Antimicrobial activity of poultry hatch baskets containing copper inserts

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

Development of copper hatch baskets and evaluation of the productive indexes of a commercial hatchery

Background: The dissemination of pathogenic microorganisms in hatcheries leads to a higher number of contaminated eggs, causing reduction in hatchability and increase of discarded chicks. Sanitation programs are crucial for maximum hatchability and chick quality. Efforts have been made to find alternative approaches to the conventional disinfectants, and surfaces with copper, which have antimicrobial properties, could assist in this process. However, the possible adverse effects of copper surfaces on chicks in hatcheries have not yet been evaluated. The present study aimed at developing hatch baskets composed of copper and evaluating the effect of these baskets on the productive indexes of a hatchery. Materials, Methods & Results: For this experiment, 3.15 kg hatch tray prototypes with 99.9% Cu (Cu11000) were developed to fit inside conventional polypropylene hatch baskets (580 × 755 × 83 mm). Six polypropylene hatch baskets (control group) and six polypropylene hatch baskets covered by 99.9% copper (Cu11000) hatch trays (test group) were evaluated during 5 hatchings. Hatched eggs and chicks remained in contact with the hatch basket surfaces for at least 72 h, corresponding to the entire period in which they were located in the hatcher. Cleaning and disinfection programs of the hatchery were not modified. The level of microbial contamination on the hatch baskets was evaluated at 6 different periods: 0 h (initial contamination after disinfection and egg transfer to the trays); 24 h, 30 h, 45 h and 60 h after the first sampling; and at the moment when chicks were removed from the hatching cabinet and transferred to the chick-holding room (> 60 h). Counting of total moulds and yeasts, mesophilic microorganisms, Enterobacteria and Escherichia coli colonies was performed. The number of hatched chicks, non-hatched eggs, and chicks discarded were registered for each hatching. Microbiologic analyses showed no growth on hatch baskets neither of the test group nor on those of the control group, regardless of the period evaluated. No significant difference was found between the two groups in the counts of hatched chicks, non-hatched eggs, and chicks discarded. Discussion: The antimicrobial efficacy of copper surfaces has been established for a variety of bacteria and fungi, including in the healthcare environment. In addition, antimicrobial resistance to copper is extremely rare because of copper’s multisite kill mechanism and other mostly nonspecific damage mechanisms. Several copper applications have been established; however, the potential adverse effects of using copper surfaces in hatcheries on chicks or on hatchery productive indexes had not yet been evaluated. The analyses performed in this study did not show microbiological growth. The results showed that the copper surface did not cause any significant adverse effects on chicks. The hatched chicks were visually healthy, and no significant difference was found between the numbers of chicks discarded in the control and test groups. Since the production parameters were not altered or impaired in the present study, it is possible to conclude that the use of copper surfaces in hatcheries can be an effective tool in the microbiological control of these environments. Nevertheless, copper alloys lend themselves to the creation of self-sanitizing surfaces that should be used as a complement and not as a substitute for standard cleaning and disinfection practices

Development of Copper Hatch Baskets and Evaluation of the Productive Indexes of a Commercial Hatchery

Background: The dissemination of pathogenic microorganisms in hatcheries leads to a higher number of contaminated eggs, causing reduction in hatchability and increase of discarded chicks. Sanitation programs are crucial for maximum hatchability and chick quality. Efforts have been made to find alternative approaches to the conventional disinfectants, and surfaces with copper, which have antimicrobial properties, could assist in this process. However, the possible adverse effects of copper surfaces on chicks in hatcheries have not yet been evaluated. The present study aimed at developing hatch baskets composed of copper and evaluating the effect of these baskets on the productive indexes of a hatchery.Materials, Methods and Results: For this experiment, 3.15 kg hatch tray prototypes with 99.9% Cu (Cu11000) were developed to fit inside conventional polypropylene hatch baskets (580 × 755 × 83 mm). Six polypropylene hatch baskets (control group) and 6 polypropylene hatch baskets covered by 99.9% copper (Cu11000) hatch trays (test group) were evaluated during 5 hatchings. Hatched eggs and chicks remained in contact with the hatch basket surfaces for at least 72 h, corresponding to the entire period in which they were located in the hatcher. Cleaning and disinfection programs of the hatchery were not modified. The level of microbial contamination on the hatch baskets was evaluated at 6 different periods: 0 h (initial contamination after disinfection and egg transfer to the trays); 24 h, 30 h, 45 h and 60 h after the first sampling; and at the moment when chicks were removed from the hatching cabinet and transferred to the chick-holding room (> 60 h). Counting of total moulds and yeasts, mesophilic microorganisms, Enterobacteria and Escherichia coli colonies was performed. The number of hatched chicks, non-hatched eggs, and chicks discarded were registered for each hatching. Microbiologic analyses showed no growth on hatch baskets neither of the test group nor on those of the control group, regardless of the period evaluated. No significant difference was found between the two groups in the counts of hatched chicks, non-hatched eggs, and chicks discarded. Discussion: The antimicrobial efficacy of copper surfaces has been established for a variety of bacteria and fungi, including in the healthcare environment. In addition, antimicrobial resistance to copper is extremely rare because of copper’s multisite kill mechanism and other mostly nonspecific damage mechanisms. Several copper applications have been established; however, the potential adverse effects of using copper surfaces in hatcheries on chicks or on hatchery productive indexes had not yet been evaluated. The analyses performed in this study did not show microbiological growth. The results showed that the copper surface did not cause any significant adverse effects on chicks. The hatched chicks were visually healthy, and no significant difference was found between the numbers of chicks discarded in the control and test groups. Since the production parameters were not altered or impaired in the present study, it is possible to conclude that the use of copper surfaces in hatcheries can be an effective tool in the microbiological control of these environments. Nevertheless, copper alloys lend themselves to the creation of self-sanitizing surfaces that should be used as a complement and not as a substitute for standard cleaning and disinfection practices

Impact of a Bacillus Direct-Fed Microbial on Growth Performance, Intestinal Barrier Integrity, Necrotic Enteritis Lesions, and Ileal Microbiota in Broiler Chickens Using a Laboratory Challenge Model

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

Atividade antimicrobiana de superfícies de cobre frente à formação de biofilmes por Salmonella enteritidis e sua potencial aplicação na indústria avícola

Salmonella é considerada um dos patógenos de maior relevância para o setor avícola, uma vez que a carne de frango representa a principal fonte de infecção desta bactéria para o homem. O sorovar S. Enteritidis é identificado como o mais frequentemente envolvido nas salmoneloses alimentares. Na indústria, o controle de biofilmes é rotineiramente realizado através da higienização com a utilização de desinfetantes, os quais apresentam toxicidade conhecida e eficácia variável, devido à evolução da resistência antimicrobiana. O cobre vem sendo aplicado como superfície de contato com poder antimicrobiano em diversos países e pode ser utilizado como um complemento das práticas padrão de limpeza e desinfecção das superfícies industriais, conferindo redução da formação de biofilmes, diminuição da carga bacteriana persistente e aumentando a segurança dos alimentos. Neste contexto, o presente estudo teve como objetivo avaliar as propriedades antimicrobianas de superfícies de cobre e suas ligas na prevenção da formação do biofilme por S. Enteritidis e avaliar o potencial de corrosão das superfícies, quando em contato com desinfetante. Para avaliação da atividade antimicrobiana, foram utilizados cupons com área de 1 cm2, compostos de cobre 99,9% (C11000), latão (C26000), cobre revestido com estanho e, como controle, aço inoxidável (AISI 316). O método utilizado foi o de diluição, com teste de suspensão de células planctônicas e contagem bacteriana realizada através do método de Drop plate, com posterior incubação em temperaturas que simulam ambientes de processamento avícola (4ºC, 12ºC e 25ºC). Para a avaliação do potencial de corrosão, foram realizados ensaios de polarização potenciostática com as mesmas superfícies. Diferenças estatísticas significativas (p<0,05) na produção de biofilme foram observadas nas superfícies de cobre e latão, quando comparadas ao aço inoxidável e às superfícies revestidas com estanho nas três temperaturas avaliadas. Não foram observadas diferenças no potencial de corrosão das superfícies quando em contato com o desinfetante cloreto de benzalcônio, nas concentrações de 100, 200 e 400ppm, em relação ao controle. Estes resultados demonstraram efetividade na redução de S. Enteritidis, sugerindo que superfícies de cobre e suas ligas podem ser utilizadas como um método antimicrobiano complementar no setor avícola para controle deste patógeno importante para a saúde pública.Salmonella is considered one of the most relevant pathogens for the poultry production chain, since poultry meat represents the main source of salmonelosis for humans. Serotype S. Enteritidis is identified as the most frequently involved in food salmonellosis. In the industry, biofilm control is routinely carried out through sanitization using disinfectants, which have known toxicity and variable efficacy, due to the evolution of antimicrobial resistance. Copper has been applied as a contact surface with antimicrobial power in several countries and can be used as a complement to standard practices of cleaning and disinfection of industrial surfaces, conferring reduction of biofilm formation, reduction of persistent bacterial load and increasing food safety. In this context, this study aimed at evaluating the antimicrobial properties of copper surfaces and their alloys in preventing biofilm formation by S. Enteritidis and to evaluate the corrosion potential of surfaces when in contact with disinfectant. To evaluate the antimicrobial activity, coupons with an area of 1 cm2, composed of 99.9% copper (C11000), brass (C26000), tin-coated copper and, as a control, stainless steel (AISI 316) were used. The method used was dilution, with planktonic cell suspension testing and bacterial counting using the Drop plate method, with subsequent incubation at temperatures that simulate poultry processing environments (4 ºC, 12 ºC and 25 ºC). Potentiostatic polarization tests, with the same surfaces were performed to evaluate the corrosion potential. Significant statistical differences (p<0,05) in the production of biofilm were observed on copper and brass surfaces when compared to stainless steel and tin-coated surfaces at the three temperatures evaluated. There were no differences in the potential for surface corrosion when in contact with the benzalkonium chloride disinfectant, at concentrations of 100, 200 and 400 ppm, compared to the control. These results demonstrated effectiveness in reducing S. Enteritidis, suggesting that copper surfaces and their alloys can be used as a complementary antimicrobial method in the poultry sector to control this important pathogen for public health

Development of copper hatch baskets and evaluation of the productive indexes of a commercial hatchery

Background: The dissemination of pathogenic microorganisms in hatcheries leads to a higher number of contaminated eggs, causing reduction in hatchability and increase of discarded chicks. Sanitation programs are crucial for maximum hatchability and chick quality. Efforts have been made to find alternative approaches to the conventional disinfectants, and surfaces with copper, which have antimicrobial properties, could assist in this process. However, the possible adverse effects of copper surfaces on chicks in hatcheries have not yet been evaluated. The present study aimed at developing hatch baskets composed of copper and evaluating the effect of these baskets on the productive indexes of a hatchery. Materials, Methods & Results: For this experiment, 3.15 kg hatch tray prototypes with 99.9% Cu (Cu11000) were developed to fit inside conventional polypropylene hatch baskets (580 × 755 × 83 mm). Six polypropylene hatch baskets (control group) and six polypropylene hatch baskets covered by 99.9% copper (Cu11000) hatch trays (test group) were evaluated during 5 hatchings. Hatched eggs and chicks remained in contact with the hatch basket surfaces for at least 72 h, corresponding to the entire period in which they were located in the hatcher. Cleaning and disinfection programs of the hatchery were not modified. The level of microbial contamination on the hatch baskets was evaluated at 6 different periods: 0 h (initial contamination after disinfection and egg transfer to the trays); 24 h, 30 h, 45 h and 60 h after the first sampling; and at the moment when chicks were removed from the hatching cabinet and transferred to the chick-holding room (> 60 h). Counting of total moulds and yeasts, mesophilic microorganisms, Enterobacteria and Escherichia coli colonies was performed. The number of hatched chicks, non-hatched eggs, and chicks discarded were registered for each hatching. Microbiologic analyses showed no growth on hatch baskets neither of the test group nor on those of the control group, regardless of the period evaluated. No significant difference was found between the two groups in the counts of hatched chicks, non-hatched eggs, and chicks discarded. Discussion: The antimicrobial efficacy of copper surfaces has been established for a variety of bacteria and fungi, including in the healthcare environment. In addition, antimicrobial resistance to copper is extremely rare because of copper’s multisite kill mechanism and other mostly nonspecific damage mechanisms. Several copper applications have been established; however, the potential adverse effects of using copper surfaces in hatcheries on chicks or on hatchery productive indexes had not yet been evaluated. The analyses performed in this study did not show microbiological growth. The results showed that the copper surface did not cause any significant adverse effects on chicks. The hatched chicks were visually healthy, and no significant difference was found between the numbers of chicks discarded in the control and test groups. Since the production parameters were not altered or impaired in the present study, it is possible to conclude that the use of copper surfaces in hatcheries can be an effective tool in the microbiological control of these environments. Nevertheless, copper alloys lend themselves to the creation of self-sanitizing surfaces that should be used as a complement and not as a substitute for standard cleaning and disinfection practices

Cutaneous Myxoma in Broiler Chicken Detected during Slaughtering

Background: A cutaneous or superficial myxoma is a benign neoplasm of dermal or subcutaneous fibroblast origin. Although rare, it has been previously described in several species, including poultry. It presents as a single node or soft mass with a gelatinous cut surface. Histopathological analysis is essential for diagnosis and to differentiate it from other mesenchymal neoplasms and inflammatory or degenerative processes. Microscopically, it consists of dermal or subcutaneous lobules of plump, stellate, or spindle-shaped, bland-looking cells embedded in a basophilic myxoid matrix. This report describes the pathological findings in a rare case of cutaneous myxoma in a 42-day-old broiler flock. Cases: During ante mortem inspection of a 42-day-old broiler flock at a slaughterhouse under the authority of the Federal Inspection Service (southern Brazil), nodular lesions or encrusted areas with yellow and black areas were observed in the head skin of less than 1% of animals. These lesions, approximately 0.5 cm in diameter, were observed on the comb, in the periocular skin region, and close to the animals' nostrils. During the breeding period, no health or epidemiological events were observed. Fragments of the lesions in the comb and periocular skin were collected and fixed in buffered 10% formalin. The samples were sent to the laboratory, routinely processed, and stained with hematoxylin and eosin and Alcian blue. Microscopically, the lesions consisted of irregular multifocal proliferation of connective tissue showing spindle cells with poorly demarcated borders and scarce cytoplasm in a slightly basophilic myxoid aspect matrix. The adjacent epidermis is compressed due to neoplastic proliferation. No areas of epithelial hyperplasia or inclusion bodies were observed. According to the pathologic description and considering its descriptive epidemiology, our main clinical suspicion was cutaneous fowl pox, a pathology characterized by the appearance of nodules in regions devoid of feathers. However, the microscopic changes observed were compatible with those described for cutaneous myxomas. In addition, the extracellular matrix was positive for Alcian Blue staining, which is an indicator of myxoma. In the present case, the SIF did not report the same macroscopic lesions in other flocks of the same origin. Discussion: Connective tissue tumors, including myxomas, occur considerably less frequently under field conditions. In addition, these neoplasms are more frequent in mature birds and are not usually described in broilers, as observed in this report. The cutaneous myxoma described in broilers is usually a sporadic neoplasm that does not cause zootechnical losses, as observed in the case report. Its etiology is unknown and has been associated with various factors, such as local trauma and foreign bodies. Some fragments of plant material from the breeding environment were microscopically detected in the encrusted areas, which may indicate previous trauma or a foreign body. Myxoma has been associated with avian leukosis virus (ALV) subgroup A, but SIF did not report the same macroscopic lesions in other flocks of the same breeder hen’s origin in the present case. Furthermore, sporadic connective tissue tumors associated with the virus occur in mature chickens but not in broilers. Myxoma lesions should be considered in the differential diagnosis of other connective tissue tumors and infectious agents that cause lesions in the skin and subcutaneous tissue. Keywords: benign neoplasia, connective tissue tumor, histopathology, poultry

Evaluation of a Solid Dispersion of Curcumin With Polyvinylpyrrolidone and Boric Acid Against Salmonella Enteritidis Infection and Intestinal Permeability in Broiler Chickens: A Pilot Study

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 &lt; 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

Evaluation of the Dietary Supplementation of a Formulation Containing Ascorbic Acid and a Solid Dispersion of Curcumin with Boric Acid against <i>Salmonella</i> Enteritidis and Necrotic Enteritis in Broiler Chickens

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