Gross Morphometry, Histomorphometry, and Immunohistochemistry Confirm Early and Persistent Jejunal Crypt Hyperplasia in Poults with Enteritis and Depressed Growth.

Phosphorylated histone 3 (PH3) and cleaved caspase 3 (CCASP3) were used to detect proliferating and apoptotic cells, respectively, in the jejunums of female sibling poults, with and without enteritis and depressed growth, from hatch to day 35. Poults that developed enteritis and depressed growth (SIB flock) were raised on a commercial farm in eastern North Carolina, whereas poults with normal growth and no enteritis (TAU flock) were raised in the Teaching Animal Unit at North Carolina State University College of Veterinary Medicine. Beginning on day 5 through day 35 and at processing, TAU poults were significantly heavier than SIB poults. Jejunal weights, relative jejunal weights, and jejunal densities were greater in SIB poults from day 10 through 35. Jejunal efficiency (body weight /jejunal length) was higher in TAU poults at day 5 and days 10 through 35. Mucosal thickness was greater in SIB poults between days 7 and 21 but greater in TAU poults at days 28 and 35. From day 7 to 35, villus-to-crypt ratios were higher for TAU poults and lower for SIB poults because hyperplastic crypts formed a greater percentage of the mucosa in SIB poults. By day 7, PH3- and CCASP3-positive cells were increased in SIB poults, showing that mucosal changes resulted from combined crypt epithelial hyperplasia and increased apoptosis of villous enterocytes. Findings in this study confirm that enteritis, in the absence of clinical signs, and depressed growth in turkey poults begins by day 7, can be identified microscopically, persists for at least 35 days, is associated with lower processing weights, and has a profound negative effect on turkey growth.

Chemoprevention of spontaneous ovarian cancer in the domestic hen.

The hen is an attractive animal model for in vivo testing of agents that thwart ovarian carcinogenesis because ovarian cancer in the domestic hen features clinical and molecular alterations that are similar to ovarian cancer in humans, including a high incidence of p53 mutations. The objective of the study was to test the potential ovarian cancer chemopreventive effect of the p53 stabilizing compound CP-31398 on hens that spontaneously present the ovarian cancer phenotype. Beginning at 79 wk of age, 576 egg-laying hens (Gallus domesticus) were randomized to diets containing different amounts of CP-31398 for 94 wk, 5 d, comprising a control group (C) (n = 144), which was fed a diet containing 0 ppm (mg/kg) of CP-31398; a low-dose treatment (LDT) group (n = 144), which was fed a diet containing 100 ppm of CP-31398; a moderate-dose treatment (MDT) group (n = 144) which was fed a diet containing 200 ppm of CP-31398; and a high-dose treatment (HDT) group (n = 144), which was fed a diet containing 300 ppm of CP-31398. Hens were killed at 174 wk of age to determine the incidence of ovarian and oviductal adenocarcinomas. Whereas the incidence of localized and metastatic ovarian cancers in the MDT and HDT groups was significantly lower (up to 77%) compared to levels in the C and LDT groups (P < 0.05), the incidence of oviductal cancer was unaffected by CP-31398. CP-31398 appears to be an effective tool for chemoprevention against ovarian malignancies, but does not appear to affect oviductal malignancies.

Bladder and bowel continence in bilateral cerebral palsy: A population study.

INTRODUCTION: The attainment of continence is an important milestone in all children, including those with disability. OBJECTIVE: To describe the age of bladder and bowel continence in children with bilateral cerebral palsy (BCP), and the association with intellectual impairment (II) and severity of motor disability. PATIENTS AND METHODS: The parents of 346 children with BCP were interviewed as part of a population-based prospective study of the children at 3, 7, and 17 years of age. The age of bladder and bowel continence by day and night was ascertained and compared with controls from the Avon Longitudinal Study of Parents and Children (ALSPAC). RESULTS: The median age for daytime bladder and bowel continence in BCP children was 5.4 years compared with 2.4 years in the controls. At 13.8 years of age, 59.4% of BCP children and 99% of controls were continent by day. In BCP children, there was no difference between the attainment of daytime bladder and bowel control. Night-time bladder and bowel control was slower and less completely attained, with 50% of BCP children continent by the age of 11.8 years compared with 3 years in control children. At 13.8 years of age, 51.9% of BCP children compared with 99.4% of controls were continent for bowel and bladder at night. Gross Motor Functional Classification Score (GMFCS) and intellectual ability (IA) (II) were strongly associated with continence attainment (P < 0.0001), but gender was not. DISCUSSION: Delayed and less complete continence attainment was noted in other clinic series of children with cerebral palsy (including hemiplegics) and children with II. Severity of motor disability (GMFCS), and II impacted on other aspects of toilet training, such as: motivation, understanding, communication, and independence skills. The presence of neurogenic bladder and bowel dysfunction can occur in all levels of GMFCS. Thus, there are many reasons that can prevent continence attainment. CONCLUSIONS: Children with BCP achieved day and night-time bladder and bowel continence more slowly and less completely than controls, with 60.8% being continent by day and 54.6% by night at the age of 17 years. The majority of BCP children who were continent by day had achieved this by the age of 5.5 years (86%). At least 88% of BCP children with GMFCS I/II and normal, specific or mild learning impairment were continent for bladder and bowel by day and night. Expectations should be shared with parents, and failure to attain expected continence should be actively investigated.

Parvovirus-associated cerebellar hypoplasia and hydrocephalus in day old broiler chickens.

Cerebellar hypoplasia and hydrocephalus were identified in day old broiler chickens showing nervous signs, impaired mobility, and diarrhea. At postmortem examination, brains of chickens were misshapen and cerebellums were smaller than normal. Microscopically, cerebellar folia were reduced in size and irregularly shaped, and the ventricles were widely distended. Affected cerebellums had focal areas along the base of folia where the internal granular cell layer had been lost, and Purkinje cells were disorganized and located within the molecular layer. Parvovirus DNA was detected by polymerase chain reaction in three of nine brains with oligonucleotide primers designed for amplification of chicken and turkey parvoviruses. On the basis of phylogenetic analyses, the detected virus was most closely related to chicken parvoviruses. These findings suggest that a chicken parvovirus might cause a neurologic disease of young chickens characterized by cerebellar hypoplasia and hydrocephalus; however, its role as the cause of the disease remains to be confirmed.

Pregnancy outcome and fibrinolytic, endothelial and coagulation markers in women undergoing uterine artery Doppler screening at 23 weeks.

BACKGROUND: Pre-eclampsia (PET) and/or fetal growth restriction (FGR) remain a major cause of maternal and fetal morbidity and mortality. In pregnancy, fibrinolysis is controlled by the maternal endothelium and placenta, both of which are central to the pathogenesis of PET/FGR. Clinically, uterine artery Doppler screening at 23 weeks is used to predict PET/FGR. An abnormal uterine artery Doppler finding is defined as early diastolic bilateral uterine artery notching (BN) in the waveform. However, about 50% of mothers with BN do not develop PET/FGR. OBJECTIVES: We investigated fibrinolytic changes and uterine artery Doppler findings in the second trimester, and related them to pregnancy outcome; in particular assessing whether fibrinolytic markers could discriminate between normal and abnormal outcome in mothers with BN. PATIENTS/METHODS: Plasma levels of tissue-type plasminogen activator (t-PA), plasminogen activator inhibitor-1 (PAI-1), plasminogen activator inhibitor-2 (PAI-2), plasmin-alpha(2) antiplasmin (PAP), D-dimers and markers of endothelial dysfunction were measured with Doppler ultrasound at 23 weeks. RESULTS: Those with BN had decreased PAP and D-dimer levels, and raised PAI-1 and thrombomodulin levels. Mothers with BN and PET/FGR had significantly increased t-PA levels and reduced PAI-2 levels. CONCLUSIONS: BN at 23 weeks of gestation is associated with increased PAI-1 levels. Within the BN group, mothers who developed PET/FGR had increased t-PA levels and decreased PAI-2 levels, although there was no net change in fibrinolysis as measured by D-dimer levels. No single fibrinolytic marker is helpful in determining pregnancy outcome in those with BN, but t-PA and PAI-2 are worthy of study in a multifactorial algorithm.

Effect of a direct-fed microbial (Primalac) on structure and ultrastructure of small intestine in turkey poults.

The effects of dietary supplementation of the direct-fed microbial (DFM) Primalac in mash or crumbled feed on histological and ultrastructural changes of intestinal mucosa was determined in 2 populations of poults; 1 with and 1 without a Salmonella spp. challenge. Three hundred thirty-six 1-d-old female Large White turkey poults were randomly distributed into 8 treatment groups with 6 replicates of 7 poults in each pen. The poults were placed on 1 of 4 dietary treatments in a 2 x 2 x 2 factorial arrangement (mash or crumble feed, with or without DFM, not-challenged or challenged at 3 d of age). The DFM groups were fed a Primalac-supplemented diet from d 1 until the last day of the experiment (d 21). At 3 d of age, 50% of the poults were challenged with 1 mL of 10(10) cfu/ mL of Salmonella spp. (Salmonella enterica serovar Typhimurium, Salmonella Heidelberg, and Salmonella Kentucky) by oral gavage. The inoculated poults were housed in a separate room from nonchallenged controls. Feed and water were provided ad libitum for all birds. At d 21, 1 poult per pen (total of 6 poults per treatment) was randomly selected and killed humanely by cervical dislocation. After necropsy, the small intestine was removed, and tissue samples from duodenum, jejunum, and ileum were taken for light and electron microscopic evaluation. The DFM birds showed increased goblet cell (GC) numbers, total GC area, GC mean size, mucosal thickness, and a greater number of segmented filamentous bacteria compared with controls. Changes in intestinal morphology as observed in this study support the concept that poultry gut health and function, and ultimately bird performance, can be improved by dietary supplementation with DFM products such as Primalac as used in this study.

Individualized risk assessment for adverse pregnancy outcome by uterine artery Doppler at 23 weeks.

OBJECTIVE: To provide individualized risk prediction of severe adverse pregnancy outcome based on uterine artery Doppler screening at 23 weeks. METHODS: Color Doppler assessment of the uterine arteries was carried out in 5121 women attending for routine care at 23 weeks in two inner-city obstetric units. The mean uterine artery pulsatility index (PI) was calculated, and the likelihood ratios in relation to PI were generated for severe adverse outcome. This was defined as fetal death, placental abruption, and delivery before 34 weeks associated with preeclampsia and birth weight less than the 10th centile. RESULTS: The likelihood of severe adverse pregnancy outcome increased quadratically with mean uterine artery PI. This relationship was not affected by maternal age, ethnicity, or parity. At a mean PI of 1.45, the 95th centile for our population, the likelihood ratio for severe adverse pregnancy outcome was 5. Cigarette smoking had an additional contribution to PI in predicting severe adverse outcome, roughly doubling the risk for a given PI. CONCLUSION: The individualized risk of severe adverse pregnancy outcome can be determined by uterine artery Doppler screening at 23 weeks and knowledge of cigarette smoking history. Such individualized risk would allow ultrasound resources and clinical follow-up to be tailored to the pregnant woman for the most appropriate use of antenatal care.

MTHFR association with arteriosclerotic vascular disease?

Complex diseases are far more common than diseases that follow simple Mendelian patterns of inheritance. Difficulties are experienced in the designing of experiments to dissect out the contribution of a single allele to a complex phenotype. We review the literature regarding a point mutation in methylenetetrahydrofolate reductase, a candidate gene for susceptibility to vascular diseases.

Comparative pathology of chickens experimentally inoculated with avian influenza viruses of low and high pathogenicity.

Pathologic changes and distribution of viral antigen as determined by immunohistochemistry were compared among 4-wk-old specific-pathogen-free chickens inoculated intratracheally with avian influenza virus (AIV) isolates of either low or high pathogenicity. Viruses of low pathogenicity, previously characterized as mildly pathogenic (MP), included A/chicken/Pennsylvania/21525/83 (H5N2) (MP-Penn) and A/chicken/Alabama/7395/75 (H4N8) (MP-Alab). Viruses of high pathogenicity included A/chicken/Pennsylvania/1370/83 (H5N2), A/chicken/Victoria/A185/85 (H7N7), and A/turkey/Ontario/7732/66 (H5N9). Extremely variable clinical signs ranging from mild respiratory distress to high mortality were present among chickens inoculated with these viruses. Chickens inoculated with highly pathogenic (HP) virus had histologic lesions of necrosis and inflammation in cloacal bursa, thymus, spleen, heart, pancreas, kidney, brain, trachea, lung, and skeletal muscle, whereas chickens inoculated with MP virus had histologic lesions most frequently in lung and trachea or lacked histologic lesions. Immunospecific staining for avian influenza viral proteins was most common in cells within heart, lung, kidney, brain, and pancreas of chicken inoculated with HP viruses, but immunospecific staining was present only and infrequently in trachea and lung of chickens inoculated with MP-Penn AIV. MP-Alab did not produce lesions nor have viral antigen in inoculated chickens but did produce serologic evidence of infection. The pattern of organ involvement and viral antigen distribution in chickens intratracheally inoculated with HP AIV isolates indicates a common capability to spread beyond the respiratory tract and confirms the pantrophic replicative, pathobiologic, and lethal nature of the viruses. However, variability in severity and lesion distribution exists between different HP AIVs. By contrast, MP viruses had the ability to replicate in respiratory or enteric tracts or both and produce lesions within the respiratory tract. These MP viruses exhibited a restricted ability to replicate or produce lesions or both in nonrespiratory or nonenteric tissues; such effects were associated with only sporadic deaths.

Diazinon toxicity in broilers.

Ten 3-day-old chicks were submitted from a flock experiencing high mortality. Necropsy revealed lacrimation, diarrhea, pleural effusion, hemorrhage and ulceration of the proventriculus, and swollen, hemorrhagic livers. Numerous yellow granules were present in the crop. Assayed crop contents contained 39 ppm diazinon [O,O-diethyl O-(2-isopropyl-4-methyl-6-pyrimidyl)phosphorothioate]. The insecticide had been applied to the litter to control fire ants. The high mortality abated after new litter was added on top of the old litter. Diazinon toxicosis was traced to ingestion of diazinon-impregnated granules and was reproduced experimentally.

Relationship of complement resistance and selected virulence factors in pathogenic avian Escherichia coli.

Complement resistance, antibiotic resistance profiles, and virulence profiles of 80 Escherichia coli isolates from the intestines of normal chickens (40 isolates) and chickens diagnosed as having colisepticemia (40 isolates) were compared. Differences were observed between the two groups for antibiotic resistance, siderophore production, presence of type 1 pili, complement resistance, motility, and size of plasmids. The systemic isolates were more likely to have siderophores and type 1 pili, and to be complement-resistant and motile than were the intestinal isolates. No differences between the two groups were observed for colicin production. Further comparison of the 10 most complement-resistant isolates from the systemic group and 10 most complement-sensitive isolates from the intestinal group revealed a correlation between an isolate's resistance to complement and its ability to kill embryos, express type 1 pili, and be motile. Virulence of avian E. coli strains appears to be correlated with complement resistance and the interaction of this resistance with the ability to produce type 1 pili and be motile.

Characteristics of Salmonella spp. and Escherichia coli isolated from broiler flocks classified as "good" or "poor" producers.

Cecal samples from 100 broiler flocks were cultured for Escherichia coli and Salmonella. Samples were selected from flocks classified as either "good" or "poor" producers by a production formula. In an attempt to identify predictors of flock productivity, isolates were studied for differences in antibiotic resistances, hemagglutination of erythrocytes, production of colicins, production of siderophores, type of hemolysis, resistance to host complement, and presence of plasmids. S. typhimurium (copenhagen) was isolated from one poor producing flock and three good producers. Salmonella isolates showed no significant differences in the parameters studied. The E. coli isolates showed significant differences only for the presence of plasmids. These data indicate that differences in host intestinal E. coli from good and poor producing flocks do not predict flock productivity.

The relationship of Cryptosporidium sp. infection of the bursa of Fabricius, intestinal tract, and respiratory system of chickens in Georgia, 1974-1988.

During 1974-1984, cryptosporidiosis was diagnosed in slightly more than 1% (63/6050 = 1.04%) of histopathology reports on chickens and turkeys made at two Georgia diagnostic laboratories. During 1985-1988, cryptosporidiosis was diagnosed in almost 6% (157/2622 = 5.99%) of histopathology reports on chickens and turkeys made at the two laboratories. Intestinal and respiratory Cryptosporidium sp. infections have increased significantly (p less than 0.01) with time. During the same period, there was no statistical pattern for bursa of Fabricius Cryptosporidium sp. infections, and there was no relationship between the incidence of bursal infections and the incidence of either respiratory or intestinal infections.

Effects of dietary aflatoxin in aflatoxin-resistant and control lines of chickens.

The resistance to a single oral dose (12 mg of aflatoxin per kg of BW) and 4 wk of dietary aflatoxin (2.5 mg per kg of feed) were investigated in chicks selected for five generations for resistance to acute aflatoxicosis (AR) and unselected control (C) chicks. The AR chicks were more resistant to a single oral dose of aflatoxin and had significantly decreased sodium pentobarbital sleeping time compared to C chicks. Four-weeks exposure to dietary aflatoxin did not result in any significant change in BW or feed conversion ratios of chicks from either the C or AR line. However, more sensitive indicators of aflatoxicosis including plasma total protein, albumin, cholesterol concentrations, and gamma glutamyl transferase activity were significantly altered in C chicks but not in AR chicks fed aflatoxin. Percentages of liver lipid and liver hyperplasia score were also significantly altered as a result of dietary aflatoxin treatment in C but not AR chicks. These data indicate that selection-associated differences exist between the C and AR lines of chickens that convey resistance to not only a single oral dose of aflatoxin but also to a more chronic dietary exposure to aflatoxin.

Effect of cold acclimation on the broiler chicks' resistance to dietary aflatoxin.

The effect of acclimation to environmental temperatures of 10 to 12 or 28 to 30 C on the resistance of broiler chicks to dietary aflatoxin was examined. Broiler chicks were acclimated from day-of-age for 2 wk to environmental temperatures of 10 to 12 or 28 to 30 C. On Day 14, a single oral dose of aflatoxin (8 mg per kg of body weight) was administered to 50 chicks in each environment. An increase in aflatoxin resistance, as assessed by survival rate, was conveyed by acclimation to cold temperatures. In each environmental chamber, a separate group of chicks was maintained for 2 additional wk, but those groups received 5 mg of aflatoxin per kg feed. By the end of the study, aflatoxicosis was characterized by: 1) a significant (P less than or equal to .05) decrease in body weight; 2) increases in spleen weight, liver weight, liver lipid, and liver dry-matter content; 3) changes in the serum levels of total protein, albumin, glucose, cholesterol, uric acid, potassium, phosphorus, iron and calcium; and 4) increased hepatic hyperplasia. Acclimation to 10 to 12 C was characterized by: 1) an increase in body weight, liver weight, spleen weight and bursa weight; 2) changes in the serum glucose and potassium levels; and 3) a decrease in glutamic-oxaloacetic transaminase activity. Significant aflatoxin by temperature interactions were evident only in serum levels of glucose and phosphorus, and in the serum activity of glutamic-oxaloacetic transaminase. These data suggest that acclimation to cool temperatures does not play a significant role in the resistance by broiler chickens to chronic aflatoxin exposure.

Marek's disease virus-induced transient paralysis in chickens: demonstration of vasogenic brain oedema by an immunohistochemical method.

The presence of vasogenic brain oedema and its distribution in Marek's disease virus (MDV)-induced transient paralysis (TP) were determined in genetically resistant and susceptible inbred White Leghorn chickens. MDV-inoculated TP-susceptible chickens with nervous signs (9 days post-inoculation) had severe vacuolation of cerebellar white matter and associated diffuse leakage of albumin and IgG. The serum protein leakage was associated morphologically with a vasculitis and intramural pseudocyst formation in the walls of blood vessels cuffed by mononuclear cells. This transient vasculitis and resulting vasogenic oedema coincided with the temporary neurological signs seen in TP-susceptible chickens. The vasculitis and vasogenic oedema were not present in brain tissue from recovered MDV-inoculated TP-susceptible chickens, MDV-inoculated TP-resistant chickens, or uninoculated control chickens from either line.

Immunosuppressive potential and pathogenicity of a recent isolate of infectious bursal disease virus in commercial broiler chickens.

At 15 days of age and in the presence of measurable levels of maternal antibody against infectious bursal disease virus serotype I (1:170 virus-neutralization geometric mean titer), a recent isolate (U-28) and a prototype virulent isolate (Edgar) of the same virus caused subclinical infections in commercial broiler chickens. Isolate U-28 caused a significant reduction in the size of the bursa of Fabricius, whereas the Edgar isolate produced splenomegaly. Both isolates reduced the serological response to Newcastle disease virus. The experimental immunosuppressive potential and pathogenicity of isolate U-28 in broiler chickens confirms the role of this virus in recent infectious bursal disease outbreaks.

Pathogenicity of recent isolates of infectious bursal disease virus in specific-pathogen-free chickens: protection conferred by an intermediate vaccine strain.

The pathogenicity of recent isolates of infectious bursal disease virus and the protection conferred against them by a commercial vaccine strain of intermediate virulence were examined in specific-pathogen-free chickens. Based on clinical signs, mortality, and macroscopic lesions in susceptible chickens, the isolates designated as A-Delmarva and U-28 were distinct from a previously known serotype I virulent isolate (Edgar). Histopathological analysis of the bursa of Fabricius did not establish differences between the field isolates. Although the vaccine strain produced some degree of bursal damage in antibody-free chickens, it was significantly less severe than the damage caused by the field isolates. The active immune response induced by vaccination was cross-protective against the pathological effects produced by the different isolates used in this study.

Marek's disease virus-induced transient paralysis in chickens. 1. Time course association between clinical signs and histological brain lesions.

Association between Marek's disease virus (MDV)-induced clinical signs of transient paralysis (TP) and brain. histological lesions (vasogenic oedema and perivascular mononuclear cell cuffs) were evaluated in TP-susceptible line G-B2 chickens in a time sequence study. The most consistent histological lesions were seen in the cerebellum. Leakage of albumin and vacuolation were parallel in development with clinical signs, but preceded the clinical signs by 6 to 12 h. During resolution of signs, a parallel decline in vacuolation, but not in extra-vascular albumin content was observed. The extravascular albumin shifted from an extracellular to intracellular location. No association was seen between the IgG leakage and vasogenic oedema or clinical signs. Perivascular mononuclear cell cuffing was statistically associated with clinical signs, but evaluation of plotted data indicated the slope for the cuffs was less than the slope for corresponding clinical signs. In addition, cuffing began 2 days prior to clinical signs. Thus, perivascular mononuclear cell cuffing was not causally associated with the TP syndrome.