Innovative use of a commercial product (Biomagic) for odor reduction, harmful bacteria inhibition, and immune enhancement in pig farm
Article information
Abstract
The global increase in livestock production has correspondingly intensified farm odors due to harmful bacteria, reduced immunity, and disease progression. In this study, we treated feces with Biomagic-Enzyme complex for 4 months to understand the relationship between farm odor, immunity against common viral diseases, immune cytokines, and changes in the microbiota. A gas meter (MultiRAE) was used to measure ammonia (NH3) and hydrogen sulfide (H2S) while odor intensity and offensiveness were characterized by the non-objective scaling method. A complete blood count was performed and plasma was obtained after blood centrifugation at 3,000 rpm for 20 minutes. The cytokine profile was evaluated using commercial kits. Microbial DNA was extracted and purified from fecal samples to analyze the microbiota. Microbial DNA and viral RNA/DNA were obtained from fecal samples and amplified to determine the expression of transmissible gastroenteritis virus (TGEV), porcine reproductive and respiratory syndrome (PRRS), and porcine circovirus type 2 (PCV2). Our results indicated that Biomagic reduced odor nuisance by decreasing ammonia levels, resulting in faint and fairly offensive odor intensity. After the enzyme treatment, Escherichia coli populations significantly reduced across all 3 farms. In contrast, beneficial Lactobacillus spp. levels remained stable, indicating the enzyme selectively targeted harmful bacteria while preserving beneficial ones. The beneficial Lachnospiraceae, Spirochaetaceae, and Bacteroidaceae were found to be higher in the third month of treatment. TGEV was not detected, while PRRS and non-pathogenic PCV2 showed a positive infection rate. In conclusion, Biomagic reduced ammonia, prevented viral infection from pig farms, and improved gut-beneficial bacteria and microbiota.
Introduction
In the Republic of Korea, pig farming plays a significant role in the country’s livestock industry, contributing to about half of its total value. However, the number of pig farms has been steadily decreasing. Despite this, the overall scale of pig rearing has significantly increased over the same period as the farms have expanded their livestock to meet the rising meat demand. This increased production has resulted in a corresponding rise in the amount of manure produced through pig farming, exacerbating the issue of odor emissions. Consequently, the number of complaints regarding livestock odor has surged, increasing from 2005 to 2017 [1]. Managing manure in this context is critical not only to reduce odor nuisances but also to enhance the overall health of the animals [2]. Strong odors can be indicative of an imbalance in the microbial ecosystem within the manure, leading to the increased proliferation of harmful bacteria and susceptibility to viral infections [3].
The global impact of greenhouse gas production and ammonia precipitation resulting from animal feces and respiration has become a growing concern [4]. Livestock emission gases, when present at high concentrations, can harm both the health and efficiency of the animals and farm labor. These emissions can contribute to environmental problems, leading to acid rain and nitrification [5]. On the other hand, the gut microbiota plays a crucial role in the overall health of pigs, influencing digestion, nutrient absorption, and the immune system [6]. A balanced gut microbiota can protect against pathogenic bacteria by competitive exclusion, production of antimicrobial compounds, and stimulation of the host’s immune responses [7]. Disruptions in the gut microbiota, often caused by the overuse of antibiotics, can lead to dysbiosis, characterized by a decrease in beneficial bacteria and an increase in harmful bacteria, which can compromise the animal's health and productivity [8]. Overall, antibiotic resistance in pig farming is a growing concern, as the overuse of antibiotics can lead to the development of multidrug-resistant pathogens. These resistant bacteria can spread from animals to humans through direct contact, environmental contamination, and the food chain, complicating treatment strategies and leading to higher morbidity and mortality rates [9]. Furthermore, the enhanced microbial activity can lead to higher levels of immune-enhancing cytokines, bolstering the pigs’ defense against viral infections such as porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) [10].
Previous research has focused on field experiments to identify the most effective way to reduce pig farm odor pollution and improve farm health [11,12]. The majority of the studies primarily assessed the effectiveness of additives (chemical and biological techniques) or mitigation structures (biological filters, manure-based liquid recharging systems, etc.) [13-18]. Recently, in growing-finishing pigs, the odor emission and gut microbiome have been reported to be controlled through arazyme treatment in conjunction with dietary carbohydrolases [19]. In contrast, this study aims to evaluate the effectiveness of a commercial product, BioMagic, in reducing odor emissions from pig manure and its broader impacts on the farm environment and animal health. BioMagic is a granulated enzyme complex containing nitrogen, phosphorus, and potassium in a typical 1:1:1 ratio and produced through the fermentation of useful ingredients extracted from fruits such as orange, lemon, and papaya (digestive enzymes such as amylase 22ut, protease 5ut, lipase 3ut, cellulase 2ut, co-enzyme, co-factors, and amino acids, etc., https://www.biomagic.com/). Specifically, we hypothesized that Biomagic would (1) reduce ammonia and hydrogen sulfide levels in pig manure, (2) decrease the intensity and offensiveness of odors, (3) modulate the fecal microbiota favorably by increasing beneficial bacteria and reducing harmful bacterial populations, (4) enhance the immune status of pigs without compromising physiological homeostasis, (5) reduce the prevalence of pathogenic viral infections. We conducted an empirical study for the first time to evaluate the effects of manure treatment with the Biomagic and analyzed pig farm gas emission, odor quality, physiological changes in blood indices, microbial disease susceptibility, microbiota analysis, and viral disease spreading. Our results indicated that Biomagic reduced ammonia, prevented viral infections, improved gut-beneficial bacteria, and enhanced the microbiota from pig farms (Fig. 1).
Graphical abstract of Biomagic was sprinkled over feces and odor intensity and offensiveness were measured by a gas meter (MultiRAE). Biomagic reduced odor nuisance by decreasing ammonia levels. Following blood collection, a complete blood count (CBC) and ELISA assays were conducted, revealing no changes in physiological parameters or cytokines levels. Microbial DNA was for microbiota analysis. Standard kits were used to extract microbial DNA and viral RNA/DNA from fecal samples, and the expression of transmissible gastroenteritis virus (TGEV), porcine reproductive and respiratory syndrome (PRRS), and porcine circovirus type 2 (PCV2) were evaluated.
Materials and Methods
Study design and study animals
Biomagic was used on the farms to improve the biodegradation of waste products and decrease environmental contamination. It is a granulated enzyme complex containing nitrogen, phosphorus, and potassium in a typical 1:1:1 ratio and produced through the fermentation of useful ingredients extracted from fruits such as orange, lemon, and papaya. It was treated with 1,000 kg feces/1 L (0.1%), once a day. This study involved a crossbreed Yorkshire and Landracer growing pigs with an initial average body weight of 60 kg (± 3.5 kg), housed at 3 different farms (n = 8) for 4 months. Seven points were measured from a 2 m point on the entrance wall of the pig pen to a 2 m point on the opposite wall, divided into 7 equal parts, and the average value was calculated. All animal care and experimental procedures were carried out according to internationally accepted guidelines on the use of laboratory animals (Institutional Animal Care and Use Committee permit no. 2022-0334) and the protocols were approved by the Animal Care Committee of the College of Veterinary Medicine, Kyungpook National University.
Gases emission measurement and non-objective odor intensity and offensiveness scaling
Before and after processing with the Biomagic, a MultiRAE gas meter (RAE Systems, Honeywell International, Inc., USA) was used to measure the representative harmful gases ammonia (NH3) and hydrogen sulfide (H2S), as well as the carbon dioxide (CO2) and oxygen (O2) concentrations. While odor intensity and offensiveness were scaled as described previously [20].
Blood collection, complete blood count, and blood plasma separation
The blood samples were collected via venepuncture from the jugular vein. The collected blood was transferred to K3-EDTA-treated tubes and stored on ice until analysis. A complete blood cell count, which includes white blood cells, red blood cells, hemoglobin, hematocrit, and mean corpuscular volume, was measured using an automatic hematology analyzer, Hemavet. After blood collection, 1 mL of each sample was dispensed into a 1.5 mL microtube and centrifuged at 3,000 rpm for 20 minutes to obtain plasma for evaluating the cytokine profile using a standard commercial kit.
Fecal bacterial disease analysis
A 99-m sterilized anaerobic diluent containing 1 g of fresh test animal feces was homogenized, followed by a 10-fold anaerobic dilution process (NaCl, 0.9 g; KCl, 0.042 g; CaCl2 6H2O, 0.048 g; NaHCO3, 0.02 g; Cysteine HCl, 0.2 g; and 400 mL of distilled water were used to select the lactic acid strains for livestock probiotics). The selection and fractionation media used are listed in Table 1 and were gradually diluted in a solid medium to determine the number of bacteria in the dilution solution. On the solid medium that was plated with incremental dilutions, aerobic or anaerobic cultures were performed. Using a GasPak system (Becton, Dickinson and Company, USA), the anaerobic culture was performed for 48 hours at 37°C (refer to Table 1).
The media and culturable methods used in the study for the selection of culturable intestinal bacteria from feces
Fecal DNA isolation and microbiome analysis
Microbial DNA was extracted and purified from fecal samples using a QIAamp DNA Stool Mini Kit (QIAGEN Inc., Germany) to analyze the microbiota from the stool samples, as previously described [21]. The DNA samples’ quality control requirement was met with an A260/A280 ratio of ≥ 1.8 and a DNA concentration of ≥ 15 ng/µL. Following the DNA extraction from the farm’s environmental feces, microbiome analysis was conducted at Kyungpook National University’s Sequencing Centre. The 16S rRNA gene sequences obtained from the fecal samples were used as the basis for microbiome profiling.
Isolation of fecal viral RNA/DNA
Viral RNA and DNA were extracted and purified from stool samples following the manufacturer’s protocol using a Patho Gene-spin RNA/DNA Extraction kit (iNTRON Biotechnology). To ensure the quality control, requirements, the extracted viral RNA and DNA were required to have an A260/A280 ratio of ≥ 1.8 and an RNA/DNA concentration of ≥ 15 ng/µL.
Evaluation of viral RNA/DNA for transmissible gastroenteritis virus, porcine reproductive and respiratory syndrome, porcine circovirus type 2
The extracted viral RNA and DNA were amplified to determine the presence of transmissible gastroenteritis virus (TGEV), porcine reproductive and respiratory syndrome (PRRS), and PCV2 using commercial kits for reverse transcription-polymerase chain reaction (RT-PCR, TGEV and PRRS) and PCR reaction (PCV2) according to the manufacturer’s protocols (LiliF TGEV, PRRS-RT-PCR, and PCV2-PCR kit, iNtRON Biotechnology). The product was run on a 1.5% electrophoresis gel to assess the presence of viral RNA and DNA.
Statistical analysis
Acquired data were subjected to a one-way analysis of variance and post hoc Dunnett’s test (SAS Institute Inc., USA) to determine the statistical significance of the observed differences. The information is presented as the mean ± standard deviation (SD). Statistical significance was defined as p < 0.05.
Results
Measurement of gas emissions from pig farms
MultiRAE gas meter was used to measure the representative harmful gases ammonia (NH3) and hydrogen sulfide (H2S) that cause odor complaints, as well as the carbon dioxide (CO2) and oxygen (O2) concentrations, which are environmental indicators. A significant reduction in ammonia concentrations was observed across the 3 farms after the application of Biomagic in August, as analyzed using repeated measures analysis of covariance (ANOVA) (Fig. 2). Farm 1 showed a significant decline in NH3 levels from August (mean ± SD, 12 ± 0.5 ppb) to December (mean ± SD, 8 ± 0.5 ppb); F (1, 4) = 22.36, p < 0.01. Farm 2’s NH3 levels halved from August (mean ± SD, 13 ± 0.7 ppb) to December (mean ± SD, 6 ± 0.5 ppb), F (1, 4) = 35.07, p < 0.001, indicating a highly significant effect. Farm 3 presented the most rapid reduction with levels decreasing from 22 ppb in August to 7 ppb in September, F (1, 4) = 45.85, p < 0.001, and stabilizing thereafter, demonstrating a substantial immediate response to Biomagic treatment. In contrast to NH3, ozone levels remained relatively constant across all 3 farms throughout the observed period. Farm 1's ozone concentration averaged around 20 ppb, while Farms 2 and 3 maintained a consistent level of approximately 25 ppb. The stability in ozone concentrations indicates that either farm operations do not significantly influence ozone levels or external factors affecting ozone concentrations are uniform across these locations. Carbon dioxide emissions demonstrated an upward trend in 2 out of the 3 farms. Farm 1 shows an increase from about 2,000 ppb to nearly 3,500 ppb by December. Similarly, CO2 emissions in Farm 2 rose from approximately 1,000 ppb to 2,000 ppb within the same period. In contrast, Farm 3 displayed a peak in CO2 emissions in September at about 2,500 ppb, followed by a reduction to below 2000 ppb by December. This variation suggests that factors such as seasonal energy use, biomass decomposition, or heating requirements could influence CO2 levels. Overall, our results demonstrated that after treatment with the enzyme complex, ammonia concentration was reduced and remained > 25 ppm, ozone concentration remained constant, CO2 remained > 3,000 ppm, and H2S was not detected. The odor intensity and offensiveness were characterized by the non-objective scaling method (Dynamic Olfactometry method) as reported previously [22]. Odor intensity and offensiveness were characterized as faint (2) and fairly offensive (3) from very strong (5) and strongly offensive (4), respectively (Table 2 and Fig. 2).
Changes in ammonia (NH3), hydrogen sulfide (H2S), oxygen (O2), and carbon dioxide (CO2) levels in pig farms before and after the application of the Biomagic. The concentrations were monitored at 7 locations, starting from a 2-meter mark on the wall near the entrance to the pig pen to a 2-meter point on the opposite wall, divided into 7 equal segments.
Scaling of odor intensity and offensiveness
Biomagic prevents changes in inflammatory markers
The blood biochemical indices (red blood cell count, white blood cell count, platelet count, hemoglobin concentration, hematocrit, and mean red blood cell volume) were measured to monitor changes in blood biochemical indices and normal homeostasis of the animals. Our results indicated no significant changes in the blood biochemical indices observed after using Biomagic (p < 0.05) (Supplementary Fig. S1). Additionally, we measured the cytokines to confirm whether it affect the pig's immune system. The blood inflammatory cytokines (interferon [IFN]-γ, interleukin [IL]-1β, IL-6, and tumor necrosis factor [TNF]-α) and immunoreactive cytokines (IL-2, IL-4, and IL-10; Supplementary Table 1) in the plasma were measured. Our results indicated that IFN-γ, IL-1β, IL-6, and TNF-α were below detectable concentrations, and no significant changes were observed in the levels of IL-2 and IL-4 (Fig. 3, Supplementary Table 1). However, IL-10 showed an increasing trend starting from the third month (November). Overall, the results indicate that the treatment with the Biomagic-Enzyme complex does not significantly affect the immune response of pigs.
Biomagic prevents changes in inflammatory markers. Blood samples were collected in a 1.5 mL microtube and centrifuged at 3,000 rpm for 20 minutes. The obtained plasma was utilized to measure blood inflammatory cytokines through a standard Enzyme-linked immunosorbent assay. Interferon (IFN)-γ, interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α were below detectable levels, there were no significant alterations were observed in the levels of IL-2, IL-4, and IL-10.
Fecal bacterial changes
The changes in the distribution of harmful and beneficial bacteria in the feces of pigs after treatment with the complex enzyme were compared from month to month. The number of bacteria in the feces was determined using selective media. Escherichia coli, Staphylococcus aureus, Clostridium, and Salmonella exhibited a decrease and then maintained a steady level, but Lactobacillus, a disease-defense bacterium, was consistently maintained at a certain level. Alternatively, Bifidobacteria showed a decrease in conjunction with E. coli, but at a certain level, they exhibited an increasing trend of recovery (Fig. 4). Changes in the number of harmful bacteria on selective media showed that after 4 months of complex enzyme treatment, the number of coliforms (MAC media) decreased in all 3 farms. S. aureus (mannitol salt [MS] medium) and Clostridium spp. (tryptone sulfite neomycin [TSN] medium) showed a decreasing trend. Salmonella spp. (SS medium) was not present in the 3 farms tested. Unusually, the level of Lactobacillus spp. (MRS medium), a beneficial bacterium often used as a probiotic remained steady. However, Bifidobacteria spp. (TOS medium) showed a decline along with E. coli, but then recovered to a steady level. These results show that the complex enzyme treatment suppressed the harmful enterobacteria while maintaining the beneficial lactobacilli.
Fecal bacterial disease analysis. Fresh feces from the test animals were quantified from a 1 g sample, and the number of bacteria was measured. Aerobic or anaerobic cultures were then performed on the solid medium that was plated with incremental dilutions. The aerobic or anaerobic cultures were conducted using a GasPak system for 48 hours at 37°C. BLA, blood agar; MAC, MacConkey; PDA, potato dextrose agar; MRS, DeMan, Rogosa and Sharpe; MACS, MacConkey with sorbitol; BG, Brilliant Green; TSN, tryptone sulfite neomycin; MS, mannitol salt.
Following the complex enzyme treatment, a marked decrease in E. coli populations was observed in all 3 farms. Initially, high counts were significantly reduced, indicating that the treatment effectively suppressed this group of harmful enterobacteria. The consistent reduction across different environmental conditions and farm management practices suggests a robust effect of the enzyme treatment against E. coli. In contrast to E. coli, the levels of Lactobacillus spp., a beneficial bacterium widely recognized for its probiotic properties, remained stable over the same period. This stability was observed despite the varying conditions across the farms, which underscores the selective efficacy of the enzyme treatment in preserving beneficial bacterial populations while targeting harmful ones.
Fecal microbiome and alpha diversity analysis
We detected 21 bacterial phyla, 390 genera, and 148 families. The top 10 of these were selected from each level for statistical evaluation of the microbiota and are shown in the representative figure (Fig. 5A). The microbiota results revealed that the relative abundance of Spirochetes, Proteobacteria, and Campylobacteria appears to be different at the phylum level. The relative abundance of the families Lachnospiraceae, Spirochaetaceae, and Bacteroidaceae appear to be different at the family level. An alpha diversity analysis was performed to determine the differences in diversity (Shannon), richness (Chao1), and evenness (Evenness) based on time (Fig. 5B). The Shannon index is a measure of diversity that considers both the richness (the number of different species) and the evenness (how evenly the individuals are distributed across those species) of a community. A higher Shannon value indicates a more diverse community, where both species richness and evenness are higher. Chao1 is an estimate of species richness. It is used to predict the total number of species in a community, including those that are rare or undetected in the sampling process. This index is especially useful for identifying whether an observed sample has captured the full diversity of a community or if there may be more species that were not sampled. Evenness measures how evenly individuals are distributed among the species present. It is calculated from the Shannon index but focuses solely on the distribution aspect, not on species richness. An evenness value close to 1 indicates that species are nearly equally abundant, while lower values indicate some species dominate over others. The same graph (parking group) showed a statistically significant difference in diversity and uniformity between the first month and third month compared to the initial period. However, in terms of abundance, there was a tendency to decrease slightly in the first month compared to the initial period, and then an increase in the third month; however, this was not statistically significant. In the beta diversity, grouping according to time (initial, 1 month, 3 months) and the presence or absence of the agent treatment (yes, no) was applied. While the samples were mixed as a whole, the samples corresponded to 1 month, initial, and 3 months. This data demonstrates that the samples are a little further apart than the samples that correspond to the month group. Applying the unweighted UniFrac method, the difference in the microbial community between the samples was confirmed. As a result, it can be seen that the microbial community improved without affecting the overall population, which remained statistically non-significant (Fig. 5B). When the complex enzyme was treated in 3 test farms for 3 months, it was confirmed that it was a very safe biomaterial because it did not affect the composition of intestinal flora, unlike antibiotics.
(A, B) Fecal microbiome and alpha diversity analysis. Microbial DNA was extracted and purified from fecal samples using a QIAamp DNA Stool Mini Kit (QIAGEN Inc., Germany) and then the microbiota was analyzed. The DNA samples quality control requirement was fulfilled with an A260/A280 ratio of > 1.8 and a DNA concentration of > 15 ng/µL. The 16S rRNA gene sequences were obtained from the fecal samples and used as the basis for microbiome profiling. The overall results indicated that the microbial community improved and the overall population remained statistically non-significant.
Evaluation of TGEV, PRRS, and PCV2 infection rates
Nucleic acid was isolated from feces samples and analyzed for TGEV, PRRS, and PCV2 using commercial assay kits. In this test system, TGEV was not detected, while PRRS and PCV2 showed a positive infection. TGEV and PRRS caused significant economic losses and deadly infection while PCV2 has been reported as non-pathogenic in swine. The infection rate for PRRS was insignificant, while the infection for PCV2 was to be 40% to 50% (Fig. 6).
Evaluation of transmissible gastroenteritis virus (TGEV), porcine reproductive and respiratory syndrome (PRRS), and porcine circovirus type 2 (PCV2) infection rates. Viral RNA and DNA were extracted and purified from stool samples following the manufacturer’s protocol using a Patho Gene-spin RNA/DNA Extraction kit (iNTRON Biotechnology, Korea). The extracted viral RNA and DNA were amplified to detect the presence of TGEV, PRRS, and PCV2 using commercial kits (LiliF TGEV, PRRS-RT-PCR, and PCV2-PCR kit, iNtRON Biotechnology, Korea). The product was analyzed on a 1.5% electrophoresis gel to confirm the presence of viral RNA and DNA.
Discussion
Ammonia and hydrogen sulfide are the main factors behind the farm odor, and they are linked to the low availability of dietary protein and increased excretion of fermentation by-products derived from proteins [23,24]. The reduction in ammonia levels and the improved odor quality observed in treated manure highlight the potential of enzyme-based treatments as a viable alternative to conventional odor management practices in pig farms (Fig. 2). Chemical compounds, which are the intermediate or final products of the microbial conversion of wasted nutrients in the feces and slurry, are responsible for the majority of the emitted odors found in livestock farms [25,26]. It is commonly known that exogenous enzymes, such as carbohydrolases and protease, efficiently break down proteins that are resistant to endogenous secretory enzymes and viscous non-starch polysaccharides, which lowers the viscosity of the digestive tract and enhances nutrient absorption [27,28]. The lack of significant changes in immunoreactive cytokine levels, alongside the observed increase in immune-enhancing cytokines, suggests that while the Biomagic-Enzyme complex does not induce an overt inflammatory response, it does enhance the immune competence of pigs (Fig. 3).
Proteobacteria play a crucial role in the carbon and nitrogen cycling in compost, and they increased following Biomagic treatment. Previous studies have reported that an increase in the Proteobacteria population has beneficial effects on growth promotion and nitrogen fixation [29]. Conversely, a decrease in Firmicutes and an increase in Bacteroidetes population from the first to the third month of supplementation generally support the concept that a lower Firmicutes/Bacteroidetes ratio has a direct correlation with weight loss and health improvement, as was observed in our results. After the enzyme treatment, E. coli populations significantly decreased across all 3 farms, demonstrating the treatment's effectiveness. In contrast, beneficial Lactobacillus spp. levels remained stable, indicating the enzyme selectively targeted harmful bacteria while preserving beneficial ones (Fig. 4).
The Lachnospiraceae family has been reported to increase metabolic activity through the coproduction of hydrogen [30]. Electrogenic species, Spirochaetaceae, were found to be higher in the third month compared to the first month and have been reported to improve methane recovery from fat, oil, and grease [31]. Similar beneficial improvements have been observed for Bacteroidaceae, which is found to be higher in our results from the third month (Fig. 5). Lachnospiraceae was also detected to be improving toward the third month. It appears that there is an increase in Treponema and Bacteroides at the genus level from the first to the third month, and these have been reported to have beneficial effects when used to treat food waste and improve growth rates [32]. The changes in microbial diversity indices such as Shannon and Chao1, observed over time with Biomagic treatment, suggest that the enzyme complex might lead to an initial disturbance in the microbial community, followed by a stabilization phase. While short-term effects show a reduction in pathogenic bacteria and stabilization of beneficial microbes, the long-term effects of Biomagic on microbial diversity and function would need further study. These results suggest that Biomagic can significantly impact the microbiota, particularly in agricultural settings, by reducing harmful bacteria and maintaining beneficial ones. This effect on microbial balance can enhance pigs' health and improve environmental conditions.
Poor environmental and rearing conditions lead to the transmission of many swine diseases. TGEV is widespread globally and significantly reduces the profitability of the pig industry [33]. TGEV is a positive-sense RNA coronavirus that targets the epithelium of the pig gut and is single-stranded [34]. Piglets infected with TGEV under 14 days of age have a mortality rate close to 100% and exhibit symptoms such as vomiting, severe diarrhea, and dehydration. PRRS, a highly contagious viral disease, includes early birth, delayed abortion, stillbirth, weak and mummified fetuses, and respiratory dysfunction in piglets and developing pigs [35]. However, PCV1 does not cause disease in pigs. In pigs that were 2 to 4 months old, PCV2 was linked in the 1990s to postweaning multisystemic wasting syndrome (PMWS). Clinical symptoms of PMWS include enlarged subcutaneous lymph nodes, wasting, diarrhea, respiratory distress, pallor, and rarely icterus [36]. The lack of significant changes in immunoreactive cytokine levels, alongside the observed increase in immune-enhancing cytokines, suggests that the immune modulation likely contributes to the reduced susceptibility to viral infections such as PRRSV and PCV2, as evidenced by the absence of TGEV and the manageable levels of PRRSV and non-pathogenic PCV2 (Fig. 6). These findings align with previous research indicating that a balanced gut microbiota and improved air quality can strengthen immune defense and reduce the prevalence of respiratory diseases in livestock [37].
The study’s results also underscore the potential of using enzyme treatments to reduce antibiotic use in pig farming. By enhancing microbial health and boosting immune function, enzyme treatments like Biomagic could decrease the reliance on antibiotics for disease prevention and treatment. This approach addresses the pressing issue of antibiotic resistance, which is exacerbated by the overuse of antibiotics in animal agriculture [38]. However, further research is needed to fully understand the long-term effects of Biomagic-Enzyme complex treatment on pig health and farm environments. Future studies should explore the mechanistic pathways through which enzyme treatments influence microbial dynamics and immune responses. Additionally, large-scale field trials are necessary to validate the efficacy and economic feasibility of implementing such treatments in commercial pig farming operations.
Notes
Author’s Contributions
Conceptualization: Rhee MH; Data curation: Akram AW, Cho HY; Formal analysis: Akram AW, Cho HY; Funding acquisition: Rhee MH; Investigation: Rhee MH; Methodology: Akram AW, Cho HY; Project administration: Rhee MH; Resources: Rhee MH, Park SC; Software: Akram AW, Cho HY; Supervision: Rhee MH; Validation: Rhee MH, Park SC, Kim SD; Visualization: Rhee MH; Writing–original draft: Akram AW, Saba E, Park SC, Lee GY, Park SC, Han YG; Writing–review & editing: Akram AW, Saba E, Park SC, Kim SD, Han YG.
Conflict of interest
The authors declare no conflict of interest.
Funding
This work was supported by an Agriculture and Fisheries R&D Revitalization Project by Gyeongsangbuk-do 2022 and National Research Foundation of Korea (NRF) grant funded by the Korea Government (MSIT) (no. 2022R1A2C1012963).
Supplementary Materials
Supplementary data are available at https://doi.org/10.14405/kjvr.20240051.
Blood inflammatory cytokines and chemokines are not affected by supplementing Biomagic
Blood biochemical indices (red blood cell [RBC] count, white blood cell count [WBC], platelet count [PLT], hemoglobin concentration [HGB], hematocrit [Ht], and mean RBC volume [MCV]) were measured using an automatic hematology analyzer.
Western blot gel bands were visualized. TGEV, transmissible gastroenteritis virus; PRRS, porcine reproductive and respiratory syndrome; PCV2, porcine circovirus type 2.