Specific Pathogen Free Turkeys Research Articles

Pathogenic evaluation of a turkey coronavirus isolate (TCoV NC1743) in turkey poults for establishing a TCoV disease model

Turkey coronavirus (TCoV) can cause a highly contagious enteric disease in turkeys with severe economic losses in the global turkey industry. To date, no commercial vaccines are available for control of the disease. In the present study, we isolated a field strain (NC1743) of TCoV and evaluated its pathogenicity in specific-pathogen-free (SPF) turkey poults to establish a TCoV disease model. The results showed that the TCoV NC1743 isolate was pathogenic to turkey poults with a minimal infectious dose at 106 EID50/bird. About 50 % of one-day-old SPF turkeys infected with the virus’s minimal infectious dose exhibited typical enteric disease signs and lesions from 6 days post-infection (dpi) to the end of the experiment (21 dpi). In contrast, fewer than 20 % of older turkeys (1- or 2-week-old) infected with the same amount of TCoV displayed enteric disease signs, which disappeared after 15–18 dpi. Although all infected turkeys, regardless of age, shed TCoV, the older turkeys shed less virus than the younger birds, and 50 % of the 2-week-old birds even cleared the virus at 21 dpi. Furthermore, the viral infection caused day-old turkeys more body-weight-gain reduction than older birds. The overall data demonstrated that the TCoV NC1743 isolate is a highly pathogenic strain and younger turkeys are more susceptible to TCoV infection than older birds. Thus, one-day-old turkeys infected with the minimal infectious dose of TCoV NC1743 could be used as a TCoV disease model to study the disease pathogenesis, and the TCoV NC1743 strain could be used as a challenge virus to evaluate a vaccine protective efficacy.

Specific-pathogen-free Turkey model for reoviral arthritis

Turkey arthritis reovirus (TARV) infections have been recognized since 2011 to cause disease and significant economic losses to the U.S. turkey industry. Reoviral arthritis has been reproduced in commercial-origin turkeys. However, determination of pathogenesis or vaccine efficacy in these turkeys can be complicated by enteric reovirus strains and other pathogens that ubiquitously exist at subclinical levels among commercial turkey flocks. In this study, turkeys from a specific-pathogen-free (SPF) flock were evaluated for use as a turkey reoviral arthritis model. One-day-old or 1-week-old poults were orally inoculated with TARV (O'Neil strain) and monitored for disease onset and progression. A gut isolate of turkey reovirus (MN1 strain) was also tested for comparison. Disease was observed only in TARV-infected birds. Features of reoviral arthritis in SPF turkeys included swelling of hock joints, tenosynovitis, distal tibiotarsal cartilage erosion, and gait defects (lameness). Moreover, TARV infection resulted in a significant depression of body weights during the early times post-infection. Age-dependent susceptibility to TARV infection was unclear. TARV was transmitted to all sentinel birds, which manifested high levels of tenosynovitis and tibiotarsal cartilage erosion. Simulation of stressful conditions by dexamethasone treatment did not affect the viral load or exacerbate the disease. Collectively, the clinical and pathological features of reoviral arthritis in the SPF turkey model generally resembled those induced in commercial turkeys under field and/or experimental conditions. The SPF turkey reoviral arthritis model will be instrumental in evaluation of TARV pathogenesis and reoviral vaccine efficacy.

Host specificity of avian metapneumoviruses

ABSTRACTTo date, four subgroups of avian metapneumoviruses have been defined (AMPV-A, B, C and D) based on genetic and antigenic differences. The extent of infection in the three principal species (turkeys, chickens and ducks) by these subgroups is, however, not well defined. Here, a series of controlled and ethically approved experimental infections were performed in specific pathogen-free turkeys, chickens and ducks with each of the four AMPV subgroups. For subgroup C, one strain isolated from turkeys in the USA (turkey AMPV-C) and one isolated from ducks in France (duck AMPV-C) were compared. Globally, these extensive experimental trials demonstrated that AMPV-A, B, turkey C and D were well adapted to Galliformes, especially turkeys; however, chickens showed limited clinical signs and differences in seroconversion and transmission. Notably, chickens did not transmit AMPV-A to contacts and were shown for the first time to be susceptible to AMPV-D. The duck AMPV-C was well adapted to ducks; however, chickens and turkeys seroconverted and were positive by virus isolation. In addition, seroconversion of contact turkeys to duck AMPV-C demonstrated horizontal transmission of this virus in a non-palmiped species under our experimental conditions. Interestingly, in chickens and turkeys, duck AMPV-C isolation was possible despite a lack of detection of viral RNA. Likewise, the turkey AMPV-C virus was well adapted to turkeys yet was also isolated from chickens despite a lack of detection of viral RNA. These results would suggest a selection for viral genetic sequences that differ from the original strain upon adaptation to a ‘non-conventional host’.

Transmission Kinetics and histopathology induced by European Turkey Coronavirus during experimental infection of specific pathogen free turkeys.

Numerous viruses, mostly in mixed infections, have been associated worldwide with poult enteritis complex (PEC). In 2008 a coronavirus (Fr‐TCoV 080385d) was isolated in France from turkey poults exhibiting clinical signs compatible with this syndrome. In the present study, the median infectious dose (ID 50), transmission kinetics and pathogenicity of Fr‐TCoV were investigated in 10‐day‐old SPF turkeys. Results revealed a titre of 104.88 ID 50/ml with 1 ID 50/ml being beyond the limit of genome detection using a well‐characterized qRT‐PCR for avian coronaviruses. Horizontal transmission of the virus via the airborne route was not observed however, via the oro‐faecal route this proved to be extremely rapid (one infectious individual infecting another every 2.5 hr) and infectious virus was excreted for at least 6 weeks in several birds. Histological examination of different zones of the intestinal tract of the Fr‐TCoV‐infected turkeys showed that the virus had a preference for the lower part of the intestinal tract with an abundance of viral antigen being present in epithelial cells of the ileum, caecum and bursa of Fabricius. Viral antigen was also detected in dendritic cells, monocytes and macrophages in these areas, which may indicate a potential for Fr‐TCoV to replicate in antigen‐presenting cells. Together these results highlight the importance of good sanitary practices in turkey farms to avoid introducing minute amounts of virus that could suffice to initiate an outbreak, and the need to consider that infected individuals may still be infectious long after a clinical episode, to avoid virus dissemination through the movements of apparently recovered birds.

Pathogenicity of Genome Reassortant Infectious Bursal Disease Viruses in Chickens and Turkeys.

Infectious bursal disease virus (IBDV) contains two genome segments (segment A/segment B) that can reassort among the viruses. Reassortant IBDVs have been identified in several countries including the United States. These reassortant viruses usually include at least one genome segment from a very virulent (vv)IBDV strain. In vivo virulence of six reassortant IBDV from the United States was assessed relative to the virulence of three frequently described IBDV pathotypes: vvIBDV (rB strain), classic virulent (cv)IBDV (STC strain), and subclinical (sc)IBDV (Del-E strain). Morbidity and mortality in 4-wk-old specific-pathogen-free (SPF) leghorns indicated that reassortant IBDV with a vv genome segment A and non-vv segment B were less pathogenic than the vv/vv rB strain but more pathogenic than the cv/cv STC strain. The sc/vv IBDV strain D6337 (sc/vv) was comparable to the STC strain in pathogenicity. Viruses with a serotype 2 (ser2) genome segment A, regardless of the type of genome segment B, did not cause clinical disease in SPF chickens or turkeys. None of the reassorted viruses caused morbidity, mortality, or gross lesions in SPF turkeys. Histopathologic lesions in the bursa of turkeys were not observed in any group except those challenged with the serotype 2 OH strain, which had a mild lymphocytic depletion. No mortality was observed in maternally immune broilers inoculated with any of the IBDV pathotypes at 1, 2, 3, and 4 wk of age. No bursal lesions were observed in any of the broiler chicken groups at 1 wk of age except for the D2712 (ser2/cv)-inoculated birds that had mild lymphocyte depletion. Based on evaluation of bursal lesion scores and IBDV reverse transcriptase-PCR on broilers challenged at 2 wk of age, the K669 (vv/ser2) virus broke through the maternal immunity while the STC, Del-E, rB, D2712 (ser2/cv), and 7741 (vv/cv) viruses did not. All viruses broke through maternal immunity in the broilers at 3 wk of age except the Del-E scIBDV and D2712 (ser2/cv) reassortant IBDVs. At 4 wk of age, maternal antibodies were very low and bursal lesions were observed in all broilers challenged with the viruses. The data indicate that genome reassortant IBDVs are less pathogenic than is the rB (vv/vv) IBDV. However, the reassortant viruses with a vv genome segment A can still cause morbidity and mortality in SPF chickens, and they were able to break through maternal immunity produced via use of commercial classic and variant vaccines at an early age. This suggests that current breeder vaccination programs may not adequately protect against the reassortant vv/ser2 and vv/cv IBDV strains.

An experimental study of the survival of turkey coronavirus at room temperature and +4°C

Turkey coronavirus (TCoV) is a gammacoronavirus (Coronaviridae, Nidovirales) responsible for digestive disorders in young turkeys. TCoV has been associated with poult enteritis complex, a syndrome that severely affects turkey production. No medical prophylaxis exists to control TCoV, therefore sanitary measures such as cleaning and disinfection are essential. It is thus important to evaluate temperatures that allow persistence of TCoV in the environment. Two series of aliquots of a suspension of a French isolate of TCoV (Fr TCoV) were stored at room temperature or +4°C for 0 to 40 days. As TCoV does not grow in cell culture, the presence of residual infectious TCoV in the stored samples was tested by inoculating embryonated specific pathogen free turkey eggs. As TCoV does not induce lesions in the embryo, virus replication in the jejunum and ileum of the embryos was detected 4 days post inoculation, using RNA extraction and a real-time reverse transcriptase-polymerase chain reaction based on the nucleocapsid gene. No surviving virus was detected after 10 days storage at +21.6±1.4°C or after 40 days storage at +4.1±1.6°C, these temperatures being representative of the mean summer and winter temperatures, respectively, in the major French poultry-producing region. The relatively short survival of the virus at room temperature should contribute to limited virus survival during summer months. However, infectious virus was still detected after 20 days storage at the cooler temperatures, a finding that suggests prolonged survival of Fr TCoV and easier transmission between poultry farms in a cool environment are possible.

The pathogenicity of avian metapneumovirus subtype C wild bird isolates in domestic turkeys

BackgroundAvian metapneumovirus subtype C (aMPV/C) causes severe upper respiratory disease in turkeys. Previous report revealed the presence of aMPV/C in wild birds in the southeast regions of the U.S.MethodsIn this study, aMPV/C positive oral swabs from American coots (AC) and Canada geese (CG) were passaged three times in the respiratory tract of specific pathogen free (SPF) turkeys and used as aMPV/C P3 virus isolates in subsequent studies.ResultsWild bird P3 isolates showed similar growth characteristics when compared to virulent aMPV/C in chicken embryo fibroblast ( CEF) cell cultures and their glycoprotein G gene sequence was closely related to the G gene of aMPV/C Colorado reference virus. Three-day-old commercial or SPF turkeys were inoculated oculonasally with wild bird aMPV/C P3 isolates. At 5 and 7 days post-inoculation (DPI), severe clinical signs were observed in both of the AC and CG virus-exposed groups. Viral RNA was detected in tracheal swabs by reverse transcriptase polymerase chain reaction (RT-PCR). In addition, immunohistochemistry showed virus replication in the nasal turbinate and trachea. All virus-exposed turkeys developed positive antibody response by 14 DPI.ConclusionsOur data demonstrate that aMPV/C wild bird isolates induced typical aMPV/C disease in the domestic turkeys.

Pathogenesis of Two Strains of Avian Paramyxovirus Serotype 2, Yucaipa and Bangor, in Chickens and Turkeys

Nine serologic types of avian paramyxovirus (APMV) have been recognized. Newcastle disease virus (APMV-1) is the most extensively characterized virus, while relatively little information is available for the other APMV serotypes. In the present study, we examined the pathogenicity of two strains of APMV-2, Yucaipa and Bangor, in 9-day-old embryonated chicken eggs, 1-day-old specific-pathogen-free (SPF) chicks, and 4-wk-old SPF chickens and turkeys. The mean death time in 9-day-old embryonated chicken eggs was more than 168 hr for both strains, and their intracerebral pathogenicity index (ICPI) was zero, indicating that these viruses are nonpathogenic in chickens. When inoculated intracerebrally in 1-day-old chicks, neither strain caused disease or replicated detectably in the brain. This suggests that the zero ICPI value of APMV-2 reflects the inability of the virus to grow in neural cells. Groups of twelve 4-wk-old SPF chickens and turkeys were inoculated oculonasally with either strain, and three birds per group were euthanatized on days 2, 4, 6, and 14 postinoculation for analysis. There were no overt clinical signs of illnesses, although all birds seroconverted by day 6. The viruses were isolated predominantly from the respiratory and alimentary tracts. Immunohistochemistry studies also showed the presence of a large amount of viral antigens in epithelial linings of respiratory and alimentary tracts. There also was evidence of systemic spread even though the cleavage site of the viral fusion glycoprotein does not contain the canonical furin protease cleavage site.

Experimental avian paramyxovirus serotype-3 infection in chickens and turkeys

Avian paramyxoviruses (APMV) are divided into nine serotypes. Newcastle disease virus (APMV-1) is the most extensively characterized, while relatively little information is available for the other APMV serotypes. In the present study, we examined the pathogenicity of two divergent strains of APMV-3, Netherlands and Wisconsin, in (i) 9-day-old embryonated chicken eggs, (ii) 1-day-old specific pathogen free (SPF) chicks and turkeys, and (iii) 2-week-old SPF chickens and turkeys. The mean death time in 9-day-old embryonated chicken eggs was 112 h for APMV-3 strain Netherlands and > 168 h for strain Wisconsin. The intracerebral pathogenicity index in 1-day-old chicks for strain Netherlands was 0.39 and for strain Wisconsin was zero. Thus, both strains are lentogenic. Both the strains replicated well in brain tissue when inoculated intracerebrally in 1-day-old SPF chicks, but without causing death. Mild respiratory disease signs were observed in 1-day-old chickens and turkeys when inoculated through oculonasal route with either strain. There were no overt signs of illness in 2-weeks-old chickens and turkeys by either strain, although all the birds seroconverted after infection. The viruses were isolated predominantly from brain, lungs, spleens, trachea, pancreas and kidney. Immunohistochemistry studies also showed the presence of large amount of viral antigens in both epithelial and sub-epithelial lining of respiratory and alimentary tracts. Our result suggests systemic spread of APMV-3 even though the viral fusion glycoprotein does not contain the canonical furin proteases cleavage site. Furthermore, there was little or no disease despite systemic viral spread and abundant viral replication in all the tissues tested.

Generation and biological assessment of recombinant avian metapneumovirus subgroup C (aMPV-C) viruses containing different length of the G gene

Genetic variation in length of the G gene among different avian metapneumovirus subgroup C (aMPV-C) isolates has been reported. However, its biological significance in virus replication, pathogenicity and immunity is unknown. In this study, we developed a reverse genetics system for aMPV-C and generated two Colorado (CO) strain-based recombinant viruses containing either the full-length G gene derived from a Canadian goose isolate or a C-terminally truncated G gene of the CO strain. The truncated short G (sG) gene encoded 252 amino acids (aa), which is 333 aa shorter than the full-length G (585 aa). The biological properties of these two recombinant G variants were assessed in Vero cells and in specific-pathogen-free (SPF) turkeys. In Vero cells, the short G variant displayed a similar level of growth dynamics and virus titers as the parental aMPV-CO strain, whereas the full-length G variant replicated less efficiently than the sG variant during the first 72 h post-infection. Both of the G variants induced typical cytopathic effects (CPE) that were indistinguishable from those seen with the parental aMPV-CO infection. In SPF turkeys, both of the G variants were attenuated and caused little or no disease signs, but the full-length G variant appeared to grow more readily in tracheal tissue than the sG variant during the first 5 days post-infection. Both G variants were immunogenic and induced a slightly different level of antibody response. These results demonstrated that the large portion (333 aa) of the extracellular domain of the viral attachment protein is not essential for virus viability in vitro and in vivo, but may play a role in enhancing virus attachment specificity and immunity in a natural host.

Evaluation of the prophylactic use of ovotransferrin against chlamydiosis in SPF turkeys

Chlamydophila ( C.) psittaci infections are highly prevalent in turkeys and the economical and public health importance of these infections has been recognized since 1950. As there are no vaccines, antibiotic treatment (tetracylines, enrofloxacine) is often needed to allow marketing of poultry. In this study, we explored the use of ovotransferrin (ovoTF), a natural anti-microbial protein, in preventing an experimental C. psittaci infection in specific pathogen free (SPF) turkeys. Turkeys were treated with aerosolized ovoTF prior to the infection. Groups 1 and 2 received a single dose of 10 and 5 mg ovoTF per turkey, respectively. Group 3 received a daily dose of 5 mg ovoTF per turkey during 12 days. Group 4 served as untreated, infected control group. Turkeys were aerosol infected using 10 6 TCID 50 of the virulent C. psittaci serovar/genotype D strain 92/1293. Birds were monitored (clinical signs, bacterial excretion) during 12 subsequent days before being necropsied. At necropsy, pathology and C. psittaci replication in various tissues was examined. A single dose of 10 mg ovoTF and a repeated daily dose of 5 mg ovoTF could not prevent the birds from becoming infected with C. psittaci, but they significantly reduced the outcome of the infection. A single dose of 5 mg ovoTF had no influence on the outcome of the infection as compared to the non-treated infected controls. Our results demonstrate the anti-chlamydial effect of ovoTF in vivo and present a base for further research on practical applications of ovoTF on turkey farms.

Pathogenesis of type 2 turkey astroviruses with variant capsid genes in 2-day-old specific pathogen free poults

The pathogenicity of three different type 2 turkey astroviruses (TAstV-2) was studied in specific pathogen free turkeys. These viruses differ based on sequence analysis of the capsid gene. Poults were inoculated at 2 days of age and examined during 14 days for clinical signs and virus shedding. All inoculated poults presented signs of enteric disease including diarrhoea and growth depression. Virus presence and shedding was detected by real-time reverse transcriptase-polymerase chain reaction from intestinal contents and cloacal swabs collected at 3, 7 and 14 days post-inoculation. Viraemia was also confirmed by this method. Common lesions observed at necropsy were dehydration; distended intestines filled with watery contents and undigested feed, and dilated caeca with foamy contents. Microscopic lesions present in the intestines consisted of mild crypt hyperplasia, villous atrophy and lymphocytic infiltration, and were most common in the jejunum. Presence of the viruses was demonstrated by immunohistochemistry and by in situ hybridization in both villi and crypt enterocytes in the jejunum and, less frequently, the duodenum, ileum and caeca. Mild lesions consisting mainly of lymphocytic infiltration were also observed in other organs including the pancreas, liver, spleen and kidneys. Mild to moderate bursal atrophy occurred in all TAstV-2-infected poults examined; however, no specific viral staining was observed in this organ or any other tissues examined apart from the intestines. In conclusion, TAstV-2 viruses with variant capsids produce a similar enteric disease in young turkeys and may also affect the immune system of the birds by causing bursal lymphoid depletion.

Pathogenicity of Turkey Astroviruses in Turkey Embryos and Poults

The pathogenicity of turkey astrovirus 2001 (TAstV2001) and turkey astrovirus 1987 (TAstV1987) in specific-pathogen-free (SPF) turkey embryos and commercial poults was investigated. The virus shedding in poults was monitored using electron microscopy (EM) and reverse transcription-polymerase chain reaction (RT-PCR) during the 14-day experimental period. Both viruses caused enteritis and growth depression in SPF turkey embryos and poults. The TAstV2001 did not induce macroscopic or microscopic lesions in thymuses and bursas of embryos or poults. No macroscopic changes were observed in thymuses and bursas of embryos and poults inoculated with TAstV1987, and no statistically significant differences in bursa weight/ body weight ratios (P > 0.05) were detected. However, TAstV1987 infection resulted in microscopic lesions in bursas but not in thymuses of infected embryos and poults. Both TAstV2001 and TAstV1987 were shed during the whole 14-day experimental period as detected by EM and RT-PCR. These findings indicated that both TAstV1987 and TAstV2001 are etiologic agents of turkey enteritis. In addition, TAstV1987 might cause impairment of the immune system of infected poults. The pathogenicity of TAstV1987 is somewhat different from TAstV2001.

Experimental Pathogenesis for Chickens, Turkeys, and Pigeons of Exotic Newcastle Disease Virus from an Outbreak in California during 2002-2003

Exotic Newcastle disease virus (NDV) isolated from chickens during the 2002-2003 California outbreak (CA exotic Newcastle disease [END] virus) was inoculated into 4-week-old specific-pathogen-free (SPF) White Leghorn chickens, 3-week-old SPF Beltsville White turkeys, 6-week-old commercial Broad Breasted White turkeys, and 10- to 20-week-old racing pigeons, and the clinicopathologic features of disease were compared. Birds were monitored clinically and euthanized sequentially with collection of tissues. Tissues were examined by histopathology, by immunohistochemistry to detect viral nucleoprotein, and by in situ hybridization to detect viral mRNA. Clinically, infected chickens and SPF turkeys showed severe depression, and all died or were euthanized because of severe clinical signs by day 5 postinoculation. In these birds, histologic lesions were widespread and virus was detected in multiple organs. All infected commercial turkeys showed mild depression, and incoordination was observed in some birds. Histologic lesions were mild, and viral distribution was limited. In pigeons, only 1 bird showed overt clinical disease, and histologic lesions and viral distribution were present in limited organs. Consequently, susceptibility to highly virulent NDV was shown to vary among chickens, SPF turkeys, commercial turkeys, and pigeons. Additionally, we have evidence of CA END virus subclinical infections that suggest pigeons could be subclinical carriers of other virulent NDV.

Pathogenesis of Newcastle Disease in Commercial and Specific Pathogen-free Turkeys Experimentally Infected with Isolates of Different Virulence

The pathogenesis of five different Newcastle disease virus (NDV) isolates representing all pathotypes was examined in commercial and specific pathogen-free (SPF) turkeys. Experimentally-infected birds were monitored clinically and euthanatized, with subsequent tissue collection, for examination by histopathology, by immunohistochemistry for the presence of NDV nucleoprotein, and by in situ hybridization for the presence of replicating virus. Clinically, the lentogenic pathotype did not cause overt clinical signs in either commercial or SPF turkeys. Mesogenic viruses caused depression in some birds. Turkeys infected with velogenic neurotropic and velogenic viscerotropic isolates showed severe depression, and neurologic signs. Histologic appearances for all strains had many similarities to lesions observed in chickens inoculated with the various isolates; that is, lesions were present predominantly in lymphoid, intestinal, and central nervous tissues. However, in general, disease among turkeys was less severe than in chickens, and turkeys could be considered a subclinical carrier for some of the isolates.

Synergy between avian pneumovirus and Ornithobacterium rhinotracheale in turkeys

The purpose of this study was to assess the possible synergism between Ornithobacterium rhinotracheale (ORT) and avian pneumovirus (APV), inoculated into turkeys via the natural route, for the reproduction of respiratory disease. Three-week-old specific pathogen free turkeys were inoculated oculonasally with either APV subtype A, ORT or both agents using two different time intervals (3 and 5 days) between APV and ORT. The birds were observed clinically on a daily basis and swabbed intratracheally at short, regular intervals. They were killed at 1, 3, 5, 8 and 15 days post single or dual inoculation and examined for gross lesions at necropsy. Samples of the turbinates, trachea, lungs, air sacs, heart, pericardium and liver were taken for bacteriological and/or histological examination. Combined APV/ORT infections resulted in overt clinical signs and a longer persistence of ORT in the respiratory tract and aggravated the macroscopic and histological lesions in comparison with the groups given single infections. In all ORT-challenged turkeys, ORT was isolated from the turbinates, trachea and lungs, but in turkeys infected with both agents ORT was frequently found in the air sacs and on a single occasion in the heart and pericardium. The time interval between APV and ORT inoculation did not have a significant effect on the outcome of the dual infection. A conspicuous important feature was the attachment of ORT to the cilia of the epithelium of the turbinates and trachea of both ORT-infected and APV/ORT-infected birds. In conclusion, the results show that ORT is able to adhere to and colonize the respiratory tract but, under the circumstances used in this study, is not capable of inducing respiratory disease without viral priming.

The clinical, pathological and microbiological outcome of an Escherichia coli O2:K1 infection in avian pneumovirus infected turkeys

The purpose of this study was to evaluate the effect of an Escherichia coli infection in avian pneumovirus (APV)-infected turkeys. One group of 2-week-old specific pathogen-free (SPF) and two groups of 3-week-old conventional (CON) turkeys were inoculated oculonasally with virulent APV subtype A alone, with E. coli O2:K1 alone or with both agents at varying intervals (1, 3, 5 or 7 days) between the two inoculations. The birds were followed clinically and examined for macroscopic lesions at necropsy. Titres of APV were determined in the turbinates, trachea, lungs and air sacs. The number of E. coli O2:K1were assessed in the turbinates, trachea, lungs, air sacs, liver and heart. In both SPF and CON turkeys, dual infection resulted in an increased morbidity and a higher incidence of gross lesions compared to the groups given single infections, especially with a time interval between APV and E. coli inoculations of 3 and 5 days. APV was isolated from the respiratory tract of all APV-infected groups between 3 and 7 days post inoculation. E. coli O2:K1 was isolated only from turkeys that received a dual infection. It was recovered from the turbinates, trachea, lungs, heart and liver. These results show that APV may act as a primary agent predisposing to E. coli colonization and invasion.

Oxygen radicals and nitric oxide production by turkey respiratory macrophages

The influence of different induction protocols on the recovery of elicited turkey respiratory macrophages (RM), and on their oxygenation activity and nitric oxide (NO) production was examined. RM were induced in three week old specific pathogen free turkeys with Sephadex G-50, Thioglycollate broth, and an emulsion of incomplete Freund’s adjuvant (IFA), supplemented either with Mycoplasma hyorhinis grown in Modified Channock broth (IFA- M. hyorhinis) or with Modified Channock broth (IFA-Broth). The RM were recovered by lavage of the lungs and air sacs and were purified by centrifugation through a Percoll suspension. Their oxygenation activity was evaluated in luminol-enhanced chemiluminescence assays, following stimulation with Zymosan A. The NO production was evaluated by incubating the RM with lipopolysaccharide (LPS) from Salmonella enteritidis for 24 or 48 hours. The number of recovered RM was slightly, but not significantly lower for Sephadex G-50 and IFA-Broth than for Thioglycollate broth and IFA- M. hyorhinis. RM elicited with Sephadex G-50 and IFA-Broth showed a significantly higher oxidative burst response to Zymosan A, compared to the Thioglycollate and IFA- M. hyorhinis elicited RM. Although all elicited RM showed a high NO production upon stimulation with LPS, no significant differences were seen in the NO production of the RM obtained following the different induction treatments. Our results point out that care should be taken when applying elicited RM for in vitro assays, as distinct levels of oxygenation activity were obtained using different induction protocols.

Chlamydia psittaci in turkeys: pathogenesis of infections in avian serovars A, B and D

At 7 days of age, 4 groups, each of twenty specific pathogen free turkeys kept in isolation units were inoculated by aerosol with the Texas Turkey strain (avian Chlamydia psittaci serovar D), strain 92 1293 (avian Chlamydia psittaci serovar D), strain 84 55 (avian Chlamydia psittaci serovar A) or strain 89 1326 (avian Chlamydia psittaci serovar B). A fifth group of 4 specific pathogen free turkeys were sham inoculated controls. At daily intervals for 10 days and then twice weekly up to 34 days post infection, one bird in each group was killed and the target tissues and cells for replication and the sequence of events of serovar A, B and D infections was examined. In these turkeys, the primary site of replication was the respiratory tract. Chlamydial replication could be detected in the respiratory tract on day 1 post inoculation (p.i.) for group A, on day 3 p.i. for group B and on day 1 to 2 p.i. for groups D 1 and D 2. Subsequently, there was chlamydaemia and localisation in the digestive tract, in one or more parenchymatous organs, in the pericardium and in the conjunctivae. Specific immunoperoxidase staining revealed chlamydiae in these organs in epithelial cells and in monomorphonuclear cells in all infected groups. The monomorphonuclear cells were identified as macrophages by double immunofluorescence staining. Chlamydiae were present in the same tissues for serovars A and D, but could not be demonstrated in proventriculus, duodenum, pancreas, ovaries and testes for serovar B. Furthermore, the intensity of replication was similar for all serovars. However, for serovar B in comparison with the other serovars, the bacteria appeared in most tissues 1 to 6 days later and the maximal replication in these tissues occurred 3 to 4 days later.

Reproduction expérimentale de mycoplasmose à Mycoplasma gallisepticum chez le poulet, le dindonneau, la poule pondeuse et l’embryon de poule

Experimental infection with Mycoplasma gallisepticum in chickens, turkeys, laying hens and cluck embryos. The Mycoplasma gallisepticum R strain is serially passaged ten times through specific pathogen free (SPF) chickens in order to exacerbate its potential pathogenicity. The recovered MGR P10 strain is inoculated in SPF chickens and turkeys, commercial laying fowl and nineteen-day-old chick embryos. The inoculation induces general and respiratory symptoms in the different hosts. Mortality occurs in the chicks hatched from inoculated embryos. A drop in egg production is observed in the laying hens. The strain is also shown to be able to spread from bird to bird.