Coxiella burnetii Infections in Small Ruminants and Humans in Switzerland

The recent Q fever epidemic in the Netherlands raised concerns about the potential risk of outbreaks in other European countries. In Switzerland, the prevalence of Q fever in animals and humans has not been studied in recent years. In this study, we describe the current situation with respect to Coxiella (C.) burnetii infections in small ruminants and humans in Switzerland, as a basis for future epidemiological investigations and public health risk assessments. Specific objectives of this cross-sectional study were to (i) estimate the seroprevalence of C. burnetii in sheep and goats, (ii) quantify the amount of bacteria shed during abortion and (iii) analyse temporal trends in human C. burnetii infections. The seroprevalence of C. burnetii in small ruminants was determined by commercial ELISA from a representative sample of 100 sheep flocks and 72 goat herds. Herd-level seroprevalence was 5.0% (95% CI: 1.6-11.3) for sheep and 11.1% (95% CI: 4.9-20.7) for goats. Animal-level seroprevalence was 1.8% (95% CI: 0.8-3.4) for sheep and 3.4% (95% CI: 1.7-6) for goats. The quantification of C. burnetii in 97 ovine and caprine abortion samples by real-time PCR indicated shedding of >10(4) bacteria/g in 13.4% of all samples tested. To our knowledge, this is the first study reporting C. burnetii quantities in a large number of small ruminant abortion samples. Annual human Q fever serology data were provided by five major Swiss laboratories. Overall, seroprevalence in humans ranged between 1.7% and 3.5% from 2007 to 2011, and no temporal trends were observed. Interestingly, the two laboratories with significantly higher seroprevalences are located in the regions with the largest goat populations as well as, for one laboratory, with the highest livestock density in Switzerland. However, a direct link between animal and human infection data could not be established in this study.

Producción ovina extensiva en la Patagonia Austral : El caso de la zona centro de Santa Cruz

El propósito del artículo consiste en analizar la meseta central de Santa Cruz en sus aspectos socioproductivos. El objetivo de la investigación, de la cual este trabajo es una elaboración parcial, se orienta a establecer la viabilidad del modelo vigente en la región -ovino extensivo-, así como las posibilidades de que las prácticas asociativas se constituyan en el eje articulador de un relanzamiento productivo. Aquí presentamos un primer análisis socioeconómico y productivo con la finalidad de definir el escenario en el que actualmente se lleva adelante la producción ovina extensiva. Incluimos una descripción de los nuevos actores que actualmente operan en el centro-norte de la provincia y, especialmente en nuestra área de estudio, destacándose entre ellos las empresas mineras transnacionales.

Análisis y pautas para el mejoramiento socioeconómico de los productores del oeste pampeano

En la zona comprendida entre las isoyetas de 500 mm a 300 mm, perteneciente a la provincia fitogeográfica del Monte, una población rural de 6.845 habitantes, 9 del total provincial, con una densidad de 0,2 hab /km2, habita los cinco departamentos del centro y extremo oeste de La Pampa, cuya superficie abarca el 38 de la provincia. Su economía de subsistencia se desarrolla sobre campos naturales, con una ganadería de bovinos, caprinos, ovinos y equinos. La población presenta un alto grado de dispersión y un significativo aislamiento respecto de los centros más poblados. El objetivo a cumplir, modificar las condiciones de vida de los "crianceros", hace necesaria la aplicación de pautas que permitan convertir estas economías de subsistencia en pequeños emprendimientos ganaderos con aceptables índices de rentabilidad mediante el uso más eficiente del medio y sus recursos. Esta estructura productiva ha de permitir una relación más solidaria entre los ganaderos, una mayor libertad en la toma de decisiones y una mayor independencia económica. La consolidación estable de este proyecto y el logro de sus objetivos redundarán en un mejor manejo del ecosistema, así como en un marcado mejoramiento de la calidad de vida de estos alejados productores rurales.

Producción ovina extensiva en la Patagonia Austral : El caso de la zona centro de Santa Cruz

El propósito del artículo consiste en analizar la meseta central de Santa Cruz en sus aspectos socioproductivos. El objetivo de la investigación, de la cual este trabajo es una elaboración parcial, se orienta a establecer la viabilidad del modelo vigente en la región -ovino extensivo-, así como las posibilidades de que las prácticas asociativas se constituyan en el eje articulador de un relanzamiento productivo. Aquí presentamos un primer análisis socioeconómico y productivo con la finalidad de definir el escenario en el que actualmente se lleva adelante la producción ovina extensiva. Incluimos una descripción de los nuevos actores que actualmente operan en el centro-norte de la provincia y, especialmente en nuestra área de estudio, destacándose entre ellos las empresas mineras transnacionales.

Análisis y pautas para el mejoramiento socioeconómico de los productores del oeste pampeano

En la zona comprendida entre las isoyetas de 500 mm a 300 mm, perteneciente a la provincia fitogeográfica del Monte, una población rural de 6.845 habitantes, 9 del total provincial, con una densidad de 0,2 hab /km2, habita los cinco departamentos del centro y extremo oeste de La Pampa, cuya superficie abarca el 38 de la provincia. Su economía de subsistencia se desarrolla sobre campos naturales, con una ganadería de bovinos, caprinos, ovinos y equinos. La población presenta un alto grado de dispersión y un significativo aislamiento respecto de los centros más poblados. El objetivo a cumplir, modificar las condiciones de vida de los "crianceros", hace necesaria la aplicación de pautas que permitan convertir estas economías de subsistencia en pequeños emprendimientos ganaderos con aceptables índices de rentabilidad mediante el uso más eficiente del medio y sus recursos. Esta estructura productiva ha de permitir una relación más solidaria entre los ganaderos, una mayor libertad en la toma de decisiones y una mayor independencia económica. La consolidación estable de este proyecto y el logro de sus objetivos redundarán en un mejor manejo del ecosistema, así como en un marcado mejoramiento de la calidad de vida de estos alejados productores rurales.

Producción ovina extensiva en la Patagonia Austral : El caso de la zona centro de Santa Cruz

El propósito del artículo consiste en analizar la meseta central de Santa Cruz en sus aspectos socioproductivos. El objetivo de la investigación, de la cual este trabajo es una elaboración parcial, se orienta a establecer la viabilidad del modelo vigente en la región -ovino extensivo-, así como las posibilidades de que las prácticas asociativas se constituyan en el eje articulador de un relanzamiento productivo. Aquí presentamos un primer análisis socioeconómico y productivo con la finalidad de definir el escenario en el que actualmente se lleva adelante la producción ovina extensiva. Incluimos una descripción de los nuevos actores que actualmente operan en el centro-norte de la provincia y, especialmente en nuestra área de estudio, destacándose entre ellos las empresas mineras transnacionales.

Análisis y pautas para el mejoramiento socioeconómico de los productores del oeste pampeano

En la zona comprendida entre las isoyetas de 500 mm a 300 mm, perteneciente a la provincia fitogeográfica del Monte, una población rural de 6.845 habitantes, 9 del total provincial, con una densidad de 0,2 hab /km2, habita los cinco departamentos del centro y extremo oeste de La Pampa, cuya superficie abarca el 38 de la provincia. Su economía de subsistencia se desarrolla sobre campos naturales, con una ganadería de bovinos, caprinos, ovinos y equinos. La población presenta un alto grado de dispersión y un significativo aislamiento respecto de los centros más poblados. El objetivo a cumplir, modificar las condiciones de vida de los "crianceros", hace necesaria la aplicación de pautas que permitan convertir estas economías de subsistencia en pequeños emprendimientos ganaderos con aceptables índices de rentabilidad mediante el uso más eficiente del medio y sus recursos. Esta estructura productiva ha de permitir una relación más solidaria entre los ganaderos, una mayor libertad en la toma de decisiones y una mayor independencia económica. La consolidación estable de este proyecto y el logro de sus objetivos redundarán en un mejor manejo del ecosistema, así como en un marcado mejoramiento de la calidad de vida de estos alejados productores rurales.

Antibody-mediated inhibition of the growth of larvae from an insect causing cutaneous myiasis in a mammalian host

Many insects feed on blood or tissue from mammalian hosts. One potential strategy for the control of these insects is to vaccinate the host with antigens derived from the insect. The larvae of the fly Lucilia cuprina feed on ovine tissue and tissue fluids causing a cutaneous myiasis associated with considerable host morbidity and mortality. A candidate vaccine antigen, peritrophin 95, was purified from the peritrophic membrane, which lines the gut of these larvae. Serum from sheep vaccinated with peritrophin 95 inhibited growth of first-instar L. cuprina larvae that fed on this serum. Growth inhibition was probably caused by antibody-mediated blockage of the normally semipermeable peritrophic membrane and the subsequent development of an impervious layer of undefined composition on the gut lumen side of the peritrophic membrane that restricted access of nutrients to the larvae. The amino acid sequence of peritrophin 95 was determined by cloning the DNA complementary to its mRNA. The deduced amino acid sequence codes for a secreted protein containing a distinct Cys-rich domain of 317 amino acids followed by a mucin-like domain of 139 amino acids. The Cys-rich domain may be involved in binding chitin. This report describes a novel immunological strategy for the potential control of L. cuprina larvae that may have general application to the control of other insect pests.

The diversity and evolutionary relationships of the pregnancy-associated glycoproteins, an aspartic proteinase subfamily consisting of many trophoblast-expressed genes

The pregnancy-associated glycoproteins (PAGs) are structurally related to the pepsins, thought to be restricted to the hooved (ungulate) mammals and characterized by being expressed specifically in the outer epithelial cell layer (chorion/trophectoderm) of the placenta. At least some PAGs are catalytically inactive as proteinases, although each appears to possess a cleft capable of binding peptides. By cloning expressed genes from ovine and bovine placental cDNA libraries, by Southern genomic blotting, by screening genomic libraries, and by using PCR to amplify portions of PAG genes from genomic DNA, we estimate that cattle, sheep, and most probably all ruminant Artiodactyla possess many, possibly 100 or more, PAG genes, many of which are placentally expressed. The PAGs are highly diverse in sequence, with regions of hypervariability confined largely to surface-exposed loops. Nonsynonymous (replacement) mutations in the regions of the genes coding for these hypervariable loop segments have accumulated at a higher rate than synonymous (silent) mutations. Construction of distance phylograms, based on comparisons of PAG and related aspartic proteinase amino acid sequences, suggests that much diversification of the PAG genes occurred after the divergence of the Artiodactyla and Perissodactyla, but that at least one gene is represented outside the hooved species. The results also suggest that positive selection of duplicated genes has acted to provide considerable functional diversity among the PAGs, whose presence at the interface between the placenta and endometrium and in the maternal circulation indicates involvement in fetal–maternal interactions.

Prolactin is not a juvenile hormone in Xenopus laevis metamorphosis

Prolactin (PRL) is widely considered to be the juvenile hormone of anuran tadpoles and to counteract the effects of thyroid hormone (TH), the hormone that controls amphibian metamorphosis. This putative function was concluded mainly from experiments in which mammalian PRL was injected into tadpoles or added to cultured tadpole tissues. In this study, we show that overexpression of ovine or Xenopus laevis PRL in transgenic X. laevis does not prolong tadpole life, establishing that PRL does not play a role in the life cycle of amphibians that is equivalent to that of juvenile hormone in insect metamorphosis. However, overexpression of PRL produces tailed frogs by reversing specifically some but not all of the programs of tail resorption and stimulating growth of fibroblasts in the tail. Whereas TH induces muscle resorption in tails of these transgenics, the tail fibroblasts continue to proliferate resulting in a fibrotic tail that is resistant to TH.

The negative feedback actions of progesterone on gonadotropinreleasing hormone secretion are transduced by the classical progesterone receptor

Progesterone (P) powerfully inhibits gonadotropin-releasing hormone (GnRH) secretion in ewes, as in other species, but the neural mechanisms underlying this effect remain poorly understood. Using an estrogen (E)-free ovine model, we investigated the immediate GnRH and luteinizing hormone (LH) response to acute manipulations of circulating P concentrations and whether this response was mediated by the nuclear P receptor. Simultaneous hypophyseal portal and jugular blood samples were collected over 36 hr: 0–12 hr, in the presence of exogenous P (P treatment begun 8 days earlier); 12–24 hr, P implant removed; 24–36 hr, P implant reinserted. P removal caused a significant rapid increase in the GnRH pulse frequency, which was detectable within two pulses (175 min). P insertion suppressed the GnRH pulse frequency even faster: the effect detectable within one pulse (49 min). LH pulsatility was modulated identically. The next two experiments demonstrated that these effects of P are mediated by the nuclear P receptor since intracerebroventricularly infused P suppressed LH release but 3α-hydroxy-5α-pregnan-20-one, which operates through the type A γ-aminobutyric acid receptor, was without effect and pretreatment with the P-receptor antagonist RU486 blocked the ability of P to inhibit LH. Our final study showed that P exerts its acute suppression of GnRH through an E-dependent system because the effects of P on LH secretion, lost after long-term E deprivation, are restored after 2 weeks of E treatment. Thus we demonstrate that P acutely inhibits GnRH through an E-dependent nuclear P-receptor system.

Structural requirements for peptidic antagonists of the corticotropin-releasing factor receptor (CRFR): Development of CRFR2β-selective antisauvagine-30

Different truncated and conformationally constrained analogs of corticotropin-releasing factor (CRF) were synthesized on the basis of the amino acid sequences of human/rat CRF (h/rCRF), ovine CRF (oCRF), rat urocortin (rUcn), or sauvagine (Svg) and tested for their ability to displace [125I-Tyr0]oCRF or [125I-Tyr0]Svg from membrane homogenates of human embryonic kidney (HEK) 293 cells stably transfected with cDNA coding for rat CRF receptor, type 1 (rCRFR1), or mouse CRF receptor, type 2β (mCRFR2β). Furthermore, the potency of CRF antagonists to inhibit oCRF- or Svg-stimulated cAMP production of transfected HEK 293 cells expressing either rCRFR1 (HEK-rCRFR1 cells) or mCRFR2β (HEK-mCRFR2β cells) was determined. In comparison with astressin, which exhibited a similar affinity to rCRFR1 (Kd = 5.7 ± 1.6 nM) and mCRFR2β (Kd = 4.0 ± 2.3 nM), [dPhe11,His12]Svg(11–40), [dLeu11]Svg(11–40), [dPhe11]Svg(11–40), and Svg(11–40) bound, respectively, with a 110-, 80-, 68-, and 54-fold higher affinity to mCRFR2β than to rCRFR1. The truncated analogs of rUcn displayed modest preference (2- to 7-fold) for binding to mCRFR2β. In agreement with the results of these binding experiments, [dPhe11,His12]Svg(11–40), named antisauvagine-30, was the most potent and selective ligand to suppress agonist-induced adenylate cyclase activity in HEK cells expressing mCRFR2β.

Direct transformation of rodent fibroblasts by jaagsiekte sheep retrovirus DNA

Jaagsiekte sheep retrovirus (JSRV) is the causative agent of ovine pulmonary carcinoma, a unique animal model for human bronchioalveolar carcinoma. We previously isolated a JSRV proviral clone and showed that it was both infectious and oncogenic. Thus JSRV is necessary and sufficient for the development of ovine pulmonary carcinoma, but no data are available on the mechanisms of transformation. Inspection of the JSRV genome reveals standard retroviral genes, but no evidence for a viral oncogene. However, an alternate ORF in pol (orf-x) might be a candidate for a transforming gene. We tested whether the JSRV genome might encode a transforming gene by transfecting an expression plasmid for JSRV [pCMVJS21, driven by the cytomegalovirus (CMV) immediate early promoter] into mouse NIH 3T3 cells. Foci of transformed cells appeared in the transfected cultures 2–3 weeks posttransfection; cloned transformants showed anchorage independence for growth, and they expressed JSRV RNA. These results indicate that the JRSV genome contains information with direct transforming potential for NIH 3T3 cells. Transfection of a mutated version of pCMVJS21 in which the orf-x protein was terminated by two stop codons also gave transformed foci. Thus, orf-x was eliminated as the candidate transforming gene. In addition, another derivative of pCMVJS21 (pCMVJS21ΔGP) in which the gag, pol (and orf-x) coding sequences were deleted also gave transformed foci. These results indicate that the envelope gene carries the transforming potential. This is an unusual example of a native retroviral structural protein with transformation potential.

Mutation in bone morphogenetic protein receptor-IB is associated with increased ovulation rate in Booroola Mérino ewes

Ewes from the Booroola strain of Australian Mérino sheep are characterized by high ovulation rate and litter size. This phenotype is due to the action of the FecBB allele of a major gene named FecB, as determined by statistical analysis of phenotypic data. By genetic analysis of 31 informative half-sib families from heterozygous sires, we showed that the FecB locus is situated in the region of ovine chromosome 6 corresponding to the human chromosome 4q22–23 that contains the bone morphogenetic protein receptor IB (BMPR-IB) gene encoding a member of the transforming growth factor-β (TGF-β) receptor family. A nonconservative substitution (Q249R) in the BMPR-IB coding sequence was found to be associated fully with the hyperprolificacy phenotype of Booroola ewes. In vitro, ovarian granulosa cells from FecBB/FecBB ewes were less responsive than granulosa cells from FecB+/FecB+ ewes to the inhibitory effect on steroidogenesis of GDF-5 and BMP-4, natural ligands of BMPR-IB. It is suggested that in FecBB/FecBB ewes, BMPR-IB would be inactivated partially, leading to an advanced differentiation of granulosa cells and an advanced maturation of ovulatory follicles.

Antiviral activities of the soluble extracellular domains of type I interferon receptors

Alternative splicing leads to the expression of multiple isoforms of the subunits (IFNAR1 and IFNAR2) of the type I IFN receptor. Here we describe two transcripts representing extracellular forms of ovine IFNAR1 and show that soluble extracellular forms of both IFNAR2 and IFNAR1, prepared in recombinant form in Escherichia coli, have antiviral (AV) activity in the absence of IFN. Exposure of Madin-Darby bovine kidney cells to the extracellular domain (R2E) of IFNAR2 at concentrations as low as 10 nM afforded complete protection against vesicular stomatitis virus and led to the rapid activation of the transcription factors ISGF3 and GAF. Although R2E can bind IFN (Kd ≈70 nM), activity was observed irrespective of whether or not ligand was present. R2E was inactive on mouse L929 cells but active on L929 cells expressing a membraneanchored, ovine/human chimeric IFNAR2 with an ovine extracellular domain. The data suggest that AV activity is conferred by the ability of soluble R2E to associate with the transfected IFNAR2 subunit rather than resident murine IFNAR1. Soluble extracellular forms of IFNAR1 have lower AV activity than R2E on Madin-Darby bovine kidney cells but are less species-specific and protect wild-type L929 cells as efficiently as the transfected cell line, presumably by interacting with one of the murine receptor subunits.