Insulin Dependant Diabetes Mellitus: Implications for Male Reproductive Function.

BACKGROUND Diabetes mellitus (DM) is increasing in men of reproductive age. Despite this, the prevalence of diabetes in men attending fertility clinics is largely unknown. Furthermore, studies examining the effects of DM on sperm fertility potential have been limited to conventional semen analysis. METHODS Conventional semen analysis (semen volume, sperm count, motility and morphology) was performed for 27 diabetic (mean age 34 +/- 2 years) and 29 non-diabetic subjects (control group, men undergoing routine infertility investigations, mean age 33 +/- 1 years). Nuclear DNA (nDNA) fragmentation was assessed using the alkaline Comet assay and mitochondrial DNA (mtDNA) deletions by Long-PCR. RESULTS Other than a small, but significant, reduction in semen volume in diabetic men (2.6 versus 3.3 ml; P <0.05), conventional semen parameters did not differ significantly from control subjects. Diabetic subjects had significantly higher mean nDNA fragmentation (53 versus 32%; P <0.0001) and median number of mtDNA deletions (4 versus 3; P <0.05) compared with control subjects. CONCLUSIONS Diabetes is associated with increased sperm nuclear and mtDNA damage that may impair the reproductive capability of these men.

Fasciola hepatica: Histological changes in the reproductive structures of triclabendazole (TCBZ)-sensitive and TCBZ-resistant flukes after treatment in vivo with TCBZ and the related benzimidazole derivative, Compound Alpha

Twenty-four shed-reared lambs were each infected orally with 250 metacercariae of Fasciola hepatica, using either the triclabendazole (TCBZ)-sensitive Cullompton isolate or the TCBZ-resistant Sligo isolate. Twelve weeks after infection the lambs were treated with TCBZ (10 mg/kg) or with the experimental fasciolicide, Compound Alpha (Cpd alpha), a benzimidazole derivative of TCBZ (15 mg/kg). The lambs were euthanised 48,72 and 96 h after TCBZ treatment, or 24, 48 and 72 h after Cpd a treatment, and flukes were collected from the liver and/or gall bladder of each animal. Untreated animals harbouring 12-week infections were euthanised 24 h after administration of anthelmintic to the treatment groups, and the untreated flukes provided control material. A semi-quantitative assessment of the degree of histological change induced by the two drugs after different times of exposure was achieved by scoring the intensity of three well-defined lesions that developed in the testes and uteri of a representative sample of flukes from each lamb. In general, it was found that in those tissues where active meiosis and/or mitosis occurred (testis, ovary, and vitelline follicles), there was progressive loss of cell content due to apparent failure of cell division to keep pace with expulsion of the mature or effete products. Further, actively dividing cell types tended to become individualised, rounded and condensed, characteristic of apoptotic cell death. Protein synthetic activity was apparently inhibited in the Mehlis' secretory cells. In the uterus, where successful formation of shelled eggs represents the culmination of a complex sequence of cytokinetic, cytological and synthetic activity involving the vitelline follicles, the ovary and the Mehlis' gland, histological evidence indicating failure of ovigenesis was evident from 24 h post-treatment onwards. The development of these lesions may be related to the known antitubulin activity of the benzimidazole class of anthelmintics, to the induction of apoptosis in cells where mitosis or meiosis has aborted due to failure of spindle formation, and to drug-induced inhibition of protein synthesis. The semi-quantitative findings indicated that Cpd a is slightly less efficacious than TCBZ itself in causing histological damage to the reproductive structures of TCBZ-sensitive flukes, and that, like TCBZ, it caused no histological damage in flukes of the TCBZ-resistant isolate. This study illustrates the potential utility of histological techniques for conveniently screening representative samples of flukes in field trials designed to validate instances of drug resistance or to test the efficacy of new products against known drug-resistant and drug-susceptible fluke isolates. It also provides reference criteria for drug-induced histopathological changes in fluke reproductive structures which may aid interpretation of TEM findings. (C) 2009 Elsevier B.V. All rights reserved.

FMRFamide-like peptides in root knot nematodes and their potential role in nematode physiology

FMRFamide-like peptides (FLPs) are a diverse group of neuropeptides that are expressed abundantly in nematodes. They exert potent physiological effects on locomotory, feeding and reproductive musculature and also act as neuromodulators. However, little is known about the specific expression patterns and functions of individual peptides. The current study employed rapid amplification of cDNA ends-polymerase chain reaction (RACE-PCR) to characterize flp genes from infective juveniles of the root knot nematodes, Meloidogyne incognita and Meloidogyne minor. The peptides identified from these transcripts are sequelogs of FLPs from the free-living nematode, Caenorhabditis elegans; the genes have therefore been designated as Mi-flp-1, Mi-flp-7, Mi-flp-12, Mm-flp-12 and Mi-flp-14. Mi-flp-1 encodes five FLPs with the common C-terminal moiety, NFLRFamide. Mi-flp-7 encodes two copies of APLDRSALVRFamide and APLDRAAMVRFamide and one copy of APFDRSSMVRFamide. Mi-flp-12 and Mm-flp-12 encode the novel peptide KNNKFEFIRFamide (a longer version of RNKFEFIRFamide found in C. elegans). Mi-flp-14 encodes a single copy of KHEYLRFamide (commonly known as AF2 and regarded as the most abundant nematode FLP), and a single copy of the novel peptide KHEFVRFamide. These FLPs share a high degree of conservation between Meloidogyne species and nematodes from other clades, including those of humans and animals, perhaps suggesting a common neurophysiological role which may be exploited by novel drugs. FLP immunoreactivity was observed for the first time in Meloidogyne, in the circumpharyngeal nerve ring, pharyngeal nerves and ventral nerve cord. Additionally, in situ hybridization revealed Mi-flp-12 expression in an RIR-like neuron and Mi-flp-14 expression in SMB-like neurons, respectively. These localizations imply physiological roles for FLP-12 and FLP-14 peptides, including locomotion and sensory perception.

The paternal genome and the health of the assisted reproductive technology child

As a number of children born by assisted reproductive technology (ART) are increasing each year across the developed world, the health of such offspring is a matter of public concern. Does the integrity of the paternal genome impact on offspring health? In societal terms, as birth rates fall, and the Western population become unsustainable, do the benefits outweigh the costs of creating and providing for this ART conceived sub population? There are little data to date to answer these questions. The long‑term health of such children has largely been ignored, and success measured only by early (pre-birth) outcomes such as embryo quality or pregnancy. However, there are powerful paradigms such as ageing and smoking that give vital clues as to the potential impact of unhealthy spermatozoa on disease risk, mental and physical health, fertility and mortality of these offspring.

Potential biomarkers on gametogenesis process and in sperm quality of european EEL (Anguilla anguilla)

Dissertação de mestrado, Aquacultura, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2015

Vascular dysfunction in children conceived by assisted reproductive technologies: underlying mechanisms and future implications.

Epidemiological studies in humans have demonstrated a relationship between pathological events during fetal development and increased cardiovascular risk later in life and have led to the so called "Fetal programming of cardiovascular disease hypothesis". The recent observation of generalised vascular dysfunction in young apparently healthy children conceived by assisted reproductive technologies (ART) provides a novel and potentially very important example of this hypothesis. This review summarises recent data in ART children demonstrating premature subclinical atherosclerosis in the systemic circulation and pulmonary vascular dysfunction predisposing to exaggerated hypoxia-induced pulmonary hypertension. These problems appear to be related to the ART procedure per se. Studies in ART mice demonstrating premature vascular aging and arterial hypertension further demonstrate the potential of ART to increase cardiovascular risk and have allowed to unravel epigenetic alterations of the eNOS gene as an underpinning mechanism. The roughly 25% shortening of the life span in ART mice challenged with a western style high-fat-diet demonstrates the potential importance of these alterations for the long-term outcome. Given the young age of the ART population, data on cardiovascular endpoints will not be available before 20 to 30 years from now. However, already now cohort studies of the ART population are needed to early detect cardiovascular alterations with the aim to prevent or at least optimally treat cardiovascular complications. Finally, a debate needs to be engaged on the future of ART and the consequences of its exponential growth for public health.

Problems of allometric scaling analysis: examples from mammalian reproductive biology.

Biological scaling analyses employing the widely used bivariate allometric model are beset by at least four interacting problems: (1) choice of an appropriate best-fit line with due attention to the influence of outliers; (2) objective recognition of divergent subsets in the data (allometric grades); (3) potential restrictions on statistical independence resulting from phylogenetic inertia; and (4) the need for extreme caution in inferring causation from correlation. A new non-parametric line-fitting technique has been developed that eliminates requirements for normality of distribution, greatly reduces the influence of outliers and permits objective recognition of grade shifts in substantial datasets. This technique is applied in scaling analyses of mammalian gestation periods and of neonatal body mass in primates. These analyses feed into a re-examination, conducted with partial correlation analysis, of the maternal energy hypothesis relating to mammalian brain evolution, which suggests links between body size and brain size in neonates and adults, gestation period and basal metabolic rate. Much has been made of the potential problem of phylogenetic inertia as a confounding factor in scaling analyses. However, this problem may be less severe than suspected earlier because nested analyses of variance conducted on residual variation (rather than on raw values) reveals that there is considerable variance at low taxonomic levels. In fact, limited divergence in body size between closely related species is one of the prime examples of phylogenetic inertia. One common approach to eliminating perceived problems of phylogenetic inertia in allometric analyses has been calculation of 'independent contrast values'. It is demonstrated that the reasoning behind this approach is flawed in several ways. Calculation of contrast values for closely related species of similar body size is, in fact, highly questionable, particularly when there are major deviations from the best-fit line for the scaling relationship under scrutiny.

An investigative study on the effects of black fly (diptera: simuliidae) sugar meals on reproductive success and parasite transmission /

Black flies are opportunistic sugar-feeders. They take sugar meals from Homopteran honeydew secretions or plant nectars, depending on availability. Homopteran honeydew secretions contain both simple and complex carbohydrates while plant nectars contain primarily simple carbohydrates. In order to determine whether honeydew secretions offer more energy than plant nectars to their insect visitors a study of wild-caught black flies was undertaken in Algonquin Provincial Park, Canada during the spring of 1 998 and 1 999. It was hypothesized that female black flies maintained on honeydew sugars will survive longer, produce more eggs and have a greater parasite vectoring potential than those maintained on artificial nectar or distilled water. Results demonstrated that: (1) host-seeking female Prosimulimfuscum/mixtum and Simulium venustum maintained on artificial honeydew did not survive longer than those maintained on artificial nectar when fed ad libitum; (2) fiiUy engorged S. venustum and Simulium rugglesi maintained on artificial honeydew did not produce more eggs than those maintained on artificial nectar when fed ad libitum; and (3) S. rugglesi did not have a greater vectoring potential of Leucocytozoon simondi when maintained on artificial honeydew as opposed to artificial nectar when fed ad libitum. However, all flies maintained on the two sugars (artificial honeydew and artificial nectar) survived longer, produce more eggs and had greater vectoring potential than those maintained on distilled water alone.

Broad reduction and adoption in ring-billed gulls (Larus Delaworensis): the potential for inergenerational conflict

Patterns of intra-clutch egg size variation and intra-clutch hatch intervals in the Ring-billed gull (Larus delawarensis) were documented during the peak nesting period of two consecutive breeding seasons, at a colony near Port Colborne, Ontario. Egg size decreased with laying order; third laid eggs were significantly smaller than first laid eggs. Hatching of the third egg was delayed from that of first and second eggs. Intraclutch egg size differences established initial size disparities among chicks at hatch. Hatch intervals further exaggerated size disparities during the early post brood completion period. Competitive asymmetries among chicks were associated with increased mortality rates among third hatched chicks despite the lack of evidence of a sibling feeding hierarchy. Fledging success in 1987 was greater than in 1988. A "brood reduction strategy" appears to have enabled parents in 1987, to obtain an extra unit of reproductive fitness, while in 1988 parents were often unable to raise the entire brood and third chicks likely represented insurance reproductive value. Experimental broods (1988) were created in which hatch intervals were double those of natural intervals. The size disparities among chicks were significantly greater than in control broods, and the pattern of mortality among chicks suggested that first chicks benefited at a cost to second and third chicks. Parents of peak experimental broods achieved a fledging success rate similar to that of control broods. Characteristics of chick adoptions were also recorded. In each study year, 9 chicks abandoned their natal territories, 6 of which were adopted. Chicks consistently established themselves into broods where they were older than resident chicks. No direct evidence of cost to foster parents, or benefits to adopted chicks was obtained, although fledging success of adopted chicks was high.

Santé reproductive et santé mentale des femmes qui ont subi la violence sexuelle en temps de conflit armé : cas de la République Démocratique du Congo

Problématique : Depuis bientôt deux décennies, la République Démocratique du Congo (RDC) est le territoire d’un conflit armé qui, selon l’International Rescue Commite, aurait occasionné plus de 3 millions de décès et autant de déplacés internes. Plusieurs rapports font également cas des nombreux actes de violence sexuelle (les viols, les mutilations, l’esclavage, l’exploitation sexuelle, etc.) commis envers les filles, les femmes et dans une moindre ampleur les hommes. S’il existe un consensus sur le côté barbare des actes de violence sexuelle liés aux conflits armés, rares sont les études qui ont évalué leurs conséquences sur la santé reproductive des survivantes surtout en termes d’issues telles que les fistules, les douleurs pelviennes chroniques (DPC), le désir de rapports sexuels, le désir d’enfant et le désir d’interruption de la grossesse issue de tels actes. Par ailleurs, même si la santé mentale des populations en zones de conflit représente un sujet d’intérêt, l’impact spécifique de la violence sexuelle liée au conflit sur la santé mentale des survivantes a été peu étudié. De plus, ces travaux s’intéressent aux effets de la violence sexuelle liée au conflit sur la santé mentale et sur la santé reproductive séparément et ce, sans évaluer les relations qui peuvent exister entre ces deux dimensions qui, pourtant, s’influencent mutuellement. Aussi, l’impact social de la violence sexuelle liée au conflit, ainsi que la contribution des normes socioculturelles aux difficultés que rencontrent les survivantes, a été peu étudié. Pourtant, l’impact social de la violence sexuelle liée au conflit peut permettre de mieux comprendre comment l’expérience d’un tel acte peut affecter la santé mentale. Enfin, aucune étude n’a évalué les effets de la violence sexuelle liée au conflit en la comparant à la violence sexuelle non liée au conflit (VSNLC). Pourtant, il est reconnu qu’à de nombreux égards, la violence sexuelle liée au conflit est bien différente de la VSNLC puisqu’elle est perpétrée avec l’intention de créer le maximum d’effets adverses pour la victime et sa communauté. Objectifs : Les objectifs poursuivis dans cette thèse visent à : 1) évaluer les effets de la violence sexuelle liée au conflit sur la santé reproductive; 2) évaluer les effets de la violence sexuelle liée au conflit sur la santé mentale en termes de sévérité des symptômes de stress posttraumatique (PTSD), de sévérité des symptômes de détresse psychologique et de probabilité de souffrir de troubles mentaux communs (TMC); 3) évaluer la contribution des troubles physiques de santé reproductive, en particulier les fistules et les douleurs pelviennes chroniques (DPC), aux effets de la violence sexuelle liée au conflit sur la santé mentale; 4) évaluer la contribution de l’état de santé mentale aux effets de la violence sexuelle liée au conflit sur le désir de rapports sexuels et le désir d’enfant; et 5) étudier l’impact de la violence sexuelle liée au conflit sur le plan social ainsi que la contribution des normes socioculturelles à ses effets adverses et la façon dont ces effets pourraient à leur tour influencer la santé des femmes et leur relation avec l’enfant issu de l’acte de violence sexuelle subi. Méthodologie : Un devis mixte de nature convergente a permis de collecter des données quantitatives auprès de l’ensemble des participantes (étude transversale) et des données qualitatives sur un nombre plus restreint de femmes (étude phénoménologique). Une étude transversale populationnelle a été conduite entre juillet et août 2012 auprès de 320 femmes âgées de 15 à 45 ans habitant quatre (4) quartiers de la ville de Goma située dans la province du Nord-Kivu en RDC. Les femmes ont été recrutées à travers des annonces faites par les responsables des programmes d’alphabétisation et de résolution de conflits implantés dans les différents quartiers par le Collectif Alpha Ujuvi, une ONG locale. Les issues de santé reproductive évaluées sont : les fistules, les DPC, le désir de rapports sexuels, le désir d’enfant et le désir d’interruption de la grossesse issue d’un acte de violence sexuelle. Les variables de santé mentale d’intérêt sont : la sévérité des symptômes de détresse psychologique, la sévérité des symptômes de PTSD et la probabilité de souffrir de TMC. Pour les analyses, l’exposition a été définie en trois (3) catégories selon l’expérience passée de violence sexuelle : les femmes qui ont vécu des actes de violence sexuelle liée au conflit, celles qui ont vécu des actes de VSNLC et celles qui ont déclaré n’avoir jamais subi d’acte de violence sexuelle au cours de leur vie. Les variables de confusion potentielles mesurées sont : l’âge, le statut matrimonial, le nombre d’enfants, le niveau d’éducation le plus élevé atteint et l’occupation professionnelle. Les mesures d’associations ont été évaluées à l’aide de modèles de régressions logistiques et linéaires simples et multiples. Des tests d’interaction multiplicative et des analyses stratifiées ont été également conduits pour évaluer l’effet potentiellement modificateur de quelques variables (âge, statut matrimonial, nombre d’enfants) sur la relation entre la violence sexuelle et les variables de santé reproductive ou de santé mentale. Ces tests ont également été utilisés pour évaluer la contribution d’une variable de santé reproductive ou de santé mentale aux effets de la violence sexuelle sur l’autre dimension de la santé d’intérêt dans cette étude. Une étude phénoménologique a été conduite dans le même intervalle de temps auprès de 12 femmes ayant participé à la partie quantitative de l’étude qui ont vécu la violence sexuelle liée au conflit et ont eu un enfant issu d’une agression sexuelle. Les sujets explorés incluent : la perception de l’acte de violence sexuelle liée au conflit vécu et de la vie quotidienne par les victimes; la perception de l’acte de violence sexuelle liée au conflit par la famille et l’entourage et leurs réactions après l’agression; la perception de la grossesse issue de l’acte de violence sexuelle par la victime; la perception de l’enfant issu de la violence sexuelle liée au conflit par la victime ainsi que son entourage; les conséquences sociales de l’expérience de violence sexuelle liée au conflit et les besoins des victimes pour leur réhabilitation. Une analyse thématique avec un codage ouvert a permis de ressortir les thèmes clés des récits des participantes. Par la suite, l’approche de théorisation ancrée a été utilisée pour induire un cadre décrivant l’impact social de l’expérience de la violence sexuelle liée au conflit et les facteurs y contribuant. Résultats : Le premier article de cette thèse montre que, comparées aux femmes qui n’ont jamais vécu un acte de violence sexuelle, celles qui ont vécu la violence sexuelle liée au conflit ont une probabilité plus élevée d’avoir une fistule (OR=11.1, IC 95% [3.1-39.3]), des DPC (OR=5.1, IC 95% [2.4-10.9]), de rapporter une absence de désir de rapports sexuels (OR=3.5, IC 95% [1.7-6.9]) et une absence de désir d’enfant (OR=3.5, IC 95% [1.6-7.8]). Comparées aux mêmes femmes, celles qui ont vécu la VSNLC ont plus de probabilité de souffrir de DPC (OR=2.3, IC 95% [0.95-5.8]) et de rapporter une absence de désir d’enfant (OR=2.7, IC 95% [1.1-6.5]). Comparées aux femmes qui ont vécu la VSNLC, celles qui ont vécu la violence sexuelle liée au conflit ont également une probabilité plus élevée d’avoir une fistule (OR=9.5, IC 95% [1.6-56.4]), des DPC (OR=2.2, IC 95% [0.8-5.7]) et de rapporter une absence de désir de rapports sexuels (OR=2.5, IC 95% [1.1-6.1]). En ce qui concerne les grossesses issues des viols, comparées aux femmes qui ont vécu la VSNLC, celles qui ont vécu la violence sexuelle liée au conflit sont plus nombreuses à souhaiter avorter (55% vs 25% pour celles qui ont vécu la VSNLC). Elles sont également plus nombreuses à déclarer qu’elles auraient avorté si les soins appropriés étaient accessibles (39% vs 21% pour celles qui ont vécu la VSNLC). Le second article montre qu’en comparaison aux femmes qui n’ont jamais subi de violence sexuelle, celles qui ont vécu la violence sexuelle liée au conflit présentent des symptômes de détresse psychologique (moyennes de score respectives 8.6 et 12.6, p<0.0001) et des symptômes de PTSD (moyennes de score respectives 2.2 et 2.6, p<0.0001) plus sévères et ont plus de probabilité d’être dépistées comme un cas de TMC (30% vs 76%, p<0.0001). De plus, comparées aux femmes qui ont vécu la VSNLC, celles qui ont vécu la violence sexuelle liée au conflit présentent des symptômes de détresse psychologique (moyennes de score respectives 10.1 et 12.6, p<0.0001) et des symptômes de PTSD (moyennes de score respectives 2.2 et 2.6, p<0.0001) plus sévères et ont plus de probabilité d’être dépistées comme un cas de TMC (48% vs 76%, p<0.001). Les valeurs minimales et maximales de score de sévérité de symptômes de détresse psychologique sont de 0/12 pour les femmes qui n’ont jamais vécu de violence sexuelle, 4/19 pour celles qui ont vécu la VSNLC et de 5/18 pour celles qui ont vécu la violence sexuelle liée au confit. En ce qui concerne la sévérité des symptômes de PTSD, les scores minimal et maximal sont respectivement de 0.36/3.22, 0.41/3.41 et 0.95/3.45. Le fait d’avoir développé une fistule ou de souffrir de DPC après l’agression sexuelle augmente la force des associations entre la violence sexuelle et la santé mentale. Les femmes qui ont subi la violence sexuelle liée au conflit et qui ont souffert de fistules présentent des symptômes de détresse psychologique et de PTSD plus sévères comparées aux femmes qui ont subi la violence sexuelle liée au conflit mais n’ont pas de fistules. Les résultats sont similaires pour les femmes qui ont subi la violence sexuelle liée au conflit et qui souffrent de DPC. Des résultats complémentaires suggèrent que le statut matrimonial modifie l’effet de la violence sexuelle sur la sévérité des symptômes de détresse psychologique, les femmes divorcées/séparées et les veuves étant celles qui ont les moyennes de score les plus élevées (respectivement 11.3 et 12.1 vs 9.26 et 9.49 pour les célibataires et les mariées). Par ailleurs, la sévérité des symptômes de détresse psychologique modifie l’association entre la violence sexuelle liée au conflit et le désir d’enfant. Le troisième article montre que, sur le plan social, l’expérience de violence sexuelle liée au conflit entraine également de lourdes conséquences. Toutes celles qui ont vécu ce type d’acte décrivent leur vie de survivante et de mère d’un enfant issu d’une agression sexuelle comme difficile, oppressive, faite de peines et de soucis et sans valeur. Plusieurs facteurs influencent la description que les victimes de violence sexuelle liée au conflit font de leur vie quotidienne, et ils sont tous reliés aux normes socioculturelles qui font de la femme une citoyenne de seconde zone, ne font aucune différence entre un viol et un adultère, condamnent les victimes de violence sexuelle plutôt que leurs agresseurs, rejettent et stigmatisent les victimes de tels actes ainsi que l’enfant qui en est issu. En réponse au rejet et au manque de considération, les femmes victimes de violence sexuelle liée au conflit ont tendance à s’isoler pour éviter les insultes et à garder le silence sur leur agression. En plus, les réactions de leur entourage/communauté ont tendance à leur faire revivre l’agression sexuelle subie, autant d’éléments qui nuisent davantage à leur réhabilitation. D’autres résultats démontrent que les enfants issus d’actes de violence sexuelle liée au conflit sont également rejetés par leur communauté, leur famille adoptive ainsi que le conjoint de leur mère, ce qui affecte davantage les survivantes. Avec leurs mères, les relations développées varient entre le rejet, la résignation et l’affection. Néanmoins, ces relations sont plus souvent tendues probablement à cause de la stigmatisation de la communauté. Conclusion: La violence sexuelle liée au conflit a des effets adverses sur la santé reproductive, la santé mentale mais également sur le plan social. Ces trois dimensions sont loin d’être isolées puisque cette étude a permis de démontrer qu’elles s’influencent mutuellement. Ceci suggère que la prise en charge des victimes de violence sexuelle liée au conflit ne doit pas se concentrer sur un aspect ou un autre de la santé mais prendre en compte l’ensemble des dimensions de la femme pour offrir une aide holistique, plus adaptée et qui sera plus efficace à long terme.

Propidium monoazide (PMA) and ethidium bromide monoazide(EMA) improve DNA array and high-throughput sequencing ofporcine reproductive and respiratory syndrome virus identification

Pan-viral DNA array (PVDA) and high-throughput sequencing (HTS) are useful tools to identify novel viruses of emerging diseases. However, both techniques have difficulties to identify viruses in clinical samples because of the host genomic nucleic acid content (hg/cont). Both propidium monoazide (PMA) and ethidium bromide monoazide (EMA) have the capacity to bind free DNA/RNA, but are cell membrane-impermeable. Thus, both are unable to bind protected nucleic acid such as viral genomes within intact virions. However, EMA/PMA modified genetic material cannot be amplified by enzymes. In order to assess the potential of EMA/PMA to lower the presence of amplifiable hg/cont in samples and improve virus detection, serum and lung tissue homogenates were spiked with porcine reproductive and respiratory virus (PRRSV) and were processed with EMA/PMA. In addition, PRRSV RT-qPCR positive clinical samples were also tested. EMA/PMA treatments significantly decreased amplifiable hg/cont and significantly increased the number of PVDA positive probes and their signal intensity compared to untreated spiked lung samples. EMA/PMA treatments also increased the sensitivity of HTS by increasing the number of specific PRRSV reads and the PRRSV percentage of coverage. Interestingly, EMA/PMA treatments significantly increased the sensitivity of PVDA and HTS in two out of three clinical tissue samples. Thus, EMA/PMA treatments offer a new approach to lower the amplifiable hg/cont in clinical samples and increase the success of PVDA and HTS to identify viruses.

Actinobacillus pleuropneumoniae induces SJPL cell cycle arrest in G2/M-phase and inhibits porcine reproductive and respiratory syndrome virus replication

Background: Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most important pathogens in the swine industry and causes important economic losses. No effective antiviral drugs against it are commercially available. We recently reported that the culture supernatant of Actinobacillus pleuropneumoniae, the porcine pleuropneumonia causative agent, has an antiviral activity in vitro against PRRSV in SJPL cells. Objectives of this study were (i) to identify the mechanism behind the antiviral activity displayed by A. pleuropneumoniae and (ii) to characterize the active molecules present in the bacterial culture supernatant. Methods: Antibody microarray analysis was used in order to point out cellular pathways modulated by the A. pleuropneumoniae supernatant. Subsequent, flow cytometry analysis and cell cycle inhibitors were used to confirm antibody microarray data and to link them to the antiviral activity of the A. pleuropneumoniae supernatant. Finally, A. pleuropneumoniae supernatant characterization was partially achieved using mass spectrometry. Results: Using antibody microarray, we observed modulations in G2/M-phase cell cycle regulation pathway when SJPL cells were treated with A. pleuropneumoniae culture supernatant. These modulations were confirmed by a cell cycle arrest at the G2/M-phase when cells were treated with the A. pleuropneumoniae culture supernatant. Furthermore, two G2/M-phase cell cycle inhibitors demonstrated the ability to inhibit PRRSV infection, indicating a potential key role for PRRSV infection. Finally, mass spectrometry lead to identify two molecules (m/z 515.2 and m/z 663.6) present only in the culture supernatant. Conclusions: We demonstrated for the first time that A. pleuropneumoniae is able to disrupt SJPL cell cycle resulting in inhibitory activity against PRRSV. Furthermore, two putative molecules were identified from the culture supernatant. This study highlighted the cell cycle importance for PRRSV and will allow the development of new prophylactic or therapeutic approaches against PRRSV.

Status and Potential of Smallholder Livestock Production Systems in Xishuangbanna, southern P.R. China

With Chinas rapid economic development during the last decades, the national demand for livestock products has quadrupled within the last 20 years. Most of that increase in demand has been answered by subsidized industrialized production systems, while million of smallholders, which still provide the larger share of livestock products in the country, have been neglected. Fostering those systems would help China to lower its strong urban migration streams, enhance the livelihood of poorer rural population and provide environmentally save livestock products which have a good chance to satisfy customers demand for ecological food. Despite their importance, China’s smallholder livestock keepers have not yet gained appropriate attention from governmental authorities and researchers. However, profound analysis of those systems is required so that adequate support can lead to a better resource utilization and productivity in the sector. To this aim, this pilot study analyzes smallholder livestock production systems in Xishuangbanna, located in southern China. The area is bordered by Lao and Myanmar and geographically counts as tropical region. Its climate is characterized by dry and temperate winters and hot summers with monsoon rains from May to October. While the regionis plain, at about 500 m asl above sea level in the south, outliers of the Himalaya mountains reach out into the north of Xishuangbanna, where the highest peak reaches 2400 m asl. Except of one larger city, Jinghong, Xishuangbanna mainly is covered by tropical rainforest, areas under agricultural cultivation and villages. The major income is generated through inner-Chinese tourism and agricultural production. Intensive rubber plantations are distinctive for the lowland plains while small-scaled traditional farms are scattered in the mountane regions. In order to determine the current state and possible future chances of smallholder livestock production in that region, this study analyzed the current status of the smallholder livestock sector in the Naban River National Nature Reserve (NRNNR), an area which is largely representative for the whole prefecture. It covers an area of about 50square kilometer and reaches from 470 up to 2400 m asl. About 5500 habitants of different ethnic origin are situated in 24 villages. All data have been collected between October 2007 and May 2010. Three major objectives have been addressed in the study: 1. Classifying existing pig production systems and exploring respective pathways for development 2. Quantifying the performance of pig breeding systemsto identify bottlenecks for production 3. Analyzing past and current buffalo utilization to determine the chances and opportunities of buffalo keeping in the future In order to classify the different pig production s ystems, a baseline survey (n=204, stratified cluster sampling) was carried out to gain data about livestock species, numbers, management practices, cultivated plant species and field sizes as well associo-economic characteristics. Sampling included two clusters at village level (altitude, ethnic affiliation), resulting in 13 clusters of which 13-17 farms were interviewed respectively. Categorical Principal Component Analysis (CatPCA) and a two-step clustering algorithm have been applied to identify determining farm characteristics and assort recorded households into classes of livestock production types. The variables keep_sow_yes/no, TLU_pig, TLU_buffalo, size_of_corn_fields, altitude_class, size_of_tea_plantationand size_of_rubber_fieldhave been found to be major determinants for the characterization of the recorded farms. All farms have extensive or semi-intensive livestock production, pigs and buffaloes are predominant livestock species while chicken and aquaculture are available but play subordinate roles for livelihoods. All pig raisers rely on a single local breed, which is known as Small Ear Pig (SMEP) in the region. Three major production systemshave been identified: Livestock-corn based LB; 41%), rubber based (RB; 39%) and pig based (PB;20%) systems. RB farms earn high income from rubber and fatten 1.9 ±1.80 pigs per household (HH), often using purchased pig feed at markets. PB farms own similar sized rubber plantations and raise 4.7 ±2.77 pigs per HH, with fodder mainly being cultivated and collected in theforest. LB farms grow corn, rice and tea and keep 4.6 ±3.32 pigs per HH, also fed with collected and cultivated fodder. Only 29% of all pigs were marketed (LB: 20%; RB: 42%; PB: 25%), average annual mortality was 4.0 ±4.52 pigs per farm (LB: 4.6 ±3.68; RB: 1.9 ±2.14; PB: 7.1 ±10.82). Pig feed mainly consists of banana pseudo stem, corn and rice hives and is prepared in batches about two to three times per week. Such fodder might be sufficient in energy content but lacks appropriate content of protein. Pigs therefore suffer from malnutrition, which becomes most critical in the time before harvest season around October. Farmers reported high occurrences of gastrointestinal parasites in carcasses and often pig stables were wet and filled with manure. Deficits in nutritional and hygienic management are major limits for development and should be the first issues addressed to improve productivity. SME pork was found to be known and referred by local customers in town and by richer lowland farmers. However, high prices and lacking availability of SME pork at local wet-markets were the reasons which limited purchase. If major management constraints are overcome, pig breeders (PB and LB farms) could increase the share of marketed pigs for town markets and provide fatteners to richer RB farmers. RB farmers are interested in fattening pigs for home consumption but do not show any motivation for commercial pig raising. To determine the productivity of input factors in pig production, eproductive performance, feed quality and quantity as well as weight development of pigs under current management were recorded. The data collection included a progeny history survey covering 184 sows and 437 farrows, bi-weekly weighing of 114 pigs during a 16-months time-span on 21 farms (10 LB and 11 PB) as well as the daily recording of feed quality and quantity given to a defined number of pigs on the same 21 farms. Feed samples of all recorded ingredients were analyzed for their respective nutrient content. Since no literature values on thedigestibility of banana pseudo stem – which is a major ingredient of traditional pig feed in NRNNR – were found, a cross-sectional digestibility trial with 2x4 pigs has been conducted on a station in the research area. With the aid of PRY Herd Life Model, all data have been utilized to determine thesystems’ current (Status Quo = SQ) output and the productivity of the input factor “feed” in terms of saleable life weight per kg DM feed intake and monetary value of output per kg DM feed intake.Two improvement scenarios were simulated, assuming 1) that farmers adopt a culling managementthat generates the highest output per unit input (Scenario 1; SC I) and 2) that through improved feeding, selected parameters of reproduction are improved by 30% (SC II). Daily weight gain averaged 55 ± 56 g per day between day 200 and 600. The average feed energy content of traditional feed mix was 14.92 MJ ME. Age at first farrowing averaged 14.5 ± 4.34 months, subsequent inter-farrowing interval was 11.4 ± 2.73 months. Littersize was 5.8 piglets and weaning age was 4.3 ± 0.99 months. 18% of piglets died before weaning. Simulating pig production at actualstatus, it has been show that monetary returns on inputs (ROI) is negative (1:0.67), but improved (1:1.2) when culling management was optimized so that highest output is gained per unit feed input. If in addition better feeding, controlled mating and better resale prices at fixed dates were simulated, ROI further increased to 1:2.45, 1:2.69, 1:2.7 and 1:3.15 for four respective grower groups. Those findings show the potential of pork production, if basic measures of improvement are applied. Futureexploration of the environment, including climate, market-season and culture is required before implementing the recommended measures to ensure a sustainable development of a more effective and resource conserving pork production in the future. The two studies have shown that the production of local SME pigs plays an important role in traditional farms in NRNNR but basic constraints are limiting their productivity. However, relatively easy approaches are sufficient for reaching a notable improvement. Also there is a demand for more SME pork on local markets and, if basic constraints have been overcome, pig farmers could turn into more commercial producers and provide pork to local markets. By that, environmentally safe meat can be offered to sensitive consumers while farmers increase their income and lower the risk of external shocks through a more diverse income generating strategy. Buffaloes have been found to be the second important livestock species on NRNNR farms. While they have been a core resource of mixed smallholderfarms in the past, the expansion of rubber tree plantations and agricultural mechanization are reasons for decreased swamp buffalo numbers today. The third study seeks to predict future utilization of buffaloes on different farm types in NRNNR by analyzing the dynamics of its buffalo population and land use changes over time and calculating labor which is required for keeping buffaloes in view of the traction power which can be utilized for field preparation. The use of buffaloes for field work and the recent development of the egional buffalo population were analyzed through interviews with 184 farmers in 2007/2008 and discussions with 62 buffalo keepers in 2009. While pig based farms (PB; n=37) have abandoned buffalo keeping, 11% of the rubber based farms (RB; n=71) and 100% of the livestock-corn based farms (LB; n=76) kept buffaloes in 2008. Herd size was 2.5 ±1.80 (n=84) buffaloes in early 2008 and 2.2 ±1.69 (n=62) in 2009. Field work on own land was the main reason forkeeping buffaloes (87.3%), but lending work buffaloes to neighbors (79.0%) was also important. Other purposes were transport of goods (16.1%), buffalo trade (11.3%) and meat consumption(6.4%). Buffalo care required 6.2 ±3.00 working hours daily, while annual working time of abuffalo was 294 ±216.6 hours. The area ploughed with buffaloes remained constant during the past 10 years despite an expansion of land cropped per farm. Further rapid replacement of buffaloes by tractors is expected in the near future. While the work economy is drastically improved by the use of tractors, buffaloes still can provide cheap work force and serve as buffer for economic shocks on poorer farms. Especially poor farms, which lack alternative assets that could quickly be liquidizedin times of urgent need for cash, should not abandon buffalo keeping. Livestock has been found to be a major part of small mixed farms in NRNNR. The general productivity was low in both analyzed species, buffaloes and pigs. Productivity of pigs can be improved through basic adjustments in feeding, reproductive and hygienic management, and with external support pig production could further be commercialized to provide pork and weaners to local markets and fattening farms. Buffalo production is relatively time intensive, and only will be of importance in the future to very poor farms and such farms that cultivate very small terraces on steep slopes. These should be encouraged to further keep buffaloes. With such measures, livestock production in NRNNR has good chances to stay competitive in the future.

Decision making and planning of children in the reproductive process of males

Reproduction is not restricted to the biological; it is also a complex and dynamic social process. The objective of this paper is to analyze the decision and planning of children as part of the reproductive process in men. Methodologically, a qualitative research was made. The analysis includes in-depth interviews with 30 parents between 20 and 45 years, of medium socio-economic level in México. The results show that the reproductive process incorporates family formation negotiated with their couple the marriage until they had economic potential. In some cases, their decision hastened because the couple was pregnant. The planning was not contemplated but when getting the news, they accept it and get involved in the process, while others see it as a conflict.

Reproductive conflict delays the recovery of an endangered social species

Evolutionary theory predicts that individuals, in order to increase their relative fitness, can evolve behaviours that are detrimental for the group or population. This mismatch is particularly visible in social organisms. Despite its potential to affect the population dynamics of social animals, this principle has not yet been applied to real-life conservation. Social group structure has been argued to stabilize population dynamics due to the buffering effects of nonreproducing subordinates. However, competition for breeding positions in such species can also interfere with the reproduction of breeding pairs. Seychelles magpie robins, Copsychus sechellarum, live in social groups where subordinate individuals do not breed. Analysis of long-term individual-based data and short-term behavioural observations show that subordinates increase the territorial takeover frequency of established breeders. Such takeovers delay offspring production and decrease territory productivity. Individual-based simulations of the Seychelles magpie robin population parameterized with the long-term data show that this process has significantly postponed the recovery of the species from the Critically Endangered status. Social conflict thus can extend the period of high extinction risk, which we show to have population consequences that should be taken into account in management programmes. This is the first quantitative assessment of the effects of social conflict on conservation.