Heparan sulfate-protein binding specificity.

Heparan sulfate (HS) represents a large class of linear polysaccharides that are required for the function of all mammalian physiological systems. HS is characterized by a repeating disaccharide backbone that is subject to a wide range of modifications, making this class of macromolecules arguably the most information dense in all of biology. The majority of HS functions are associated with the ability to bind and regulate a wide range of proteins. Indeed, recent years have seen an explosion in the discovery of new activities for HS where it is now recognized that this class of glycans functions as co-receptors for growth factors and cytokines, modulates cellular uptake of lipoproteins, regulates protease activity, is critical to amyloid plaque formation, is used by opportunistic pathogens to enter cells, and may even participate in epigenetic regulation. This review will discuss the current state of understanding regarding the specificity of HS-protein binding and will describe the concept that protein binding to HS depends on the overall organization of domains within HS rather than fine structure.

Wing shape and size of the western corn rootworm (Coleoptera: Chrysomelidae) is related to sex and resistance to soybean-maize crop rotation.

The western corn rootworm, Diabrotica virgifera virgifera LeConte, is a major pest of maize in the United States and more recently, Europe. Understanding the dispersal dynamics of this species will provide crucial information for its management. This study used geometric morphometric analysis of hind wing venation based on 13 landmarks in 223 specimens from nine locations in Illinois, Nebraska, Iowa, and Missouri, to assess whether wing shape and size differed between rotated and continuously grown maize where crop rotation-resistant and susceptible individuals are found, respectively. Before assessing differences between rotation-resistant and susceptible individuals, sexual dimorphism was investigated. No significant difference in wing (centroid) size was found between males and females; however, females had significantly different shaped (more elongated) wings compared with males. Wing shape and (centroid) size were significantly larger among individuals from rotated maize where crop-rotation resistance was reported; however, cross-validation of these results revealed that collection site resistance status was an only better than average predictor of shape in males and females. This study provides preliminary evidence of wing shape and size differences in D. v. virgifera from rotated versus continuous maize. Further study is needed to confirm whether wing shape and size can be used to track the movement of rotation-resistant individuals and populations as a means to better inform management strategies.

Infectious bursal disease virus as a surrogate for studies on survival of various poultry viruses in compost.

Infectious bursal disease virus (IBDV) is resistant to many environmental stresses and often persists on farms for months. This study investigated survival of a vaccine strain of IBDV in the bursa of Fabricius and splenic tissue from experimentally infected chickens and in splenic tissue and manure that had been inoculated with the virus. The specimens buried in compost were contained within nylon mesh bags, and the tissues were enclosed within the abdominal cavity of chicken carcasses. Extracts of composted specimens were inoculated into Vero cell cultures, and real-time reverse transcriptase PCR was used to quantify the virus in the cultures. By day 7 in compost, the temperature had been slightly above 55 C for 2.6 days and IBDV had been inactivated in specimens that had been inoculated with virus but had survived in tissues that had been taken from infected chickens. By day 14, the temperature had been above 55 C for 8.8 days and the virus was inactivated in all specimens. The results suggest that composting of poultry carcasses and manure would help to break the cycle of infection with IBDV and that the virus could be valuable as a surrogate for predicting the inactivation of less resistant viruses during composting.

Under-report and underdiagnosis of chronic respiratory diseases in an African country.

BACKGROUND: Chronic respiratory diseases (CRD) are greatly underestimated. The aim of this study was to assess the burden associated with reported CRD and chronic obstructive pulmonary disease, as defined on the basis of various standardized criteria, by estimating their point prevalence in a sample of individuals attending the Primary Health Care (PHC) level and Emergency Room (ER) Departments in Cape Verde (CV) archipelago. The second aim of the study was to identify factors related to airways obstruction and reported CRD in this population. METHODS: A cross-sectional study was carried out in CV during 2 weeks. Outpatients aged more than 20 years seeking care at PHC level and ER answered a standardized questionnaire and were subjected to spirometry, independently of their complaint. Two criteria for airways obstruction were taken into account: forced expiratory volume (FEV(1)) <80% of the predicted value and FEV(1)/forced vital capacity (FVC) ratio <0.70. RESULTS: A total of 274 individuals with a satisfactory spirometry were included. 22% of the individuals had a FEV(1) < 80%. Individuals older than 46 years had a higher risk of having airways obstruction. Asthma diagnosis (11%) had a clear association with airways obstruction. Smoking was a risk factor for a lower FEV(1). Working in a dust place and cooking using an open fire were both related to chronic bronchitis and asthma diagnosis. CONCLUSION: Under-report and underdiagnosis of chronic respiratory conditions seem to be a reality in CV just as in other parts of the world. To improve diagnosis, our results reinforce the need of performing a spirometry.

Survival of avian influenza and Newcastle disease viruses in compost and at ambient temperatures based on virus isolation and real-time reverse transcriptase PCR.

In four composting experiments, survival of avian influenza (AI) and Newcastle disease (ND) viruses was assessed by virus isolation in embryonated chicken eggs (ECEs) and by real-time reverse transcriptase-polymerase chain reaction. Specimens contained in nylon mesh bags consisted of 20-g samples of chicken manure, used litter, or feed that had been inoculated with allantoic fluid containing an AI virus (H6N2, Expt. 1) or an ND vaccine virus (Expt. 2). Other specimens consisted of 20-g samples of infected ECEs that had been homogenized and mixed with corn silage. As a control, allantoic fluid diluted in phosphate-buffered saline was contained in sealed vials. Except for the feed, in which the AI virus was inactivated soon after the specimen was inoculated, on day 0 the specimens buried in compost or placed outside at ambient temperatures contained at least 5.0 log10 of virus and 7.7 log10 of viral RNA. By day 7, temperatures in compost ranged from 50 C to 65 C, and viruses had been killed in all specimens in bags. In comparison, viruses in sealed vials remained viable to day 10. Viral RNA in mesh-bag specimens had been degraded to nondetectable levels by day 10, but it was still detected in sealed vials on day 21. In specimens that were held at ambient temperatures (13 C-28 C), the viruses in mesh-bag specimens were inactivated by day 21, but their RNA was still detected. In comparison, the viruses in sealed vials survived to day 21. In Expts. 3 and 4, viruses were inactivated in carcass specimens and in whole ECEs during composting. In an in vitro experiment, the time required for a 1-log10 reduction of viruses was significantly shorter (P < 0.05) in water extracts from compost than in phosphate buffers at temperatures of 25 C to 45 C. This study provided evidence that microbial activity during composting contributed to the rapid killing of AI and ND viruses and to the degradation of their viral RNA.

Estrous cycle regulates activation of hippocampal Akt, LIM kinase, and neurotrophin receptors in C57BL/6 mice.

Estradiol modulates dendritic spine morphology and synaptic protein expression in the rodent hippocampus, as well as hippocampal-dependent learning and memory. In the rat, these effects may be mediated through nongenomic steroid signaling such as estradiol activation of the Akt and LIM kinase (LIMK) pathways, in addition to genomic signaling involving estradiol upregulation of brain-derived neurotrophic factor expression (BDNF). Due to the many species differences between mice and rats, including differences in the hippocampal response to estradiol, it is unclear whether estradiol modulates these pathways in the mouse hippocampus. Therefore, we investigated whether endogenous fluctuations of gonadal steroids modulate hippocampal activation of the Akt, LIMK, and the BDNF receptor TrkB in conjunction with spatial memory in female C57BL/6 mice. We found that Akt, LIMK, and TrkB were activated throughout the dorsal hippocampal formation during the high-estradiol phase, proestrus. Cycle phase also modulated expression of the pre- and post-synaptic markers synaptophysin and post-synaptic density 95. However, cycle phase did not influence performance on an object placement test of spatial memory, although this task is known to be sensitive to the complete absence of ovarian hormones. The findings suggest that endogenous estradiol and progesterone produced by the ovaries modulate specific signaling pathways governing actin remodeling, cell excitability, and synapse formation.

Development of methods for detection and quantification of avian influenza and Newcastle disease viruses in compost by real-time reverse transcription polymerase chain reaction and virus isolation.

Composting has been used for disposal of poultry carcasses and manure following outbreaks caused by avian influenza virus (AIV) and Newcastle disease virus (NDV), but methods are needed to test for survival of the viruses in compost to ensure biosecurity. Methods developed in the present study include extracting viruses from compost and purifying viral RNA. The extracted viruses were detected by virus isolation using embryonated chicken eggs, and the purified RNA was detected by real-time reverse transcription PCR (RRT-PCR). The virus isolation and the RRT-PCR methods were evaluated with 3 compost preparations that were produced from chicken manure mixed with corn silage, wood shavings, or wheat straw. The detection limits of both methods were 1,700 and 1,000 embryo lethal doses of AIV and NDV per gram of compost, respectively. The copy number of viral RNA quantified by RRT-PCR was highly correlated with the amount of virus in compost. The results suggested that the RRT-PCR method may be used as an alternative to the virus isolation method for rapid detection and accurate quantification of AIV and NDV in compost.

Antibody response of chickens to serotype 1, 2, or 3 Marek's disease vaccines based on ELISA with infected cells as antigen.

An enzyme-linked immunosorbent assay (ELISA) was applied to evaluate the antibody response of commercial White Leghorn chickens to vaccination against Marek's disease (MD) at hatch (day 0) with serotype-1 (Rispens), -2 (SB-1), or -3 (turkey herpesvirus, HVT) vaccine virus and to challenge on day 21 with MD virus. Antigens for the test were whole chicken embryo fibroblast cells infected with Rispens, SB-1, or HVT. The chickens were progeny of stock that had been vaccinated with HVT, and on day 21 the nonvaccinated group had higher levels of maternal antibodies to HVT than to other antigens (P < 0.05). Only SB-1 vaccine had induced antibodies by day 21, and this was detected only against homologous antigens. On day 49, all three vaccines had induced higher levels of antibodies to homologous than to heterologous antigens. Marek's Disease virus (MDV) induced antibodies to all three antigens, but challenging vaccinated chicks did not significantly increase levels of antibodies on day 81 to any of the three antigens. It was concluded that an ELISA using whole cells as antigens would have potential value for monitoring the antibody response induced by MD vaccines and virulent MDV.

Economic analysis of dynamic management strategies utilizing. Transgenic corn for control of western corn rootworm (Coleoptera: Chrysomelidae).

We studied management strategies for western corn rootworm, Diabrotica virgifera virgifera LeConte, using transgenic corn, Zea mays L., from both a biological and an economic perspective. In areas with and without populations adapted to a 2-yr rotation of corn and soybean (rotation-resistant), the standard management strategy was to plant 80% of a cornfield (rotated and continuous) to a transgenic cultivar each year. In each area, we also studied dynamic management strategies where the proportion of transgenic corn increased over time in a region. We also analyzed management strategies for a single field that is the first to adopt transgenic corn within a larger unmanaged region. In all areas, increasing the expression of the toxin in the plant increased economic returns. In areas without rotation-resistance, planting 80% transgenic corn in the continuous cornfield each year generated the greatest returns with a medium toxin dose or greater. In areas with alleles for rotation-resistance at low initial levels, a 2-yr rotation of nontransgenic corn and soybean, Glycine max (L.) Merr., may be the most economical strategy if resistance to crop rotation is recessive. If resistance to crop rotation is additive or dominant, planting transgenic corn in the rotated cornfield was the most effective strategy. In areas where rotation-resistance is already a severe problem, planting transgenic corn in the rotated cornfield each year was always the most economical strategy. In some cases the strategies that increased the proportion of transgenic corn in the region over time increased returns compared with the standard strategies. With these strategies the evolution of resistance to crop rotation occurred more rapidly but resistance to transgenic corn was delayed compared with the standard management strategy. In areas not managed by a regional norm, increasing the proportion of transgenic corn and increasing toxin dose in the managed field generally increased returns. In a sensitivity analysis, among the parameters investigated, only density-dependent survival affected the results.

A continuous analyzer for monitoring respiratory gases and expired radioactivity in clinical studies.

A system for the continuous monitoring of gas exchange as well as 14CO2 from metabolic studies is described. The analyzer incorporates modifications over previously published systems. These modifications include accurate control of analyzer pressure for stable baselines during the long periods of measurement, accurate control of temperature, the measurement of oxygen consumption during high oxygen patient therapy, the measurement of oxygen consumption of patients on a mechanical respirator, and describes a unique flux system to calibrate the analyzer detectors. The closed system provides expired breath data for O2, CO2, and 14CO2 on critically ill patients for long periods of time without stressing the patient and without using a face mask or mouthpiece. These data allow for the calculation of daily energy expenditures of hypermetabolic patients and for the assessment of various metabolic responses with 14C substrates.

Detection of avian leukosis virus in albumen of chicken eggs using reverse transcription polymerase chain reaction.

A reverse transcriptase polymerase chain (RT-PCR) assay was developed to detect avian leukosis retrovirus (ALV) in egg albumen. Eggs of Single Comb White Leghorns were from a commercial breeder (stock F) and from a pathogen-free flock (stock N). RT-PCR was undertaken on isolated RNA from 20 unfertilized egg samples using seven sets of primers that correspond to the ALV gp85 envelope glycoprotein which determines the ALV subgroup classification. An ELISA assay for ALV gs antigen of egg albumen was positive for all stock F birds tested and negative for all stock N birds. Virus isolation was undertaken by inoculating egg albumen, feather pulp, or blood from five stock F chickens onto cultures of chicken embryo fibroblasts (C/E). IFA analysis of the inoculated C/E cultures indicated that all stock F birds tested contained infectious ALV. For the virus-positive stock F chickens, RT-PCR analyses using primers designed to detect all ALV subgroups detected ALV in 15/15 (100%) egg albumen samples, while primers designed to detect subgroup A ALV were positive for 12/15 (80%) egg albumen samples. RT-PCR products were not detected from five egg albumen samples from five stock N chickens by any primer sets. Direct sequencing using primers specific for subgroup A ALV verified the viral subgroup in the RT-PCR amplification products. The combined use of RT-PCR and direct sequencing of the RT-PCR product provides a new approach for identifying ALV-infected poultry.

Serum ionized calcium, serum and intracellular phosphate, and serum parathormone concentrations in acute malaria.

Previous studies have shown depressed serum corrected calcium and phosphate concentrations in acute falciparum malaria. To characterize malaria-associated disturbances in mineral homoeostasis further, serum ionized calcium and intracellular phosphate were measured in 18 patients (10 with falciparum malaria, 8 with vivax malaria) and 10 healthy controls. Six patients (4 falciparum, 2 vivax) had admission serum ionized calcium concentrations below the absolute control range (< 1.15 mmol/litre) and a further six (3 falciparum, 3 vivax) developed ionized hypocalcaemia during treatment. The patients with falciparum malaria had the lowest values at presentation (median [95% confidence intervals in brackets]: 1.17 [1.12-1.23] vs. 1.20 [1.18-1.24] mmol/litre in controls, P = 0.035) in the presence of depressed simultaneous serum parathormone concentrations (1.2 [0.6-1.9] vs. 1.6 [1.1-2.6] pmol/litre; P = 0.05). Admission serum phosphate concentrations were lower in the malaria patients (P = 0.007 vs. controls), especially in those with falciparum malaria (0.85 [0.7-1.1] vs. 1.2 [1.1-1.3] mmol/litre in controls; P = 0.002); patients with falciparum malaria also had significantly lower intracellular phosphate than controls (0.74 [0.58-0.90] vs. 0.88 [0.66-1.04] mmol/litre red cells; P = 0.047). There was a weak association between serum corrected and ionized calcium in the malaria patients (rs = 0.31, n = 18, P > 0.1), but serum and intracellular phosphate correlated significantly (rs = 0.71, n = 17, P < 0.001) with a regression line slope of 0.49 and intercept of 0.27 mmol/litre of red cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Relationship between egg size and subgroup J avian leukosis virus in eggs from broiler breeders.

Hatching eggs from three broiler breeder flocks that had experienced losses from myeloid leukosis were tested for infection with avian leukosis virus of subgroup J (ALV-J). Sufficient eggs were positive in two flocks to relate infection to egg weight. Allantoic fluid, embryonic tissue and yolk were collected after 18 days of incubation. The albumen and allantoic fluid were tested by enzyme-linked immunosorbent assay (ELISA) for group-specific (gs) antigen and all specimens were inoculated onto cell cultures to test for virus by immunofluorescence assay. Virus detected was identified as ALV-J by polymerase chain reaction techniques. The percentage of eggs that tested positive for gs antigen and virus was higher in those that weighed under 60 g than in heavier eggs (P < 0.01). In one flock, antibody to ALV-J was detected by ELISA in yolk from 14 and 43% of the eggs that tested positive or negative for virus, respectively. Testing the same eggs for antigen, virus and antibody should be useful for establishing the status of infection of the hens.

Platelet-activating factor and lipid metabolism in acute malaria.

Platelet-activating factor (PAF) contributes to a range of pathophysiological responses in severe illness. To examine PAF metabolism in acute malaria, venous blood was drawn from 10 untreated adults with falciparum malaria, from 8 with untreated vivax malaria, and from 10 controls. Plasma lyso-PAF, produced from PAF through the enzyme acetylhydrolase (AH), was bioassayed, after acetylation to PAF, by platelet [14C]-serotonin release. AH activity was measured by hydrolysis of [3H]-acetyl-PAF. Amounts of plasma lyso-PAF were lower in falciparum (median [range] 24 [9-221] ng/ml) and vivax (35 [7-236] ng/ml) infected patients than in controls (399 [212-504] ng/ml; P < 0.01), and correlated significantly with serum total and HDL-cholesterol (P < 0.001). Plasma AH activities were similar in the control, falciparum and vivax malaria groups. These data suggest that in malaria plasma lyso-PAF values fall together with other blood lipids, but independently of changes in AH activity. This may reflect a generalised decrease in lipid and phospholipid synthesis. However, the reduction of plasma lyso-PAF concentrations in our patients was much greater than that of serum lipoproteins. This is consistent with conversion of lyso-PAF to PAF or with increased PAF-receptor interactions. These two possibilities would have pathophysiological implications.

Chronic myocarditis and circulatory syndrome in a White Leghorn strain induced by an avian leukosis virus: light and electron microscopic study.

Specific-pathogen-free white leghorn chickens were inoculated at 1 day of age with avian leukosis virus (ALV, RAV-1). All chickens in Expt. 1, killed 33 or 64 days postinoculation, had focal chronic lymphocytic or lymphoplasmacytic myocarditis. Among those held beyond 33 days, eight of 22 developed lesions in the myocardium that resulted in a chronic circulatory syndrome (CCS) typical of right-sided heart failure. Chickens in Expt. 2 were held for 210 days, and 21% of 125 developed CCS. In Expt. 2, ALV particles were found by electron microscopy in myocardium of 100%, 72%, and 89% of inoculated chickens that developed CCS, lymphoid leukosis, or that had no gross lesions, respectively. These findings were in accord with the immunoperoxidase staining of tissue sections for group-specific antigen of ALV. In areas of extensive virus replication, there were often abnormal virus particles and also round bodies, which may have been remnants of host-cell membranes formed in the budding process. In contrast to findings in hearts, the spleens were usually negative for virus and viral antigen.

Cell-mediated and humoral immune responses in chickens infected with Salmonella typhimurium.

The cellular immune responses of leghorn and New Hampshire chickens following oral challenge at 4 weeks of age with Salmonella typhimurium were measured using assays for lymphocyte proliferative responses to mitogens and cytotoxic activity of natural killer (NK) cells. In addition, the humoral immune response to Newcastle disease virus (NDV) vaccine was assessed in S. typhimurium-infected chickens. At 8 and 20 days after infection with S. typhimurium, lymphocyte proliferative responses to various mitogens were significantly higher in S. typhimurium-infected chickens than in uninfected birds of the same breeds. Cytotoxic activity of NK cells was also significantly increased in infected birds on these days. Thirteen days after S. typhimurium infection, infected NDV-vaccinated chickens had higher antibody titers in response to NDV vaccination than uninfected vaccinated birds. NDV titers did not differ among groups at any other time of testing. These results show that both cellular and humoral immune functions are activated in the first 3 weeks following infection of 4-week-old chickens with S. typhimurium.

Distribution of lymphoid leukosis virus and p27 group-specific antigen in tissues from laying hens.

In order to gain insight into transmission and pathogenesis of infection, specimens from laying hens that had been naturally exposed to lymphoid leukosis virus (LLV) were tested for group-specific antigen (gsa) of the virus by immunofluorescence (IF), complement fixation (CF), and the enzyme-linked immunosorbant assay (ELISA). Electron microscopic examinations determined the distribution of C-type virus particles in tissues, and the phenotypic-mixing test served as a biological assay for exogenous LLV. The IF gsa was found in all organs tested, and fluorescence was usually found where virus particles were concentrated. In the oviduct and intestine, IF gsa was frequently at the border of the lumina and in the connective tissue associated with basal membranes of glands. In skin, the antigen was detected in smooth muscle, in feather pulp, and in basal epidermal cells of developing feathers. Results of various tests on Ottawa strains of chickens were usually in agreement. For example, among hens that shed gsa into egg albumen, only the viremic hens were consistently positive for IF gsa in both spleens and oviducts. Geometric mean CF titers of antigen were respectively five- and 23-fold higher in spleens and oviducts from viremic hens than in those from nonviremic hens. These findings suggest that the gsa was associated with exogenous virus infection. In Cornell S strain hens that had not been exposed to LLV, gsa was detected in splenic tissue by CF and ELISA but not by the IF test. This gsa was presumed to be of endogenous origin.

Studies on genetic and vaccination-induced resistance of chickens to lymphoid tumor transplants 2. Transmissible lymphoid tumor of Olson.

Transmissible lymphoid tumor (TLT) was inoculated in wing webs of five-week-old chickens of 6 strains. About half of the chickens of each strain had been vaccinated with turkey herpesvirus (HVT) one week before challenge in the wing web with TLT. Tumors which developed at the site of inoculation usually reached maximum size within 2 weeks and then regressed. In some chickens, however, tumors developed in visceral organs and caused death in the 2nd through 5th weeks postinoculation. Comparisons among strains of chickens in Expt. 1 revealed no differences in mortality. Vaccination with HVT reduced mortality and also the incidence of wing-web tumors (WWT) in all strains of chickens. A lymphoid leukosis virus and a Marek's disease (MD) virus of low virulence were detected in preparations of TLT, and it is suggested that the immunity induced by vaccination may have been directed against tumor antigens associated with MD virus.

Influence of Marek's disease virus strain AC-1 on cellular immunity in birds carrying endogenous viral genes.

The effects of Marek's disease virus (MDV) strain AC-1 on humoral and cellular immune responses were investigated in two lines of chickens segregating for the endogenous viral (ev) genes ev-6 and ev-12. All birds were vaccinated at 14 days of age against Newcastle disease virus (NDV). At 3 wk of age, 48 birds per line received an intraperitoneal injection of MDV (AC-1 isolate), and 24 were injected with saline. Birds of each group were killed on days 5, 7, 12, and 14 postinfection. Data were first analyzed for each day of testing. Results showed that, for variable measured, treatment effects were similar on days 5 and 7, and on days 12 and 14. Therefore, day 5 and day 7 data, and day 12 and day 14 data were pooled and analyzed. In MDV-infected chickens, proliferative lymphocyte responses to mitogens were suppressed (P < 0.001) after the first and second week of infection, whereas responses to NDV antigen were enhanced (P < 0.001) after the first week and then reduced (P < 0.01) by the end of the second week when compared to uninfected birds. The percentage of CD4+ T cells was higher (P < 0.01), and the percentage of cells expressing major histocompatibility complex (MHC) class II antigens was lower (P < 0.001), in MDV-infected chickens than in uninfected birds. The cytotoxic activity of natural killer (NK) cells was also enhanced (P < 0.001) in MDV-infected birds when compared to uninfected birds. Antibody responses to NDV were not different among groups, and the presence of ev-6 or ev-12 genes did not influence the immune response parameters measured in both infected and uninfected chickens. In conclusion, a marked increase in the CD4+ T lymphocyte population occurred in the early stage of MDV infection in all chickens regardless of the presence of ev genes, whereas the number of cells expressing MHC class II antigen was severely reduced.

Lymphoid leukosis: interrelations among virus infections in hens, eggs, embryos, and chicks.

Hens from a commercial source were selected because they were infected with lymphoid leukosis virus (LLV). LLV was detected in vaginal swabs from 17 viremic hens and from 27 of 44 hens that were not viremic. All hens that were positive on the vaginal swab test (VST) produced one or more eggs with virus in albumen or in embryos, whereas in comparable tests, virus was detected only in eggs from 5 of 17 hens that were negative on VST. Congenital transmission of LLV was erratic and neither the VST nor tests for virus in egg albumen prior to incubating eggs identified all hens that transmitted infection. For example, 14 hens negative on VST produced 50 eggs negative for virus in albumen and yet one of the embryos from these eggs was infected. Eggs from other hens had infectious virus in albumen and about half of the embryos from these were infected. Tests for virus in cloacal swabs from one-day-old chicks were as sensitive as tests on embryos for detecting congenital transmission. Titers of LLV in the meconium of congenitally infected chicks were as high as 10(7) infectious units per ml. The cloacal swab test should be a valuable adjunct to the VST and tests on egg albumen in programs designed to eradicate lymphoid leukosis from chickens.