Recovery of Campylobacter from segments of the reproductive tract of broiler breeder hens

Three groups of >60-wk-old broiler breeder hens were assessed for the presence of Campylobacter within segments of the reproductive tract. In the first group, after stunned, the hens were bled, scalded, and defeathered, the reproductive tracts were aseptically excised from 18 hens, six from each of three adjacent floor pens that were feces positivefor Campylobacter. The reproductive tract segments (infundibulum, magnum-isthmus, shell gland, vagina, and cloaca) were pooled by pen. In the second group, 10 individual hens were sampled from the pens; the reproductive tract was divided into the following segments: magnum, isthmus, shell gland, vagina, and cloaca. For the third group, hens were obtained from two commercial farms that had been determined to be feces positive for Campylobacter, and the reproductive tract was divided into five segments, as described for the second group. Segments of the reproductive tract were placed into sterile plastic bags and suspended 1:3 (w/v) in Bolton enrichment broth, and serial dilutions were plated (0.1 ml) onto Campy-Cefex agar. The agar places were incubated at 42 C for 24 hr in a microaerobic atmosphere. In group 1, the pooled reproductive tract segments for hens from pen A were Campylobacter positive for the shell gland, vagina, and cloaca; hens from pen B were positive for the cloaca only; and hens from pen C were positive for the magnum-isthmus and doaca. In the second group, 9 of 10 cloaca samples were Campylobacter positive. Commercial hens in group 3 had campylobacter-positive cloaca samples (12/12), vagina (10/12), shell gland (7/12), isthmus (2/12), and magnum (4/12). Campylobacter colonization of the reproductive tract of the hen could enable vertical transmission of Campylobacter from the hen to the chick.     

Field testing the influence of sperm competition based on sperm mobility in breeder turkey toms

1. Commercial reproduction of turkeys relies on pooling of semen from multiple males for inseminations. Understanding how sperm characteristics influence paternity under commercial breeding conditions is important to improving production efficiency. 2. The objective of this study was to evaluate progeny production of individual toms following commercial practices of pooling semen to determine if sperm mobility influences progeny production in field conditions. 3. A total of 104 toms were evaluated for sperm mobility. A subset of 10 toms were housed together and semen was collected, pooled and used to inseminate hens (n = 28). Hens were inseminated at 30 weeks of age and weekly thereafter. 4. Ejaculates from each tom were evaluated on two separate days for sperm mobility. Semen from each tom was diluted and layered upon 6% (wt/vol) Accudenz solution. The sperm suspension was incubated at 41 degrees C for 5 min and absorbance was measured with a spectrophotometer. 5. Toms were ranked by absorbance and categorised as high or low if mobility score was +/- 1 SD from the flock mean (average). 6. For parentage determination, DNA was extracted from tom, hen and poult blood. Poult parentage (n = 276) was determined at one day of age or at 14 weeks by analysis of marker genotypes that were generated by polymerase chain reaction (PCR) amplification of genomic DNA with selected microsatellite markers. 7. Sperm mobility differed across males with absorbance values ranging from 0.147 to 0.366. 8. Findings demonstrate differences in poult production among individual toms when semen from multiple males was pooled and inseminated. Toms classified as high, average and low produced 55, 41 and 4% of the offspring, respectively. 9. It appears that sperm mobility is a trait that influences sperm competition among toms under field conditions where sperm numbers inseminated from individual toms are not controlled or constant and that toms with low sperm mobility produce few offspring.     

The Reproductive Tract of the Turkey Hen (A Biometrical Study)

A biometrical study of the reproductive tracts of 173 hens was carried out after slaughter at 53 weeks of age.

Mean weights and total lengths of the reproductive tracts of four strains of actively laying turkey hens were 265 - 279.5 grams and 80.9 - 84.5 centimetres. Mean measurements in centimetres of oviduct anatomical divisions were: infundibulum 10.9 - 11.7, magnum 44.5 - 45.2, isthmus 12.6 - 14.5, uterus 9.6 - 10.8, and vagina 2.6 - 3.5.

     

Localization of immunoglobulins in the chicken oviduct

Studies were made on the distribution of lymphoid tissues and immunoglobulin (Ig: IgA, IgG and IgM)-containing cells (cIg: cIgA, cIgG and cIgM) and the localization of immunoglobulins (Igs) in the oviducal walls of laying hens. Lymphocyte accumulations were occasionally observed, located mainly in the middle infundibulum and in the regions from the isthmus to the vagina. The number of cIgG significantly predominated over that of cIgA or cIgM in the mucosal connective tissue of the magnum and the isthmus. In contrast, in the regions other than the magnum and the isthmus, these three types of cIg were fewer in number. Igs were localized in some superficial epithelial cells (SECs) and glandular cells (GlCs) of the oviduct. Many IgG-containing SECs were found in the infundibulum, the isthmus, and the cranial and major uterus. IgA- or IgM-containing SECs were rare throughout the oviduct. Three types of Ig-containing GlCs were numerously found in the magnum, though lymphocyte accumulations were scarce there. In the isthmus, many IgG-containing GlCs were found, while IgA- or IgM-containing GlCs were rarely observed. Ig-containing GlCs in the magnum were considerably decreased in number after the egg passage. The results suggest that the maternal Igs are transferred to the egg mainly through GlCs in the magnum of the chicken oviduct, and that the oviducal lymphoid tissues have little relationship to the passive immunity.     

Localisation of MHC class II,lymphocytes and immunoglobulins in the oviduct of laying and moulting hens

1. Our aim was to determine the presence and numbers of immunocompetent cells in the oviduct of laying and moulting hens. Immunocompetent cells were localised by immunocytochemistry throughout the entire length of the oviduct.

2. In laying birds, MHC class II + cells were observed in the subepithelial and middle part of the stroma of all oviducal segments and the mucosal epithelium of the infundibulum and vagina. CD3 + cells were also localised in subepithelial and middle part of stroma as well as in mucosal epithelium of each oviducal segment. Bu‐lb + and IgG + cells were also observed in the epithelium and subepithelial and middle part of the stroma of all oviducal segments, though stroma of the magnum, isthmus and uterus contained few Bu‐lb + cells. IgA + cells were observed only in the mucosal epithelium of the magnum in small numbers.

3. In moulting hens, there were few numbers of immunocompetent cells in the mucosal epithelium of each oviducal segment, although CD3 + cells were observed in the infundibulum and vagina. In the subepithelial stroma, the populations of MHC class II + cells in the infundibulum, magnum and uterus, CD3 + cells in the infundibulum and vagina, as well as IgG + cells in each oviducal segment except for isthmus were smaller than in laying hens. In contrast, the number of immunocompetent cells in the middle part of stroma of moulting hens were equal to or greater than in laying hens.

4. These results suggest that the oviducal immune function is active in the surface tissues of the mucosa in laying hens, whereas it is reduced in moulting hens.     

Ultrastructural study of infectious bronchitis virus infection in infundibulum and magnum of commercial laying hens

The infundibulum and magnum of the oviduct were examined in hens in full lay which were infected with two Australian strains of infectious bronchitis virus (IBV). The ultramicroscopic changes in the infundibulum and magnum were compared with control hens which had eggs at different positions in the oviduct. The ciliated and granular cells of the surface epithelia and secretory epithelial cells of the tubular glands were the target cells of IBV. No pathological changes were recorded during 2-8 days post-infection (p.i.). Patchy loss of cilia occurred at 10-14 days p.i. Between 16 and 24 days p.i., there was no cilia loss and lymphoid nodules were observed in the muscularis layer of the infundibulum and magnum of some hens from both infected groups. Virus particles were detected mostly in the rough endoplasmic reticulum (RER) and Golgi complex between 10 and 12 days p.i. Cytopathology was noticed in various cell organelles between the 10th and 14th days p.i. There was an increase in RER deposits in infected cells, irrespective of egg position in the oviduct. The magnum was more affected than the infundibulum. Cellular changes were more severe in the infundibulum and magnum of T-infected hens as compared to N1/88-infected hens. Eggs with watery whites which were laid by infected hens could be attributed to cytopathological changes in the granular epithelial cells and tubular gland epithelial cells of the magnum resulting in reduced synthesis of albumen proteins. IBV can cause pathology in parts of the fully functional oviduct which may persist up to the 30th day p.i. However, both the challenge strains of IBV can cause a small number of hens to cease production. Loss of cilia in both the infundibulum and magnum pose a potential threat of secondary bacterial infection and also may affect fertility in breeder hens.     

Recovery of Campylobacter jejuni in feces and semen of caged broiler breeder roosters following three routes of inoculation

We previously reported the recovery of Campylobacter (naturally colonized) from the ductus deferens of 5 of 101 broiler breeder roosters, and four of those five positive roosters had previously produced Campylobacter-positive semen samples. Those results prompted further evaluation to determine if inoculation route influenced the prevalence or level of Campylobacter contamination of semen, the digestive tract, or reproductive organs. Individually caged roosters, confirmed to be feces and semen negative for Campylobacter, were challenged with a marker strain of Campylobacter jejuni either orally using 1.0 ml of a diluted cell suspension (log(10)4.3 to 6.0 cells), by dropping 0.1 ml of suspension (log(10)5.3 to 7.0 cells) on the everted phallus immediately after semen collection or by dip coating an ultrasound probe in the diluted cell suspension (log(10)4.3 to 6.0 cells) and then inserting the probe through the vent into the colon. Six days postinoculation, individual feces and semen samples were again collected and cultured for Campylobacter. Seven days postinoculation, roosters were killed, the abdomen aseptically opened to expose the viscera, and one cecum, one testis, and both ductus deferens were collected. The samples were then suspended 1:3 (weight/volume) in Bolton enrichment broth for the culture of Campylobacter. Samples were also directly plated onto Cefex agar to enumerate Campylobacter. Campylobacter was recovered 6 days after challenge from feces in 82% of samples (log(10)4.1 colony-forming units [CFU]/g sample), 85% of semen samples (log(10)2.9 CFU/ml), and on the seventh day postchallenge from 88% of cecal samples (log(10)5.8 CFU/g sample). Campylobacter was not directly isolated from any testis sample but was detected following enrichment from 9% (3/33) of ductus deferens samples. Roosters challenged with Campylobacter orally, on the phallus, or by insertion of a Campylobacter dip-coated ultrasound probe were all readily colonized in the ceca and produced Campylobacter-positive semen and feces on day 6 after challenge. The low prevalence of recovery of Campylobacter from the ductus deferens samples and failure to recover from any testis sample suggests that semen may become Campylobacter positive while traversing the cloaca upon the everted phallus. The production of Campylobacter-positive semen could provide a route in addition to fecal-oral for the horizontal transmission of Campylobacter from the rooster to the reproductive tract of the hen.     

Brain and skull lesions resulting from use of percussive bolt, cervical dislocation by stretching, cervical dislocation by crushing and blunt trauma in turkeys

Three experiments were conducted to assess brain damage resulting from percussive bolt shooting and cervical dislocation by crushing (neck crushing) in turkey hens (mean [se] bodyweight 11.4 [0.1] kg); percussive bolt shooting and blunt trauma in turkey toms (13.1 [0.2] kg); and percussive bolt shooting, blunt trauma and cervical dislocation by stretching (neck stretching) in broiler turkeys (3.9 [0.3] kg). Brain and skull damage were assessed using macroscopic and microscopic evaluations and CT. Macroscopic subcutaneous haemorrhage was significantly greater with the percussive bolt in all three experiments (hens P=0.01, toms P=0.02, broilers P=0.0003), and skull fractures were more severe for toms (P<0.0001) and broilers (P=0.03) killed with the percussive bolt versus blunt trauma. In a subsample of turkeys, microscopic brain damage was present in all turkeys killed by percussive bolt shooting (five hens, 10 toms and four broilers) and blunt trauma (nine toms and three broilers), but only in one of four turkeys killed by neck crushing and one of four turkeys killed by neck stretching. Percussive bolt shooting and blunt trauma most likely caused death by directly disrupting brain function, whereas neck stretching and neck crushing probably resulted in death from cerebral hypoxia and ischaemia.     

Dam and Sire Effects on Sperm Penetration of the Perivitelline Layer of Eggs Produced by Two Strains of Commercial Turkeys

The reproductive performance of 2 commercial turkey breeder lines was examined using reciprocal crosses between sires and dams of each line. One line had been selected using artificial inseminations performed at biweekly intervals, whereas the second line had been selected using inseminations performed at weekly intervals. The hypothesis was proposed that sires and dams of the 2 lines differ because of different abilities for sperm to penetrate the inner perivitelline layer (IPVL) and fertilize eggs.Fertilized eggs to hatch poults for the experiment were obtained from the primary breeders and were incubated using conditions recommended by the industry. Hens (n = 72) and toms (n = 15) from each strain were identified and reared in preparation for a reproductive cycle using commercially accepted standards. Beginning just prior to the onset of egg production and at weekly intervals thereafter, half of the hens were inseminated with semen from males of the same line, whereas the remaining half received semen from the opposite line. Eggs were collected from the pens daily and set in incubators to determine fertility and embryo survival. At biweekly intervals 3 eggs per pen were used for counting sperm penetration holes in the IPVL. Data were collected for fertility, hatchability, and time that embryos died for each of the pens. Dam and sire affected IPVL penetration independently. A dam by sire interaction influenced fertility, whereas hatchability was affected only by dam. Thus, it is concluded that selection of dam and sires for commercial breeders alters IPVL sperm penetration ability of hens as well as egg-binding ability of sires.     

Prolonged survival of fowl spermatozoa in the oviducal tissues in organ culture

1. When fowl spermatozoa were incubated at 41 °C with tissues from the infundibulum, magnum, isthmus, shell gland, uterovaginal junction and vagina, their motility, assessed at room temperature (20 to 25 °C), was maintained for 4 to 12 d.

2. The survival period was much longer for spermatozoa incubated with the tissues from the infundibulum and uterovaginal junction (with sperm‐host glands, 11 to 12 d) than for those incubated with tissues from the other regions (lacking host glands, 4 to 7 d).

3. In the tissues of the infundibulum and uterovaginal junction, spermatozoa entered the sperm‐host glands and were more closely associated with the epithelial cells than they were in tissues from other regions.     

Genotype analyses of Campylobacter isolated from the gastrointestinal tracts and the reproductive tracts of broiler breeder roosters

Campylobacter is considered to be the leading bacterial etiologic agent of acute gastroenteritis in humans. Evidence implicates poultry as a major source of the organism for human illness; however, the pathways involved in Campylobacter contamination of poultry flocks, horizontal transmission and/or vertical transmission, remain unclear. Recent evidence implicates breeders as a potential source for Campylobacter contamination of the subsequent broiler offspring. In this investigation, Campylobacter isolated from feces, cloacal swabs, ceca, semen, and vas deferens of 12 breeder broiler roosters were genotyped by both flagellin A short variable region (flaA SVR) DNA sequence analysis and repetitive element (rep)-polymerase chain reaction (PCR). In 9 of 12 roosters, Campylobacter was isolated from multiple sites sampled. Comparison of multiple isolates obtained from individual roosters revealed variable results. In five of the nine roosters, all Campylobacter isolated demonstrated closely related flaA SVR DNA sequences as well as rep-PCR patterns; isolates from these roosters were collected from both the gastrointestinal and the reproductive tracts or from the gastrointestinal tract alone. The remaining four roosters had Campylobacter that were distinct by both typing methods. Isolates from two of these four roosters originated from both the gastrointestinal and the reproductive tracts. Isolates from the remaining two roosters originated from only the reproductive tract. Comparisons of all Campylobacter isolates recovered from a distinct sample type within either the reproductive tract or the gastrointestinal tract (feces, semen, cloaca, vas deferens, or ceca) were quite diverse. No relationship between the genotypes and the sample type could be ascertained. Further investigation is needed to determine the route of contamination and if the presence of Campylobacter within the rooster leads to contamination of the broiler offspring via the fertilized egg.     

A Novel Bedding Material Made from Cotton Waste,Gypsum, and Old Newsprint for Rearing Turkeys

Two experiments and 3 field trials were conducted to examine the usefulness of a novel bedding material for rearing Large White commercial turkeys. The control bedding was pine shavings (PS) in both experiments and all trials. The novel bedding, aGroChips (AC), is a chopped paper product made from cotton lint waste, gypsum, and old newsprint following a proprietary paper manufacturing process. In both experiments, hens and toms were reared according to typical industry techniques. In the first experiment, use of AC resulted in significantly (P <0.05) heavier toms and hens. In the second, the toms brooded and reared on AC were significantly (P <0.05) heavier than those brooded and reared on PS, whereas toms brooded on one bedding and then reared on the other were intermediate in weight. There were no differences in final cumulative FCR or carcass yield in either experiment. Three field trials were conducted with Large White commercial turkey hens in which the hens were brooded either on PS or AC, with both groups reared on PS. There was a mean increase of 0.2 kg in BW, a decrease (improvement) of 0.03 in FCR, and an increase of 3,200 kg per trial for AC-brooded birds (based on 16,000 hens placed per brooder house). A hard, dry litter crust was observed in the AC houses. With subsequent testing, further changes in the manufacturing process to create a hard, durable pellet may result in a more usable and useful bedding material.     

The relation between ultrasonographic observations in the oviduct and plasma progesterone, luteinizing hormone and estradiol during the egg laying cycle in ostriches

In this study we investigated the temporal relationship between ovulation, egg formation, oviposition and the changes in plasma concentrations of progesterone, luteinizing hormone and estradiol-17beta during the egg laying cycle in farmed ostriches. In 10 egg-producing birds, transcutaneous ultrasound scanning was performed at 3h intervals and blood sampling at hourly intervals during a period of at least 48h (one egg laying cycle). In hens (n=8) that ovulated during the observational period, the ovulated egg was first detected 2h after oviposition; thus, ovulation occurred shortly after oviposition in all birds. During the period between two consecutive ovipositions, the developing egg remained for 9h in the proximal part (infundibulum, magnum or isthmus) and for 39h in the distal part of the oviduct (uterus). In ovulating hens, plasma progesterone concentrations showed a characteristic and consistent profile: from basal levels of around 0.1ng/ml concentrations started to increase 12h before oviposition, reached an average maximum of 3.5ng/ml at 3h before oviposition and returned to basal levels 3h and 30min after oviposition. Changes in plasma luteinizing hormone and estradiol-17beta concentrations showed comparable patterns of elevation and decline relative to the timing of oviposition and ovulation. However, variation in their individual basal concentrations was generally larger and peak values were less conspicuous than those of progesterone. In non-ovulating hens (n=2) neither progesterone, nor luteinizing hormone nor estradiol-17beta showed elevations to peak concentrations before oviposition. These data demonstrate that during the egg laying cycle of ostriches, events such as ovulation, egg development and oviposition evolve according to a rather strict time schedule, and that progesterone, luteinizing hormone and estradiol-17beta reach peak concentrations shortly before ovulation. Additionally, our findings also show that on-farm ultrasound scanning is a useful technique to discriminate between ovulating and non-ovulating hens.     

Content of retinol and retinyl esters in blood plasma,liver, kidney and reproductive organs of Japanese quails

Due to its importance in many physiological processes such as cell proliferation and differentiation, vitamin A plays a key role in reproduction. The present study examines the content and distribution of retinol and retinyl esters in the blood plasma, liver, kidney, ovary and oviduct (infundibulum, magnum, isthmus and uterus) of the laying Japanese quail. (1) The results show that the stage of egg laying had no influence on the level of vitamin A (retinol or retinyl esters) in plasma, kidney and liver. (2) The results further indicate that in the oviduct there are quantitative and qualitative differences in the concentration of retinol and retinyl esters, but that these differences are not altered by the stage of egg formation. (3) The highest levels of vitamin A in the isthmus and uterus were associated with a predominance of retinyl esters (palmitate and stearate); sections with lower total levels of vitamin A (infundibulum, magnum) had retinol as the more dominant form of vitamin A. (4) Changes in the ratio of retinol to retinyl esters in the various sections of the avian oviduct might point to metabolic differences. The storage of vitamin A might therefore be the predominant function of the uterus and isthmus; in the infundibulum and magnum, where vitamin A is predominantly present as retinol, vitamin A serves rather as a precursor for the modulation of the cellular metabolism of these structures.     

Transaminase and phosphomonoesterase activities in different regions of the oviduct epithelium of laying Japanese quail (Coturnix coturnix japonica)

1. Activities of alkaline (ALP) and acid phosphomonoesterases (ACP) and aspartic (GOT) and alanine (GPT) aminotransferases in the glandular epithelia of the infundibulum, magnum, isthmus and shell gland of the laying quail were measured when the egg was in the shell gland. 2. The activities of all the enzymes were significantly (P less than 0.001) greater in shell-gland than in isthmus and magnum. 3. Gel-electrophoretic analysis of proteins showed that the magnal epithelia had the same proteins as the albumen of the laid egg.     

The influence of substance P on oviductal,duodenal and blood pressure in the anaesthetized domestic hen

In 13 anaesthetized hens in the peak phase of their first laying year the influence of intravenously injected substance P (SP), 1–10 g/animal, on oviductal pressure, duodenal pressure, blood pressure and heart rate has been studied within 5 h of oviposition. The neuropeptide induced a significant pressure increase in the different segments of the oviduct (infundibulum, magnum, isthmus, uterus and vagina) as well as in the duodenum. Blood pressure revealed a distinct biphasic response: a short period of hypotension accompanied by a tachycardia and a more pronounced and sustained hypertension, inducing a subsequent bradycardia. The complexity of the observed effects demonstrates the overall impact of intravenously administered SP on the anaesthetized hen.     

Effects of Metabolizable Energy and Amino Acid Levels on Turkey Performance from Hatch to Marketing

Two experiments were conducted with growing turkeys from hatch to market examining the performance at various dietary energy and amino acid (AA) levels as well as the relationship between energy and essential amino acids at these energy levels. The results of the first experiment indicated that increasing ME alone improved feed efficiency (FE); however, it did not affect BW in either toms or hens. In addition, increasing ME alone resulted in better regulation of caloric intake (CI) in toms than in hens. The caloric efficiency (CE) (BW/CI) improved with the higherenergy diets. In a second experiment both AA and ME levels were changed. Using 95% of NRC amino acid levels depressed performance, whereas turkeys receiving 105 and 110% of the recommended AA levels had improved BW and FE until 6 wk. From 7 to 19 wk, increasing dietary AA levels decreased feed intake (FI). Increasing both AA and ME together did not, in all cases, produce the same effect as altering their levels independently.     

Macroscopic anatomy of the reproductive tract of the reproductively quiescent female emu (Dromaius novaehollandiae)

Three reproductively quiescent female emus (Dromaius novaehollandiae) were embalmed with 10% formalin solution. The reproductive tract was dissected and described. The reproductive tract consists of an ovary and oviduct situated on the left side of the abdominal cavity. The left ovary is dark brown to black in colour with follicles covering the ventral surface. The ovary is located medial to the spleen and closely associated with the ventral surface of the cranial and middle lobes of the left kidney. The oviduct is a relatively straight tube that extends from the level of the cranial extent of the left ilium to the caudal border of the left pubic bone. The oviduct is grossly divided into the infundibulum, magnum, isthmus, uterus and vagina using variations in the mucosal fold pattern.     

Testosterone concentration in the seminal plasma of cocks

Testosterone concentrations in the seminal plasma of cocks ranged from 0.46 to 5.05 ng/ml and were substantially lower than in blood plasma. No significant variation was noted in seminal plasma testosterone concentrations during the light phase of the day, whereas the concentration in blood declined over this period. Spermatozoal concentration and seminal testosterone decreased in the third sample of the semen collected sequentially at 3 h intervals. Testosterone concentrations in seminal plasma (1.57 +/- 0.17 ng/ml) and in the semen from the ductus deferens (1.34 +/- 0.24 ng/ml) were not significantly different.