 14.6% and 10.2% (p˂0.05), respectively, while circulating immune complexes increased by 26.3%. The nonspecific resistance indicators in the experimental group of hens exhibited an upward trend: the phagocytic index, bactericidal activity, and lysozyme activity rose by 11.5%, 12.0%, and 13.9%, respectively. The clinical and biochemical findings of the trial indicate that the suggested composition of probiotic bacteria is non-harmful to laying hens. The observational data demonstrate no adverse effect on egg production.

The importance of probiotic feed additives in diets and programs for raising farm poultry, as well as the effectiveness of their use to improve the quality and safety of poultry products, has been analyzed. Existing standard compound feeds contain an insufficient amount of individual mineral and biologically active substances, as well as substances of a preventive nature that would support the body's immune system and have a preventive effect on the processes occurring in the gastrointestinal tract. The most important task at present is the creation and practical application of such feed mixtures that would be maximally absorbed by the body to ensure its vital functions, have preventive properties and contribute to the restoration of the macroorganism's own microflora. Probiotics actually form the intestinal microbiocenosis, produce biologically active substances and create unfavorable conditions for the development of pathogenic and conditionally pathogenic microflora, positively affect the digestibility of feed nutrients, improve metabolism and absorption of nutrients and reduce feed costs per unit of product. Therefore, today probiotic feed additives are positioned as a promising, safe alternative to antibiotics. Excessive uncontrolled use of antibiotics and the development of related problems, including antibiotic resistance, have prompted the world community to gradually completely abandon their use in animal and poultry feeding as growth stimulants. The prospect of using probiotics is a natural and effective alternative to traditional methods of poultry farming. In the final stage, the action of probiotics improves the overall functional state of the body, normalizes the functioning of various links of immune defense, optimizes physiological indicators, increases metabolism, increases productivity, and reduces feed costs. Probiotic feed additives have a significant impact on feed and water intake, feed efficiency and conversion, and egg weight. The obvious advantage of probiotics is that they are mono or mixed cultures of live microorganisms that benefit the health of the bird, improving welfare when administered in small quantities.