Probiotic microorganisms detoxify ochratoxin A in both a chicken liver cell line and chickens.

BACKGROUND: The administration of probiotics and prebiotics (synbiotics) is a promising method for detoxification of ochratoxin A (OTA) in animals. The aim of this study was to investigate the ability of five probiotic strains of lactic acid bacteria (LAB) and one Saccharomyces cerevisiae yeast strain, from three different synbiotics for poultry, to detoxify OTA. In addition, we also investigated the genotoxicity of faecal water (FW) of chickens after administering OTA and/or synbiotics for 42 days. RESULTS: All tested LAB and yeast strains had the ability to detoxify OTA by significant (P < 0.05) reducing its concentration (by 31.3-47.7% and 31.9%, respectively, after 24 h incubation) and genotoxicity (by 22.6-51.8% and 52.7%, respectively). Synbiotics composed of four and five probiotic strains significantly (P < 0.05) decreased FW genotoxicity of chicks, after exposure to OTA, to the level seen in the control group (21.8% ± 1.7%) and were more effective than synbiotics composed of three probiotic strains (31.5%). CONCLUSION: These results showed that there was a beneficial effect of the synbiotics on the gastrointestinal tract of animals. Furthermore, synbiotic preparations containing four or five of tested strains can be considered as preventive agents in the contamination of poultry with OTA. © 2019 Society of Chemical Industry.

3D chick embryo chorioallantoic membrane model as an in vivo model to study morphological and histopathological features of feline fibrosarcomas.

BACKGROUND: The chick embryo chorioallantoic membrane (CAM) model is well described in human medicine as a cost-effective, easy to perform preclinical oncological model for observing pro- and antiangiogenic response, tumor biology and metastasis. The main objective of this article was to present the modification of the CAM assay in order to evaluate tumor growth from two feline fibrosarcoma cell lines (FFS1, FFS3) and describe their morphological and histopathological features. RESULTS: The authors described morphological and histopathological features of two feline fibrosarcoma cell lines (FFS1 and FFS3) grown on the CAM. Tumors from the FFS1 cell line showed high malignancy (grade III), while tumors from the FFS3 cell line were grade II. Proliferation markers (Ki-67 and PCNA) were determined and the positive correlation between PCNA and tumor grade (r = 0.8247; p < 0.001) was demonstrated, as opposed to Ki-67. CONCLUSIONS: The results obtained indicate that PCNA may be helpful to evaluate the tumor grade, better than Ki-67, for feline fibrosarcomas. However, further investigations of proliferation marker, in bigger number of feline spontaneous fibrosarcomas and feline fibrosarcomas grown on the CAM from different cell lines, are needed to confirm these observations.

Doxorubicin Conjugated to Glutathione Stabilized Gold Nanoparticles (Au-GSH-Dox) as an Effective Therapeutic Agent for Feline Injection-Site Sarcomas-Chick Embryo Chorioallantoic Membrane Study.

Feline injection-site sarcomas are malignant skin tumours with a high local recurrence rate, ranging from 14% to 28%. The treatment of feline injection-site sarcomas includes radical surgery, radiotherapy and/or chemotherapy. In our previous study it has been demonstrated that doxorubicin conjugated to glutathione-stabilized gold nanoparticles (Au-GSH-Dox) has higher cytotoxic effects than free doxorubicin for feline fibrosarcoma cell lines with high glycoprotein P activity (FFS1, FFS3). The aim of the present study was to assess the effectiveness of intratumoural injection of Au-GSH-Dox on the growth of tumours from the FFS1 and FFS3 cell lines on chick embryo chorioallantoic membrane. This model has been utilized both in human and veterinary medicine for preclinical oncological studies. The influence of intratumoural injections of Au-GSH-Dox, glutathione-stabilized gold nanoparticles and doxorubicin alone on the Ki-67 proliferation marker was also checked. We demonstrated that the volume ratio of tumours from the FFS1 and FFS3 cell lines was significantly (p < 0.01) decreased after a single intratumoural injection of Au-GSH-Dox, which confirms the positive results of in vitro studies and indicates that Au-GSH-Dox may be a potent new therapeutic agent for feline injection-site sarcomas.

Supporting Measures to Improve Biosecurity within Italian Poultry Production.

This paper describes the selection and validation of supporting measures (SMs) aimed at enhancing biosecurity compliance within Italian poultry farms. A tailored methodology, based on a stakeholders' survey involving farmers and advisors, included a virtual farm tour, group discussion, and farmer coaching. Virtual farm tours and group discussions were delivered during two meetings targeting meat and egg production stakeholders, separately. Coaching was validated in 26 pilot farms (PFs) by assessing farmers' attitudes towards change (i.e., ADKAR®) and farms' biosecurity score (i.e., Biocheck.UgentTM) before and after a minimum six-month period. A total of 20 out of 26 farmers agreed to implement at least one action plan (AP). Full implementation of the agreed APs was observed in ten farms, while others only partially implemented (n = 7) or did not implement (n = 3) the improvement. Most APs focused on enhancing house hygiene locks (n = 7), followed by bacterial auto-control after cleaning and disinfection (n = 4). Scoring tools indicated minimal or no variations in farmers' attitudes towards change and farm biosecurity. Virtual farm tours and group discussions were found to be effective in fostering interaction and facilitating the exchange of experiences and knowledge among farmers and stakeholders of poultry production. Coaching indicated that farmers might prefer implementing minor changes possibly influenced by time and cost constraints associated with structural interventions. These limitations could have also impacted the scores of the farmer/farm. The findings of this study provide a foundation for further application of SMs to improve biosecurity in Italian poultry farms.

Factors Affecting Poultry Producers' Attitudes towards Biosecurity.

Poultry producers' attitudes towards biosecurity practices were assessed by using the ADKAR® (Awareness, Desire, Knowledge, Ability, and Reinforcement) behavioral change model. Conventional poultry producers (n = 155) from different production types including broilers (n = 35), layers (n = 22), breeders (n = 24), turkeys (n = 19), ducks (n = 23), free-range broilers (n = 11), free-range layers (n = 11), and hatcheries (n = 10) from seven European countries were scored for each ADKAR element (1 = total absence to 5 = perfect fulfilment). Each country performed selected interventions (e.g., coaching, participatory meetings, etc.) to improve biosecurity compliance. After the interventions, significant change was observed in three of the four attitude elements. The overall mean scores (x¯ ± SD) obtained during the initial assessment (n = 130) were 4.2 ± 0.6 for Awareness, 4.1 ± 0.7 for Desire, 3.8 ± 0.8 for Knowledge, and 4.0 ± 0.7 for Ability, whereas after intervention, the scores were A = 4.3 ± 0.6, D = 4.2 ± 0.7, K = 4.1 ± 0.7, and Ab = 4.1 ± 0.7. The Reinforcement component was only evaluated after the change and obtained a score of 3.7 ± 0.7 on average. Identifying the elements influencing poultry producers and their behavior related to farm management decisions was useful in guiding our educational interventions to effectively change their behavior.

Development and use of Biocheck.UGent™ scoring system to quantify biosecurity in conventional indoor (turkey, duck, breeder) and free-range (layer and broiler) poultry farms.

To assess and enhance the application of biosecurity measures in poultry farming, an objective measurement tool (Biocheck.UGent™) was already available for broiler and layer. This study describes the development, validation and application of a risk-based weighted scoring tool for breeder, turkey, duck, free-range layer and free-range broiler production. In collaboration with an expert panel (n= 38), five different questionnaires were developed, following the format of the existing Biocheck.UGent scoring tools. Weights were attributed to external (7-9 subcategories) and internal (3-4 subcategories) biosecurity categories, as well as to the corresponding individual questions within each subcategory. The biosecurity measures were prioritized and weighed based on their relative importance in preventing disease transmission. Upon completion of the questionnaire, and upload of all answers to the Biocheck.UGent website, the algorithm generates a biosecurity score varying between ''0'' which equals the total absence of any biosecurity measure up to ''100'' which refers to full application of all biosecurity measures. The final scoring systems are available online (https://biocheckgent.com/en) for free and have been used to assess biosecurity in 70 breeders, 100 turkeys, 23 ducks, 16 free-range broilers, and 15 free-range layer farms originating from 12 countries so far. On average, the overall biosecurity score (mean ± std. dev) was 78 ± 7 % for breeders, 73 ± 11 % for turkeys, 71 ± 8 % for ducks, 73 ± 8 % for free-range layers and 70 ± 13 % for free-range broilers. There were significant differences (p < 0.05) in biosecurity (both at the overall and subcategory levels) across different poultry types. The overall farm biosecurity score for breeders was significantly higher than that for turkey (p <.001) and duck production (p = 0.001). External biosecurity levels were highest in breeders in comparison to turkeys (p < 0.001), ducks (p = 0.008) and broiler free-range (p = 0.005). There was a notable difference in internal biosecurity levels between duck and turkey (p = 0.041) production as well. The study contributed to the poultry biosecurity database which allows benchmarking of the biosecurity levels of the users' farm results to national or international averages, indicating room for improvement and aiding to motivate stakeholders to enhance their biosecurity levels.

Biosecurity implementation on large-scale poultry farms in Europe: A qualitative interview study with farmers.

Biosecurity is an essential tool for rearing healthy animals. Biosecurity measures (BMs) are well known in poultry production, but it is difficult to assess actual implementation on farms. The aims of this qualitative study were (1) to provide an overview of biosecurity implementation according to poultry farmers in Europe; and (2) to better understand the reported reasons and potential obstacles for not implementing the measures. In seven European Union Member States, 192 farmers (118 under contract with a company and 68 independents) working in seven different categories of poultry production were interviewed on 62 BMs to determine the frequency of implementation and the reasons for non-implementation. Most of the replies (n = 7791) concerning BM implementation were reported by the farmers as "always" implemented (81%), statistically higher for breeders (87%) and layers (82%) and lower for independent farms versus farms under contract with a company (79.5% and 82.5%, respectively). Regardless the poultry production category, the most frequently implemented BMs declared by the farmers were daily surveillance of birds, rodent control and feed storage protection. Standard hygiene practices were also mentioned as high-implementation measures for most production categories, with some deficiencies, such as rendering tank disinfection after each collection and, for meat poultry, disinfection of the feed silo and bacterial control of house cleaning and disinfection between each cycle. The entry of vehicles and individuals onto poultry farms, especially during critical points of eggs collection for breeders and layers, as well as the presence of other animals, such as the "all in/all out" practice, particularly in layers and ducks, were also reported as the least commonly practiced measures. The main reasons for not implementing the measures (n = 1683 replies) were low awareness and poor knowledge of the expected benefits of biosecurity ("no known advantages" 14%, and "not useful" 12%), the lack of training ("not enough training" 5% and "advice" 7%), lack of time (19%), and financial aspects (17%). Despite the good overall biosecurity mentioned by the farmers, these findings highlight certain deficiencies, suggesting room for improvement and the need for targeted and tailored support of poultry farmers in Europe.

Tissue tropism of Ornithobacterium rhinotracheale in chickens determined by culture and nucleic acid detection.

The aim of the present study was to evaluate the ability of Ornithobacterium rhinotracheale (ORT) to colonize chosen organs of chicks infected intratracheally (group A1), or intravenously (group A2), with the use of bacteriological methods and PCR. The bacteriological methods enabled to reisolate ORT bacteria from trachea and lungs of the birds from group A1 only on day 3 and 6 after infection. The PCR technique additionally detected the bacterial genetic material in these organs on the 9th day after infection, and gave positive results in the samples from air sacs until the 6th day of the experiment. In birds infected intravenously (A2) ORT was reisolated from liver on day 3 and from spleen on day 3 and 6 after infection, whereas the reisolation from the tibiotarsal joint occurred during the entire experimental period. PCR enabled to detect the bacterial DNA in the liver, spleen and lungs of chickens until the 9th day after infection and in case of tibiotarsal joint during the whole time of the study.

Silica-Calcite Sedimentary Rock (Opoka) Enhances the Immunological Status and Improves the Growth Rate in Broilers Exposed to Ochratoxin A in Feed.

Mycotoxins, such as Ochratoxin A (OTA), originating from fungi like Aspergillus and Penicillium, represent serious health hazards to poultry. The use of mycotoxin-adsorbing feed additives can reduce these risks. Opoka, a porous transitional rock, shows promise as one of these additives. This study is the first to examine the effect of Opoka administered with OTA on zootechnical parameters and the immune response of chickens. A 42-day investigation examined the impact of 1% of Opoka supplementation in feed on OTA-challenged broiler chickens. Seventy-two chickens were allocated into three groups of twenty-four individuals each: a control group, an OTA-exposed (2 mg/kg feed) group, and an OTA (2 mg/kg feed) plus 1% of Opoka group. Growth and blood parameters were monitored at predetermined intervals, and comprehensive biochemical, hematological, and cytometric analyses were conducted. The study showed that OTA exposure had a negative impact on chicken weight gain. However, adding Opoka to the diet improved weight gain, indicating its potential as a protective agent. Chickens fed with Opoka also had an increased white blood cell count, which suggests an improved immune response and elevated glucose and cholesterol concentrations. These findings indicate that Opoka may be useful in mitigating health complications caused by OTA exposure in broilers.

Monitoring biosecurity in poultry production: an overview of databases reporting biosecurity compliance from seven European countries.

Compliance with required on-farm biosecurity practices reduces the risk of contamination and spread of zoonotic and economically important diseases. With repeating avian influenza epidemics in the poultry industry, the need to monitor and improve the overall level of biosecurity is increasing. In practice, biosecurity compliance is assessed by various actors (e.g., academic, private and public institutions), and the results of such assessments may be recorded and gathered in databases which are seldom shared or thoroughly analyzed. This study aimed to provide an inventory of databases related to the assessment of biosecurity in poultry farms in seven major poultry-producing European countries to highlight challenges and opportunities associated with biosecurity data collection, sharing, and use. The institutions in charge of these databases were contacted and interviewed using a structured questionnaire to gather information on the main characteristics of the databases and the context of their implementation. A total of 20 databases were identified, covering the gamut of poultry species and production types. Most databases were linked to veterinary health authorities or academia, and to a lesser extent interbranch organizations. Depending on the institutions in charge, the databases serve various purposes, from providing advice to enforcing regulations. The quality of the biosecurity data collected is believed to be quite reliable, as biosecurity is mostly assessed by trained farm advisors or official veterinarians and during a farm visit. Some of the databases are difficult to analyze and/or do not offer information concerning which biosecurity measures are most or least respected. Moreover, some key biosecurity practices are sometimes absent from certain databases. Although the databases serve a variety of purposes and cover different production types, each with specific biosecurity features, their analysis should help to improve the surveillance of biosecurity in the poultry sector and provide evidence on the benefits of biosecurity.

The effect of synbiotic preparations on the intestinal microbiota and her metabolism in broiler chickens.

The aim of the research was to determine the effect of newly elaborated synbiotic preparations on the count of dominant intestinal microorganisms, on the profile of fatty acids (short chain - SCFA and branched chain - BCFA), the lactic acid produced and the performance of chickens. The studies determined the composition of the dominant intestinal microbiota with use of the culture method. The fatty acid profile was also determined using the high-performance liquid chromatography method (HPLC). Moreover, the performance of chickens was determined such as the daily cumulative mortality rate, the feed conversion ratio (FCR) and the European Production Efficiency Factor (EPEF). It was found that synbiotics had a beneficial effect on parameters of the performance of chickens, and also resulted in increase in the count of beneficial bacteria and to the restriction in growth of potential pathogens in the gastrointestinal tract. Synbiotics caused an increase in the concentration of lactic acid and SCFA and a decrease in the concentration of BCFA in the broiler's excreta. These results showed a beneficial effect of the tested synbiotics on the intestinal microbiota, their metabolism and the performance of broiler chickens. The elaborated synbiotics can be successfully used as feed additives for broiler chickens.

Comparative Effects of Using New Multi-Strain Synbiotics on Chicken Growth Performance, Hematology, Serum Biochemistry and Immunity.

In this study, the influence of new multi-strain synbiotics on chicken growth performance, hematology, serum biochemistry and immunity was explored. Each synbiotic preparation (A, B and C) comprised three, four or five strains of Lactobacillus sp., respectively, as well as S. cerevisiae and inulin. All strains used in the synbiotics originated from wild-type strains from animal farms in Poland. Six groups of chickens, ROSS 308 line, were fed with three different synbiotics at a dose of 0.5 g/1 kg of feed. Body weight, as well as the biochemical and hematological parameters of the animals in each study group, were determined on the 7th, 14th, 28th and 42nd day of life. Body weight on day 42 differed between groups and was the highest in control group. This group also had the highestfeed conversion ratio (FCR) value. All measured biochemical parameters were in the normal ranges for poultry; however, we observed a lower alkaline phosphatase (AP) concentration on day 7 in the groups fed with synbiotics, which correlated with a lower level of triglycerides in those groups. The aspartate transaminase (AST) concentration was significantly lower in all groups on day 42 in comparison with the control. On day 7, the control group showed the highest concentration of Ca, K and P. Other parameters did not differ significantly throughout the experiment. All groups showed a similar tendency of increase in the red blood cells (RBC) count according to the age of the birds. Every white blood cells (WBC) population showed differences in the proportions between T and B lymphocytes. The T cell and monocyte counts increased until day 28 in all groups. The results showed that our newly developed synbiotic formulas do not have any unfavorable influence on chicken health and may modulate immune response and biochemical parameters. However, this hypothesis needs to be evaluated in future experiments.

Effects of synbiotics on the gut microbiota, blood and rearing parameters of chickens.

The aim of the study was to assess the safety of three newly developed synbiotic preparations in feeding chickens. The study was carried out on 84 Specific Pathogen Free chickens, between 1st and 21st day of their life. Animals were divided into four groups fed ad libitum: three with an addition of synbiotics A, B or C and a control group (feed with no additives). Synbiotics contained Lactobacillus, Saccharomyces cerevisiae and inulin (prebiotic). Rearing parameters were determined taking into account undesirable clinical and sectional symptoms, daily mortality and body weight of birds. In addition, chicken blood parameters were determined based on haematological, biochemical and serological tests. Samples of the intestinal content and of the excreta constituted a material for research aimed at determination of the dominant chicken gut microbiota. On the basis of the conducted investigation, it was found that synbiotics met the basic requirements for this type of formulas regarding the safety of use and had a positive effect on the health of chickens. Therefore, further research is being carried out on the application of these formulas in large-scale production. The next step of the research will be the comparison of effects of synbiotics with commercial probiotics.

Experimental tumor growth of canine osteosarcoma cell line on chick embryo chorioallantoic membrane (in vivo studies).

The chick embryo chorioallantoic membrane (CAM) model is extensively used in human medicine in preclinical oncological studies. The CAM model has several advantages: low cost, simple experimental approach, time saving and following "3R principles". Research has shown that the human osteosarcoma cell lines U2OS, MMNG-HOS, and SAOS can form tumors on the CAM. In veterinary medicine, this has been described only for feline fibrosarcomas, feline mammary carcinomas and canine osteosarcomas. However, in case of canine osteosarcomas, it has been shown that only non-adherent osteosarcoma stem cells isolated from KTOSA5 and CSKOS cell lines have the ability to form microtumors on the CAM after an incubation period of 5 days, in contrast to adherent KTOSA5 and CSKOS cells. In the presented study, we have proven that the commercial adherent canine osteosarcoma cell line (D-17) can form vascularized tumors on the CAM after the incubation period of 10 days.

The response of mute swans (Cygnus olor, Gm. 1789) to vaccination against avian influenza with an inactivated H5N2 vaccine.

BACKGROUND: Recent epidemics of highly pathogenic avian influenza (HPAI) produced an unprecedented number of cases in mute swans (Cygnus olor) in European countries, which indicates that these birds are very sensitive to the H5N1 virus. The HPAI outbreaks stirred a debate on the controversial stamping-out policy in populations of protected bird species. After preventive vaccination had been approved in the European Union, several countries have introduced vaccination schemes to protect poultry, captive wild birds or exotic birds in zoos against HPAI. The aim of this study was to investigate the immune response of wild mute swans to immunization with an inactivated AI H5N2 vaccine approved for use in poultry. The serological responses of mute swans were assessed by comparison with racing pigeons (Columba livia), a species which is characterized by different susceptibility to infection with the H5N1 HPAI virus and plays a questionable role in the ecology of influenza (H5N1) viruses. RESULTS: Swans were vaccinated once or twice at an interval of 4 weeks. The humoral immune response was evaluated by hemagglutination inhibition (HI) and NP-ELISA. The lymphocyte blast transformation test was used to determine the cell-mediated immune response. Higher values of the geometric mean titer (GMT) and 100 % seroconversion (HI ≥32) were noted in double vaccinated swans (1448.2) than in single vaccinated swans (128.0) or in double vaccinated pigeons (215.3). Significant differences in HI titers were observed between swans and pigeons, but no variations in ELISA scores were noted after the booster dose. Immunization of swans had no effect on the proliferative activity of lymphocytes. CONCLUSIONS: The inactivated H5N2 vaccine was safe and immunogenic for mute swans and pigeons. Vaccination may have practical implications for swans kept in zoos, wildlife parks or rehabilitation centers. However, challenge studies are needed to prove the efficacy of the H5N2 AI vaccine.

Histomonosis - an existing problem in chicken flocks in Poland.

Histomonosis (histomoniasis, blackhead), beside coccidiosis, belongs to the most important parasitic protozoan diseases in poultry. So far Histomonas meleagridis infections with varied mortality rates have been mainly diagnosed in young turkeys. Recently an increasing number of cases have been reported in chicken flocks in Europe resulting in economic losses. It is thought that this situation is predominantly caused by a complete withdrawal of the effective antihistomonals in the EU. Authors listed the selected outbreaks of histomonosis in 10 chicken flocks originated from different farms of 4 regions in Poland: 8 broiler breeder flocks (at mean age of 33 weeks) and 2 commercial layers flocks (at mean age of 38 weeks). This study reported here naturally occurring case of H.meleagridis infection in commercial broiler breeder (BB) flock line ROSS 308 at the age of 16 weeks. We showed acute form of infection with characteristic necrotic foci in the liver, and ulcerative typhilitis. Beside the liver and caeca, the multiple histomonads, lymphoid tissue depletion and heavy destruction in the bursa of Fabricius were observed. Additionally, the absence of systemic diffuse histomonads and lack of Heterakis gallinarum, caecal worm eggs in faecal samples were noted. PCR technique enabled to detect the presence of H.meleagridis genetic material in the investigated tissue samples. Authors indicate that histomonosis can be considered as re-emerging infectious diseases in chicken flocks of intensive production system.