Yolkin, a Polypeptide Complex from Egg Yolk, Affects Cytokine Levels and Leukocyte Populations in Broiler Chicken Blood and Lymphoid Organs after In Ovo Administration.

Yolkin is a polypeptide complex isolated from hen egg yolk that exhibits immunomodulating properties. The aim of the present study was to determine whether in-ovo-delivered yolkin affects leukocyte populations and cytokine levels in broiler chickens. The experiment was carried out on eggs from Ross 308 broiler breeder birds. Yolkin was administered in ovo on the 18th day of incubation, once, at the following three doses: 1, 10, or 100 µg/egg. The immunological parameters were assessed in 1-, 7-, 14-, 21-, 28-, 35-, and 42-day-old birds kept under farming conditions and routinely vaccinated. The leukocyte populations were determined in the thymus, spleen, and blood. The cytokine (IL-1ß, IL-2, IL-6, and IL-10) levels were determined in the plasma of the broiler chickens. Each experimental group included eight birds. The most pronounced effect of yolkin was an increase in the population of T cells, both CD4+ and CD8+, mainly in the blood. This effect on the lymphocyte subsets may be valuable regarding chicken immune responses, mainly against T-dependent antigens, during infection or after vaccination.

Erysipelas Outbreaks in Flocks of Geese in Poland--Biochemical and Genetic Analyses of the Isolates.

Erysipelothrix rhusiopathiae causes erysipelas in many vertebrate species. Severe disease outbreaks have been noted in many poultry species--chickens, ducks, emus, pheasants, pigeons, and geese. This article describes the biochemical and genetic analyses of six E. rhusiopathiae strains isolated from geese for meat production. The isolates came from birds kept in different poultry houses on one farm, and were collected during two erysipelas outbreaks. We analyzed and compared the isolates by random amplified polymorphic DNA with the use of NK6 primer and pulsed-field gel electrophoresis, with the restriction enzyme SmaI. Biochemical examination was performed with API Coryne test. Analyses showed that the three strains isolated during the first outbreak differed, whereas the three isolates from the second outbreak were identical to one another, but distinct from the isolates from the first outbreak. The results of biochemical and genetic analyses of E. rhusiopathiae strains isolated from geese suggest different sources of infection for the erysipelas outbreaks.

Hemorrhagic nephritis and enteritis in a goose flock in Poland--disease course analysis and characterization of etiologic agent.

Hemorrhagic nephritis enteritis of geese (HNEG) is an epizootic viral disease caused by infection with goose hemorrhagic polyomavirus (GHPV) that affects domestic geese. This study describes the epizootic analysis, laboratory diagnosis, and molecular characterization of GHPV isolates associated with HNEG cases in Poland. HNEG symptoms persisted in infected flocks for 2 wk with a 32% mortality rate. Primary gross lesions included hemorrhaging of the kidneys, intestines, and lungs. Histopathologic examination confirmed HNEG and identified that the causative agent was similar to other GHPV isolates and identical to the Toulouse 2008 isolate.

Impact of early posthatch feeding on the immune system and selected hematological, biochemical, and hormonal parameters in broiler chickens.

Under commercial conditions, chicks hatch within a 24 to 48 h window, a period known as the hatching window. Subsequently, they undergo various treatments before finally being transported to the broiler farm. These procedures may delay the chicks' access to food and water, sometimes receiving them as late as 72 h after hatching. Previous studies have indicated that fasting during this initial period is detrimental, leading to impaired body growth, compromised immune system response, and hindered muscle development. The objective of this study was to assess the impact of early posthatch feeding on immune system organs and selected hematological, biochemical, and hormonal parameters. The experiment utilized Ross 308 broiler eggs incubated under typical commercial hatchery conditions. The experimental group's eggs were hatched in HatchCare hatchers (HC) with immediate access to feed and water, while the control group's eggs were hatched under standard conditions (ST). Thirty chickens from each group were assessed on the 1st (D1), 7th (D7), 21st (D21), and 35th (D35) day after hatching. On D1, the HC group exhibited lower hemoglobin, hematocrit, and total serum protein values, suggesting that early access to water prevents initial dehydration in newborn chicks. Conversely, the ST group showed a stress reaction on D1 due to feed deprivation, leading to an almost 2-fold higher serum corticosterone concentration compared to the HC group. However, this increase did not result in a significant change in the heterophil/lymphocyte ratio. Furthermore, the HC group displayed an increase in triglyceride concentration and a decrease in HDL concentration on D1. On D7, the HC group exhibited an increased relative weight of the bursa and a higher CD4+ cell number in the cecal tonsil (CT), indicating a more rapid development of these organs resulting from early stimulation of the gastrointestinal tract. However, early feeding did not influence the numbers of Bu-1+, CD4+, and CD8+ cells or the germinal center (GC) areas in the spleen. In conclusion, early feeding contributes to the welfare of newborn chicks by reducing dehydration and stress levels and stimulating the development of gut-associated lymphoid tissue.

Antimicrobial Resistance of Erysipelothrix rhusiopathiae Strains Isolated from Geese to Antimicrobials Widely Used in Veterinary Medicine.

The aim of this study was to determine the antibiotic resistance of E. rhusiopathiae when isolated from clinical outbreaks of erysipelas in geese to antimicrobials commonly used in poultry production. All isolates were susceptible to amoxicillin alone or with clavulanic acid, with MIC values ranging from 0.016 to 0.125 µg/mL. Ninety-six percent of isolates were fully sensitive to penicillin G (MIC 0.125-0.5 µg/mL). All isolates were fully or moderately sensitive to erythromycin (MIC 0.125-0.5 µg/mL). Most E. rhusiopathiae isolates proved resistant to fluoroquinolones (76.6% of isolates were resistant to enrofloxacin, with MIC values ranging from 0.064 to 32 µg/mL, and 68% were resistant to norfloxacin, with MIC values ranging from 0.094 to 96 µg/mL), and tetracyclines (61.7% of isolates were resistant to doxycycline, with MIC values ranging from 0.25 to 64 µg/mL, and 63.8% were resistant to tetracycline, with MIC values ranging from 0.38 to 256 µg/mL). Point mutations in the gyrA gene (responsible for fluoroquinolone resistance) and the presence of the tetM gene (responsible for tetracycline resistance) were noted in most of the resistant isolates. Multidrug resistance, defined as resistance to at least one substance in three or more antimicrobial classes, was not observed.

Prevalence of Blastocystis in Geese Reproductive Flocks.

Blastocystis is a unicellular, anaerobic protozoan that has a low specificity for the hosts, and it could be a zoonosis. There are not many data about the occurrence of Blastocystis in bird species, and this study aimed to check the prevalence of Blastocystis infection in reproductive geese flocks. The result obtained showed that a parasite was present in 46.5% of tested flocks. The extensiveness of the Blastocystis invasion in reproductive geese flocks was low because the genetic material of parasites was found only in 7.48% of samples. There was no correlation between the infection and the bird's age or the flock size. The data obtained showed that geese could be the source of infections in humans who have contact with carriers of the infection.

Molecular Analysis of the Heterakis dispar Population in Domestic Geese Based on the ITS1-5.8rRNA-ITS2 Fragment.

Heterakidosis is a parasitic infection in birds caused by the cecal parasite Heterakis spp. The most common species in geese is H. dispar, the largest avian heterakids species. Because of a scarcity of data concerning the H. dispar population, the aim of this study was the genetic analysis of Heterakis dispar isolated from geese flocks based on the ITS1-5.8rRNA-ITS2 fragment. Among the 71 H. dispar specimens isolated from 20 geese flocks, six haplotypes were determined (A, B, C, D, E, and F). The four nucleotide substitutions were noted in both ITS fragments, and all of them were transitions between adenine and guanine, or thymine and cytosine. The most frequently noted haplotype was type A (45%), followed by type B (18.3%), type C and D (11.3%), type E (8.5%), and F (5.6%). Infection with nematodes from different haplotype groups was noted in 30% of the flocks, with type A being the most prevalent, followed by types B, D, or E to make up 100%. This study represents the first H. dispar population analysis based on the ITS1-5.8rRNA-ITS2 fragment.

Early Post-Hatch Nutrition Influences Performance and Muscle Growth in Broiler Chickens.

The poultry industry is under pressure to produce safe and good quality meat in the welfare conditions. Many areas such as genetics, biosecurity, and immunoprophylaxis were improved, and hatchery is one of the areas in which welfare could be improved for better production output. The aim of the study was to investigate the effect of early post-hatch nutrition providing body weight and muscle development in broiler chickens. The experiment involving two groups (chicken hatched with access to water and feed in the hatcher, and chicken without feed and water in hatcher) was replicated three times, and the body weights and breast-muscle index of the randomly chosen 30 chickens per group in each term were measured on the 1st, 7th, 21st, and 35th day of life. The breast-muscle sample was taken for genetic examination (the expression of the myoD, myoG, and MRF4 genes) and histological examination. The results showed that the positive effect of early nutrition was observed on the seventh day of bird life with higher expression of myoG and MRF4 and higher body weight of the birds. The positive effect of early nutrition on the diameter of the breast-muscle fibers was visible on days 21 and 35 of chicken life. The average final body weight in groups with early access to food and water was 5% higher than in groups hatched under classic conditions. Conclusions: early feeding in the hatcher improves performance and muscle growth in broiler chickens.

Tetratrichomoniasis in the Geese Flock-Case Report.

Infections caused by tetratrichomonas are commonly observed in geese. Most cases are subclinical, and the clinical form of the disease manifests itself with a greater mortality and the presence of caseous content in ceca. We describe the case of tetratrichomoniasis in a geese flock caused by Tetratrichomonas gallinarum, with the genetic analysis of the isolate being based on the fragments of 18SrRNA and ITS1-5.8rRNA-ITS2.

Comparative Characteristics and Pathogenic Potential of Escherichia coli Isolates Originating from Poultry Farms, Retail Meat, and Human Urinary Tract Infection.

The pathogenicity of many bacterial strains is determined by the acquisition of virulence genes and depends on many factors. The aim of this study was to analyse the phylogenetic background, virulence patterns, and drug susceptibility of 132 E. coli isolates tested in the context of the ExPEC (Extraintestinal Pathogenic E. coli) pathotype and the correlation of these features with bacterial isolation source: food (retail meat), poultry farms (AFEC-Avian Faecal E. coli), and patients with UTI (urinary tract infection) symptoms. The drug-susceptibility results of tested E. coli isolates obtained indicate that the resistance profile-ampicillin/tetracycline/trimethoprim+sulfamethoxazole/ciprofloxacin (AMP/TE/SXT/CIP)-was most frequently observed. The multidrug resistance (MDR) phenotype was found in 31.8% of isolates from poultry farms, 36.8% of strains isolated from food, and 20% of clinical samples. The greatest similarity of virulence profiles applied to isolates derived from poultry farms and food. Most of the AFEC from poultry farms and food-derived isolates belonged to commensals from phylogroups A and B1, while among the isolates from patients with UTI symptoms, the most common was the B2 phylogroup. The collective analysis showed similarity of the three studied groups of E. coli isolates in terms of the presented patterns of antimicrobial resistance, while the virulence profiles of the isolates studied showed great diversity. The phylogroup analysis showed no similarity between the poultry/food isolates and the UTI isolates, which had significant pathogenic potential.

Uncommon Non-Candida Yeasts in Healthy Turkeys-Antimicrobial Susceptibility and Biochemical Characteristic of Trichosporon Isolates.

The microbiota of the gastrointestinal tract of humans and animals is inhabited by a diverse community of bacteria, fungi, protozoa, and viruses. In cases where there is an imbalance in the normal microflora or an immunosuppression on the part of the host, these opportunistic microorganisms can cause severe infections. The study presented here evaluates the biochemical and antifungal susceptibility features of Trichosporon spp., uncommon non-Candida strains isolated from the gastrointestinal tract of healthy turkeys. The Trichosporon coremiiforme and Trichosporon (Apiotrichum) montevideense accounted for 7.7% of all fungi isolates. The biochemical tests showed that Trichosporon coremiiforme had active esterase (C4), esterase-lipase (C8) valine arylamidase, naphthol-AS-BI phosphohydrolase, α-galactosidase, and ß-glucosidase. Likewise, Trichosporon montevideense demonstrated esterase-lipase (C8), lipase (C14), valine arylamidase, naphthol-AS-BI phosphohydrolase, α-galactosidase, and ß-glucosidase activity. T.coremiiforme and T. monteviidense isolated from turkeys were itraconazole resistant and amphotericin B, fluconazole, and voriconazole susceptible. Compared with human isolates, the MIC range and MIC values of turkey isolates to itraconazole were in a higher range limit in both species, while MIC values to amphotericin B, fluconazole, and voriconazole were in a lower range limit. Furthermore, the obtained ITS1-5.8rRNA-ITS2 fragment sequences were identical with T. coremiiforme and T. montevideense sequences isolated from humans indicating that these isolates are shared pathogens.

Prevalence of Heterakis Infection in Parental Flocks of Geese.

The prevalence of Heterakis infection in reproductive geese from 56 flocks was investigated between February 2015 and November 2019 by using both anatomopathologic and coproscopic examinations. The nematodes were microscopically examined, and 37.5% (21/56) of the flocks were infected with Heterakis nematodes, mostly with Heterakis dispar (35.7%). In one particular flock, a Heterakis gallinarum infection was noted. In two flocks, Heterakis infection was identified only during coproscopic examination. The infection susceptibility of the geese depended on the bird's age-in the first reproductive season, 50% of the flocks were positive; in the second season, 42% were positive; and in the third season, 30% were positive. The number of nematodes in each necropsied goose varied from 8 to 216 individuals. During the 5 years of epidemiologic analysis, the infection occurrence decreased from 66.7% to 11%.

E-test Determination of Antifungal Susceptibility of Candida Species Isolated from Turkeys.

INTRODUCTION: Candida species are a natural component of the intestinal tract microflora, but in favourable conditions they can cause superficial, mucosal, or even systemic candidiasis. Poultry production might be a source of human drug-resistant yeast infections, including Candida spp. The limited data concerning the antifungal susceptibility of poultry Candida isolates prompted us to carry out research to determine the susceptibility of isolates from turkey intestinal tracts. METHOD AND MATERIALS: The beak cavity, crop and cloaca were swabbed of 580 turkeys from 58 flocks in western Poland. The susceptibility tests were conducted using the E-test method with amphotericin B, fluconazole, itraconazole, and voriconazole on 52 isolates of C. albicans, C. catenulata, C. glabrata, C. palmioleophila, C. rugosa, C. krusei and C. lusitaniae. RESULTS: All isolates were susceptible to voriconazole. According to the MIC values obtained for amphotericin B and fluconazole, all Candida spp. isolates were classified as susceptible according to the described breakpoints except for C. krusei, which was the only isolate that was amphotericin B-, fluconazole- and itraconazole-resistant. The susceptibility to itraconazole varied: 11 of the Candida isolates were susceptible (21.1%), 29 were dose-dependently susceptible (55.8%), and 12 isolates were resistant (23.1%). CONCLUSION: There are few resistant strains of Candida in turkeys, and the drug resistance varies. When Candida passes from turkeys to humans, there is a wide range of antifungal treatment options.

The Prevalence of Tetratrichomonas spp. in Reproductive Geese Flocks.

Tetratrichomonas is a genus of parasites that usually inhabits the lower digestive tract, especially the cecum, of various bird species. The infection might lead to birds' death, but in many cases it could be asymptomatic or with mild clinical signs which might be not observed. Subclinical infections can be undiagnosed, leading to production losses. To investigate the prevalence of Tetratrichomonas spp. in geese, 23 cloacal swabs were taken from each of 43 flocks of reproductive geese from five major geese production provinces in Poland after first, second, or third laying season. The obtained swabs were placed in culture medium for propagation of the parasite. All cultures were screened microscopically before PCR was applied to detect the parasites' DNA. After cultivation, the presence of genetic material of Tetratrichomonas was found in 430 out of 989 samples, which correspond to 38 (88.4%) of 43 flocks. The study shows how the number of laying seasons and the size of the flock in which the birds were kept affects the distribution of protozoa of the genus Tetratrichomonas in geese reproductive flocks in Poland.

Molecular Identification and Phylogenetic Analysis of Heterakis dispar Isolated from Geese.

PURPOSE: Heterakidosis is a common parasitic infection caused in domestic birds by Heterakis species: Heterakis gallinarum, H. isolonche, and H. dispar. Among them, the best described species is H. gallinarum, noted mainly in gallinaceous birds. In waterfowl, H. dispar is the predominant species. The variations in morphology and host specificity qualify H. dispar as a different species, but the phylogenetic relationships between heterakids were unclear for a long time, because of a lack of H. dispar sequences. METHODS: The authors provided the molecular data for H. dispar and analyzed the obtained sequences of the partial 18S rRNA gene and region ITS1-5.8SrRNA-ITS2 with the homological sequences. RESULTS: The 18S rRNA PCR product of H. dispar was about 800 bp, and the ITS-5.8S-ITS2 PCR product was about 920 bp, noticeably smaller size compared to H. gallinarum product. The BLAST analysis of H. dispar 18S sequence showed a 99% similarity with the sequences of Heterakis gallinarum and Ascaridia galli, 98% with A. nymphii, but only 94% with the sequence of Heterakis sp. Our ITS sequence of H. dispar was almost identical to the H. isolonche isolate, there is only one nucleotide of difference among the 943 sites analyzed. It also showed a lower similarity to the ITS sequences of H. gallinarum (88%), H. spumosa (87%), and H. dahomensis (87%). CONCLUSIONS: In our phylogenetic analysis, it is the first attempt at the reconstruction of relationships within this superfamily Heterakoidea based on 18S rDNA and ITS region.

The Prevalence of Yeast and Characteristics of the Isolates from the Digestive Tract of Clinically Healthy Turkeys.

This study was undertaken to explore the issue of yeast species prevalence in colonizing the gastrointestinal tract of healthy turkeys. The samples were collected from the beak cavity, crop, and cloaca of 5-wk-old turkeys, cultured and classified using morphological, biochemical, and genetic analysis based on ITS1-5.8rRNA-ITS2 fragment sequencing. Out of all the samples, 12.4% were yeast positive. The highest number of strains, 50% of the total collected, were isolated from the crop, with 30.8% coming from the beak cavity and 19.2% from the cloaca. The most frequently isolated yeast belonged to Candida species (88.5%), followed by Trichosporon (7.7%) and Rhodotorula (3.8%). The most prevalent species was Candida catenulata (30.7%), followed by Candida albicans (21.7%), Candida palmioleophila (17.4%), Candida rugosa (17.4%), and Candida glabrata (8.7%). The present study, showing the prevalence of yeast species of the gastrointestinal tract, is a stepping stone to investigating the physiological mycobiota of turkeys' gastrointestinal tract.

Ascaridia galli isolates with ITS1-5.8rRNA-ITS2 fragment homologous to Ascaridia columbae.

Ascaridia (A.) galli is one of the most commonly occurring nematodes in poultry worldwide, often in hens and broiler chickens. The infection with Ascaridia galli in free-range chickens was even 70%. There is not much information about A. galli genetic features. The present study was conducted to assess the genetic diversity of A. galli isolated from hens in Poland by analyzing the nucleotide sequence of the region ITS1-5.8rRNA-ITS2 and to define its homology within the family Ascaridiidae. Adult A. galli were collected from the intestines of naturally infected hens from two flocks of free-run laying hens from the Wielkopolska region in Poland. From all parasites an identical ITS1-5.8rRNA-ITS2 sequence was obtained, which was homologous in 99% with A. columbae (JQ995321.1) sequence. The high homology sequences of A. galli (KX683286) from Poland and A. columbae (JQ995321.1) isolate from the USA, support the observations of other authors suggesting that A. galli and A. columbae might be closely related. It is the first whole ITS1-5.8rRNA-ITS2 of A. galli in the GenBank database, so there is not enough data for detailed phylogenetic analysis of A. galli. Detailed genetic analysis is necessary to get better insight into the birds' Ascaridia species.

Development of Adenosine Deaminase-Specific IgY Antibodies: Diagnostic and Inhibitory Application.

Adenosine deaminase (ADA) is currently used as a diagnostic marker for tuberculous pleuritis. Although ADA has been suggested as a potential marker for several types of cancer, the importance of each of ADA isoforms as well as their levels and enzymatic activities in tumors need to be further investigated. Herein we developed avian immunoglobulin Y highly specific to human ADA via hens immunization with calf adenosine deaminase. The obtained antibodies were used for the development of a sensitive double-egg yolk immunoglobulin (IgY) sandwich ELISA assay with an ADA detection limit of 0.5 ng/ml and a linearity range of up to 10 ng/ml. Specific, affinity-purified IgYs were able to recognize human recombinant ADA and ADA present in human cancer cell lines. In addition, antigen-specific IgY antibodies were able to inhibit catalytic activity of calf ADA with an IC50 value of 47.48 nM. We showed that generated IgY antibodies may be useful for ADA detection, thus acting as a diagnostic agent in immunoenzymatic assays.

Adjuvant-dependent immunogenicity of Staphylococcus aureus Efb and Map proteins in chickens.

The avian IgY antibodies generated in hens and isolated from egg yolk have gained in popularity as they present an alternative source of antibodies for diagnostic as well as therapeutic applications. One of the advantages of IgY technology are the large amounts of produced antibodies from a single animal combined with their high reactivity representing an attractive alternative for mammalian antibodies. Despite many known protocols for the immunization of chickens, the administration of new antigens often requires additional modification such as antigen dose or use of an adjuvant in order to elicit a significant immune response. We investigated the immunogenicity of three Staphylococcus aureus antigens including two extracellular proteins Map and Efb and one selected Efb105-124 epitope conjugated to KLH that were administered to the animals. Additionally, the immunization protocol included two adjuvant systems: Freund's complete adjuvant and Emulsigen-D. The results demonstrated a high immunostimulatory potency of Freund's complete adjuvant, especially in case of Efb compared to the immune response elicited by Emulsigen-D. However, after immunization with the KLH-Efb105-124 conjugate, the obtained antibodies showed similar reactivity regardless of adjuvant system used with the only exception being their avidity.

Pharmacokinetics of repeated sodium salicylate administration to laying hens: evidence for time dependent increase in drug elimination from plasma and eggs.

Salicylates were the first non-steroid anti-inflammatory drugs (NSAIDs) to be used in any species and are still widely used in humans and livestock. However, the data on their pharmacokinetics in animals is limited, especially after repeated administration. Evidence exist that in chickens (Gallus gallus) salicylate (SA) may induce its own elimination. The aim of this study was to investigate salicylate pharmacokinetics and egg residues during repeated administration of sodium salicylate (SS) to laying hens. Pharmacokinetics of SA was assessed during 14 d oral administration of SS at daily doses of 50 mg/kg and 200 mg/kg body weight to laying hens. On the 1st, 7th and 14th d a 24 h-long pharmacokinetic study was carried out, whereas eggs were collected daily. Salicylate concentrations in plasma and eggs were determined using high-performance liquid chromatography with ultraviolet detection and pharmacokinetic variables were calculated using a non-compartmental model. Mean residence time (MRT), minimal plasma concentration (Cmin, C16h) and elimination half-life (T1/2el) of SA showed gradual decrease in layers administered with a lower dose. Total body clearance (ClB) increased. Layers administered with the higher dose showed a decrease only in the T1/2el. In the low dose group, SA was found only in the egg white and was low throughout the experiment. Egg whites from the higher dose group showed initially high SA levels which significantly decreased during the experiment. Yolk SA levels were lower and showed longer periods of accumulation and elimination. Repeated administration of SS induces SA elimination, although this effect may differ depending on the dose and production type of a chicken. Decreased plasma drug concentration may have clinical implications during prolonged SS treatment.