The natural course of otitis media with effusion in infants who failed universal newborn hearing screening: a retrospective cohort study.

OBJECTIVES: To analyse the natural course of infants with otitis media with effusion who failed universal newborn hearing screening and to explore the appropriate observation period. METHODS: This retrospective cohort analysis included infants with otitis media with effusion who failed universal newborn hearing screening every 3 months for 12 months. RESULTS: The average recovery time of the 155 infants was 7.08 ± 0.32 months after diagnosis. Multivariate Cox regression analysis confirmed that frequent reflux, maxillofacial deformities and initial hearing status were independent factors affecting recovery. Moreover, the cumulative recovery of most infants with mild hearing loss and infants with moderate hearing loss accompanied by frequent reflux was significantly higher at six months after diagnosis than at three months. CONCLUSION: For most infants with mild hearing loss, as well as those with moderate hearing loss accompanied by frequent reflux, the observation period can be extended to six months after diagnosis.

Knowledge and management of WEEE by independent repairers in the city of Bouake in a context of health and environmental risk, 2022

Studies on waste management in Côte d'Ivoire are mostly focused on household and solid waste. It is appropriate to take a sustained look at the management of electrical and electronic equipment waste to better guide policies in this area. This work is a cross-sectional study covering the period from April 1, 2022 to June 30, 2022. The aim was to determine the knowledge and practices of the independent electrical and electronic repairers of the city of Bouaké in terms of electrical and electronic equipment waste (WEEE) management. A comprehensive sample of 307 people was interviewed by means of a questionnaire. The repairers interviewed had already heard of WEEE in 47.2% of cases. They had a good level of knowledge of WEEE in 85.5% of cases. In the final model in multivariate analysis, only the possession of a radio set at home had a significant influence on knowledge of WEEE. Repairers were unaware of the existence of D3E management legislation in 60.7% of cases; apart from humans (84.8%) who were most exposed to the dangers of inappropriate management of WEEE, other entities (air, soil, water, animals) would be polluted according to 36.6, 35.2, 4.1, and 3.4% of repairers, respectively. The most feared type of harm reported is injury (95.65%). Subjects dumped WEEE in a municipal landfill in 32.4% of cases. Sale, empowerment of pre-collection agents, and handing over to a recycling professional, cumulatively represent the most common practice (66.9%).

Knowledge and attitudes of health workers in bouake, Cote d'ivoire on zika virus disease in the context of a global epidemic

Since January, 2012, the Pacific region has faced a heavy burden of concurrent epidemics of dengue, chikungunya, and zika virus infections. In 2016, WHO developed a global response strategic framework to ensure that zika virus is a priority and accelerated area of public health research. This study conducted in Bouaké (Côte d'Ivoire) is part of this framework. The main objective was to assess the knowledge and attitudes of health workers working there on the zika virus disease in order to consider a better preparation and response to a possible epidemic in Côte d'Ivoire. Cross-sectional study covering the period from October 2016 to March 2017 was used here. The sampling was comprehensive and included interviews with 258 persons. Subjects were interviewed using a questionnaire edited and adapted from the CAP questionnaire developed by WHO in 2016. People with prior knowledge of the zika virus disease represented 66.3% of the health workers surveyed. Their level of knowledge was insufficient in 83.5% of cases. Their attitudes were good in 51.5% of cases. In the authors' final model, the exercise structure which was adjusted to the level of education and the corporation significantly influenced health workers' attitudes toward illness. Health workers in public settings appeared to have a better attitude compared to their private colleagues (adjusted OR = 4.88; CI: 2.37-10.03; p-value:0.000). The zika virus disease has attracted the attention of the medical community during the 2014-2016 period. This attention, while mitigated by the West African Ebola virus epidemic, deserves to be highlighted.

A resonance suppression method in platform style inertial reference unit via particle swarm optimization notch filter.

With the development of laser communication, remote sensing imaging, and other technologies, an inertial reference unit (IRU) plays an essential part in the line-of-sight (LOS) stabilization system used for acquiring, pointing, and tracking targets. The IRU provides a stable reference beam to realize accurate LOS pointing under external disturbances. Compared with the frame style IRU, the platform style IRU (PIRU) can achieve a higher bandwidth and better precision. However, mechanical resonance is introduced by a flexure hinge inevitably in the PIRU, which affects the performance of the LOS stabilization system. In this paper, an open-loop dynamic model of PIRU is established. Identification experiments are carried out with results indicating a 28.7 dB resonance peak at 27.07 Hz in the x axis and a 30.3 dB resonance peak at 26.59 Hz in the y axis. An asymmetric notch filter is used to suppress the resonance peak to achieve a higher control bandwidth. A fitness function is designed to represent the effect of resonance suppression. A particle swarm optimization algorithm is used to search for an optimal solution of the fitness function to obtain the parameters of the asymmetric notch filter. Experimental results show that the resonance peak is reduced by 97.88% and the system bandwidth reaches 159.31 Hz.

A tale of three kingdoms: members of the Phylum Nematoda independently acquired the detoxifying enzyme cyanase through horizontal gene transfer from plants and bacteria.

Horizontal gene transfer (HGT) has played an important role in the evolution of nematodes. Among candidate genes, cyanase, which is typically found only in plants, bacteria and fungi, is present in more than 35 members of the Phylum Nematoda, but absent from free-living and clade V organisms. Phylogenetic analyses showed that the cyanases of clade I organisms Trichinella spp., Trichuris spp. and Soboliphyme baturini (Subclass: Dorylaimia) represent a well-supported monophyletic clade with plant cyanases. In contrast, all cyanases found within the Subclass Chromadoria which encompasses filarioids, ascaridoids and strongyloids are homologous to those of bacteria. Western blots exhibited typical multimeric forms of the native molecule in protein extracts of Trichinella spiralis muscle larvae, where immunohistochemical staining localized the protein to the worm hypodermis and underlying muscle. Recombinant Trichinella cyanase was bioactive where gene transcription profiles support functional activity in vivo. Results suggest that: (1) independent HGT in parasitic nematodes originated from different Kingdoms; (2) cyanase acquired an active role in the biology of extant Trichinella; (3) acquisition occurred more than 400 million years ago (MYA), prior to the divergence of the Trichinellida and Dioctophymatida, and (4) early, free-living ancestors of the genus Trichinella had an association with terrestrial plants.

The Identification of Haemonchus Species and Diagnosis of Haemonchosis.

Diagnosis is often equated with identification or detection when discussing parasitic diseases. Unfortunately, these are not necessarily mutually exclusive activities; diseases and infections are generally diagnosed and organisms are identified. Diagnosis is commonly predicated upon some clinical signs; in an effort to determine the causative agent, identification of genera and species is subsequently performed. Both identification and diagnosis play critical roles in managing an infection, and involve the interplay of direct and indirect methods of detection, particularly in light of the complex and expanding problem of drug-resistance in parasites. Accurate and authoritative identification that is cost- and time-effective, based on structural and molecular attributes of specimens, provides a foundation for defining parasite diversity and changing patterns of geographical distribution, host association and emergence of disease. Most techniques developed thus far have been grounded in assumptions based on strict host associations between Haemonchus contortus and small ruminants, that is, sheep and goats, and between Haemonchus placei and bovids. Current research and increasing empirical evidence of natural infections in the field demonstrates that this assumption misrepresents the host associations for these species of Haemonchus. Furthermore, the capacity of H. contortus to utilize a considerably broad spectrum of ungulate hosts is reflected in our understanding of the role of anthropogenic forcing, the 'breakdown' of ecological isolation, global introduction and host switching as determinants of distribution. Nuanced insights about distribution, host association and epidemiology have emerged over the past 30years, coincidently with the development of increasingly robust means for parasite identification. In this review and for the sake of argument, we would like to delineate the diagnosis of haemonchosis from the identification of the specific pathogen. As a foundation for exploring host and parasite biology, we will examine the evolution of methods for distinguishing H. contortus from other common gastrointestinal nematodes of agriculturally significant and free-ranging wild ruminants using morphological, molecular and/or immunological methods for studies at the species and genus levels.

Characterization of the Eimeria maxima sporozoite surface protein IMP1.

The purpose of this study was to characterize Eimeria maxima immune-mapped protein 1 (IMP1) that is hypothesized to play a role in eliciting protective immunity against E. maxima infection in chickens. RT-PCR analysis of RNA from unsporulated and sporulating E. maxima oocysts revealed highest transcription levels at 6-12h of sporulation with a considerable downregulation thereafter. Alignment of IMP1 coding sequence from Houghton, Weybridge, and APU-1 strains of E. maxima revealed single nucleotide polymorphisms that in some instances led to amino acid changes in the encoded protein sequence. The E. maxima (APU-1) IMP1 cDNA sequence was cloned and expressed in 2 different polyHis Escherichia coli expression vectors. Regardless of expression vector, recombinant E. maxima IMP1 (rEmaxIMP1) was fairly unstable in non-denaturing buffer, which is consistent with stability analysis of the primary amino acid sequence. Antisera specific for rEmaxIMP1 identified a single 72 kDa protein or a 61 kDa protein by non-reducing or reducing SDS-PAGE/immunoblotting. Immunofluorescence staining with anti-rEmaxIMP1, revealed intense surface staining of E. maxima sporozoites, with negligible staining of merozoite stages. Immuno-histochemical staining of E. maxima-infected chicken intestinal tissue revealed staining of E. maxima developmental stages in the lamnia propia and crypts at both 24 and 48 h post-infection, and negligible staining thereafter. The expression of IMP1 during early stages of in vivo development and its location on the sporozoite surface may explain in part the immunoprotective effect of this protein against E. maxima infection.

Inhibition of CD23-mediated IgE transcytosis suppresses the initiation and development of allergic airway inflammation.

The epithelial lining of the airway tract and allergen-specific IgE are considered essential controllers of inflammatory responses to allergens. The human low affinity IgE receptor, CD23 (FcɛRII), is capable of transporting IgE or IgE-allergen complexes across the polarized human airway epithelial cell (AEC) monolayer in vitro. However, it remains unknown whether the CD23-dependent IgE transfer pathway in AECs initiates and facilitates allergic inflammation in vivo, and whether inhibition of this pathway attenuates allergic inflammation. To this end, we show that in wild-type (WT) mice, epithelial CD23 transcytosed both IgE and ovalbumin (OVA)-IgE complexes across the airway epithelial barrier, whereas neither type of transcytosis was observed in CD23 knockout (KO) mice. In chimeric mice, OVA sensitization and aerosol challenge of WT/WT (bone-marrow transfer from the WT to WT) or CD23KO/WT (CD23KO to WT) chimeric mice, which express CD23 on radioresistant airway structural cells (mainly epithelial cells) resulted in airway eosinophilia, including collagen deposition and a significant increase in goblet cells, and increased airway hyperreactivity. In contrast, the absence of CD23 expression on airway structural or epithelial cells, but not on hematopoietic cells, in WT/CD23KO (the WT to CD23KO) chimeric mice significantly reduced OVA-driven allergic airway inflammation. In addition, inhalation of the CD23-blocking B3B4 antibody in sensitized WT mice before or during airway challenge suppressed the salient features of asthma, including bronchial hyperreactivity. Taken together, these results identify a previously unproven mechanism in which epithelial CD23 plays a central role in the development of allergic inflammation. Further, our study suggests that functional inhibition of CD23 in the airway is a potential therapeutic approach to inhibit the development of asthma.

Detection and survival of Toxoplasma gondii in milk and cheese from experimentally infected goats.

The consumption of unpasteurized goat cheese and goat's milk has been suggested as a risk factor for toxoplasmosis in humans. In the present study, detection and survival of Toxoplasma gondii in milk and cheese was studied by bioassay in mice (milk) and in cats (cheese). Eight goats were inoculated orally with 300 to 10,000 oocysts of T. gondii strain TgGoatUS26. Milk samples were collected daily up to 30 days postinoculation and bioassayed in mice and cats. For mouse bioassay, 50 ml of milk samples were centrifuged, and the sediment was inoculated subcutaneously into mice. Mice were tested for T. gondii infection by seroconversion and by the demonstration of parasites. By mouse bioassay, T. gondii was detected in milk from all eight goats. The T. gondii excretion in milk was intermittent. For cat bioassay, 400 ml (100 ml or more from each goat) of milk from four goats from 6 to 27 days postinoculation were pooled daily, and cheese was made using rennin. Ten grams of cheese was fed daily to four cats, and cat feces were examined for oocyst shedding. One cat fed cheese shed oocysts 7 to 11 days after consuming cheese. Attempts were made to detect T. gondii DNA in milk of four goats; T. gondii was detected by PCR more consistently, but there was no correlation between detection of viable T. gondii by bioassay in mice and T. gondii DNA by PCR. Results indicate that T. gondii can be excreted in goat's milk and can survive in fresh cheese made by cold-enzyme treatment. To prevent transmission to humans or animals, milk should not be consumed raw. Raw fresh goat cheese made by cold-enzyme treatment of unpasteurized milk also should not be consumed.

Fetal death in cows experimentally infected with Neospora caninum at 110 days of gestation.

Neospora caninum is a major cause of abortion in cattle, but the reasons why some animals abort and not others remain unclear. Most of the N. caninum experimental primary infections in cattle late in gestation, after 120 days of pregnancy, result in birth of full-term congenitally infected fetuses. In the present study, the distribution of parasites and pathogenesis of infection in both dams and fetuses after inoculation with 10(7) culture derived tachyzoites of N. caninum NC-Illinois cattle strain at 110 days of gestation were analyzed at 3 weeks, 6 weeks and 9 weeks after infection (WAI) in eight Angus heifers. One dam from the group euthanized at 6 WAI had a dead fetus at necropsy. Extensive lesions were observed in the placenta and tachyzoites were detected in both the placenta and the fetus. The fetus was seropositive and had high IFN-gamma g production in fetal fluids. Another fetus, still alive when euthanized at 3 WAI, had severe lesions and high IFN-gamma production and a similar fate could have been expected if the experimental period would have been longer. Lesions in the placenta of the remaining six dams that had live fetuses at necropsy were mild. In those dams, the fetal and maternal placentas had not separated and contained focal areas of placentitis at the materno-fetal junction. Transplacental infection took place on all fetuses based on detection of parasitic DNA in fetal tissues. The present study shows that experimental N. caninum infection of naïve dams after 110 days of pregnancy can lead to fetal death. The results suggest that the severity of placental lesions and the strong IFN-gamma response in some fetuses, possibly as part of the immune response trying to control the high parasitemia, might, in fact, be the cause of their death.

Neosporosis in Beagle dogs: clinical signs, diagnosis, treatment, isolation and genetic characterization of Neospora caninum.

Clinical neosporosis was diagnosed in a litter of five pups born to a Beagle bitch from Virginia, USA. Four of the pups developed limb weakness starting at 4 weeks of age. The dogs were suspected to have neosporosis based on clinical signs and empirically treated with Clindamycin (75 mg, oral, twice daily, total 150 mg) starting at 9 weeks of age and the dosage was doubled at 13 weeks of age. Antibodies to Neospora caninum were detected in sera of the dam and pups when first tested serologically at the age of 4 months. The owner donated the pup with the worst clinical signs and the dam for research; both dogs were euthanized. Viable N. caninum was isolated in gamma interferon gene knock out (KO) mice and in cell culture from the pup killed at 137 days of age. Tissue cysts, but no tachyzoites, were found in histological sections of brain and muscles. The isolate was also identified as N. caninum by PCR and sequence analysis and designated NC-9. N. caninum was neither isolated by bioassay in KO mice nor found in histological sections of tissues of the bitch. Clinical signs in the remaining three pups improved considerably after a 6-month treatment with Clindamycin; N. caninum antibody titers were still persistent in these pups at 23 months of age. Results indicate that medication with Clindamycin can improve clinical condition but not eliminate N. caninum infection.

Establishment of Neospora caninum antigen-specific T cell lines of primarily CD4 T cells.

Neosporosis is an important cause of pregnancy loss in cattle worldwide. Protective immunity against Neospora caninum infection may include both cell-mediated (CMI) and humoral immune responses. This study was to establish short-term antigen-specific T cell lines composed of primarily CD4(+)T cells from peripheral blood lymphocytes (PBL) of infected cows, which may be used to identify immunodominant antigens for the development of N. caninum vaccines. Crude N. caninum tachyzoite antigen was prepared from in vitro derived N. caninum tachyzoites. Multiple T cell lines were established and maintained for 11 weeks by weekly re-stimulation with N. caninum antigen and antigen-presenting cells. All cell lines responded highly to antigen between weeks 3 and 11. Phenotypically, these cells were composed primarily of CD4(+)T cells between weeks 2-8, with a gradual expansion of gamma/delta(+)T cells thereafter. The results indicate that N. caninum-specific T cell lines can be established and maintained without exogenous T cell growth factors and may be used to identify N. caninum antigens. This research will enhance our understanding of bovine CMI to neosporosis and may facilitate development of a proven neosporosis vaccine.

Age-related innate immune response in calves to Babesia bovis involves IL-12 induction and IL-10 modulation.

There is a strong innate immunity in calves to infection with Babesia bovis. Interleukin (IL)-12 and IL-10 have been shown in vitro to be important immunoregulatory cytokines. Here we demonstrate in vivo that the protective innate response in young calves to infection with virulent B. bovis involves the early appearance of IL-12 and interferon-gamma (IFN-gamma) transcripts in the spleen. In contrast, IL-12 and IFN-gamma mRNA expression in the spleens of adult cattle that succumbed to the infection was delayed and depressed and occurred within the context of IL-10 expression. Also in contrast with calves, there was no detectable antibody response before death in adults. A vigorous CD8+ T-cell expansion occurred in the spleens of both calves and adults.

The age-related immunity in cattle to Babesia bovis infection involves the rapid induction of interleukin-12, interferon-gamma and inducible nitric oxide synthase mRNA expression in the spleen.

Young calves possess a strong innate immunity against Babesia bovis infection that lasts for approximately 6 months after birth and is abrogated with the removal of the spleen. This immunity is characterized as cellular involving a soluble mediator. Nitric oxide has been implicated by virtue of its babesiacidal affects in vitro, but questioned to be as effective in vivo, due to its ability to downregulate type-1 immunity. Spleen cells were obtained from 4-month-old calves and adult steers and processed for monitoring cytokine and inducible nitric oxide synthase (iNOS) mRNA expression during the response to initial B. bovis infection. The data provided evidence of a transient role for nitric oxide in innate immunity, characterized by brief iNOS induction in the spleen of calves that was not detectable in the spleens of adults. The iNOS message followed the early induction of interleukin (IL)-12 and interferon (IFN)-gamma message in calves. The induction of IL-12 and IFN-gamma message in adults was delayed until IL-10 message was induced. Transformation growth factor-beta mRNA expression levels were greater in spleen cells from adults early in infection and then declined, whereas expression levels increased in spleen cells from calves later in the infection process. Together, the data support the concept of 'first come, first serve' cytokine influence over cellular activities, the importance of a type-1 response in the control of an initial infection and the need for tight regulation in order to prevent pathology associated with over production of nitric oxide and inflammatory cytokines.

Prp-c and Prp-Sc at the fetal-maternal interface.

Scrapie is a naturally occurring prion (PrP) disease causing a fatal neurodegenerative disorder in sheep and goats. Previous studies suggest that scrapie is transmitted naturally through exposure to the scrapie agent in wasted placentas of infected ewes. This study determined the distribution and biochemical properties of PrP cellular (PrP-C) and the distribution of PrP scrapie (PrP-Sc) in reproductive, placental, and selected fetal tissues and fetal fluids in sheep. Glycosylated, N-terminally truncated, proteinase K-sensitive PrP-C with apparent molecular masses of 23-37 kDa was present in reproductive, placental, and fetal tissues and fetal fluids. PrP-C was low or undetectable in intercotyledonary chorioallantois, amnion, urachus, amniotic fluid, and fetal urine. In pregnant ewes, cotyledonary chorioallantois, allantoic fluid, and caruncular endometrium contained higher levels of PrP-C than did intercaruncular endometrium, myometrium, oviduct, ovary, fetal bladder, or fetal kidney. Caruncular endometrial PrP-C was up-regulated during pregnancy. Despite the wide distribution of PrP-C in reproductive, placental, and selected fetal tissues and fetal fluid, PrP-Sc was detected only in caruncular endometrium and cotyledonary chorioallantois of pregnant scrapie-infected ewes. The embryo/fetus may not be exposed to scrapie in utero because it is separated physically from PrP-positive allantois and chorioallantois by PrP-negative amnion.

Interleukin-12 as an adjuvant promotes immunoglobulin G and type 1 cytokine recall responses to major surface protein 2 of the ehrlichial pathogen Anaplasma marginale.

Anaplasma marginale is a tick-transmitted pathogen of cattle closely related to the human ehrlichiae, Ehrlichia chaffeensis and the agent of human granulocytic ehrlichiosis (HGE). These pathogens have in common a structurally conserved outer membrane protein (OMP) designated the major surface protein 2 (MSP-2) in A. marginale and HGE and OMP-1 in E. chaffeensis. Protective immunity against ehrlichial pathogens is believed to require induction of gamma interferon (IFN-gamma) and opsonizing immunoglobulin (Ig) subclasses directed against OMP epitopes that, in concert, activate macrophages for phagocytosis and killing. Because interleukin-12 (IL-12) acts as an adjuvant for protein immunization to induce IFN-gamma and protective immunity against intracellular pathogens, we hypothesized that as an adjuvant with MSP-2, IL-12 would augment type 1 recall responses to A. marginale. IL-12 was coadsorbed with MSP-2 to alum and shown to significantly enhance IFN-gamma production by lymph node cells (LNC) and LNC-derived CD4(+) T-cell lines from immunized calves following recall stimulation with A. marginale. LNC proliferation and IL-2 production were also enhanced in IL-12-treated calves. Elevated recall proliferative responses by peripheral blood mononuclear cells were still evident 9 months after immunization. Serum IgG levels were consistently increased in IL-12 immunized calves, predominantly due to higher IgG1 responses. The results support the use of IL-12 coadsorbed with OMP of ehrlichial pathogens in alum to amplify both antibody and type-1 cytokine responses important for protective immunity.

Comparative effects of interleukin-12 and interleukin-4 on cytokine responses by antigen-stimulated memory CD4+ T cells of cattle: IL-12 enhances IFN-gamma production, whereas IL-4 has marginal effects on cytokine expression.

Interleukin-12 (IL-12) and IL-4 are important immunoregulatory cytokines that determine the fate of naive T cells during antigen priming in mice and also influence cytokine synthesis by differentiated murine and human T cells. The roles of these cytokines in regulating the differentiation and effector function of bovine T cells are less well studied. We investigated the ability of human IL-12 and bovine IL-4 to modify cytokine expression by antigen-stimulated T cells from cattle immune to the protozoal parasites Babesia bovis and Babesia bigemina or reactive with Mycobacterium bovis purified protein derivative. Peripheral blood mononuclear cells (PBMC) were cultured with specific antigen and IL-4 or IL-12 for 1 week. Then viable lymphoblasts consisting of predominantly CD4+ T cells were restimulated with antigen and antigen-presenting cells (APC) with or without cytokine. Cell lines were cultured for several weeks, and following restimulation with antigen and APC in the absence of exogenous cytokine, the cell lines were analyzed for proliferation, interferon-gamma (IFN-gamma) production, and expression of IL-2, IL4-, IL-10, or IFN-gamma transcript levels using a quantitative competitive RT-PCR. IL-12 and IL-4 had no effect on the composition of CD4, CD8, or gammadelta T cells in the cell lines or on the level of antigen-induced proliferation. IL-12 stimulated enhanced levels of IFN-gamma protein and transcript expression in all cell lines, with no consistent effects on IL-2 or IL-4 expression. In two B. bovis-specific cell lines, IL-12 suppressed IL-10 expression. IL-4 had no consistent effect on expression of any cytokine. These results indicate the use of IL-12 as an adjuvant to enhance type 1 cytokine responses in cattle during antigen priming.

Upregulation of interleukin-4 and IFN-gamma expression by IFN-tau, a member of the type I IFN family.

Trophoblast interferon-tau (IFN-tau) is a new member of the type I IFN family that is produced in large quantities by the ruminant conceptus. Like other type I IFN, IFN-tau inhibits viral replication and activates natural killer (NK)-mediated cytotoxicity. In mice and humans, type I IFN enhances type 1 T helper (Th) cell responses, but the effects of type I IFN, including IFN-tau, on cytokine expression by bovine Th cells have not been described. The present study determined the effects of IFN-tau on interleukin-4 (IL-4), IFN-gamma, and IL-10 expression by antigen-specific, CD4+ T cell lines derived from cattle immune to either Babesia bovis, Babesia bigemina rhoptry-associated protein-1, or Anaplasma marginale. IFN-tau upregulated IFN-gamma secretion and steady-state levels of IFN-gamma and IL-4 mRNA by cell lines cultured for 3-6 weeks. In contrast, the steady-state levels of IL-10 mRNA were either not changed or inhibited at these times. Similar effects were obtained with human IFN-alpha. Comparison of the quantities of IFN-gamma, IL-4, and IL-10 transcripts in IFN-tau-treated or IFN-alpha-treated cultures revealed that even though IFN-gamma was the predominant cytokine expressed by all T cell lines, both IFN-gamma and IL-4 steady-state transcript levels were upregulated by a comparable degree. Thus, these studies demonstrate that IFN-tau is an immunomodulatory cytokine that promotes enhanced IL-4 and IFN-gamma responses by effector T cells but not, strictly speaking, Thl-biased responses in cattle. These results indicate the potential use of this cytokine as an adjuvant in ruminants to boost cell-mediated immune responses.

Intrauterine infusion of bacterial lipopolysaccharide (LPS) prior to mating has no adverse effect on fertility, fetal survival and fetal development.

Lipopolysaccharide (LPS, endotoxin) is a component of the cell wall of gram-negative bacteria and a potent inducer of severe inflammatory reactions. In mice, systemically administered LPS induces fetal resorption and increases fetal mortality. However, effects of intrauterine LPS on fertility, fetal survival and development have not been reported. In the present study, pigs were used to determine the effect of intrauterine infused LPS on fertility, fetal survival and development. Prior to mating, gilts received intrauterine infusion of either a single dose of saline or increasing doses of LPS in saline using an insemination catheter. On day 30 of pregnancy, gilts were hysterectomized and litter size, fetal length, number of corpora lutea (CL), ovarian and placental weights, and allantoic and amniotic fluid volumes were recorded. Blood progesterone levels from days 10-30 of pregnancy were also determined. Results indicated that intrauterine infusion of LPS had no adverse effects on blood progesterone levels, fertility, fetal survival or fetal development. Intrauterine injection of LPS did cause an increase in fetal weight and amniotic fluid volume (P < 0.05). These results suggest that sperm, oocytes and gametes are tolerant of local LPS challenge and, to some extent, this mechanism protects gametes and conceptuses from maternal response to mating introduced bacteria and their potential endotoxins.

Babesia bovis: common protein fractions recognized by oligoclonal B. bovis-specific CD4+ T cell lines from genetically diverse cattle.

CD4+ helper T cells are believed to be important for inducing protective immunity against Babesia bovis through the production of cytokines, including IFN-gamma, that will provide help to B lymphocytes for IgG production and activate macrophages to become parasiticidal. To provide maximum protection in an outbred population, an effective vaccine against B. bovis should contain antigens that would elicit an IFN-gamma response and would be recognized by cattle with diverse genetic backgrounds. To identify potentially protective "universal" T helper (Th) cell antigens, fractions of homogenized B. bovis merozoites were tested for the ability to stimulate proliferation of oligoclonal CD4+, IFN-gamma-producing T cell lines derived from four immune animals previously shown to differ in major histocompatibility complex class II expression. Homogenized B. bovis merozoites were separated by denaturing continuous flow electrophoresis (CFE) on 15, 10, and 7.5% polyacrylamide gels into fractions containing proteins ranging from <14.5 to approximately 95 kDa. Eighteen of 280 CFE fractions elicited anamnestic proliferative responses in all T cell lines tested. Nine of these cross-stimulatory fractions contained proteins of <14.5 to 24.5 kDa, and the remaining ones contained proteins with estimated molecular weights of 30, 31.5, 44.5, 49, 49.5, 54, 62, 72, and 82 kDa. Immunoblot analysis showed that four cross-stimulatory fractions contained a predicted known B. bovis antigen of similar molecular size. Previous studies had demonstrated that fractionated merozoite proteins stimulatory for CD4+ Th cell clones had apparent molecular weights similar to those present in 7 of the 18 stimulatory fractions. In the present study, two Th cell clones responded to cross-stimulatory CFE fractions, underscoring the potential to use both oligoclonal and monoclonal Th cell lines to identify commonly recognized polypeptides as potential vaccine antigens.