Acute newcastle viral infection of the upper respiratory tract of the chicken. II. The effect of diets deficient in vitamin A on the pathogenesis of the infection.

Keratotic and squamous changes characteristic of vitamin A deficiency were minimal even in chicks which were malnourished and growth stunted and had no vitamin A in their diet. However, when these chicks were infected with Newcastle disease virus (NDV), keratotic changes appeared, most markedly in areas regenerating after infection. In chicks raised on full nutrient diets lacking only vitamin A, keratotic changes appeared in several areas of nasal mucosa but were absent from the mucosa of the inner (under) surface of the maxillary turbinate. Following NDV infection, such changes did appear in the inner lining epithelia. It is suggested that depletion of vitamin A causes regenerating epithelial cells to keratinize. Other effects of combined lack of vitamin A plus NDV infection were exhaustion of lymphoid cells from cranial bone marrow and exhaustion of lymphoid cell systems locally from the nose and paranasal glands.     

Immunohistochemical and serological investigation of experimental Ornithobacterium rhinotracheale infection in chickens

Immunohistochemical techniques were used to prove that Ornithobacterium rhinotracheale was the causative agent of lesions in the air sacs and lungs in chickens, but only after infection with Newcastle Disease virus (NDV). At first, the bacteria attached to the epithelium of the air sacs. Subsequently, they infiltrated the air sacs, and caused thickening of the air sacs, the formation of oedematous and granulomatous tissue, and accumulation of macrophages. The infection peaked at 5 to 9 days, after which recovery was seen. In the lungs, some areas with bronchially-associated lymphoid tissue were affected. The other organs investigated were shown not to be affected. In the absence of NDV infection, aerosol exposure of chickens to O. rhinotracheale only resulted in minimal and temporary microscopic air sac lesions. No O. rhinotracheale cells or fragments could be detected at any time point later than 2 days post-exposure. In spite of the absence of visible lesions, chickens exposed to O. rhinotracheale without prior NDV infection reacted serologically. The duration and the titre of this immune response was indistinguishable from that obtained in chickens exposed after NDV infection. Thus, infection with O. rhinotracheale appears to be restricted to the respiratory tract, with lesions only evident in birds previously infected with NDV, even though a strong serological response can be established in the absence of prior viral infection.     

Early occurrence of apoptosis in lymphoid tissues from chickens infected with strains of Newcastle disease virus of varying virulence

Newcastle disease virus (NDV), the causative agent of Newcastle disease, is a prevalent problem in the poultry industry and often the cause of severe economic loss. There are many strains of the virus and these have varying virulence. The most virulent strains cause systemic lesions of lymphoid tissues, with necrosis and severe lymphoid depletion. Less virulent strains do not cause as much necrosis, but may predispose to secondary infection with other pathogens. Apoptosis or programmed cell death, has been demonstrated to play a role in the pathogenesis of other paramyxovirus infections, notably those caused by measles and canine distemper viruses. To investigate the role of apoptosis in lymphoid organs during NDV infection, immunohistochemistry for determination of expression of caspase-3, a marker of imminent apoptosis, was performed on formalin-fixed paraffin wax-embedded tissues (spleen, thymus, caecal tonsils and bursa of Fabricius) from 4-week-old chickens infected with NDV strains of varying virulence 2 days previously. The amount of apoptosis was proportional to the severity of the clinical disease elicited by the strains.     

Vitamin A levels and immunity in humans

In animal studies, vitamin A deficiency induces a shift from type 2 (humoral) to type 1 (cellular) cytokines; there are no similar data for humans. Control of human immunodeficiency virus (HIV) and Mycobacterium tuberculosis infections requires type 1 cytokine (cellular) immunity. These infections and vitamin A deficiency are highly prevalent in Africa. We therefore examined the interactions among serum vitamin A levels, immune parameters, HIV infection status, Mycobacterium bovis BCG vaccine scarring (as an indicator of a type 1 cytokine profile), and clinical findings for 70 hospitalized children in Malawi, Africa. Directly conjugated monoclonal antibodies and flow cytometry were used to assess cell-specific cytokine production by peripheral blood monocytes and lymphocyte subpopulations. The statistical techniques employed included nonparametric statistics and logistic regression analyses. Thirty percent of the participants had severe vitamin A deficiency (<10 microg/dl), 34% had moderate deficiency (10 to <20 microg/dl), and 36% had normal levels (> or = 20 microg/dl). Vitamin A levels were lower for HIV-positive than for HIV-negative children (median, 10 and 17 microg/dl, respectively). Vitamin A-deficient children (<20 microg/dl) were more likely than non-vitamin A-deficient children to have higher proportions of natural killer (NK) cells (median, 8.3 and 5.2%, respectively) and lower ratios of interleukin-10-producing monocytes to tumor necrosis factor alpha-producing monocytes after induction (median, 1.0 and 2.3, respectively). Vitamin A-deficient children were also more likely than non-vitamin A-deficient children to exhibit respiratory symptoms (47% versus 12%) and visible BCG vaccine scars (83% versus 48%), which are indicative of a type 1 response to vaccination. Vitamin A status did not vary with gender, age, incidence of malaria parasitemia, blood culture positivity, or rates of mortality (6% of vitamin A-deficient children died versus 20% of non-vitamin A-deficient children). Lower vitamin A levels were associated with a relative type 1 cytokine dominance and proportionately more NK cells, both of which may be somewhat beneficial to persons who are exposed to HIV, M. tuberculosis, or other type 1 pathogens.     

Vitamin A deficiency predisposes to Staphylococcus aureus infection.

We have investigated the consequences of vitamin A deficiency in a rat model of T-cell-dependent and superantigen-mediated Staphylococcus aureus arthritis. After intravenous inoculation of enterotoxin A-producing staphylococci, the vitamin-A-deficient rats showed a decreased weight gain compared with the paired fed controls despite equal food consumption. The control rats developed arthritis in the first few days after bacterial inoculation, with a peak frequency at day 5, and then gradually recovered; however, the frequency of arthritis 18 days after bacterial inoculation was 86% among the vitamin A-deficient rats and 44% among the control rats. During this period, 3 of 10 deficient rats and 1 of 10 control rats died. Further in vitro analysis revealed that T-cell responses to S. aureus were significantly higher in the vitamin A-deficient rats than in the control animals. In contrast, B-cell reactivity, measured as immunoglobulin levels, autoantibody levels, and specific antibacterial antibody levels in serum, did not differ between the groups. Interestingly, the innate host defense mechanisms against S. aureus were also profoundly affected by vitamin A deficiency. Thus, despite a larger number of circulating phagocytic cells in the vitamin-A-deficient group, the capacity to phagocytize and exert intracellular killing of S. aureus was significantly decreased in comparison with the control rats. Furthermore, serum from the vitamin A-deficient rats inoculated with Staphylococcus aureus displayed decreased complement lysis activity. Our results suggest that the increased susceptibility to S. aureus infection observed in the vitamin-A-deficient rats is due to a concerted action of antigen-specific T-cell hyperactivity, impaired function of the phagocytes, and decreased complement activity.     

Pathogenesis of infectious bursal disease in embryonally bursectomised chickens

Embryonally bursectomised and nonbursectomised chickens were infected with infectious bursal disease virus (IBDV) at 36 days of age. Neither clinical signs nor gross lesions were observed in the infected, bursectomised (IB) chickens. No significant changes were observed in carcass, thymus or spleen weights of IB and noninfected bursectomised chickens. A mild lymphocytic necrosis and depletion were found in the spleen, thymus and caecal tonsil of the IB chickens. Neither precipitating nor serum neutralising antibodies were detected in the sera but IBDV was reisolated from the spleen and thymus. Infected, nonbursectomised (IN) chickens developed severe depression with diarrhoea and high mortalities. Haemorrhages were found in the muscles of the breast and thigh, proventriculus and intestines. Significant changes were observed between the carcass, thymus and bursa weights of the IN and noninfected, nonbursectomised chickens. There was severe lymphatic necrosis of the bursa, thymus, spleen and caecal tonsil. Both precipitating and neutralising antibodies were detected in the sera and the virus was reisolated from the bursa, thymus and spleen. It is concluded that the bursa of Fabricius is not essential for the establishment of an IBDV infection but is required for the clinical infection.     

Vitamin A Levels and Immunity in Humans

In animal studies, vitamin A deficiency induces a shift from type 2 (humoral) to type 1 (cellular) cytokines; there are no similar data for humans. Control of human immunodeficiency virus (HIV) and Mycobacterium tuberculosis infections requires type 1 cytokine (cellular) immunity. These infections and vitamin A deficiency are highly prevalent in Africa. We therefore examined the interactions among serum vitamin A levels, immune parameters, HIV infection status, Mycobacterium bovis BCG vaccine scarring (as an indicator of a type 1 cytokine profile), and clinical findings for 70 hospitalized children in Malawi, Africa. Directly conjugated monoclonal antibodies and flow cytometry were used to assess cell-specific cytokine production by peripheral blood monocytes and lymphocyte subpopulations. The statistical techniques employed included nonparametric statistics and logistic regression analyses. Thirty percent of the participants had severe vitamin A deficiency (<10 μg/dl), 34% had moderate deficiency (10 to <20 μg/dl), and 36% had normal levels (≥20 μg/dl). Vitamin A levels were lower for HIV-positive than for HIV-negative children (median, 10 and 17 μg/dl, respectively). Vitamin A-deficient children (<20 μg/dl) were more likely than non-vitamin A-deficient children to have higher proportions of natural killer (NK) cells (median, 8.3 and 5.2%, respectively) and lower ratios of interleukin-10-producing monocytes to tumor necrosis factor alpha-producing monocytes after induction (median, 1.0 and 2.3, respectively). Vitamin A-deficient children were also more likely than non-vitamin A-deficient children to exhibit respiratory symptoms (47% versus 12%) and visible BCG vaccine scars (83% versus 48%), which are indicative of a type 1 response to vaccination. Vitamin A status did not vary with gender, age, incidence of malaria parasitemia, blood culture positivity, or rates of mortality (6% of vitamin A-deficient children died versus 20% of non-vitamin A-deficient children). Lower vitamin A levels were associated with a relative type 1 cytokine dominance and proportionately more NK cells, both of which may be somewhat beneficial to persons who are exposed to HIV, M. tuberculosis, or other type 1 pathogens.     

Pathology of lymphoid organs in chickens fed a diet deficient in zinc.

An experiment was conducted, including flow cytometry, to study the pathology of the lymphoid organs and peripheral blood T lymphocytes in zinc (Zn)-deficient chickens. One hundred 1-day-old broiler chickens were randomly divided into two groups and fed on diets with 100 mg/kg Zn (controls) or Zn-deficient diets (Zn, 23.63 mg/kg) for 7 weeks. The weight and growth index of the bursa of Fabricius, thymus and spleen were significantly reduced (P<0.05 or P<0.01) in Zn-deficient birds when compared with those of control broilers. The G0/G1 phase of the cell cycle of the bursa, thymus and spleen was much higher (P<0.01), and the S, G2+M phases and proliferating index lower (P<0.05 or P<0.01) in Zn-deficient broilers than in the controls. The acid alpha-naphthyl acetate esterase-positive ratio of the peripheral blood T lymphocytes and the CD4 and CD8 numbers were markedly reduced (P<0.05 or P<0.01), and the CD4/CD8 ratio increased. Histopathologically, lymphocytes of lymphoid organs were depleted and the reticular cells of the thymus were also degenerate or necrotic in the Zn-deficient birds. The results demonstrate that Zn deficiency seriously inhibited the development of lymphoid organs, impaired the progression of lymphocytes from the G0/G1 phase to the S phase, and caused pathological injury in the lymphoid organs. The results also showed that the effect of Zn deficiency on the primary lymphoid organs occurred earlier than on the secondary lymphoid organs. The effect of Zn deficiency was greatest on the bursa of Fabricius, followed by the thymus, and then the spleen. Potential mechanisms underlying the observations are discussed.     

Local immunity in the respiratory tract of the chicken. I. Transudation of circulating antibody in normal and virus-infected birds.

Three-week-old chickens were given sheep erythrocytes or bovine serum albumin intravenously. Seven days later their tears and saliva possessed low levels of antibody to those antigens. Concurrent infection with lentogenic Newcastle disease virus (NDV) caused a significant increase in transuded antibody in those fluids. In chickens with circulating antibody to NDV, induced by parenterally administered inactivated vaccine, respiratory infection with heterologous infectious bronchitis virus resulted in limited transudation of anti-NDV. In contrast, the tears, saliva and tracheal fluid of non-vaccinated chickens undergoing primary infection with NDV acquired considerable levels of specific anti-NDV. The difference between the two groups is attributed to locally synthesized antibody.     

Aberrant T-cell function in vitro and impaired T-cell dependent antibody response in vivo in vitamin A-deficient rats.

We have previously reported that vitamin A deficiency resulted in a reduced IgA antibody response to cholera toxin (CT) after per-oral immunization. In the present investigation we have studied the in vivo and in vitro immune response in vitamin A-deficient rats to two parenterally applied antigens, beta-lactoglobulin (beta-LG) and picrylsulphonic acid (TNP)-Ficoll. The serum IgG and IgM antibody responses to the T-cell dependent antigen beta-LG were significantly lower in the vitamin A-deficient rats than in the pair-fed control rats. No such differences were seen with the IgG and IgM responses to the T-cell independent antigen TNP-Ficoll. However, the biliary IgA and the serum IgE antibodies against both antigens were decreased in the vitamin A-deficient rats. In vitro lymphocyte stimulation with concanavalin A (Con A) or beta-LG gave higher T-cell proliferation rates in the vitamin A-deficient than in the control rats. Interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) levels in supernatants from Con A-stimulated mesenteric lymph node cells were also higher in the vitamin A-deficient rats, while IL-6 levels were decreased, which is consistent with an up-regulated Th1 activity. Proliferation studies on purified accessory cells and T cells from the deficient and the control rats, mixed in different combinations, showed that the T cells, but not the accessory cells, were disturbed in the vitamin A-deficient rats. Despite the increased T-cell activity in vitro the vitamin A-deficient rats had a lower delayed-type hypersensitivity (DTH) reaction than the pair-fed control rats. In conclusion, the increased IL-2 and IFN-gamma levels may reflect an up-regulation of Th1 cell function, while the decreased IgA, IgE and IL-6 levels indicate a suppression of Th2 cells. The disturbed T-lymphocyte function is manifested in vivo as a decreased DTH reaction and suppressed antibody production, the latter possibly due to a lack of B-cell switching and proliferation factors in vitamin A-deficient rats.     

Infectious bursal disease: distribution and persistence of the virus in inoculated chickens. Study on the transmission of the disease

The distribution, concentration and persistence of infectious bursal disease virus (IBDV) in the lymphoid organs of inoculated chickens, and its persistence in contaminated premises was examined. The virus only multiplied in the bursa of Fabricius where it induced degeneration and necrosis of the lymphoid cells. It persisted for 10 days in this organ and the highest viral concentrations were observed between the 4th and 8th day following inoculation. The virus was found at a low concentration in the spleen and thymus only during the viraemia phase. The inoculated chickens shed virus in the excreta during the first days of infection. The disease was transmitted to other chickens by direct contact with birds which had been inoculated 4, 10 and 14 days previously with IBDV. Litter on which infected chickens had been reared had a high level of infectivity for 30 days after removal from the chickens and still had some infectivity after 60 days. The long life of the virus in an infected house explains its persistence on infected farms and its transmission to successive flocks.     

Differential effects of thyroxine on immune development and autoimmune thyroiditis in the obese strain chicken

The effects of dietary thyroxine (T4) supplementation for specific periods on the early development of the primary lymphoid organs and spontaneous autoimmune thyroiditis (SAT) was examined in the Obese (OS) strain of chicken. Effects of the treatments on concentrations of serum growth hormone (GH) and testosterone were also determined. All treatment groups were examined at 6 weeks. T4 supplementation did not affect serum testosterone or GH concentrations. However T4 given for the first three weeks resulted in significantly increased bursa weights, no change in thymic weights, significantly decreased lymphoid infiltration of the thyroid and reduced thyroglobulin autoantibody levels (TgAAb). T4 supplementation for the full six weeks resulted in no change in bursal weight, significantly increased thymic weight, significantly decreased lymphoid infiltration of the thyroid, and reduced TgAAb. These results suggest that the effects of T4 supplementation on SAT and immune development are dependent on the interval during which it is administered and that testosterone and GH probably do not mediate these effects.     

Immunopathology of chickens infected in vivo and at hatching with the avian osteopetrosis virus MAV.2–0

The immunological capacity of chickens infected with MAV.2–0, an avian osteopetrosis virus, was studied both morphologically and functionally. Infection of 11 to 12-day-old embryos with a high (0.58 × 10⁶-1.2 × 10⁶ plaque-forming units; PFU) and an intermediate (5.8 × 10⁴ PFU) dose of virus resulted in severe stunting, as manifested by lower body and lymphoid organs (bursa, thymus and spleen) weights. The bursa and spleen of osteopetrotic chickens were poorly developed, the thymus partially necrotized; all lymphoid organs had fewer lymphocytes than controls, and there were far fewer germinal centers in the spleen of infected birds, as compared to controls. The humoral and cellular immunity of these osteopetrotic chickens was significantly suppressed, as assessed by (a) the plaque-forming cell response against sheep red blood cells (SRBC) in the spleen; (b) circulating antibody responses against SRBC, Brucella abortus and human γ-globulins (HGG); (c) the delayed hypersen-sitivity reaction against HGG; and (d) mitogenic responsiveness of peripheral blood and spleen lymphocytes to concanavalin A, phytohemagglutinin M and pokeweed mitogen. A few birds, infected as 11-day embryos with the lowest concentration of virus (2.9 × 10⁴ PFU) had no palpable bone lesions, the lymphoid organs had normal histology, and immune responses were quite similar to those in uninfected control birds. Osteopetrotic chickens, infected within 48 h after hatching (5.8 × 10⁵ PFU), had normal IgM-class antibody responses against all antigens studied (SRBC, Brucella, HGG), whereas IgG and/or IgA responses tended to be lower than those observed in the normal controls. These findings, together with the regularly organized small lymphoid follicles in the bursa, indicate a late affection of B cell development, whereas in the birds infected at 11–12 days of incubation, an almost total arrest of B cell development was observed. T cell functions of birds infected at hatching were suppressed to the same extent as those of in ovo infected birds indicating susceptibility of the T cell lineage to MAV.2–0 also at later stages of development.     

Impaired mucosal antibody response to cholera toxin in vitamin A-deficient rats immunized with oral cholera vaccine.

To investigate the importance of vitamin A in the ability to respond to oral antigen administration, rats were fed a vitamin A-free diet. The animals were immunized perorally three times with a mixture of cholera toxin (CT) and a commercial cholera vaccine. The total immunoglobulin A (IgA) concentration as well as the specific IgA anti-CT antibody levels in serum and bile was significantly lower in the vitamin A-deficient animals than in the paired fed controls (animals that were fed a normal commercial diet in an amount equal to the amount the deficient animals consumed), while the levels of total and specific anti-CT IgG were not affected to the same extent by the vitamin A deficiency. The number of IgA anti-CT antibody-producing cells in the mesenteric lymph nodes after immunization was also significantly lower in the vitamin A-deficient rats than in the control rats. Supplementation of the diet with retinyl palmitate restored the ability to mount an IgA antibody response to the antigen, since the level of specific IgA anti-CT antibodies in relation to the total IgA concentration was as high in the vitamin A-supplemented group as in the paired fed control group. Restricted diet intake by itself did not affect the ability to respond adequately to the antigen since there was no difference in IgA anti-CT antibody level between paired fed rats and those being fed ad libitum. Assessment of transforming growth factor beta in cell cultures revealed no difference between vitamin A-deficient and paired fed animals. In summary, vitamin A deficiency resulted in a decreased number of IgA-producing cells, decreased IgA production, and a reduced ability to respond with IgA antibodies to the oral cholera vaccine.     

Effects of photoperiods on the weights of bursa of Fabricius and thymus in Japanese quail

Effects of photoperiods on the growth of the bursa of Fabricius and the thymus were investigated in male (1-8 weeks after hatching) and female (5-7 weeks after hatching) Japanese quail. In males from hatching to 4 weeks of age, the bursal growth was not affected by photoperiod. However, the growth pattern was different between long and short days after 4 weeks of age, i.e., the bursal weight under long days (LD 16:8) increased rapidly until 6 weeks of age and regressed thereafter, whereas it continued to increase under short days (LD 8:16), keeping a constant ratio to body weight. The growth of the thymus showed a similar pattern to that of the bursa of Fabricius. The age for the thymus to reach the maximum weight under LD 16:8 was almost the same as that for the bursa. In LD 8:16, thymus weight increased until 6 weeks and thereafter remained constant. In females, bursa of Fabricius and thymus weights decreased from 6 to 7 weeks under LD 16:8, which was coincided well with the rapid oviduct growth, whereas the organ weights increased under short days. Thus, it was clearly shown that the growth pattern of lymphoid organs is affected by photoperiods in both male and female Japanese quail.     

Chemical bursectomy of chickens with colchicine applied to the anal lips.

To induce chemical bursectomy, 30 microliter colchicine dissolved in saline solution (1 mg/ml) was applied on the anal lips of White Leghorn chickens once daily for four consecutive days after hatching. Histologic characteristics of the bursa of Fabricius, spleen, thymus, cecal tonsils, and rectal wall were studied 1-7 days after hatching. Total necrosis of the lymphoid cells and the follicle-associated epithelium in the bursa was observed during the four days of colchicine application. The bursal stroma remained unchanged, and only minor changes were found in the interfollicular surface epithelium. After colchicine application ceased, some regeneration of the epithelium, as evidenced by small epithelial buds, was found. At the end of the observation period the epithelial buds were often covered by the follicle-associated epithelium, which was capable of phagocytizing carbon. However, practically no lymphoid repopulation was seen in the buds. Since this method of colchicine application had no direct effect on other lymphoid organs or on the survival or weight of the chickens, this bursectomy model seems to be a new tool for use in studies of bursal function.     

Studies on acquired immunity to coccidiosis in bursaless and thymectomized fowls

Fowls hatched from embryos inoculated in ovo with testosterone between the sixth and ninth day of incubation hatched without a detectable bursa of Fabricius. These fowls failed to develop antibodies as the result of repeated infection with Eimeria tenella and levels of serum immune globulin were usually markedly reduced or undetectable. There were very few pyroninophilic cells in the caeca or spleen and secondary foci in the spleen and caecal lymphoid tissue were either not detected or were very reduced in number. The spleen and thymus weights were significantly reduced by testosterone treatment.

Nevertheless, these fowls were successfully immunized, so that they resisted infection when challenged with viable oocysts of E. tenella.

Active E. tenella infection of normal, susceptible, control fowls significantly reduced the thymus weight and increased the bursa weight. Severe haemorrhage into the caecal lumen of infected fowls resulted in lowered blood erythrocyte and lymphocyte counts and a reduction in the total serum protein.

Antibodies, capable of lysing sporozoites, were detected in normal fowls after immunization and in normal susceptible fowls 5 days after initial infection.

Complete surgical thymectomy was attempted within the first 1½ hours after hatching. However, this was only exceptionally complete and about 10 per cent of thymic tissue was detectable at subsequent post-mortem 56 days later. Thymectomized birds produced antibody, pyroninophilic cells and secondary spleen foci indistinguishable from normal control immunized chickens. However, there was a significant reduction in the number of small lymphocytes in the blood. Although thymectomized fowls were successfully immunized against E. tenella infection, there was an indication, shown by daily oocyst discharge determinations that thymectomized fowls were less resistant during immunization than normal fowls. However, both groups of fowls, when challenged, were fully immune. The significance of partial thymectomy and the time of thymectomy in the fowl in relation to the acquisition of resistance is discussed.

The results support our previous observations which have failed to demonstrate a significant role for humoral antibody in the mediation of resistance to E. tenella.

     

Immunosuppressive effects of Marek's disease virus (MDV) and herpesvirus of turkeys (HVT) in broiler chickens and the protective effect of HVT vaccination against MDV challenge

Much of the impact of Marek's disease in broiler chickens is considered to be due to immunosuppression induced by Marek's disease virus (MDV). The present study evaluates the effects of an Australian isolate of pathogenic MDV (strain MPF 57) and a non-pathogenic vaccinal strain of herpesvirus of turkeys (HVT) (strain FC 126) on the immune system of commercial broiler chickens for 35 days following challenge at days 0 or 3 of age. It also investigates the extent of protection provided by HVT vaccine against MDV-induced immunosuppression. Immune system variables, including relative lymphoid organ weight, blood lymphocyte phenotype (CD45+/CD3+, putatively T, and CD45+/LC+, putatively B) and antibody production following vaccination against infectious bronchitis (IB) at hatch, were used to assess the immune status of chickens. Immunosuppression was also assessed by susceptibility to secondary challenge with pathogenic Escherichia coli on day 29 post-MDV challenge. MDV infection reduced the weight of the thymus and bursa of Fabricius, the numbers of circulating T lymphocytes and B lymphocytes, and IB antibody titre. The timing of these effects varied. MDV infection greatly increased susceptibility to E. coli infection. HVT alone caused mild depletion of T and B lymphocytes but no effect on immune organ weight or IB titre. Vaccination with HVT provided good protection against most of the immunosuppressive effects of MDV but not against MDV-induced growth impairment and reduced responsiveness to IB vaccination, suggesting that recent Australian strains of MDV may be evolving in virulence to overcome the protective effects of HVT.     

Marek's disease in broiler chickens: Effect of route of infection and herpesvirus of turkey-vaccination status on detection of virus from blood or spleen by polymerase chain reaction,and on weights of birds,bursa and spleen

The polymerase chain reaction (PCR) has recently emerged as an additional tool for the monitoring and diagnosis of Marek's disease. We investigated a number of factors that may influence the interpretation of PCR results in commercial broiler chickens including the effects of route of infection and herpesvirus of turkeys (HVT)-vaccination status. We also investigated the suitability of peripheral blood lymphocytes (PBL) and spleen as tissues for Marek's disease virus (MDV) detection. HVT-vaccinated and unvaccinated commercial broiler chickens were challenged or not challenged with virulent MDV either by intraperitoneal injection or inhalation of feather dust containing the virus. Blood and spleen samples were collected at weekly intervals to day 35 post-infection for PCR of spleen or PBL. Live weight and lymphoid organ weights were also measured. Spleen and PBL were found to provide similar sensitivity of detection of MDV with a small advantage in favour of spleen. In terms of the timing of detection of MDV, intraperitoneal challenge broadly mimicked natural challenge via inhalation, although infection of birds by inhalation of infective feather dust resulted in slightly later but more complete detection of MDV in challenged birds. Vaccination with HVT delayed the detection of MDV by approximately 10 to 14 days and did not protect against the reduced growth observed in challenged chickens at day 35 post-challenge.