Identifying selection strategies based on the practices and preferences of small ruminant farmers to improve the sustainability of their breeding systems.

Small ruminant farming is of socio-economic and environmental importance to many rural communities around the world. The SMARTER H2020 project aims to redefine genetic selection criteria to increase the sustainability of the sector. The objective of this study was to analyse the selection and breeding management practices of small ruminant producers and breeders, linked with socio-technical elements that shape them. The study is based on farm surveys using semi-structured interviews conducted in five countries (France, Spain, Italy, Greece, and Uruguay) across 272 producers and breeders of 13 sheep and goat breeds, and 15 breed × system combinations. The information was collected in four sections. The first and second sections dealt with general elements of structure and management of the system and the flock/herd. The third section focused on selection and breeding management practices: criteria for culling and replacement of females, selection criteria for males, use of estimated breeding values and global indexes, and preferences for indexing new traits to increase the sustainability of their system. The fourth section aimed to collect socio-technical information. We used a data abstraction method to standardise the representation of these data. A mixed data factor analysis followed by a hierarchical ascending classification allowed the characterisation of three profiles of selection and breeding management: (1) a profile of producers (n = 93) of small flocks/herds, with little knowledge or use of genetic selection and improvement tools (selection index, artificial insemination, performance recording); these farmers do not feel that new traits are needed to improve the sustainability of their system. (2) a profile of producers (n = 34) of multibreed flocks/herds that rely significantly on grazing; they are familiar with genetic tools, they currently use AI; they would like the indexes to include more health and robustness characteristics, to make their animals more resistant and to increase the sustainability of their system. And (3) a profile of producers or breeders (n = 145) of large flocks/herds, with specific culling criteria; these farmers are satisfied with the current indexes to maintain the sustainability of their system. These results are elements that can be used by private breeding companies and associations to support the evolution of selection objectives to increase the resilience of animals and to improve the sustainability of the small ruminant breeding systems.

Effect of temporary weaning and creep feeding on calf growth and the reproductive efficiency of their Hereford dams.

OBJECTIVE: The objective was to test if creep feeding (CF) improves the average daily gain (ADG) and weaning weight of calves submitted to temporary weaning (TW) and if the combination of CF and TW improves conception and pregnancy rates of cows. METHODS: Primiparous (n = 74) and primiparous and multiparous (n = 104) cows grazing native grasslands were used in experiment 1 and 2; respectively. The experimental design was in plots divided into complete random blocks with two replications. The CF was the big plot and TW the small plot, thus four experimental groups were formed: i) -CF-TW (n = 21 and 27); ii) -CF+TW (n = 16 and 24); iii) +CF-TW (n = 20 and 26); iv) +CF+TW (n = 17 and 27) with cow-calf pairs for experiments 1 and 2; respectively. Nose plate application for TW had a duration of 14 and 15 days for experiment 1 and 2: respectively. In experiment 1, calves were fed at 1% of live weight for 112 days using a commercial supplement with 18.4% crude protein. In experiment 2, the supplementation lasted 98 days, and was carried out with corn dried distillers grains with soluble (DDGS) at 40% of the potential intake on a daily basis. RESULTS: The TW reduced ADG during the TW period and the following 14 days, but the negative effect of TW was maintained until the final weaning only in experiment 2. The CF increased ADG during TW period in both experiments. The TW promoted an earlier conception of the dams (12 days in -CF treatment and 19 days in +CF treatment, p<0.01) and CF increased pregnancy rate in experiment 1, being the effects not consistent between experiments. CONCLUSION: The CF consistently promoted an increase in ADG during the period of TW and increased final weaning weight of calves, therefore it is economically viable.

Long-term effect of early nutrition on endocrine parameters and liver and endometrial gene expression of the members of the somatotrophic axis in Hereford heifers.

This study compared the effect of different management systems on endocrine parameters, and gene expression of members of the somatotrophic axis in the liver and endometrium of beef heifers. Twenty-two 709-days-old heifers submitted to Early Weaning (EW, n = 8), Traditional Weaning (TW, n = 7) and TW plus creep feeding (TW+CF, n = 7) were used. Animals were synchronized with two prostaglandin (PG) injections at 11-day interval (Oestrus = Day 0). Blood samples were collected daily for progesterone (P4) determination, and endometrial and liver biopsies on Days 7 and 16 for transcript determination of members of the somatotrophic axis. Progesterone concentrations were greater on Days 15 and 16 (p < .02) of the cycle in TW+CF than TW and EW heifers. On Day 7, TW+CF heifers expressed greater liver total growth hormone receptor transcripts than TW heifers (p = .05) and greater insulin-like growth factor (IGF)-binding protein 3 mRNA than both EW and TW groups (p < .05). On Days 7 and 16, TW+CF expressed more endometrial IGF1 mRNA than the other groups (p < .01). We conclude that increasing the plane of nutrition of nursing calves may have a long-term effect on the functioning of the somatotrophic axis both in the liver and in the endometrium.

Detection and genetic characterization of Porcine circovirus type 3 in Italy.

Porcine circovirus type 3 (PCV3) was recently proposed as a new porcine circovirus. It has been described by researchers in the USA and China and associated with porcine dermatitis and nephropathy syndrome, reproductive failure and systemic inflammation disease. The study reports the occurrence of the new virus in Italy. PCV3 was detected in the tissues of foetuses and stillborn piglets coming from two farms located in the Po Valley. The genome sequences of the two Italian strains share 99.7% to 97.8% of nucleotide identity with those available in GenBank. Results strengthen the hypothesis of PCV3 as a new emerging porcine circovirus, widespread all over the world. It follows the urgency of investigating in depth epidemiology and pathogenicity associated with this new virus.

Isolation and molecular identification of Vibrio spp. by sequencing of 16S rDNA from seafood, meat and meat products in Libya.

The genus Vibrio includes several food-borne pathogens that cause a spectrum of clinical conditions including septicemia, cholera and milder forms of gastroenteritis. Several Vibrio spp. are commonly associated with food-borne transmission including Vibrio cholerae, Vibrio parahemolyticus, and Vibrio vulnificus. Microbiological analysis for enumeration and isolation of Vibrio spp. were carried out for a total of 93 samples of seafood, meat and meat products from different geographic localities in Libya (Tripoli, Regdalin, Janzour and Tobruk). Vibrio spp. were detected by conventional cultural and molecular method using PCR and sequencing of 16S rDNA. Out of the 93 cultured samples only 48 (51.6%) yielded colonies on Thiosulfate Citrate Bile Salt agar (TCBS) with culture characteristics of Vibrio spp. More than half (n=27) of processed seafood samples (n=46) yielded colonies on TCBS, while only 44.6 % of samples of meat and meat products showed colonies on TCBS. Among cultured seafood samples, the highest bacterial count was recorded in clam with a count of 3.8 ×10(4) CFU\g. Chicken burger samples showed the highest bacterial count with 6.5 ×10(4) CFU\g. Molecular analysis of the isolates obtained in this study, showed that 11 samples out of 48 (22.9%) were Vibrio spp. Vibrio parahemolyticus was isolated from camel meat for the first time. This study is an initial step to provide a baseline for future molecular research targeting Vibrio spp. foodborne illnesses. This data will be used to provide information on the magnitude of such pathogens in Libyan seafood, meat and meat products.

BET protein inhibition shows efficacy against JAK2V617F-driven neoplasms.

Small molecule inhibition of the BET family of proteins, which bind acetylated lysines within histones, has been shown to have a marked therapeutic benefit in pre-clinical models of mixed lineage leukemia (MLL) fusion protein-driven leukemias. Here, we report that I-BET151, a highly specific BET family bromodomain inhibitor, leads to growth inhibition in a human erythroleukemic (HEL) cell line as well as in erythroid precursors isolated from polycythemia vera patients. One of the genes most highly downregulated by I-BET151 was LMO2, an important oncogenic regulator of hematopoietic stem cell development and erythropoiesis. We previously reported that LMO2 transcription is dependent upon Janus kinase 2 (JAK2) kinase activity in HEL cells. Here, we show that the transcriptional changes induced by a JAK2 inhibitor (TG101209) and I-BET151 in HEL cells are significantly over-lapping, suggesting a common pathway of action. We generated JAK2 inhibitor resistant HEL cells and showed that these retain sensitivity to I-BET151. These data highlight I-BET151 as a potential alternative treatment against myeloproliferative neoplasms driven by constitutively active JAK2 kinase.

Influenza A pandemic (H1N1) 2009 virus outbreak in a cat colony in Italy.

In April 2009, a novel H1N1 influenza A virus (pH1N1) was recognized as the cause of the flu pandemic in humans. Here, we report the isolation of pH1N1 virus from the lung homogenates of two cats, which died after severe respiratory symptoms. The cats belonged to a cat colony consisting of 90 caged cats and were found dead following a 2-week period of respiratory and gastrointestinal diseases in the colony. During the outbreak, 25 cats died and 50% of the animal colony showed anorexia, depression, respiratory and gastrointestinal symptoms. Histological examination of the lungs of the two tested cats displayed lesions centred on terminal airways with epithelial bronchiolar hyperplasia and alveolar necrosis. Influenza A virus was detected in the lung tissues by immunohistochemistry and real-time RT-PCR (rRT-PCR). Partial sequences of haemagglutinin (HA) genes and complete sequences of neuraminidase (NA) genes of the two isolates displayed high similarity to the pH1N1 viruses circulating in humans (99% for HA gene and 100% for NA gene). To determine whether the pandemic virus had circulated among cats, serum samples and pharyngeal swabs were collected from 38 cats of the colony. Serum samples were tested by ELISA to detect antibodies against pH1N1 nucleoprotein and by hemagglutination-inhibition test, while pharyngeal swabs were examined by pH1N1 specific rRT-PCR. Twenty-one (55%) of the tested cats carried antibodies against the isolated strain and two swabs were positive for the presence of pH1N1 RNA. Our results confirm that the pH1N1 virus was able to infect cats and raise the hypothesis of the circulation of the virus within the colony being due to cat-to-cat transmission. The case reported here provides, to the best of the authors' knowledge, the first description of the pH1N1 infection involving numerous cats that lived in a restricted area with limited contact with humans.

Molecular characterization and phylogenetic analysis of VP1 of porcine enteric picornaviruses isolates in Italy.

Porcine enterovirus (PEV), Porcine Teschovirus and Porcine sapelovirus, belonging to the family Picornaviridae, are ubiquitous and mainly cause asymptomatic infections in pigs. In this study, a total of 40 Italian porcine picornavirus isolates were characterized by sequencing the capsid VP1-encoding gene. This procedure turned out to be a useful diagnostic tool for the molecular identification of porcine enterovirus, teschovirus and sapelovirus strains and for the study of molecular epidemiology and evolution of these viruses confirming the possibility of correlating virus genotype to serotype.

Co-existence of classical scrapie and Nor98 in a sheep from an Italian outbreak.

Nor98 is an atypical scrapie strain characterized by a molecular pattern and brain distribution of the pathological prion protein (PrP(Sc)) different from classical scrapie. In Italy, 69 atypical cases have been identified so far and all were characterized as Nor98 strain. In this paper we report an unusual case in a sheep which showed immunohistochemical and molecular features of PrP(Sc) different from the other atypical cases. The sheep was from an outbreak where the index and the other four cases were affected by classical scrapie. Histopathological, immunohistochemical and Western blot analyses on the brain of the unusual case revealed the simultaneous presence of pathological features characteristic of Nor98 and classical scrapie. Interestingly, the prevalent disease phenotype in the brainstem was classical scrapie-like, while in the cerebral cortex and cerebellum the Nor98 phenotype was dominant. The sub-mandibular lymph node was positive and showed a PrP(Sc) molecular pattern referable to classical scrapie. The PrP genotype was AL(141)RQ/AF(141)RQ. Taken together, the occurrence of classical scrapie in the outbreak, the PrP genotype, the involvement of different cellular targets in the brain and the pathological and molecular PrP(Sc) features observed suggest that this unusual case may result from the co-existence of Nor98 and classical scrapie.

Mycobacterium porcinum strains isolated from bovine bulk milk: implications for Mycobacterium avium subsp. paratuberculosis detection by PCR and culture.

In this study, the isolation of 52 mycobactin-independent fast growing mycobacteria from 631 bulk milk samples (8.2%), is reported. These strains, isolated during a bulk milk survey for Mycobacterium avium subsp. paratuberculosis (Map), strongly affected Map detection both by PCR and by culture, as they gave a positive IS900 PCR signal and resulted to totally inhibit the growth of Map when spotted on HEYM slants already inoculated with 200 microl of 10-fold dilutions containing from 5 x 10 to 5 x 10(3)Map cells/ml. 16S rRNA gene sequencing, using the MicroSeq 500 16S rDNA Bacterial Sequencing Kit (Applied Biosystems), was performed on a subset of six strains, identifying Mycobacterium porcinum with 100% homology in all six cases. The 52 strains were characterized by PCR-restriction fragment length polymorphism (RFLP) analysis of the hsp65 gene, which confirmed the identification of M. porcinum for all the isolates. Using specific primers designed on the Map-IS900 sequence and on the M. porcinum sequence determined in this study, a 1385bp sequence from the M. porcinum genome was characterized. This IS900-like sequence showed 82% homology with Map IS900. From our findings the following results emerged: (a) any culture showing one or more M. porcinum colonies represents a potential "false negative" result and should therefore be considered as contaminated; (b) IS900-like elements could be more widespread than was previously thought; (c) IS900 PCR positive results should be interpreted cautiously, as confirmed by the evidence that the primer pair used in this study resulted not to be specific.

Interpersonal problem areas and onset of panic disorder.

BACKGROUND: Numerous studies indicate that stressful life events are key precipitants of psychological disturbances. Severe stress often precedes the onset or exacerbation of illness in vulnerable individuals and may be of primary importance in the genesis of some mental disorders. Several authors have suggested that major life events play a role in the development of panic-related symptoms. The aim of this study was to examine the presence of stressful life events, in particular events focused in the interpersonal psychotherapy problem areas (grief, role disputes, role transitions, interpersonal deficits), in patients suffering from panic disorder. METHODS: We interviewed 55 patients suffering from panic disorder, with or without agoraphobia, in accordance with the diagnostic criteria specified in MINI PLUS. The panic attack profile was evaluated with the Panic Attack and Anticipatory Anxiety Scale. We assessed the ability to adapt to and derive satisfaction from the social environment with the Social Adjustment Scale Self Report and interpersonal problems with Interpersonal Questionnaire. RESULTS: Using the Interpersonal Questionnaire we found that all subjects had had relevant interpersonal problems in the year preceding the onset of PD: 92.7% had experienced a role transition, 85.5% interpersonal deficits, 74.5% a role dispute and 38.2% had suffered the loss of a relative or significant other. These results were confirmed by Paykel's scale, on which the whole sample reported a high frequency of life events in the 6 months before onset of illness. These preliminary data suggest a rationale for the therapeutic strategies of interpersonal psychotherapy in individuals with panic disorder.

Interleukin-10 modulates neuronal threshold of vulnerability to ischaemic damage.

Interleukin-10 (IL-10) is a powerful suppressor of cellular immune responses, with a postulated role in brain inflammation. First, we have evaluated the role of this cytokine in ischaemic brain damage using IL-10 knockout (IL-10-/-) mice. The middle cerebral artery (MCA) was occluded in either IL-10-/- or wild-type animals of corresponding strain (C57Bl/6) and age. Infarct volume was assessed 24 h later in serial brain sections. Brain infarct produced by MCA occlusion was 30% larger in the IL-10-/- than in wild-type mice (21. 8 +/- 1.2 vs. 16.9 +/- 1.0 mm3, respectively; P < 0.01; Student's t-test). To further characterize these findings, studies were extended to in vitro models. Primary neuronal cortical cultures derived from IL-10-/- animals were more susceptible to both excitotoxicity and combined oxygen-glucose deprivation compared with cell cultures from wild-type mice. Moreover, when added to the culture medium, recombinant murine IL-10 (0.1-100 ng/mL) exerted a concentration-dependent prevention of neuronal damage induced by excitotoxicity in both cortical and cerebellar granule cell cultures taken from either strain. The accordance of in vivo and in vitro data allows us to suggest a potential neuroprotective role of IL-10 against cerebral ischaemia when administered exogenously or made available from endogenous sources.

Chromosome segregation from cell hybrids. VII. Reverse segregation from karyoplast hybrids suggests control by cytoplasmic factors.

Using human and Chinese hamster established lines as cell parents, we constructed hamster-human cell hybrids and human cell - hamster karyoplast hybrids. The cell hybrids retained one or two sets of hamster chromosomes and lost most of the human chromosomes. The karyoplast hybrids, however, retained a full set of human chromosomes and lost most of the Chinese hamster chromosomes. This reverse segregation pattern implies that cytoplasmic factors are major determinants of the direction of chromosome segregation.

Use of a cell hybrid test system to demonstrate that benomyl induces aneuploidy and polyploidy.

We have monitored the segregation of a single human chromosome in a human-Chinese hamster hybrid cell line, EUBI, following exposure to benomyl. We found a dose-dependent increase in frequency of aneuploidy, but a much more marked induction of polyploidy was noted at the highest benomyl concentration. We confirm the usefulness of this assay for determining genetic risk associated with human exposure to environmental chemicals.

Foreign bodies of the biliary tract. Endoscopic management.

In the period 1978-1984, 23 patients underwent endoscopic intervention for foreign bodies of the biliary tract. The patients are subdivided into three groups: the first group consists of 11 cases in which the foreign bodies were suture threads, either simple or as a nidus for gallstones; the second group consists of six patients with a sump syndrome of the biliary tract; the third group includes six patients who retained drainage tubes or stents after a biliary tract operation. In our series, endoscopic extraction was performed as a first-choice procedure. The high success rate may favor endoscopy as a low-morbidity, low-mortality approach and as an alternative to a relaparotomy.

Gastrointestinal hemorrhage from an aortoduodenal fistula: endoscopic considerations.

A case of gastrointestinal hemorrhage from a secondary aortoduodenal fistula, treated with removal of the prosthesis and reconstruction of duodenal and aortic wall, is reported. The endoscopic findings that provided a preoperative diagnosis are discussed.

Bile-acid binding to isolated rat liver plasma membranes. Failure to find a specific binding site.

Bile-acid transport across the hepatocyte is an active Na-dependent process and specific putative binding sites on liver plasma membrane have been described. We studied cholic-acid binding to isolated liver plasma membranes over a wide concentration range. We found no saturability of binding over a concentration range of 0.001-2000 microM. We did demonstrate saturability of binding over a concentration range (1-30 nM) at pH 6.0 and 4 degrees C, but the saturability was an artifact caused by the increasing concentration of ethanol necessary to avoid precipitation of cholic acid at pH 6.0, 4 degrees C. We were unable to eliminate specific binding by heating the plasma membrane for 3 h at 37 degrees C, as other authors have. Although we were able to show a structural specificity of the bile-acid uptake site on the isolated liver cells, we were unable to demonstrate a specific saturable bile-acid binding site on isolated liver plasma membranes.