Significant alterations of intestinal symbiotic microbiota induced by intraperitoneal vaccination mediate changes in intestinal metabolism of NEW Genetically Improved Farmed Tilapia (NEW GIFT, Oreochromis niloticus).

BACKGROUND: After millions of years of coevolution, symbiotic microbiota has become an integral part of the host and plays an important role in host immunity, metabolism, and health. Vaccination, as an effective means of preventing infectious diseases, has been playing a vital role in the prevention and control of human and animal diseases for decades. However, so far, minimal is known about the effect of vaccination on fish symbiotic microbiota, especially mucosal microbiota, and its correlation with intestinal metabolism remains unclear. METHODS: Here we reported the effect of an inactivated bivalent Aeromonas hydrophila/Aeromonas veronii vaccine on the symbiotic microbiota and its correlation with the intestinal metabolism of farmed adult Nile tilapia (Oreochromis niloticus) by 16S rRNA gene high-throughput sequencing and gas chromatography-mass spectrometry metabolomics. RESULTS: Results showed that vaccination significantly changed the structure, composition, and predictive function of intestinal mucosal microbiota but did not significantly affect the symbiotic microbiota of other sites including gill mucosae, stomach contents, and stomach mucosae. Moreover, vaccination significantly reduced the relative abundance values of potential opportunistic pathogens such as Aeromonas, Escherichia-Shigella, and Acinetobacter in intestinal mucosae. Combined with the enhancement of immune function after vaccination, inactivated bivalent Aeromonas vaccination had a protective effect against the intestinal pathogen infection of tilapia. In addition, the metabolite differential analysis showed that vaccination significantly increased the concentrations of carbohydrate-related metabolites such as lactic acid, succinic acid, and gluconic acid but significantly decreased the concentrations of multiple lipid-related metabolites in tilapia intestines. Vaccination affected the intestinal metabolism of tilapia, which was further verified by the predictive function of intestinal microbiota. Furthermore, the correlation analyses showed that most of the intestinal differential microorganisms were significantly correlated with intestinal differential metabolites after vaccination, confirming that the effect of vaccination on intestinal metabolism was closely related to the intestinal microbiota. CONCLUSIONS: In conclusion, this paper revealed the microbial and metabolic responses induced by inactivated vaccination, suggesting that intestinal microbiota might mediate the effect of vaccination on the intestinal metabolism of tilapia. It expanded the novel understanding of vaccine protective mechanisms from microbial and metabolic perspectives, providing important implications for the potential influence of vaccination on human intestinal microbiota and metabolism. Video Abstract.

An investigation into an outbreak of pancytopenia in cats in the United Kingdom.

BACKGROUND: In spring 2021 increasing numbers of cats presenting with severe pancytopenia were noted in United Kingdom (UK). OBJECTIVE: To describe process and outcome of the investigation performed into the outbreak of pancytopenia in cats. ANIMALS: Five hundred and eighty client owned cats that presented with severe bi- or pancytopenia of unknown cause. METHODS: Real-time data collection was performed by an online registration forum available to all veterinary surgeons in UK. Data collected included demographics, clinicopathological findings, diagnostic testing, dietary and drug history, outcome and COVID household status. Mycotoxicological feed analysis was performed on feed samples of 3 diets frequently mentioned in the database and 3 control diets. RESULTS: Five hundred and eighty cats presented to 378 veterinary practices were included for analysis. Case fatality rate was 63.3%. Dietary history was available for 544 (93.8%) cats, of which 500 (86%) were fed 1 of 3 diets (which were recalled midinvestigation). 54 (9.3%) cats were not fed a recalled product, with diet information unknown in 26 (4.5%) cats. Analysis of feed samples revealed concentrations of hematotoxic trichothecene T-2/HT-2 mycotoxins greater than recommended by the European Commission in 5/7 recalled diet samples but in none of control diet samples. The trichothecene mycotoxin diacetoxyscirpenol (DAS) was detectable in all recalled diet samples but not in any of control samples. CONCLUSION AND CLINICAL IMPORTANCE: Contaminated-feed induced trichothecene mycotoxicosis should be considered as a differential diagnosis for pancytopenia in cats.

Production efficiency of animal feed obtained from food waste in Japan.

Converting food waste into animal feed is highly useful for tackling the problem of food waste, which is particularly severe in developed countries. This study quantified the inefficiencies in converting food waste into animal feed and identified the sources of inefficiencies through a data envelopment analysis (DEA) of the monthly input-output data of two food waste-based animal feed producers in Japan. Our empirical analysis revealed that the producers of animal feed obtained from food waste (especially those treating food waste from retail and service industries) demonstrated inefficiencies in production technology and scale; moreover, expanding the production scale and improving the quality of food waste could enhance production efficiency. Based on the empirical results, specific policy implications were provided for the widespread use of animal feed obtained from food waste in Japan and elsewhere, globally. Furthermore, it was suggested that the COVID-19 pandemic contributed to a severe reduction in the production efficiency of animal feed producers treating food waste obtained from retail and service industries.

Growth performance, carcass and meat quality, bone strength, and immune response of broilers fed low-calcium diets supplemented with marine mineral complex and phytase.

Influence of marine mineral complex (CeltiCal) as a partial substitute for limestone on growth efficiency, carcass traits, meat quality, bone strength, calcium (Ca) retention, and immune response was investigated in broilers fed low-Ca diets with or without phytase (PHY) addition for a 35-d trial period. A total of 300 one-day-old Ross 308 straight-run broilers were randomly allocated to: T1 (positive control), recommended Ca levels + PHY; T2 (negative control), 0.2% below the recommended Ca levels + PHY; T3, 0.1% below the recommended Ca levels + 0.2% CeltiCal + PHY; T4, 0.2% below the recommended Ca levels + 0.4% CeltiCal + PHY; T5, 0.2% below the recommended Ca levels + 0.4% CeltiCal. PHY was added at 500 phytase units/kg diets. Each dietary treatment had 10 replications of 6 chicks each. Results revealed that production efficiency factor was greater for T4 compared to T2 and T5 during 22-35 d and for T1, T3, and T4 compared to T2 during 0 to 35 d (P < 0.05). Feed conversion ratio was lower for T3 and T4 compared to T2 and T5 during 0 to 35 d (P < 0.05). T4 had a greater (P < 0.05) dressing percentage than T2, which had a lighter (P < 0.01) small intestinal relative weight than all other treatments. Breast meat temperature at 15 min postmortem was highest for T1 and lowest for T3 (P < 0.001). Breast meat pH was greater for T1 compared to T5 at 15 min postmortem and for T3 compared to T4 at 24 h postmortem (P < 0.05). T5 had a lower breast meat redness than all other treatments at 15 min postmortem and then T1 and T3 at 24 h postmortem (P < 0.01). Tibia and femur weights were greater (P < 0.05) for T3, T4, and T5 compared to T2, which had the lowest tibia ash content (P < 0.05) and femur geometric properties (P < 0.001). Greater antibodies to infectious bronchitis virus (P < 0.01) and Ca retention (P < 0.001) were observed for T3 and T4 in comparison to T2. Based on the findings of this research, CeltiCal can adequately replace a considerable portion of limestone in broiler reduced-Ca diets containing PHY.

Mycotoxins in Maize Silage from China in 2019.

Animal feed (including forage and silage) can be contaminated with mycotoxins. Here, 200 maize silage samples from around China were collected in 2019 and analyzed for regulated mycotoxins, masked mycotoxins (deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, and deoxynivalenol-3-glucoside), and emerging mycotoxins (beauvericin, enniatins, moniliformin, and alternariol). Deoxynivalenol and zearalenone were detected in 99.5% and 79.5% of the samples, respectively. Other regulated mycotoxins were detected in fewer samples. The highest deoxynivalenol and zearalenone concentrations were 3600 and 830 µg/kg, respectively. The most commonly detected masked mycotoxin was 15-acetyldeoxynivalenol, which was detected in 68.5% of the samples and had median and maximum concentrations of 61.3 and 410 µg/kg, respectively. The emerging mycotoxins beauvericin, alternariol, enniatin A, enniatin B1, and moniliformin were detected in 99.5%, 85%, 80.5%, 72.5%, and 44.5%, respectively, of the samples but at low concentrations (medians <25 µg/kg). The samples tended to contain multiple mycotoxins, e.g., the correlation coefficients for the relationships between the concentrations of beauvericin and deoxynivalenol, deoxynivalenol and zearalenone, and zearalenone and beauvericin were 1.0, 0.995, and 0.995, respectively. The results indicated that there needs to be more awareness of the presence of one or more masked and emerging mycotoxins in maize silage in China.

Different methods of eubiotic feed additive provision affect the health, performance, fermentation, and metabolic status of dairy calves during the preweaning period.

BACKGROUND: This study aimed to evaluate whether different methods of providing eubiotic feed additives to neonatal calves, during the preweaning period, can improve the calves' health, performance, ruminal fermentation, and metabolic status. Forty-four (3-day-old) Holstein-Friesian dairy calves (22 female and 22 male) were divided into four treatment groups for the duration of the 8-week trial. The eubiotic feed additive consisted of a combination of probiotic Lactobacillus spp. (multiple-strains at a dose of 250 mg/calf/day) and phytobiotics containing rosmarinic acid, as the main bioactive compound (at a dose of 50 mg/calf/day). The groups were named: CON (control, without eubiotic in either the milk replacer or the starter feed), MR (eubiotic in the milk replacer), SF (eubiotic in the starter feed), MRS (eubiotic in both the milk replacer and the starter feed). The individual intake of starter feed and the fecal scores were measured daily, and body weight and biometric measurements were taken weekly until calves were 56 days of age. Blood samples were collected on day 3 and then every 14 days to determine concentrations of insulin-like-growth-factor-I, ß-hydroxybutyrate, non-esterified fatty acids, and blood urea nitrogen. Ruminal fluid was collected on days 28 and 56 for short-chain fatty acids, NH3-N, and pH measurements. RESULTS: The body weight of the calves of the MR treatment group was higher compared to all other groups on days 28 and 56. Including the eubiotic feed additive in the milk replacer increased average daily gain, starter intake, and total dry matter intake from day 29 to day 56 and the overall experimental period compared to the CON group. The calves with MR treatment had lower fecal scores from days 3 to 28, a number of parasite oocysts/cysts per gram of feces on day 28, and the occurrences of fecal consistency scores of 3 (mild diarrhea) and 4 (severe diarrhea) were 3.2 and 3.0 times lower, respectively, compared with the CON group. The MR group had higher ruminal concentrations of short-chain-fatty-acids, propionate, and butyrate on day 56 than the CON group. Adding eubiotics into milk replacer resulted in the highest concentrations of blood insulin-like-growth-factor-I and ß-hydroxybutyrate from days 29 to 56 and the overall experimental period. CONCLUSION: The addition of eubiotic feed additives into the milk replacer can improve health, performance, ruminal fermentation, and biochemical blood indices in dairy calves during the preweaning period.

Effect of Dietary L-Threonine and Toxin Binder on Performance, Blood Parameters, and Immune Response of Broilers Exposed to Aflatoxin B1.

To evaluate the effect of L-Threonine (L-Thr) and Mycofix® Plus (MP) on aflatoxicosis, an experiment with a 3-way ANOVA model was carried out with 8 replicates and 640 birds. Treatments included two levels of L-Thr (100% and 125% of the requirements, Cobb 500, Cobb-Vantress), Aflatoxin B1 (AFB1) (0, 500 ppb), and MP (0, 1 g/kg). As the main effects showed, AFB1 decreased breast meat yield and carcass percentage (p < 0.001), serum urea, antibody titer against infectious bronchitis virus (IBV), and bone density (p < 0.05), while it increased the plasma concentrations of glucose and alkaline phosphatase (ALP) (p < 0.05). Mycofix Plus improved the grower feed intake (FI), tibia fresh weight, and body weight (BW) to bone weight (p < 0.05). L-Threonine increased the grower FI, breast meat yield, serum aspartate transaminase (AST), and glutathione peroxidase (GPX) (p < 0.05). There were positive interactions with breast meat yield, cholesterol, lactate dehydrogenase (LDH), and IBV titer. Of the treatments used, the combination of L-Thr and MP without AFB1 improved breast meat and carcass percentage. L-Threonine and MP significantly improved IBV titer in birds challenged with AFB1 (p < 0.001). In conclusion, L-Thr and MP were beneficial to improve immunity.

Adaptation of fuel selection to acute decrease in voluntary energy expenditure is governed by dietary macronutrient composition in mice.

In humans, exercise-induced thermogenesis is a markedly variable component of total energy expenditure, which had been acutely affected worldwide by COVID-19 pandemic-related lockdowns. We hypothesized that dietary macronutrient composition may affect metabolic adaptation/fuel selection in response to an acute decrease in voluntary activity. Using mice fed short-term high-fat diet (HFD) compared to low-fat diet (LFD)-fed mice, we evaluated whole-body fuel utilization by metabolic cages before and 3 days after omitting a voluntary running wheel in the cage. Short-term (24-48 h) HFD was sufficient to increase energy intake, fat oxidation, and decrease carbohydrate oxidation. Running wheel omission did not change energy intake, but resulted in a significant 50% decrease in total activity and a ~20% in energy expenditure in the active phase (night-time), compared to the period with wheel, irrespective of the dietary composition, resulting in significant weight gain. Yet, while in LFD wheel omission significantly decreased active phase fat oxidation, thereby trending to increase respiratory exchange ratio (RER), in HFD it diminished active phase carbohydrate oxidation. In conclusion, acute decrease in voluntary activity resulted in positive energy balance in mice on both diets, and decreased oxidation of the minor energy (macronutrient) fuel source, demonstrating that dietary macronutrient composition determines fuel utilization choices under conditions of acute changes in energetic demand.

Quantification of soya-based feed ingredient entry from ASFV-positive countries to the United States by ocean freight shipping and associated seaports.

African swine fever virus (ASFV) can survive in soya-based products for 30 days with T ½ ranging from 9.6 to 12.9 days in soya bean meals and soya oil cake. As the United States imports soya-based products from several ASFV-positive countries, knowledge of the type and quantity of these specific imports, and their ports of entry (POE), is necessary information to manage risk. Using the data from the International Trade Commission Harmonized Tariff Schedule website in conjunction with pivot tables, we analysed imports across air, land and sea POE of soya-based products from 43 ASFV-positive countries to the United States during 2018 and 2019. In 2018, 104,366 metric tons (MT) of soya-based products, specifically conventional and organic soya bean meal, soya beans, soya oil cake and soya oil were imported from these countries into the United States via seaports only. The two largest suppliers were China (52.7%, 55,034 MT) and the Ukraine (42.9%, 44,775 MT). In 2019, 73,331 MT entered the United States and 54.7% (40,143 MT) came from the Ukraine and 8.4% (6,182 MT) from China. Regarding POE, 80.9%-83.2% of soya-based imports from China entered the United States at the seaports of San Francisco, CA, and Seattle, WA, while 89.4%-100% entered from the Ukraine via the seaports of New Orleans, LA, and Charlotte, NC. Analysis of five-year trends (2015-2019) of the volume of soya imports from China indicated reduction over time (with a noticeably sharp decrease between 2018 and 2019), and seaport utilization was consistent. In contrast, volume remained high for Ukrainian soya imports, and seaport utilization was inconsistent. Overall, this exercise introduced a new approach to collect objective data on an important risk factor, providing researchers, government officials and industry stakeholders a means to objectively identify and quantify potential channels of foreign animal disease entry into the United States.

Use of Integrative Interactomics for Improvement of Farm Animal Health and Welfare: An Example with Fescue Toxicosis.

Rapid scientific advances are increasing our understanding of the way complex biological interactions integrate to maintain homeostatic balance and how seemingly small, localized perturbations can lead to systemic effects. The 'omics movement, alongside increased throughput resulting from statistical and computational advances, has transformed our understanding of disease mechanisms and the multi-dimensional interaction between environmental stressors and host physiology through data integration into multi-dimensional analyses, i.e., integrative interactomics. This review focuses on the use of high-throughput technologies in farm animal research, including health- and toxicology-related papers. Although limited, we highlight recent animal agriculture-centered reports from the integrative multi-'omics movement. We provide an example with fescue toxicosis, an economically costly disease affecting grazing livestock, and describe how integrative interactomics can be applied to a disease with a complex pathophysiology in the pursuit of novel treatment and management approaches. We outline how 'omics techniques have been used thus far to understand fescue toxicosis pathophysiology, lay out a framework for the fescue toxicosis integrome, identify some challenges we foresee, and offer possible means for addressing these challenges. Finally, we briefly discuss how the example with fescue toxicosis could be used for other agriculturally important animal health and welfare problems.

Antimicrobial multidrug resistance in the era of COVID-19: a forgotten plight?

BACKGROUND: Antimicrobial resistance (AMR) is a growing global problem to which the ongoing COVID-19 pandemic may further contribute. With resources deployed away from antimicrobial stewardship, evidence of substantial pre-emptive antibiotic use in COVID-19 patients and indirectly, with deteriorating economic conditions fuelling poverty potentially impacting on levels of resistance, AMR threat remains significant. MAIN BODY: In this paper, main AMR countermeasures are revisited and priorities to tackle the issue are re-iterated. The need for collaboration is stressed, acknowledging the relationship between human health, animal health and environment ("One Health" approach). Among the stated priorities, the initiative by the European Medicines Regulatory Network to further strengthen the measures in combatting AMR is highlighted. Likewise, it is asserted that other emerging health threats require global collaboration with the One Health approach offering a valuable blueprint for action. CONCLUSION: The authors stress the importance of an integrated preparedness strategy to tackle this public health peril.

Reviewing the risk of feed as a vehicle for swine pathogen transmission.

BACKGROUND: While porcine biological hazards have had the potential to be transmitted through feed and feed mills for decades, the emerging threat of foreign animal disease has elevated the concern that these may enter or be transmitted throughout the domestic swine herd via a feed vehicle. OBJECTIVE: The goal of this review was to describe the current classification for emerging porcine biological pathogen transmission through the feed supply chain so resources can be best directed towards those of highest risk. METHODS: By assessing the pathogen severity to pigs and the probability of pathogen transmission through feed, an overall risk can be established using a hazard analysis matrix. RESULTS: There is negligible risk for feed-based transmission of a transmissible spongiform encephalopathy, Trichinella spiralis, Toxoplasma gondii, Salmonella Choleraesuis, Salmonella spp. except Choleraesuis and I 4,[5],12:i:-, porcine deltacoronavirus, Senecavirus A, mammalian orthoreovirus 3, foot and mouth disease virus, classical swine fever virus or Chinese pseudorabies virus. However, the combined severity and probability of Salmonella enterica serotype I 4,[5],12:i:-, porcine epidemic diarrhoea virus and African swine fever virus warrant a moderate risk characterization for transmission through the US feed supply chain. CONCLUSIONS: This risk can be maintained below critical status by minimizing the likelihood that a pathogen can enter the feed supply chain, such as by excluding high-risk ingredients from facilities, extending biosecurity to mills, and considering proactive mitigation strategies. In reality, all these actions may be necessary to prevent the detrimental transmission of porcine biological hazards into the US swine herd through the feed supply chain.

Zinc source modulates zootechnical characteristics, intestinal features, humoral response, and paraoxonase (PON1) activity in broilers.

The current experiment was performed to find the potential effect of inorganic and organic forms of zinc (Zn) on growth, intestinal histomorphology, immune response, and paraoxonase (PON1) activity in broiler. In this experiment, a total of 450 broiler chickens were assigned to four experimental and control groups. The birds received organic Zn at the rate of 50 mg/kg (OZ-50) and 60 mg/kg (OZ-60) or inorganic Zn at the rate of 50 mg/kg (IZ-50) and 60 mg/kg (IZ-60) for an experimental period of 30 days. Significantly (P < 0.05) higher feed consumption, body weight, feed conversion ratio, and production efficiency factor (PEF) were recorded in OZ-50. Similarly, antibody titer against infectious bronchitis (IB) and PON1 activity was higher (P < 0.05) in OZ-50 compared with the control group. In addition, significantly (P < 0.05) higher villus dimensions and goblet cell count were recorded for the group OZ-50 compared with other treatments. It was concluded that the organic form of Zn was superior in improving the growth, histological features of intestines, humoral response, and PON1 activity in broiler.

Effect of dietary medium-chain fatty acids on nursery pig growth performance, fecal microbial composition, and mitigation properties against porcine epidemic diarrhea virus following storage.

An experiment was conducted to evaluate the effect of dietary medium-chain fatty acid (MCFA) addition on nursery pig growth performance, fecal microbial composition, and mitigation of porcine epidemic diarrhea virus (PEDV) following storage. A total of 360 pigs (DNA 400 × 200, Columbus, NE; initially 6.7 ± 0.07 kg) were randomized to pens (5 pigs per pen) on the day of weaning (approximately 20 d of age), allowed a 6-d acclimation, blocked by BW, and randomized to dietary treatment (9 pens per treatment). All MCFA (Sigma-Aldrich, St. Louis, MO) were guaranteed ≥98% purity, including hexanoic (C6:0), octanoic (C8:0), and decanoic (C10:0) acids. Treatment diets were formulated in 2 phases (7 to 11 and 11 to 23 kg BW) and formulated to meet or exceed NRC requirement estimates. Treatments (n = 8) were a dose response including 0%, 0.25%, 0.5%, 1.0%, and 1.5% added MCFA blend (1:1:1 ratio C6:0, C8:0, and C10:0), as well as treatments with individual additions of 0.5% C6:0, C8:0, or C10:0. Fecal samples were collected from pigs fed control and 1.5% MCFA blend diets on days 0 and 14 and analyzed using 16s rDNA sequencing. Following feed manufacture, feed was stored in bags at barn temperature and humidity for 40 d before laboratory inoculation with PEDV. Subsamples of retained feed were inoculated with PEDV to achieve a titer of 104 TCID50/g and separate sample bottles were analyzed on 0 and 3 d post-inoculation (dpi). Overall, ADG and ADFI were increased (linear, P ≤ 0.010) and feed efficiency (G:F) improved (linear, P = 0.004) with increasing MCFA blend. Pigs fed 0.5% C8:0 had greater (P = 0.038) ADG compared with pigs fed the control diet, and G:F was improved (P ≤ 0.024) when pigs were fed 0.5% C6:0, 0.5% C8:0, or 0.5% C10:0 compared with control. An inclusion level × day interaction was observed (quadratic, P = 0.023), where PEDV Ct values increased (quadratic, P = 0.001) on 0 dpi with increasing levels of MCFA blend inclusion and also increased on 3 dpi (linear, P < 0.001). Fecal microbial diversity and composition were similar between control and 1.5% MCFA blend. In summary, the use of MCFA in nursery pig diets improves growth performance, provides residual mitigation activity against PEDV, and does not significantly alter fecal microbial composition.

Do different vaccination regimes affect the growth performance, immune status, carcase characteristics and meat quality of broilers?

1. A vaccination regime is a schedule for the administration of vaccines which may vary according to country or even by farm. This study aimed to measure the production and health performance of broilers treated with different vaccination regimes. 2. A total of 108 Cobb 500 broiler birds were randomly divided into three treatment groups, with six replicates consisting of six birds per replicate. Each treatment group was administered with different vaccination regimes against Newcastle Disease (ND), Infectious Bronchitis (IB) and Infectious Bursal Disease (IBD). Treatment 1 (T1) broilers were vaccinated against ND+IB and IBD on days 7 and 14 of age, respectively (control); Treatment 2 (T2) broilers were vaccinated against ND+IB on days 3 and 7 of age, and IBD on day 14; and Treatment 3 (T3) broilers were vaccinated against ND+IB on days 7 and 21 and IBD on day 14. Throughout the 42-day study period, data and samples were collected to determine the growth performance, immune status, carcase characteristics and meat quality. 3. There were significant differences (P < 0.05) on growth performance (body weight, body weight gain, feed intake and cumulative feed conversion ratio), white blood cell count (heterophils percentage, lymphocytes percentage and heterophils to lymphocytes ratio), carcase characteristics (kill-out weight, de-feathered weight, dressing percentage, drumsticks and gastrointestinal tract weight) and meat quality (cooking loss and drip loss) between treatments. T1 broilers showed better growth, white blood cell count, carcase characteristics and meat quality compared to T2 and T3 broilers. 4. Based on findings from the current work, vaccination against ND+IB and IBD on days 7 and 14 proved to be the best vaccination regime for broiler production, due to the better production performance and health status of broilers.

Effects of medium chain fatty acids as a mitigation or prevention strategy against porcine epidemic diarrhea virus in swine feed.

Feed has been shown to be a vector for viral transmission. Four experiments were conducted to: 1) determine if medium chain fatty acids (MCFA) are effective mitigants when applied to feed both pre- and post-porcine epidemic diarrhea virus (PEDV) inoculation measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR), 2) evaluate varying levels and combinations of MCFA measured by qRT-PCR, and 3) evaluate selected treatments in bioassay to determine infectivity. In exp. 1, treatments were arranged in a 2 × 2 + 1 factorial with main effects of treatment (0.3% commercial formaldehyde [CF] product, Sal CURB [Kemin Industries, Inc.; Des Moines, IA], or 1% MCFA blend (Blend) of 1:1:1 C6:C8:C10 [PMI, Arden Hills, MN]) and timing of application (pre- or post-inoculation with PEDV) plus a positive control (PC; feed inoculated with PEDV and no treatment). All combinations of treatment and timing decreased detectable PEDV compared with the PC (P < 0.05). Pre-inoculation treatment elicited decreased magnitude of PEDV detection (cycle threshold value) compared with post-inoculation (P = 0.009). Magnitude of PEDV detection was decreased for CF compared with Blend (P < 0.0001). In exp. 2, pre-inoculation treatments consisted of: 1) PC, 2) 0.3% CF, 3 to 5) 0.125% to 0.33% C6:0, 6 to 8) 0.125% to 0.33% C8:0, 9 to 11) 0.125% to 0.33% C10:0, and 12 to 15) 0.125% to 0.66% C5:0. Treating feed with 0.33% C8:0 resulted in decreased (P < 0.05) PEDV detection compared with all other treatments. Increasing concentration of each individual MCFA decreased PEDV detectability (P < 0.042). In exp. 3, pre-inoculation treatments consisted of: 1) PC, 2) 0.3% CF, 3 to 7) 0.25% to 1% Blend, 8 to 10) 0.125% to 0.33% C6:0 + C8:0, 11 to 13) 0.125% to 0.33% C6:0 + C10:0, and 14 to 16) 0.125% to 0.33% C8:0 + C10:0. Treating feed with CF, 0.5% Blend, 0.75% Blend, 1% Blend, all levels of C6:0+C8:0, 0.25% C6:0 + 0.25% C10:0, 0.33% C6:0 + 0.33% C10:0, 0.25% C8:0 + 0.25% C10:0, or 0.33% C8:0 + 0.33% C10:0 elicited decreased detection of PEDV compared with PC (P < 0.05). Increasing concentration of each MCFA combination decreased PEDV detectability (linear, P < 0.012). In exp. 4, feed was treated pre-inoculation with: 1) no treatment (PC), 2) 0.3% CF, 3) 0.5% Blend, or 4) 0.3% C8:0 and analyzed via qRT-PCR and bioassay. Adding 0.5% Blend or 0.3% C8:0 resulted in decreased PEDV compared with PC and only PC resulted in a positive bioassay. Therefore, MCFA can decrease detection of PEDV in feed. Further, inclusion of lower levels of MCFA than previously evaluated are effective against PEDV.