Efficacy of feed additives to reduce the effect of naturally occurring mycotoxins fed to turkey hen poults reared to 6 weeks of age.

Corn with naturally occurring aflatoxin (AF), wheat with naturally occurring doxynivalenol (DON), and barley with naturally occurring zearalenone (ZEA) were used to make rations for feeding turkey hen poults to 6 weeks of age. Control rations with equal amounts of corn, wheat, and barley were also fed. The control rations did contain some DON while both sets of rations contained ZEA. Within each grain source, there were 4 treatments: the control ration plus 3 rations each with a different feed additive which were evaluated for the potential to lessen potential mycotoxin effects on bird performance and physiology. The additives were Biomin BioFix (2 lb/ton), Kemin Kallsil (4 lb/ton), and Nutriad UNIKE (3 lb/ton). The mycotoxin rations reduced poult body weight (2.31 vs. 2.08 ± 0.02 kg) and increased (worsened) poult feed conversion (1.47 vs. 1.51 ± 0.01) at 6 wk. Feeding the poults the mycotoxin feed also resulted in organ and physiological changes typical of feeding dietary aflatoxin although a combined effect of AF, DON, and ZEA which cannot be dismissed. The feed additives resulted in improved feed conversion to 6 wk in both grain treatment groups. The observed physiological effect of feeding the additives was to reduce relative gizzard weight for both groups and to lessen the increase in relative kidney weight for the birds fed the mycotoxin feed. In conclusion, the feed additives used in this study did alleviate the effect of dietary mycotoxins to some degree, especially with respect to feed conversion. Further studies of longer duration are warranted.

Direct fed microbial supplementation repartitions host energy to the immune system.

Direct fed microbials and probiotics are used to promote health in livestock and poultry; however, their mechanism of action is still poorly understood. We previously reported that direct fed microbial supplementation in young broilers reduced ileal respiration without changing whole-body energy expenditure. The current studies were conducted to further investigate the effects of a direct fed microbial on energy metabolism in different tissues of broilers. One hundred ninety-two 1-d-old broiler chicks (16 chicks/pen) were randomly assigned to 2 dietary groups: standard control starter diet (CSD) and CSD plus direct fed microbial (DFMD; 0.3%) with 6 pens/treatment. Body weight, feed consumption, whole-body energy expenditure, organ mass, tissue respiration rates, and peripheral blood mononuclear cell (PBMC) ATP concentrations were measured to estimate changes in energy metabolism. No differences in whole body energy expenditure or BW gain were observed; however, decreased ileal O(2) respiration (P < 0.05) was measured in DFMD fed broilers. In contrast, the respiration rate of the thymus in those broilers was increased (P < 0.05). The PBMC from DFMD fed broilers had increased ATP concentrations and exhibited increased ATP turnover (P < 0.01). To determine if the increased energy consumption by PBMC corresponded with an altered immune response, broilers were immunized with sheep red blood cells (SRBC) and assayed for differences in their humoral response. The DFMD-fed broilers had a faster rate of antigen specific IgG production (P < 0.05) and an increase in total IgA (P < 0.05). Collectively, these data indicate that supplementation with the direct fed microbial used in this study resulted in energy re-partitioning to the immune system and an increase in antibody production independent of changes in whole body metabolism or growth performance.

Comprehensive analysis of commercially available mouse antichicken monoclonal antibodies for cross-reactivity with peripheral blood leukocytes from commercial turkeys.

In the United States, turkey production contributes approximately $14.4 billion to the US economy; however, the number of reagents specifically developed to study the immune system of this economically important species is limited. To compensate for this, laboratories focused on the turkey system have each empirically tested various chicken-specific reagents for cross-reactivity with turkeys. The result is a patchwork of reports using different genetic lines and different ages, and in many cases, leading to inconsistent conclusions about the cross-reactivity of the reagents tested. In the current study, we investigated a large panel of commercially available monoclonal antibodies specific for chicken leukocyte markers for their ability to specifically recognize the turkey homolog of their respective ligand using 2 different genetic lines of commercial turkeys. The results of these studies identify 8 chicken-specific monoclonal antibodies (F21-21, F21-2, CT4, EP96, 3-298, AV7, c264, and AV6) as demonstrating strong evidence for cross-reactivity with turkey peripheral blood mononuclear cells from both commercial lines, 3 of which (F21-2, EP96, and c264), to our knowledge, have not previously been reported. In addition, characterization of the anti-CD8α monoclonal antibody 3-298 provides evidence that turkeys, like chickens, have a relatively high percentage of CD4CD8 double-positive T-cells in circulation and have at least 5 alleles of the CD8α gene. Collectively, the results from these experiments strengthen our understanding of the turkey immune system, its relative level of conservation with the chicken system, and adds to the list of reagents that can be reliably used to assess immune responses in commercial turkeys.

Maternal antibody transfer to broiler progeny varies among strains and is affected by grain source and cage density.

Two experiments were conducted to examine the effects of broiler breeder dietary grain source and cage density on maternal antibody (MatAb) transfer to progeny in 2 genetic strains (A and B). Broiler breeders were assigned to 16 litter floor pens and fed either corn- or wheat-based diets. Breeders were administered 4 live vaccines against Newcastle disease virus (NDV). At 23 wk of age, pullets and cocks, which reflected the full BW distribution from each treatment, were moved to a cage breeder house and placed at 1 or 2 hens/cage. Breeders were artificially inseminated at 44 wk (experiment 1) and 52 wk of age (experiment 2). Eggs were collected for 8 d, incubated, and placed in individual pedigree bags at d 19 of incubation. Blood samples from 5 chicks per treatment combination were collected at hatch in both experiments. Spleen and bursa were collected from the same chicks for histomorphometry analyses in experiment 2. In the second experiment, 12 chicks per treatment were placed in cages. Progeny were provided diets based on the same grain (corn or wheat) as their parents. Serum samples were collected at 5, 9, and 13 d of age and analyzed for anti-NDV MatAb. Data were analyzed as a 2 × 2 × 2 factorial design considering strain, dietary grain source, and cage density as main factors. Interaction effects were observed in breeders and progeny. Experiment 1 showed that strain A chicks had lower levels of MatAb when hens were housed at 2 hens/cage rather than 1 hen/cage. The MatAb levels of strain B chickens were not affected by cage density in either experiment. Experiment 2 demonstrated similar effects of cage density on MatAb levels and the area of bursa follicles for both strains. Progeny of breeders fed corn-based diets had smaller spleen white pulp only when hens were housed at 2 hens/cage compared with 1 hen/cage. The results of these experiments suggest that breeder strain and cage-density conditions affected MatAb transfer to progeny and embryo development of spleen and bursa.

Broiler embryo bone development is influenced by incubator temperature, oxygen concentration and eggshell conductance at the plateau stage in oxygen consumption.

1. Four experiments were conducted to evaluate the effects of temperature (TEM) and oxygen (O(2)) concentrations during the last 4 d of incubation on bone development. Fertile eggs from two strains were obtained that either exhibited Low or High eggshell conductance (G). 2. Four experimental cabinets provided either four TEM (36, 37, 38 or 39 degrees C) or four O(2) concentrations (17, 19, 21 or 23% O(2)). Data were analysed as a 2 x 2 factorial design. In the fourth experiment, two temperatures (36 and 39 degrees C), two O(2) concentrations (17 and 23%) and the same Low and High G strains were evaluated in a 2 x 2 x 2 factorial design. 3. Body weights (BW) and residual yolks were obtained, both legs were dissected. Femur, tibia and shank weights, length and thickness were recorded. Relative asymmetry (RA) of each leg section was calculated. 4. The results indicated that elevated TEM during incubation increased RA between the two legs, mainly in the Low G strain. Chickens at the lowest O(2) concentrations had lighter and shorter tibias, lighter shanks, and increased RA of femur length compared to chickens in the 23% O(2). In the fourth experiment no interactions were observed between O(2) and TEM. High TEM depressed BW of Low G broilers, but no significant effect of treatments was observed on BW of High G broilers. Nevertheless, the high TEM or low O(2) independently caused reduced femur and tibia weights and length, shank length and thickness, and both low O(2) and high TEM together increased RA in shank weight. 5. These results suggest that late incubation conditions affect long bone development in broilers.

Direct-fed microbial PrimaLac and salinomycin modulate whole-body and intestinal oxygen consumption and intestinal mucosal cytokine production in the broiler chick.

The current study investigated whole-body O2 consumption, intestinal O2 consumption, and intestinal inflammation status through mucosal cytokine production on broiler chicks fed the direct-fed microbial PrimaLac. One hundred twenty 1-d-old broiler chicks were randomly assigned to 1 of 3 experimental diets: standard starter diet (control), standard starter diet with added salinomycin (SAL), and standard starter diet with added PrimaLac (DFM). Birds were housed in 2 separate rooms, the control and SAL treatments in one room and the DFM in another. Intact ileal and cecal samples were collected on d 19, 20, and 21 after measuring whole-body O2 consumption using indirect calorimetry. The O2 up-take of ileal tissue was measured using an in vitro O2 monitor. Analysis of intestinal immune status of broilers was measured by the relative differences in mRNA of both pro- and antiinflammatory cytokines: interleukin-(IL) 1beta, IL-6, and IL-10 using real-time reverse transcription-PCR. Broilers exhibited a 6 to 16% decrease in whole-body energy expenditures and up to a 47% decrease (P<0.05) in ileal energy expenditures in the DFM group compared with other treatments. The reverse transcription-PCR data demonstrated that DFM consortium numerically altered both pro- and antiinflammatory cytokines within the ileum of 19-d posthatch broilers. These data suggest that direct-fed microbials like PrimaLac increase metabolic efficiency via changes in intestinal physiology and metabolism.

Microarchitecture and spatial relationship between bacteria and ileal, cecal, and colonic epithelium in chicks fed a direct-fed microbial, PrimaLac, and salinomycin.

Direct-fed microbials (DFM) could serve as a potential alternative to the feeding of antibiotics in poultry production. In this study, the effects of providing a DFM were compared with the feeding of salinomycin on intestinal histomorphometrics, and microarchitecture was examined. Broiler chicks (n=18 per treatment; trials 1 and 2) were fed a standard starter diet (control), control+PrimaLac (DFM; 0.3% wt/wt), and control+salinomycin (SAL; 50 ppm) from hatch to 21d. The birds were euthanized on d 21, and the ileal, jejunal, cecal, and colon tissues were dissected. Samples were examined by light microscopy (jejunum and ileum; trial 1) and scanning electron microscopy (ileum, cecum, and colon; trial 2). Feeding of the DFM increased intestinal muscle thickness (P<0.05) up to 33% compared with the control treatment. The DFM group also had increased villus height and perimeter (P=0.009 and 0.003, respectively) in jejunum. Segmented filamentous-like bacteria were less numerous in DFM-treated chicks than in the control chicks. Very few segmented filamentous-like bacteria were found near other microbes in the ileum. The DFM chicks had a larger number of bacteria positioned over or near goblet cells and in intervilli spaces. Bacteria in the colon were observed to be attached primarily around and within the crypts. Mucous thickness was less, and the density of bacteria embedded in the mucous blanket appeared to be lower in DFM-treated animals than in the control in all intestinal segments. The birds fed SAL had fewer bacteria and enterocytes in the ileum than in the control-and DFM-treated birds, and they had thicker and fewer microvilli. Because gastrointestinal track colonization by the DFM organisms can prevent the attachment of pathogens to the epithelium, spatial relationships, in this study, demonstrate the functionality of DFM and probiotics in preventing disease. It also supports previous observations that the feeding of salinomycin may alter intestinal function.

Examining the cellular pathways involved in influenza virus induced apoptosis.

Apoptosis is essential in many physiological processes including wound healing and development of the immune response. Apoptosis also plays an important role in the pathogenesis of many infectious diseases including those caused by viruses. Influenza viruses induce apoptosis in cells that are permissive for viral replication and cells that do not support viral replication. The cellular pathways involved in influenza virus induced apoptosis are currently ill defined. Previous studies suggest that influenza virus infection increased the expression of the Fas antigen in HeLa cells, and that Fas antigen is partially involved in apoptosis. In these studies we examined the cellular pathways involved in avian influenza virus induced apoptosis in two cell lines that support productive viral replication: Madin-Darby canine kidney cells (MDCK) and mink lung epithelial (Mv1Lu) cells.

Localization of astrovirus in experimentally infected turkeys as determined by in situ hybridization.

Twenty-one 3-day-old turkey poults from British United Turkeys of America were orally inoculated with a recently characterized astrovirus, TAstV-2, isolated from turkeys with poult enteritis and mortality syndrome. At 1, 2, 3, 4, 5, 7, and 9 days postinfection (dpi), three inoculated birds were euthanatized, and tissues (intestines, spleen, bursa, and thymus) were collected immediately into 10% neutral buffered formalin. Inoculated birds were diarrheic by 3 dpi, and frothy feces persisted throughout the experimental period. Histologically, there was only slight evidence of enteric damage, which was characterized by mild epithelial necrosis, lamina propria infiltrates, minimal villus atrophy, and mild crypt hyperplasia. In situ hybridization, using a negative sense digoxigenin-labeled riboprobe to the capsid gene of TAstV-2, revealed viral RNA in intestinal epithelial cells at the basal margins of the villi, in distal small intestine, and in cecum at 2 dpi, with subsequent extension to epithelium of the large intestine and proximal small intestine (3-5 dpi). Minimal virus remained by 9 dpi.

Inactivation of an astrovirus associated with poult enteritis mortality syndrome.

Outbreaks of poult enteritis mortality syndrome (PEMS) continue to cause financial losses to the turkey industry. Clinically, PEMS is defined by mortality profiles, diarrhea, flock unevenness, and immunosuppression. PEMS is a very difficult disease to control and prevent. Depopulation of PEMS-affected flocks and thorough cleaning of the contaminated housing have failed to prevent infection (disease) in subsequent flock placements. The relationship of PEMS to other enteric disease complexes of young turkeys is unknown, partly because the causative agent of PEMS remains unknown. Recently, we isolated a unique astrovirus strain from the thymus and intestines of PEMS-infected poults. This strain is molecularly and serologically distinct from the astrovirus that circulated in turkeys in the 1980s. Mammalian astroviruses are very resistant to inactivation. In these studies, we examined the stability of partially purified PEMS-associated astrovirus to inactivation with heat, laboratory disinfectants, and commercial disinfectants used in commercial turkey houses in an embryonated egg model system. Similar to mammalian astroviruses, the PEMS-associated astrovirus is resistant to inactivation by heat, acidification, detergent treatment, and treatment with phenolic, quaternary ammonium, or benzalkonium chloride-based products. Only treatment with formaldehyde, beta-propriolactone, or the peroxymonosulfate-based product Virkon S completely inactivated the astrovirus in the embryo model. These studies provide an alternate means to potentially control at least one virus associated with PEMS through the use of specific disinfectants.

Development of an RT-PCR diagnostic test for an avian astrovirus.

Astroviruses are small round viruses that cause enteric disease in the young of several species. Detection and diagnosis of astrovirus infection in non-human hosts relies heavily on electron microscopy and fluorescent antibody tests. Recently, our laboratory isolated and sequenced an avian astrovirus from poult enteritis mortality syndrome affected turkeys. These studies describe the development of RT-PCR methods, which specifically detect regions of the viral capsid and polymerase genes, and demonstrate their use in detecting astrovirus infection in commercial turkey flocks.

Identifying agent(s) associated with poult enteritis mortality syndrome: importance of the thymus.

Poult enteritis mortality syndrome (PEMS), a highly infectious disease of young turkeys, causes serious financial losses to the turkey industry. Clinically, PEMS is defined by mortality profiles, diarrhea, growth depression, and immunosuppression. Although many viruses, bacteria, and parasites are found in PEMS-infected birds, the inciting agent remains unknown. Experimentally, PEMS can be reproduced by exposing naïve poults to the intestinal contents from infected birds. Previous reports suggest that extraintestinal tissues fail to reproduce the disease. Histopathologic examination of tissues from PEMS-infected poults suggested that the thymus exhibited the earliest signs of pathology. On the basis of these observations, we hypothesized that the thymus harbors an agent(s) involved in PEMS. In these studies, naïve turkey poults were orally inoculated with a bacteria-free filtrate composed of either the intestines and feces or the thymus from PEMS-infected birds and were monitored for clinical signs of PEMS. Poults exposed to a filtrate composed solely of the thymus from PEMS-infected birds exhibited diarrhea, growth depression, mortality, pathology, and, most importantly, immunosuppression similar to poults exposed to the intestinal filtrate. The results of this study suggest that the thymus of infected birds harbors the agent(s) that can reproduce a PEMS-like disease in turkey poults.

Molecular characterization of an avian astrovirus.

Astroviruses are known to cause enteric disease in several animal species, including turkeys. However, only human astroviruses have been well characterized at the nucleotide level. Herein we report the nucleotide sequence, genomic organization, and predicted amino acid sequence of a turkey astrovirus isolated from poults with an emerging enteric disease.

Interleukin-12 overcomes paclitaxel-mediated suppression of T-cell proliferation.

The antineoplastic agent paclitaxel (TAXOL) is a potent inhibitor of tumor cell division and a useful chemotherapeutic for the treatment of refractory ovarian and breast carcinoma. Multiple immune system actions have been ascribed to paclitaxel, including the capacity to induce macrophage antitumor cytotoxic molecule production. However, T-cells are susceptible to paclitaxel's cytostatic functions, and no studies have investigated the effects of direct paclitaxel administration on lymphocyte function in the tumor-bearing host (TBH). Because paclitaxel is currently used as an antitumor chemotherapeutic agent and tumor growth alters leukocyte functions, we assessed T-cell function following chemotherapeutic-type paclitaxel treatment. Paclitaxel administration significantly compromised the proliferative capacity of both normal host and TBH lymphocytes in vitro. Although tumor growth impaired T-cell interferon-gamma (IFN-gamma) production, paclitaxel treatment did not alter IFN-gamma. We speculate that the immunostimulatory cytokine interleukin-12 (IL-12), which promoted T-cell activation and proliferation, was capable of reversing paclitaxel-mediated immunosuppression. Exogenous IL-12 fully reconstituted proliferative reactivity and enhanced IFN-gamma production by both normal host and TBH lymphocytes in vitro. Collectively, these data suggest that chemotherapeutic paclitaxel regimens impart significant but reversible inhibition of lymphocyte populations, and IL-12 may be a useful ancillary immunotherapeutic to overcome paclitaxel-induced modulation of lymphocyte activities.

Phenylmercury and its mobilization in the organism by a metal complex-forming substance: 2,3-dimercapto-1-propane sodium sulfonate.

Workers handling dressing machines for seed treatment with the product Agronal, containing a phenylmercury chloride fungicide, were exposed to high concentrations of phenylmercury dust in the working environment. Urine analyses for mercury result in concentration of up to 0.1 mg Hg/l of urine. After administration of a complex-forming substance-Unitol (2,3-dimercapto-1-propane sodium sulfonate)-a higher urinary excretion of mercury occurred. The amount of mercury excreted confirmed its deposit in the organism. It was speculated that subjective complaints by workers handling dressing machines could be caused by high exposure to phenylmercury. This suggestion cannot, however, be fully accepted because the complaints were not necessarily specific for mercury only, but could also have been caused by factors of nontoxic origin, such as stress at the workplace, discontent with work and environmental hygiene conditions, conflicts and alcoholism. Most probably, it was a case of interpotentiation of the effects of toxic and non-toxic nature.

Semen quality and gonadotropin levels in patients operated upon for cryptorchidism.

In a long-term follow-up study we compared preoperative testicular position, age at orchiopexy, and morphology of testicular biopsies investigated at orchiopexy to sperm analysis results, testicular volume, and serum follicle-stimulating hormone and luteinizing hormone levels at follow-up in 46 men with a history of surgical correction of cryptorchidism in childhood. They had undergone orchiopexy at age 1.5-12.0 years, 24 for unilateral and 22 for bilateral cryptorchidism. Eleven (46%) of the 24 patients with unilateral and 7 (32%) of the 22 with bilateral undescended testes had a normal sperm analysis, whereas in men with impaired spermiogenesis oligo-asthenozoospermia was the predominant feature. Three (13.6%) patients with bilateral cryptorchidism showed azoospermia, but none of the patients with unilateral cryptorchidism did. Neither the age at orchiopexy nor the average germ-cell count per cross-sectioned seminiferous tubule of testicular biopsies examined at orchiopexy correlated significantly with subsequent sperm analysis results or gonadotropin levels at follow-up. The pretreatment testicular position ("testicular position value") has a high prognostic value for prediction of subsequent sperm analysis results and serum gonadotropin levels.

Playground accidents.

To study the causes and sequelae of playground accidents we analysed, in a retrospective study, 374 playground accidents. Questionnaires were sent to the parents and 103 parents (28%) provided detailed information on the playground accidents of their children. Thirty-one percent of the children injured in playground accidents sustained fractures of the extremities or concussion of the brain. Swings, slides, climbing frames, metal bars and merry-go-round accidents accounted for 71% of the 338 playground equipment-related accidents, whereas 36 accidents (10%) occurred without the use of playground equipment. The majority of these accidents were caused by children fighting. Rope-plank-type swings were frequently involved in backward falling accidents and 86% of the slide accidents were fall accidents. Climbing frames should not be taller than 1.6 m. Further efforts are mandatory to create and maintain playgrounds which help children develop their skills with a minimal risk to injury.