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SUMMARY Commercial poultry hatcheries provide ideal conditions for the multiplication and spread of microorganisms. Formaldehyde is widely used as a disinfectant; however, it is harmful to human health and can cause abnormal morphology in chicks. An alternative microbiological control is the use of copper, a metallic antimicrobial agent for contact surfaces. The antimicrobial efficacy of copper surfaces has been established in healthcare environments. However, its use in the poultry chain is still limited. This study aimed to compare the antimicrobial activity of common polypropylene hatch baskets with hatch baskets composed of copper (polypropylene hatch baskets covered by copper hatch baskets; polypropylene hatch baskets with solid copper plates on the bottom; polypropylene hatch baskets covered by copper hatch baskets and with solid copper plates on the bottom). To simulate a hatching environment with high contamination, the eggs and hatching cabinet were not fumigated. Microbiological analysis of the hatching cabinet environmental, surface of hatch basket, and fluff were performed. The results indicated that the absence of bottom holes resulted in a higher volume of organic matter that interfered with the copper's antimicrobial activity. The presence of copper in the hatch baskets did not decrease microbial contamination under the conditions evaluated, confirming that the metal should only be used as a complement to standard hygiene and not as a substitute for surface disinfectants. Further analysis will evaluate the antimicrobial activity of hatch baskets composed only of copper and the ability of nanoparticles to remove the biofilms formed by bacteria isolated from the poultry environment.

As a consequence of developing antimicrobial resistance to disinfectants, copper, which exhibits antimicrobial activity, has been studied as a possible alternative to the use of stainless steel surfaces. The aim was to evaluate the antimicrobial activity of copper surfaces in preventing biofilm formation by Salmonella Enteritidis and to determine their corrosive capacity. Strains of S. Enteritidis were incubated at 4 °C, 12 °C, and 25 °C with 1 cm2 coupons of electrolytic copper (99.9% Cu), brass (70% Cu), copper coated with tin, and stainless steel (control). A planktonic cell-suspension assay was used, followed by serial dilutions and bacterial counts. The corrosion test was performed with two disinfectants: benzalkonium chloride and sodium hypochlorite (100, 200, and 400 ppm). There was a significant reduction in biofilm production (log10 CFU cm−2) on the copper (2.64 at 4 °C, 4.20 at 12 °C, 4.56 at 25 °C) and brass (2.79 at 4 °C, 3.49 at 12 °C, 4.55 at 25 °C) surfaces compared to the control (5.68 at 4 °C, 5.89 at 12 °C, 6.01 at 25 °C). The antimicrobial surfaces showed uniform corrosion similar to that of surfaces generally used. These results demonstrated the effectiveness of copper surfaces in reducing S. Enteritidis and suggest they can be used as a complementary antimicrobial to control for this pathogen.

Two experimental models were conducted to evaluate and compare the effect of ascorbic acid (AA) or curcumin formulated in a solid dispersion (SD-CUR) as prophylactic or therapeutic alternatives to prevent or control S. Enteritidis (SE) infection in broiler chickens. In the prophylactic model, dietary administration of AA showed a significant reduction in SE counts in crop compared to the positive control (PC) group (p &lt, 0.05), whereas in cecal tonsils (CT), SD-CUR significantly reduced SE recovery. Superoxide dismutase (SOD) activity was significantly higher in chickens supplemented with AA or SD-CUR, and total intestinal IgA levels were significantly lower in both treatments when compared to the PC group. Serum fluorescein isothiocyanate-dextran (FITC-d) levels were reduced by SD-CUR compared to PC, while AA presented significantly lower total aerobic bacteria. In the therapeutic model, only the dietary administration of AA significantly decreased SE in crop and CT on days 3 and 10 post-challenge. FITC-d levels were significantly lower in both treated groups in comparison to PC, but IgA levels were significantly reduced only by AA. The results suggest that dietary AA and SD-CUR have different modes of action to reduce SE intestinal colonization in two different challenge models in broiler chickens.

Decreases in the use of antibiotics and anticoccidials in the poultry industry have risen the appearance of necrotic enteritis (NE). The purpose of this study was to evaluate the effect of a Bacillus direct-fed microbial (DFM) on growth performance, intestinal integrity, NE lesions and ileal microbiota using a previously established NE-challenged model. At day-of-hatch, chicks were randomly assigned to three different groups: Negative control (NC), Positive control (PC) challenged with Salmonella Typhimurium (day 1), Eimeria maxima (EM, day 13) and Clostridium perfringens (CP, day 18–19), and Bacillus-DFM group (DFM) challenged as the PC. Body weight (BW) and body weight gain (BWG) were measured weekly. Total feed intake (FI) and feed conversion ratio (FCR) were evaluated at day 21. Liver samples were collected to assess bacterial translocation and blood samples were used to measure superoxide dismutase (SOD) and fluorescein isothiocyanate-dextran (FITC-d). Intestinal contents were obtained for determination of total IgA and microbiota analysis. NE lesion scores (LS) were performed at day 21. Chickens consuming the DFM significantly improved BW and had a numerically more efficient FCR compared to PC at day 21. Additionally, there were no significant differences in FCR between the DFM group and NC. Furthermore, the DFM group showed significant reductions in LS, IgA and FITC-d levels compared to the PC. However, there were no significant differences in SOD between the groups. The microbiota analysis indicated that the phylum Proteobacteria was significantly reduced in the DFM group in comparison to PC. At the genus level, Clostridium, Turicibacter, Enterococcus, and Streptococcus were reduced, whereas, Lactobacillus and Bacillus were increased in the DFM group as compared to PC (p < 0.05). Likewise, the DFM significantly reduced CP as compared to PC. In contrary, no significant differences were observed in bacterial composition between NC vs. DFM. In addition, beta diversity showed significant differences in the microbial community structure between NC vs. PC, and PC vs. DFM. These results suggest that the dietary inclusion of a selected DFM could mitigate the complex negative impacts caused by NE possibly through mechanism(s) that might involve modulation of the gut microbiota.

The objectives of the present study were to evaluate the antimicrobial effect of boric acid (BA) on Salmonella enteritidis colonization, intestinal permeability, total intestinal IgA levels, and cecal microbiota composition in broiler chickens. For this purpose, sixty day-old-chicks were randomly allocated to one of two groups: 1) non-treated control group and 2) 0.1% (w/w) BA in feed. All chicks were challenged with 104 cfu of S. Enteritidis per bird at 1-d old. On day 3 and 10 post-challenge, samples of crop and ceca-cecal tonsils (CCT) were aseptically collected to evaluate overall aerobic and lactic acid bacteria, as well as S. Enteritidis recovery. Blood and intestinal samples were also collected to evaluate intestinal permeability and total intestinal IgA production, respectively. On day 10 post-challenge, the concentration of S. Enteritidis in crop and CCT significantly decreased (P

In the present study, in vitro assays were conducted to evaluate the solubility of curcumin (CUR) alone or with polyvinylpyrrolidone (PVP) at different pH, as well as its permeability in Caco-2 cells. Results confirmed that the solid dispersion of CUR with PVP (CUR/PVP) at a 1:9 ratio, significantly increased (P < 0.05) solubility and permeability compared to CUR alone. Then, the antimicrobial activity of CUR/PVP, boric acid (BA), and a combination of 0.5% CUR/PVP and 0.5% BA (CUR/PVP-BA) against Salmonella Enteritidis (SE) was determined using an in vitro digestion model that simulates crop, proventriculus, and intestine. The results revealed that in the proventriculus and intestinal compartments significant reductions of SE were observed in all the experimental treatments, but 1% BA eliminated SE in the intestinal compartment and CUR/PVP-BA showed a synergistic effect on antimicrobial activity against SE. To complement these findings, two independent in vivo trials were conducted to determine the effect of 0.1% CUR/PVP; 0.1% BA; or the combination of 0.05% CUR/PVP (1:9 ratio) and 0.05% BA (CUR/PVP-BA) on the antimicrobial activity against SE, intestinal permeability and inflammatory responses in broiler chickens. BA at 0.1% had no significant in vivo effects against SE. However, the combination of 0.05% BA and 0.05% CUR/PVP and 0.05% BA was sufficient to reduce crop and intestinal SE colonization in broiler chickens in two independent trials, confirming the synergic effect between them. A similar antimicrobial impact against SE intestinal colonization was observed in chickens treated with 0.1% CUR/PVP at a 1:9 ratio, which could be due to the increase in solubility of CUR by PVP. Furthermore, 0.1% CUR/PVP reduced the intestinal permeability of FITC-d and total intestinal IgA, as well as increase the activity of SOD when compared to control, while, CUR/PVP-BA only decreased SOD activity. Further studies to confirm and expand the in vivo results obtained in this pilot study, adding intestinal microbial commensal groups and more inflammatory biomarkers to get a complete description of the effects of BA and CUR deserves further investigation.

International audience; The aim of the present study was to evaluate the effect of a commercial Bacillus direct-fed microbial (DFM) on aflatoxin B1 toxic effects, performance, and biochemical and immunologic parameters in broiler chickens. Ninety 1-day-old Cobb 500 male broiler chicks were raised in floor pens for a period of 21 days. Chicks were neck-tagged, individually weighed, and randomly allocated to one of three groups: Negative control (basal feed), aflatoxin B1 (basal feed + 2 ppm AFB1), and DFM (basal feed + 2 ppm AFB1 + Bacillus direct-fed microbial). Each group had three replicates of 10 chickens (n = 30/group). Body weight and body weight gain were calculated weekly, while feed intake and feed conversion ratio were determined when broilers were 21 days old. On day 21, all chickens were bled, gastrointestinal samples were collected, and spleen and bursa of Fabricius were weighed. This study confirmed that 2 ppm of AFB1 causes severe detrimental effects on performance, biochemical parameters, and immunologic parameters, generating hepatic lesions in broiler chickens (P < 0.05). However, it was also observed that DFM supplementation provided beneficial effects that might help to improve gut barrier function, anti-inflammatory and antioxidant activities, as well as humoral and cellular immunomodulation. The results of the present study suggest that this Bacillus-DFM added at a concentration of 10(6) spores/gram of feed can be used to counteract the negative effects that occur when birds consume diets contaminated with AFB1, showing beneficial effects on performance parameters, relative organ weights, hepatic lesions, immune response, and serum biochemical variables. The addition of this Bacillus-DFM might mitigate and decrease aflatoxicosis problems in the poultry industry, improving food security, alleviating public health problems, and providing economic benefits. Future studies are needed to fully elucidate the specific mechanisms by which this Bacillus-DFM counteracts the toxic effects of aflatoxin B1.

Two experiments were conducted to evaluate the effect of the prophylactic or therapeutic administration of a 0.1% mixture containing ascorbic acid and a solid dispersion of curcumin with polyvinylpyrrolidone and boric acid (AA-CUR/PVP-BA) against Salmonella Enteritidis (S. Enteritidis) in broiler chickens. A third experiment was conducted to evaluate the impact of the dietary administration of 0.1% AA-CUR/PVP-BA in a necrotic enteritis (NE) model in broiler chickens. The prophylactic administration of 0.1% AA-CUR/PVP-BA significantly decreased S. Enteritidis colonization in cecal tonsils (CT) when compared to the positive control group (PC, p &lt, 0.05). The therapeutic administration of 0.1% AA-CUR/PVP-BA significantly reduced the concentration of S. Enteritidis by 2.05 and 2.71 log in crop and CT, respectively, when compared with the PC on day 10 post-S. Enteritidis challenge. Furthermore, the serum FITC-d concentration and total intestinal IgA levels were also significantly lower in chickens that received 0.1% AA-CUR/PVP-BA. Contrary, the PC group showed significantly higher total intestinal IgA levels compared to the negative control or AA-CUR/PVP-BA groups in the NE model. However, 0.1% AA-CUR/PVP-BA showed a better effect in reducing the concentration of S. Enteritidis when compared to the NE model. Further studies with higher concentration of AA-CUR/PVP-BA into the feed to extend these preliminary results are currently being evaluated.

To evaluate the effect of cellulosic polymers (CEL) and curcumin (CUR) on aflatoxin B1 (AFB1) toxic effects on performance, and the biochemical and immunological parameters in broiler chickens, 150 one-day-old male broiler chicks were randomly allocated into five groups with three replicates of 10 chickens per pen: Negative Control (feed), AFB1 (feed + 2 ppm AFB1), CUR (feed + 2 ppm AFB1 + Curcumin 0.2%), CEL (feed + 2 ppm AFB1 + 0.3% Cellulosic polymers), and, CEL + CUR (feed + 2 ppm AFB1 + 0.3% Cellulose polymers + 0.2% Curcumin). Every week, body weight, body weight gain, feed intake, and feed conversion ratio were calculated. On day 21, liver, spleen, bursa of Fabricius, and intestine from five broilers per replicate per group were removed to obtain relative organ weight. Histopathological changes in liver, several biochemical biomarkers, antibody titers, and muscle and skin pigmentation were also recorded. Dietary addition of 0.3% CEL and 0.2% CUR separately significantly diminished some of the toxic effects resulting from AFB1 on performance parameters, relative organs weight, histopathology, immune response, and serum biochemical variables (P &lt, 0.05), however, the combination of CUR and CEL showed a better-integrated approach for the management of poultry health problems that are related with the consumption of AFB1, since they have different mechanisms of action with different positive effects on the responses of broiler chickens.

Since 2008, when the US Environmental Protection Agency (EPA) registered copper and its alloys as an antimicrobial agent for contact surfaces, research has demonstrated their antimicrobial activity. The aim of this study was to evaluate the efficacy of antimicrobial copper against bacteria and fungi isolated from commercial poultry hatcheries in order to develop a microbiological control alternative in these environments. Samples were collected from the surfaces of hatcher baskets from two hatcheries. Mesophilic microorganisms and fungi/yeasts were isolated and standardized in concentration of 105 cells/mL. Four copper plates and four stainless steel plates were completely immersed for one minute in bacteria and fungi/yeasts solutions and left to dry for a day at room temperature. Subsequently, samples were collected from the metal plates with the aid of sterile swab and delimiter. These samples were planted onto Plate Count Agar (for mesophilic culture) and Sabouraud Dextrose Agar (for fungi and yeast culture) and incubated at 36°C for 48 hours and at 25°C for 5-7 days, respectively. After incubation, the colonies recovered from the plates were counted according to IN 62 of the Brazilian Ministry of Agriculture. Almost all contamination was eliminated from the surface of copper plates in a single day, while the stainless steel plates proved to be innocuous to the screened microorganisms. Copper, as a contact surface, proved to have important antimicrobial action on bacteria, fungi and yeasts common to hatcheries.