Cecal and hepatic granulomas in chickens associated with Heterakis gallinarum infection

Heterakis gallinarum infection was detected in the ceca of chickens with cecal and hepatic granulomas found at slaughter. The chickens were diagnostic submissions from four small backyard flocks in Saskatchewan. Detailed dissection of some cecal granulomas in one flock and pepsin digestion of cecal granulomas in the three other flocks demonstrated the presence of the parasite within granulomas in three of the flocks. A possible causal relationship between Heterakis gallinarum and the cecal and hepatic granulomas is discussed.     

Detection of chicken anemia virus DNA in embryonal tissues and eggshell membranes

Chicken infectious anemia virus (CIAV) is a ubiquitous and highly resistant virus of chickens that causes anemia and death in chicks less than 3 wk of age and immunosuppression in chickens older than 3 wk of age. The production of specific-pathogen-free eggs free of CIAV is essential for research and vaccine production. Currently, flocks are screened for CIAV by antibody tests to ensure freedom from CIAV infection. Recent evidence, however, indicates that chickens may carry and vertically transmit CIAV DNA independently of their antibody status. In this study, we tested embryos and eggshell membrane residues by nested polymerase chain reaction (PCR) as a sensitive method of detecting CIAV DNA. CIAV DNA could be detected in the blastodisks and semen obtained from antibody-positive and -negative chickens. Examination of different tissues between 18 and 20 days of incubation indicated that many but not all organs of individual embryos were positive. The lymphoid organs and gonads had the highest incidence of CIAV DNA, which was significantly different (P < 0.05) from the incidence in the liver. Eggshell membrane samples from embryos or newly hatched chicks were an excellent noninvasive source for the detection of CIAV DNA, identifying significantly more positive embryos than did pooled lymphoid organs. The use of dexamethasone injections as a method to improve the detection of carrier birds did not result in an increase of vertical transmission or cause seroconversion in the treated hens. A combination of testing eggshell membrane residues at hatch and periodic testing of blood DNA by nested PCR can be used to identify chickens carrying CIAV DNA and may be used to eradicate carrier birds.     

Naturally acquired antibodies against Clostridium perfringens epsilon toxin in goats

Clostridium perfringens type D-producing epsilon toxin is a common cause of death in sheep and goats worldwide. Although anti-epsilon toxin serum antibodies have been detected in healthy non-vaccinated sheep, the information regarding naturally acquired antibodies in ruminants is scanty. The objective of the present report was to characterize the development of naturally acquired antibodies against C. perfringens epsilon toxin in goats. The levels of anti-epsilon toxin antibodies in blood serum of goat kids from two different herds were examined continuously for 14 months. Goats were not vaccinated against any clostridial disease and received heterologous colostrums from cows that were not vaccinated against any clostridial disease. During the survey one of these flocks suffered an unexpectedly severe C. perfringens type D enterotoxemia outbreak. The results showed that natural acquired antibodies against C. perfringens epsilon toxin can appear as early as 6 weeks in young goats and increase with the age without evidence of clinical disease. The enterotoxemia outbreak was coincident with a significant increase in the level of anti-epsilon toxin antibodies.     

Isolation and some characteristics of a subgroup J-like avian leukosis virus associated with myeloid leukosis in meat-type chickens in the United States.

Several subgroup J-like avian leukosis viruses (ALV-Js) were isolated from broiler breeder (BB) and commercial broiler flocks experiencing myeloid leukosis (ML) at 4 wk of age or older. In all cases, diagnosis of ML was based on the presence of typical gross and microscopic lesions in affected tissues. The isolates were classified as ALV-J by 1) their ability to propagate in chicken embryo fibroblasts (CEF) that are resistant to avian leukosis virus (ALV) subgroups A and E (C/AE) and 2) positive reaction in a polymerase chain reaction with primers specific for ALV-J. The prototype strain of these isolates, an isolate termed ADOL-Hc1, was obtained from an adult BB flock that had a history of ML. The ADOL-Hc1 was isolated and propagated on C/AE CEF and was distinct antigenically from ALV of subgroups A, B, C, D, and E, as determined by virus neutralization tests. Antibody to ADOL-Hc1 neutralized strain HPRS-103, the prototype of ALV-J isolated from meat-type chickens in the United Kingdom, but antibody to HPRS-103 did not neutralize strain ADOL-Hc1. On the basis of both viremia and antibody, prevalence of ALV-J infection in affected flocks was as high as 87%. Viremia in day-old chicks of three different hatches from a BB flock naturally infected with ALV-J varied from 4% to 25%; in two of the three hatches, 100% of chicks that tested negative for virus at hatch had evidence of viremia by 8 wk of age. The data document the isolation of ALV-J from meat-type chickens experiencing ML as young as 4 wk of age. The data also suggest that strain ADOL-Hc1 is antigenically related, but not identical, to strain HPRS-103 and that contact transmission of ALV-J is efficient and can lead to tolerant infection.     

Isolation of Campylobacter from circulating blood of commercial broilers

Campylobacter spp. are present in organs and tissues of broiler chickens but the dissemination route is unclear. The aim of the current study was to determine Campylobacter prevalence within circulating blood of commercial broilers. Broilers were acquired from 19 flocks originating from three commercial poultry processing companies. Using aseptic blood collection techniques, 5 ml of circulating blood was collected from each bird and the sample analyzed for Campylobacter. The Campylobacter colonization status of each bird was determined by aseptically sampling and analyzing the ceca. Campylobacter was recovered from 58% (11/19) of flocks sampled. From the 248 total birds sampled, 12% and 46% of the birds had Campylobacter in the blood and ceca, respectively. This study documents Campylobacter prevalence in the circulating blood of commercially raised broilers. Campylobacter presence in the circulatory system may indicate the path used by the organism for rapid dissemination to organs and tissues. From a processing viewpoint, Campylobacter presence in circulating blood of market-age broilers may increase the likelihood of cross-contamination between birds during slaughter.     

Failure of viable nonculturable Campylobacter jejuni to colonize the cecum of newly hatched leghorn chicks

Campylobacter jejuni cells entered the viable but nonculturable (VBNC) state upon suspension in sterile water. Cell viability was determined with tetrazolium violet. VBNC cells suspended in water for 7, 10, or 14 days were given, by gavage, to day-of-hatch leghorn chickens. The ceca of control and challenged birds were examined for the presence of campylobacteria by conventional microbiological methods at 1 wk and 2 wk after challenge inoculation and by polymerase chain reaction methods at 1 wk after challenge. We did not find culturable Campylobacter cells in the ceca. Neither was Campylobacter DNA found in cecal samples. Therefore, VBNC cells did not revert to the culturable colonizing form, nor did VBNC cells persist within the cecal environment.     

A study of the epidemiology of infectious bursal disease in poultry in Queensland, 1976–1979

The epidemiology of infectious bursal disease (IBD) was studied by serology and sometimes by visual examination of the bursa of Fabricius in poultry flocks in Queensland during 1976–1979.
Ten flocks, each of approximately 30,000 meat breeding chickens, were surveyed. All chickens had maternally-derived antibody against IBD virus (IBDV) at hatching and active antibody was not detected while the chickens were brooded on rearing farms. When distributed to breeding farms, 7 of the flocks developed antibody when 11 to 25 weeks of age. The remaining 3 flocks were vaccinated by infection of 10% of the birds and within 4 weeks more than 80% of the chickens had developed precipitating antibody to IBDV.
Blood samples of 20 to 30 broiler chickens were collected at slaughter (7 to 9 weeks of age) from each of 312 broiler flocks raised on 37 contract farms. While the samples from 21 flocks were without detectable antibody to IBDV, all serum samples for 263 flocks contained antibody. The ratio of bursal weight to bodyweight was significantly lower in birds from 144 flocks having antibody to IBDV than in birds from 10 flocks that were without detectable antibody. In sequential studies, IBDV antibody became demonstrable in 27 of 30 flocks when the chickens were one to 6 weeks of age and was accompanied by bursal atrophy.
Serological investigation of 4 flocks of layer breeding chickens on a multi-age farm at approximately monthly intervals resulted in antibody to IBDV being detected at every examination.
Serological tests and bursal examinations were carried out weekly in 2 flocks each of 4000 layer chickens between one and 20 weeks of age. Serum antibody developed in one flock at 4 weeks of age and in the other at 17 weeks of age. In both flocks, bursal atrophy occurred concurrently with the development of antibody.     

Seroprevalence of avian influenza virus, infectious bronchitis virus, reovirus, avian pneumovirus, infectious laryngotracheitis virus, and avian leukosis virus in Nigerian poultry

Eight poultry farms in Nigeria, including chickens from nine breeder, 14 broiler, 28 pullet, 11 layer, and three cockerel flocks, were tested for antibody seroprevalence to the following poultry viruses of potential economic importance: infectious bronchitis virus (IBV), avian reovirus, avian pneumovirus (APV), infectious laryngotracheitis virus (ILTV), avian influenza virus (AIV), and avian leukosis virus (ALV). Serum samples were collected between 1999 and 2004 and were tested for antibodies using commercial enzyme-linked immunosorbent assay (ELISA) kits. Seroprevalence was very high for IBV (84%); intermediate for reovirus (41%), APV (40%), and ILTV (20%); and very low for ALV (<5%) antibodies. By commercial ELISA, the seroprevalence of antibodies against AIV was, in some flocks, up to 63%. However, more specific assays did not confirm AIV antibodies, indicating that all flocks tested were free of avian influenza antibodies. Birds seemed to be first infected by IBV (at about 7 wk of age), then by reovirus at 12 wk, before they became infected by APV (week 25) and ILTV (week 30). This is the first report of serological evidence of the above viruses in West Africa. Further studies are necessary to assess economic losses due to these avian viruses and the costs and benefits of countermeasures.     

Evidence of cross-contamination by Campylobacter spp. of broiler carcasses using genetic characterization of isolates

Campylobacter is recognized as one of the leading cause of gastroenteritis worldwide, and is frequently isolated from the small intestines and ceca microflora of chickens. Twenty-one out of 81 Campylobacter-positive poultry flocks were selected to evaluate the genetic diversity of Campylobacter isolates and to study the distribution of genotypes among flocks. Campylobacter isolates recovered from chicken carcasses and ceca were analyzed by pulsed-field gel electrophoresis (PFGE). Little diversity was found among Campylobacter strains isolated from a given carcass, with a maximum of 2 different genotypes being present. However, at flock level, as many as 4 different profiles were observed. Typing of strains showed that most strains isolated from ceca were similar to those isolated from corresponding broiler carcasses. A total of 39 different macrorestriction profiles were observed, with evidence of Campylobacter cross-contamination among broiler flocks in Quebec slaughterhouses. Surprisingly, some flocks shared related genotypes both with and without sharing similar rearing practices. Existence of such cross-contamination must be considered to in developing strategies to control Campylobacter in chickens, and to avoid bacteria contamination of noncolonized flocks. Further typing studies of Campylobacter found in hatcheries, farm environment, and crates or trucks in Quebec might be helpful in elucidating the kinetics of broiler chicken Campylobacter contamination.     

An acute form of transient paralysis induced by highly virulent strains of Marek's disease virus

A novel syndrome was observed after inoculation of 3-wk-old chickens with highly virulent Marek's disease virus (MDV) strains. This syndrome was characterized by the acute onset of neurologic signs including flaccid paralysis of neck and limbs 9-10 days postinoculation, typically resulting in death 1-3 days after the onset of clinical signs. Most affected birds died, and spontaneous recovery was rare. Few if any gross tissue changes were found. Histologic brain lesions included acute vasculitis with vasogenic edema and perivascular cuffing. The syndrome was influenced by the virus strain and dose and by chicken strain and B haplotype and was prevented by vaccination with turkey herpesvirus. Chickens up to 18 wk of age were susceptible. On the basis of clinical signs and histopathology, the syndrome was determined to be an acute form of transient paralysis (TP); its more acute nature and virtual lack of spontaneous recovery differentiated this syndrome from classical TP. Affected birds were viremic, and brains were positive for viral DNA by polymerase chain reaction assays, but these tests were also positive in inoculated chickens without clinical signs and may have limited value for diagnosis. Although acute TP should occur only rarely in Marek's disease-vaccinated commercial flocks, this syndrome may be important in laboratory studies, where it could interfere with pathogenesis trials. Finally, acute TP appears to be one component in the pathogenesis of the early mortality syndrome, a previously described immunodepressive disease induced by inoculation of 1-day-old chicks with highly virulent MDV.     

Implication of coprophagy in pathogenesis of chicken botulism

Oral administration of 1 x 10(7) viable spores of Clostridium botulinum type C killed the chickens kept on a board floor to allow them coprophagy, whereas the same dose of the spores failed to develop symptoms in those kept on a wire-net floor not to allow them coprophagy. Type C toxin was detected in the cecal droppings of the chickens of both the groups after feeding the spores and also in serum of symptomatic as well as asymptomatic chickens kept on a board floor. Thus, coprophagy, by which chickens ingest type C toxin (C1 L toxin) and the bacterial cells, seems to be a prerequisite for development of chicken botulism.     

Influence of host lineage on cecal colonization by Campylobacter jejuni in chickens

The resistance to cecal colonization by Campylobacter jejuni was assessed by challenging three crossbred stocks of commercially available broiler chickens. These three stocks, designated A, B, and C, were related as follows: Offspring from four pedigreed grandparent flocks were used as progenitors. Stock B was derived by cross-breeding grandparent 1 with grandparent 3. Stocks A and C were crossbreeds from grandparents 1 and 2 and grandparents 3 and 4, respectively. Campylobacter jejuni were gavaged into 48-hour-old chicks, using the same levels of challenge dose for each of the different chicken stocks. Six days post-challenge, the birds were sacrificed, and cecal contents were plated onto Campylobacter-selective media. Results from two replicate trials with three isolates of C. jejuni indicated that chicken stock A was colonized in only two of 60 ceca, stock B in six of 60, and stock C in 19 of 60 chicken ceca. Statistical analysis of these data indicate that resistance to cecal colonization by C. jejuni was significantly (P less than 0.05) influenced through chicken host lineage.     

Fluid accumulation in the ligated intestinal loop and histopathological changes of the intestinal mucosa caused by Clostridium botulinum C2 toxin in the pheasant and chicken

Fluid accumulation was induced in ligated intestinal loops of both pheasants and chickens injected with crude Clostridium botulinum C2 toxin. Necrosis of the epithelium and lamina propria of the duodenum and jejunum occurred in both species of bird after intraduodenal administration of crude C2 toxin. The severity of such reactions depended upon the dose and the period after administration of C2 toxin up to eight hours, and such reactions were suppressed specifically with rabbit anti-C2 toxin. These results confirm that C2 toxin is one of the causes of diarrhoea in avian botulism.     

The influence of physical and environmental variables on the in vitro attachment of Salmonella typhimurium to the ceca of chickens

This investigation was designed to study the effect of exposure time, pH, age of bird, and native intestinal microflora on the in vitro attachment of Salmonella typhimurium to the ceca of chickens. Ceca were surgically removed from chickens immediately after euthanasia, and the interiors were exposed to S. typhimurium as intact ceca with contents, intact ceca rinsed free of contents, or inverted rinsed ceca. Attachment of S. typhimurium was slightly higher in washed than in unwashed ceca. Neither pH nor age of chicks affected attachment of the organism to ceca. There was no difference in attachment of salmonellae to inverted washed ceca after 1, 2, 3, 5, and 10 min exposure, but a one log difference was noted between 10 min and 30 sec.     

Comparison of a live attenuated Salmonella Enteritidis vaccine candidate secreting Escherichia coli heat-labile enterotoxin B subunit with a commercial vaccine for efficacy of protection against internal egg contamination by Salmonella in hens

This study compared a new live attenuated Salmonella Enteritidis vaccine candidate secreting Escherichia coli heat-labile enterotoxin B subunit (SE-LTB) with a commercial Salmonella Enteritidis (SE) vaccine for efficacy of protection against SE infection in laying hens. Chickens were divided into 3 groups of 20 each. Group A chickens were inoculated orally with phosphate-buffered saline and served as controls, group B chickens were inoculated orally with the vaccine candidate, and group C chickens were inoculated intramuscularly with a commercial vaccine, the primary inoculation in groups B and C being at 10 wk of age and the booster at 16 wk. Groups B and C showed significantly higher titers of plasma immunoglobulin G, intestinal secretory immunoglobulin A, and egg yolk immunoglobulin Y antibodies compared with the control group, and both vaccinated groups showed a significantly elevated cellular immune response. After virulent challenge, group B had significantly lower production of thin-shelled and/or malformed eggs and a significantly lower rate of SE contamination of eggs compared with the control group. Furthermore, the challenge strain was detected significantly less in all of the examined organs of group B compared with the control group. Group C had lower gross lesion scores only in the spleen and had lower bacterial counts only in the spleen, ceca, and ovary. These findings indicate that vaccination with the SE-LTB vaccine candidate can efficiently reduce internal egg and internal organ contamination by Salmonella and has advantages over the commercial vaccine.     

Susceptibility of mink to Clostridium botulinum type E toxin

Experiments aiming at elucidation of the toxicity of Clostridium botulinum type E for mink are described. The observations indicate that amounts in the order of 2 x10^s intraperitoneal MLD (mice) or approximately 200 MLD per g of type E toxin will kill a mink after oral administration. The symptoms observed in the animals were atypical as there was an unusually short period between administration of the toxin and the onset of symptoms and deaths of the animals. Similar results were obtained when Clostridium botulinum type E toxin was fed to Swiss mice. When mice were protected by subcutaneous injections of type E antitoxin prior to feeding the animals survived without showing any symptoms.Subcutaneous injection of type E toxin in amounts of the order of 2 x10^s intraperitoneal MLD (mice) killed mink, and typical symptoms of botulism were observed. This quantity corresponds to ap-proximately 2 intraperitoneal MLD (mice) per g.Comparison is made with previous observations obtained in similar experiments made with Clostridium botulinum type C toxin. It is shown that mink arc substantially less susceptible to type E than to type C toxin when the toxins are given by mouth. On this basis previous results in reports on outbreaks of botulism in mink caused by Clostridium botulinum type E may be regarded as questionable.     

An outbreak of type C botulism in broiler chickens.

An outbreak of type C botulism involving three-week-old broiler chickens on deep litter is described. No direct source of toxin was found. Cl botulinum type C was distributed widely in the litter, and several thousand per gram were demonstrated both in the litter and in the intestinal contents of chicken.     

Recent increase of Marek's disease in Korea related to the virulence increase of the virus

The incidence of Marek's disease (MD), an important neoplastic disease of chickens, suddenly increased in 1997 in Korea. Most MD cases of this country were detected in chickens over 20 wk of age. Five MD viruses were isolated from field flocks in which severe MD losses had occurred, and one of the viruses was studied to compare its pathotype with the prototype JM strain. The isolate KOMD-IC induced severe depression not only in body weight but also in relative bursal weight, and the depression by KOMD-IC was more severe than that induced by JM strain. In addition, the incidence of MD tumor caused by KOMD-IC was higher than that caused by the JM strain. The protective capacity of several MD vaccines was studied against challenge with KOMD-IC. The protective levels of several MD vaccines such as herpesvirus of turkeys (HVT), HVT plus SB1, and Rispens were usually lower against challenge with KOMD-IC than those challenged with JM strain, even if the chickens vaccinated with serotype 1 were not completely protected against challenge with KOMD-IC. The above results indicate that the virulence of KOMD-IC isolated recently was increased, and the increase of MD outbreak in Korea may be related to the virulence increase of the virus. Various MD vaccine programs were applied to reduce MD loss to a broiler breeder farm where severe MD loss had occurred. Serotype 1 vaccine could dramatically decrease the mortality due to MD, and the best results were obtained from the flocks vaccinated with bivalent vaccine of Rispens and HVT.     

Scanning electron microscopy of the cecal mucosa in Eimeria-tenella-infected and uninfected chickens.

Four major types of surface conformation were observed in the ceca of uninoculated control chickens. Spatulate villi were found in the cecal neck region, low ridges in the region where the neck expands, protruding collarlike structures in the mid cecal pouch, and flattened collars in the distal portion. In ceca infected with Eimeria tenella, there was some erosion and sloughing of the mucosal cells. These lesions were slight in the neck region, more severe in the dilated portion, most severe in the midregion, and moderate in the distal area. Oocysts were observed in the mucosal tissue of the cecal pouch.