Mastitis pathogens and antibiotic resistance in beef cows in Switzerland.

INTRODUCTION: Mastitis in beef cows has not been studied as extensively as mastitis in dairy cows, and data from Switzerland are lacking. Various studies have shown a similar pathogen spectrum as in dairy cows, which could not be confirmed in this study. To gather initial data from Switzerland, milk samples from 297 lactating beef cows from 31 herds from the Engadin Valley in the Canton of Grisons were examined bacteriologically. At least one major or minor mastitis pathogen was recovered from at least one individual-quarter or composite sample from 33 % of all cows. The most common major mastitis pathogens were Staphylococcus aureus (8,4 % of cows), Pasteurella multocida (4,1 %), Streptococcus uberis (2 %) and Streptococcus dysgalactiae (1,7 %). Sixteen percent of the cows had at least one blind quarter, but only 32 % of these had been previously detected by the owners. In the second part of the study, milk samples from beef cows with mastitis were examined bacteriologically; the cows originated from various parts of Switzerland and had been presented for veterinary treatment. Pasteurella multocida (22 %) and Staphylococcus aureus (21 %) were the most common pathogens isolated. Antibiograms using microtitration and disk diffusion testing were generated for the Staphylococcus aureus, Pasteurella multocida and Streptococcus uberis strains from both parts of the study. Fifty-six percent of the Staphylococcus aureus strains were resistant to penicillin G. Our results showed that bacteriological examination of a milk sample aids in the diagnosis and allows specific treatment of mastitis in beef cows; this may be further improved with antibacterial susceptibility testing. Our preliminary data for the resistance patterns of mastitis pathogens in beef cows will facilitate evidence-based treatment strategies.

INTRODUCTION: Les mammites chez les vaches mères n'ont pas été étudiées de manière aussi approfondie que chez les vaches laitières et les données concernant la Suisse font défaut. Diverses études ont montré un spectre pathogène similaire à celui des vaches laitières, ce qui n'a pas pu être confirmé dans cette étude. Pour rassembler les premières données en Suisse, des échantillons de lait de 297 vaches mères provenant de 31 troupeaux de la vallée de l'Engadine dans le canton des Grisons ont été examinés bactério- logiquement. Au moins un agent pathogène majeur ou mineur de mammite a été retrouvé dans au moins un quartier ou dans un échantillon composite chez 33 % de toutes les vaches. Les agents pathogènes majeurs de mammite les plus courants étaient Staphylococcus aureus (8,4 % des vaches), Pasteurella multocida (4,1 %), Streptococcus uberis (2 %) et Streptococcus dysgalactiae (1,7 %). Seize pour cent des vaches avaient au moins un quartier sec mais cela n'avait été détectés auparavant par les propriétaires que dans seulement 32 % des cas. Dans la deuxième partie de l'étude, des échantillons de lait provenant de vaches mères atteintes de mammites ont été examinés sur le plan bactériologique; les vaches provenaient de diverses régions de Suisse et avaient été présentées pour un traitement vétérinaire. Pasteurella multocida (22 %) et Staphylococcus aureus (21 %) étaient les agents pathogènes les plus fréquemment isolés. Des antibiogrammes utilisant des tests de microtitration et de diffusion sur disque ont été générés pour les souches de Staphylococcus aureus, Pasteurella multocida et Streptococcus uberis des deux parties de l'étude. Cinquante-six pour cent des souches de Staphylococcus aureus étaient résistantes à la pénicilline G. Nos résultats montrent que l'examen bactériologique d'un échantillon de lait facilite le diagnostic et permet un traitement spécifique des mammites chez les vaches mères; ceci peut être encore amélioré par des tests de sensibilité aux antibactériens. Nos données préliminaires sur les profils de résistance des agents pathogènes de mammites chez les vaches de boucherie faciliteront les stratégies de traitement fondées sur des faits.

Efecto de la dibucaína sobre la SERCA del músculo pterigoideo interno / Drug action of dibucaine on the SERCA from medial pterygoid muscle

Objetivo: Determinar el efecto del anestésico local di-bucaína sobre las principales isoformas de la SERCA (calcio ATPasa de retículo sarco-endoplásmico) pre-sentes en músculo pterigoideo interno. Métodos: Se aislaron por centrifugación diferencial membranas de retículo sarcoplásmico de pterigoideo interno de conejo neozelandés macho (n=5). Se separaron las isoformas SERCA1a, 2a y 2b por cromatografía de afinidad. Se determinó in vitro la actividad enzimá-tica en presencia de diferentes concentraciones de dibucaína (0-90 mM) por el método de Fiske y Subba-row, realizando 5 experimentos por duplicado y en paralelo para cada isoforma. Se calculó la media y ES de la CI50 (mM) del anestésico para cada isofor-ma y éstas se compararon por ANOVA de una vía (p<0,05), y prueba Student-Newman-Keuls de com-paraciones múltiples. Resultados: Dibucaína inhibió la actividad enzimática en función de su concentra-ción en las tres isoformas en estudio. Las CI50 fueron: SERCA1a 20,02 ± 0,64 mM, SERCA2a 15,03 ± 0,52 mM y SERCA2b 16,00 ± 0,51 mM y resultaron signi-ficativamente diferentes (F2,27 = 11,08, p<0,001). La prueba post hoc identificó diferencias significativas entre SERCA1a y 2a, 1a y 2b. El efecto inhibitorio re-sultó significativamente mayor sobre las isoformas 2a y 2b, cuya presencia es sustancialmente mayor en músculos masticadores. Conclusión: La dibucaína inhibe a la SERCA de pterigoideo interno a concen-traciones menores que las usadas en clínica médica (29 mM). Es un anestésico local con potencial efecto miotóxico derivado de la inhibición de la SERCA (AU)

Aim: To test the effect of the local anesthetic dibu-caine on the main isoforms of the SERCA (sarco-endosplasmic reticulum calcium-ATPase) in medial pterygoid muscle. Methods: Sarcoplasmic reticulum membranes from male New Zealand rabbits (n=5) were isolated from medial pterygoid muscle by ul-tracentrifugation. The isoforms SERCA1a, 2a and 2b were separated using high affinity chromatography. In vitro enzymatic activity determinations were per-formed in the presence of different dibucaine con-centrations (0-90 mM) using the colorimetric method described by Fiske & Subbarow. Five assays in dupli-cate and run in parallel were performed for each of the isoforms. Mean and SEM of the IC50 (mM) for the effect of the anesthetic on each isoform were calcu-lated and compared by one-way ANOVA (p<0.05), and Student-Newman-Keuls multiple comparisons test. Results: Dibucaine inhibited the enzymatic activity in a concentration-dependent manner for the three studied isoforms. The IC50 values were: SERCA1a 20.02 ± 0.64 mM, SERCA2a 15.03 ± 0.52 mM and SER-CA2b 16.00 ± 0.51 mM. The values were significantly different (F2.27 = 11.08, p<0.001). The post hoc test revealed significant differences between SERCA1a and 2a, 1a and 2b. The inhibitory effect was signifi-cantly higher on 2a and 2b isoforms, whose presence is substantially higher in masticatory muscles. Con-clusion: Dibucaine inhibits SERCA in medial pterygoid muscle at concentrations lower than those used in clinical medicine (29 mM). It is a potentially myotoxic local anesthetic whose toxic effect may derive from SERCA inhibition (AU)

Pathogen dependent effects of high amounts of oxytocin on the bloodmilk barrier integrity during mastitis in dairy cows.

INTRODUCTION: The reduction of antibiotic use in food producing animals becomes increasingly important. Therefore, suitable alternatives for mastitis treatment in dairy cows have to be considered. Oxytocin (OT) induces milk ejection and hence supports milk removal from infected mammary quarters. Beyond udder emptying, the injection of very high dosages of OT causes increased somatic cell counts (SCC) in milk and enables the transfer of immunoglobulins (Ig) from blood into milk through a reduced blood-milk barrier integrity. The aim of the present study was to investigate pathogen-specific changes of SCC, the blood derived milk components lactate dehydrogenase (LDH), serum albumin (SA), and IgG in milk of cows suffering from mastitis caused by different pathogens treated with two intravenous injections of high dosages of OT (100 IU). Milk samples from 184 dairy cows from different farms were collected on day 1 (day of clinical examination and mastitis diagnosis) and on days 2, 3, 14, and 28. Bacteriological examination (day 1) identified involved pathogens. Cows were randomly assigned to treatment (OT injections on days 1 and 2) or control group (no OT). Independently of the assigned experimental group, cows received the common therapy protocol of the veterinary practice after sample collection if the general condition was affected. Milk SCC, LDH, SA, and IgG changed specifically depending on involved pathogens. Highest values of all three parameters were measured in mastitis caused by Streptococcus uberis. Changes were less pronounced with other Streptococci spp., Staphylococci spp. or Corynebacterium bovis. Oxytocin treatment did not affect any of the studied parameters independent of the involved pathogen. Only in quarters infected with Staphylococci other than Staphylococcus aureus a decreased SCC and increased IgG concentrations in quarters, where no pathogens were detected, were observed. Thus, high dosage OT administration is obviously not suitable as a stand-alone mastitis treatment in dairy cows.

INTRODUCTION: La réduction de l'utilisation d'antibiotiques chez les animaux destinés à l'alimentation devient de plus en plus importante. Par conséquent, des alternatives appropriées au traitement des mammites chez les vaches laitières doivent être envisagées. L'ocytocine (OT) induit l'éjection du lait et favorise donc l'élimination du lait des quartiers infectés. Au-delà de la vidange de la mamelle, l'injection de doses très élevées d'OT entraîne une augmentation du nombre de cellules somatiques (CSC) dans le lait et permet le transfert d'immunoglobulines (Ig) du sang vers le lait grâce à une réduction de l'intégrité de la barrière sang-lait. Le but de la présente étude était d'étudier les changements spécifiques aux agents pathogènes du CSC, les composants du lait dérivés du sang que sont la lactate déshydrogénase (LDH) et l'albumine sérique (SA) ainsi que les IgG dans le lait de vaches souffrant de mammites causées par différents agents pathogènes traités par deux injections intraveineuses de doses élevées d'OT (100 UI). Des échantillons de lait de 184 vaches laitières de différentes exploitations ont été prélevés au jour 1 (jour de l'examen clinique et diagnostic de mammite) et aux jours 2, 3, 14 et 28. L'examen bactériologique (jour 1) a identifié les agents pathogènes impliqués. Les vaches ont été assignées au hasard au traitement (injections d'OT les jours 1 et 2) ou au groupe témoin (pas d'OT). Indépendamment du groupe auquel elles étaient attribuées, les vaches ont reçu le protocole thérapeutique usuel du cabinet vétérinaire après le prélèvement de l'échantillon si leur état général était affecté. Le CSC, la LDH, la SA et les IgG du lait ont varié spécifiquement en fonction des agents pathogènes impliqués. Les valeurs les plus élevées des trois paramètres ont été mesurées dans les mammites causées par Streptococcus uberis. Les changements étaient moins prononcés avec d'autres Streptococci spp., Staphylococci spp. ou Corynebacterium bovis. Le traitement à l'ocytocine n'a affecté aucun des paramètres étudiés indépendamment de l'agent pathogène impliqué. On a uniquement observé, dans les mammites causées par des staphylocoques autres que Staphylococcus aureus, une diminution du CSC et, dans les mammites où aucun agent pathogène n'a été détecté, une augmentation des concentrations d'IgG dans les quartiers. Ainsi, l'administration d'OT à forte dose n'est pas appropriée comme traitement unique des mammites chez les vaches laitières.

Synaptotagmin 13 is neuroprotective across motor neuron diseases.

In amyotrophic lateral sclerosis (ALS) and spinal muscular atrophy (SMA), spinal and lower brainstem motor neurons degenerate, but some motor neuron subtypes are spared, including oculomotor neurons (OMNs). The mechanisms responsible for this selective degeneration are largely unknown, but the molecular signatures of resistant and vulnerable motor neurons are distinct and offer clues to neuronal resilience and susceptibility. Here, we demonstrate that healthy OMNs preferentially express Synaptotagmin 13 (SYT13) compared to spinal motor neurons. In end-stage ALS patients, SYT13 is enriched in both OMNs and the remaining relatively resilient spinal motor neurons compared to controls. Overexpression of SYT13 in ALS and SMA patient motor neurons in vitro improves their survival and increases axon lengths. Gene therapy with Syt13 prolongs the lifespan of ALS mice by 14% and SMA mice by 50% by preserving motor neurons and delaying muscle denervation. SYT13 decreases endoplasmic reticulum stress and apoptosis of motor neurons, both in vitro and in vivo. Thus, SYT13 is a resilience factor that can protect motor neurons and a candidate therapeutic target across motor neuron diseases.

Back to the origins: Human brain organoids to investigate neurodegeneration.

Neurodegenerative disorders represent a high burden in terms of individual, social and economical resources. No ultimate therapy has been established so far; human brain morphology and development can not be entirely reproduced by animal models, and genomic, metabolic and biochemical differences might contribute to a limited predictive power for human translation. Thus, the development of human brain organoid models holds a wide potential to investigate the range of physiological and pathological features that characterise the early onset of the degeneration. Moreover, central nervous system development has gained a crucial role in the study of the pathogenesis of neurodegenerative disorders. Premature alterations during brain maturation have been related to late disease manifestations; genetic mutations responsible for neurodegeneration have been found in genes highly expressed during neural development. Elucidating the mechanisms triggering neuronal susceptibility to degeneration is crucial for pathogenetic studies and therapeutic discoveries. In the present work, we provide an overview on the current applications of human brain organoids towards studies of neurodegenerative diseases, with a survey on the recent discoveries and a closing discussion on the present challenges and future perspectives.

The Sensitivity of Euro-Atlantic Regimes to Model Horizontal Resolution.

There is growing evidence that the atmospheric dynamics of the Euro-Atlantic sector during winter is driven in part by the presence of quasi-persistent regimes. However, general circulation models typically struggle to simulate these with, for example, an overly weakly persistent blocking regime. Previous studies have showed that increased horizontal resolution can improve the regime structure of a model but have so far only considered a single model with only one ensemble member at each resolution, leaving open the possibility that this may be either coincidental or model dependent. We show that the improvement in regime structure due to increased resolution is robust across multiple models with multiple ensemble members. However, while the high-resolution models have notably more tightly clustered data, other aspects of the regimes may not necessarily improve and are also subject to a large amount of sampling variability that typically requires at least three ensemble members to surmount.

Staphylococcus aureus related to bovine mastitis in Switzerland: Clonal diversity, virulence gene profiles, and antimicrobial resistance of isolates collected throughout 2017.

Staphylococcus aureus can be associated with subclinical, acute, chronic, and toxic cases of bovine intramammary infections, leading to considerable financial losses for the dairy industry in Switzerland and worldwide. In addition, milk products are one of the most common food categories implicated in staphylococcal food poisoning in humans. Detailed information on the population structure, as well as the virulence and resistance characteristics of Staph. aureus originating from bovine mastitis milk is needed to allow for source attribution and risk assessment of Staph. aureus in a food poisoning context and to improve therapeutic approaches in cattle. Our objective was to assess the population structure, phenotypic resistance patterns, and virulence and resistance gene profiles of Staph. aureus isolates from bovine mastitis milk in Switzerland. To this end, 58 Staph. aureus strains were characterized. The DNA microarray was used to test for the presence or absence of virulence and resistance genes. In addition, minimum inhibitory concentrations of various antimicrobial agents were determined by microdilution. To assess the population structure of the isolates, we determined clonal complexes (CC) using DNA microarray hybridization profiles and performed multilocus sequence typing and spa typing. The strains were assigned to 7 clonal complexes, 10 sequence types, and 11 spa types, with CC705 (43%), CC97 (33%), and CC20 (12%) representing the most common lineages and t529 (43%) and t267 (21%) representing the most common spa types. Only 1 isolate was assigned to CC8, a clonal lineage linked to high within-herd prevalence of mastitis. A total of 14% (n = 8) of strains were classified as resistant to penicillin, and 1 strain each was classified as oxacillin and pirlimycin resistant. Although no clinical breakpoints are available for the combination of kanamycin/cefalexin, growth of all strains was inhibited by the lowest combination of kanamycin/cefalexin concentrations tested (4 µg/mL of kanamycin and 0.4 µg/mL of cefalexin). One strain assigned to CC20, ST389, and t2094 exhibited resistance to penicillin, oxacillin, and pirlimycin as well as intermediate susceptibility to erythromycin and high minimum inhibitory concentration for several antimicrobial agents, for which no breakpoints were available.

In Vivo Transient and Partial Cell Reprogramming to Pluripotency as a Therapeutic Tool for Neurodegenerative Diseases.

In theory, human diseases in which a specific cell type degenerates, such as neurodegenerative diseases, can be therapeutically addressed by replacement of the lost cells. The classical strategy for cell replacement is exogenous cell transplantation, but now, cell replacement can also be achieved with in situ reprogramming. Indeed, many of these disorders are age-dependent, and "rejuvenating" strategies based on cell epigenetic modifications are a possible approach to counteract disease progression. In this context, transient and/or partial reprogramming of adult somatic cells towards pluripotency can be a promising tool for neuroregeneration. Temporary and controlled in vivo overexpression of Yamanaka reprogramming factors (Oct3/4, Sox2, Klf4, and c-Myc (OSKM)) has been proven feasible in different experimental settings and could be employed to facilitate in situ tissue regeneration; this regeneration can be accomplished either by producing novel stem/precursor cells, without the challenges posed by exogenous cell transplantation, or by changing the epigenetic adult cell signature to the signature of a younger cell. The risk of this procedure resides in the possible lack of perfect control of the process, carrying a potential oncogenic or unexpected cell phenotype hazard. Recent studies have suggested that these limits can be overcome by a tightly controlled cyclic regimen of short-term OSKM expression in vivo that prevents full reprogramming to the pluripotent state and avoids both tumorigenesis and the presence of unwanted undifferentiated cells. On the other hand, this strategy can enhance tissue regeneration for therapeutic purposes in aging-related neurological diseases as well. These data could open the path to further research on the therapeutic potential of in vivo reprogramming in regenerative medicine.

Milk somatic cell count, lactate dehydrogenase activity, and immunoglobulin G concentration associated with mastitis caused by different pathogens: A field study.

INTRODUCTION: The aim of this study is to analyze how somatic cell counts (SCC), immunoglobulin G (IgG), and lactate dehydrogenase (LDH) interact dependent on the mastitis causing pathogen. Milk samples from 152 quarters were collected on 2 Swiss dairy farms equipped with automatic milking systems. Bacteriological culturing was performed and SCC, LDH activity and IgG concentrations were measured in each sample. Correlations and regressions among SCC, LHD, and IgG were calculated after grouping by the pathogen type (control, S. aureus, C. bovis, coagulase-negative Staphylococcus and S. uberis). All the mastitis causing pathogens were gram-positive bacteria (except for 3 cases with E. coli). In this study, the SCC and LDH were affected by the pathogen group. However, only in the S. uberis group the IgG concentration was higher than in the controls. All studied variables were positively correlated among each other. SCC and LDH were the highest correlated parameters in the control, S. aureus, C. bovis and coagulase- negative Staphylococcus groups. Only in the S. uberis group the correlation between LDH and IgG was higher than the correlation between SCC and LDH. The regression coefficients for SCC and LDH differed between groups whereas regression coefficients for SCC and IgG, and for LDH and IgG were similar in all groups. Because cases with E. coli infection were so rare, we could not include these cases in the statistical evaluation. Based on these few cases E. coli (n=3) seemed to cause a much higher increase of IgG and LDH than the infection with gram-positive bacteria. This study shows that the suitability of LDH as a marker for IgG transfer is dependent on the pathogen. The use of LDH in combination with SCC may be used as a marker to differentiate between gram-positive and gram-negative bacteria, but does not allow differentiating the immune response between different gram-positive bacteria.

INTRODUCTION: Dans le présent travail, on a étudié les liens entre le nombre de cellules somatiques (somatic cell count, SCC), les immunoglobulines G (IgG) et la lactate déshydrogénase en fonction des pathogènes à l'origine de la mammite. On a prélevé des échantillons de lait provenant de 152 quartiers dans deux exploitations suisses équipées d'un robot de traite. Outre la mesure du SCC, des IgG et de la LDH, on a effectué un examen bactériologique des échantillons. Après classement selon les types de pathogènes (contrôle, S. aureus, C. bovis, Straphylocoques coagulase négatifs, S. uberis), on a calculé les corrélations et régressions entre SCC, LDH et IgG. Tous les pathogènes de mammite trouvés étaient des bactéries Gram positif; seuls trois quartiers étaient infectés par E. coli. Tant le SCC que la LDH étaient différents selon le type de pathogène mais les IgG n'étaient élevées par rapport aux contrôles que dans les quartiers dans lesquels on a mis en évidence S.uberis. Toutes les variables examinées étaient corrélées positivement entre elles. En ce qui concerne les contrôles, S. aureus, C. bovis et les Straphylocoques coagulase négatifs, la corrélation entre SCC et LDH était particulièrement élevée. Ce n'est que dans le groupe des S. uberis que la corrélation entre LDH et IgG était plus élevée qu'entre SCC et LDH. Les coefficients de régression entre SCC et LDH étaient différents selon les types de pathogènes, alors que la régression entre SCC et IgG de même qu'entre LDH et IgG ne présentait pas de différence suivant les groupes. Vu leur petit nombre (n=3), les quartiers infectés par E. coli n'ont pas pu être pris en compte dans l'exploitation statistique. On constate toutefois que, dans tous les quartiers infectés par E. coli, on mesure des valeurs nettement plus élevées d'IgG et de LDH que dans ceux infectés par des bactéries Gram positif. Cette étude montre que l'utilisabilité de la LDH comme marqueur du transfert des IgG du sang au lait dépend du type de pathogène. La relation entre SCC et LDH pourrait servir de marqueur pour la différenciation entre les infections à Gram positifs ou négatifs; par contre il ne semble pas possible de l'utiliser pour faire une différence entre les diverses bactéries à Gram positifs.

[Clinical epidemiology of cryptosporidiosis in calves]. / Klinische Epidemiologie der Kälber- Cryptosporidiose.

INTRODUCTION: The aim of this study was to determine whether there is an association between Cryptosporidium infections in calves and immunological factors, as well as farm-related factors or the application of the anti-cryptosporidiosis drug Halofuginone. From January to June 2010, 63 cow-calf-pairs from 20 different farms near Zürich, Switzerland have been investigated. Each cowcalf- pair was visited three times within the first 6 weeks of life to collect data of the farm and animals, as well as blood, faecal, colostral and milk samples. An ELISA using sporozoite antigen was developed for the specific detection of anti-Cryptosporidium-IgG in blood- and colostral serum. The IgG concentration in the bloodand colostral serum was determined using radial immuno diffusion test (RID). White blood cell isolation and differential blood cell counts and California Mastitis Test were performed. Bacteriological studies on quarter-milk-samples were carried out. Cryptosporidium oocysts were diagnosed with the modified Ziehl-Neelsen staining, other protozoa with the SAFC method and Eimeria oocysts and helminth eggs were diagnosed with the combined sedimentation/floatation test. ELISAs were performed for the detection of rota- and coronavirus, E. coli F5 and Cryptosporidium spp. in bovine feces (bio-X Diagnostics®, Belgium). The highest prevalence of Cryptosporidium oocysts was 54.0% and found 7 to 20 days post natum, whereas 47.1% were suffering from diarrhea. The transfer of total IgG with the colostrum and the humoral immunity of the calf could not prevent any infection with Cryptosporidium, but the severity of the diarrhea symptoms decreased with increasing total IgG concentrations. Calves housed in open sheds showed significantly more often diarrhea, i. e. they shed more Cryptosporidium oocysts during the first 4 days and 7 to 20 days post natum, respectively. Halofuginone (Halocur®) is approved for prophylaxis against cryptosporidiosis, but it showed no effect on the excretion of Cryptosporidium oocysts in the present study.

INTRODUCTION: Le but de la présente étude était d'étudier s'il existe un rapport entre l'apparition de cryptosporidies et des facteurs immunologiques, des facteurs liés à l'exploitation ainsi qu'à l'usage d'halofuginone. De janvier à juin 2010, on a examiné 63 paires mère-veau provenant de 20 exploitations du canton de Zürich. Au cours de 6 semaines on a effectué, à des moments choisis, trois visites. A ces occasions, des données relatives à l'exploitation ainsi que des échantillons de sang, de selles, de colostrum respectivement de lait ont été collectés. On a développé un test ELISA avec des antigènes de sporozoïtes pour mettre en évidence la présence IgG anti-cryptosporidies dans le sang et dans le colostrum. La concentration en IgG dans le colostrum et dans le sérum a été mesurée avec un test d'immunodiffusion radiale (RID). En outre on a réalisé une image sanguine différentielle des vaches et des veaux et effectué un test de Schalm chez les vaches. Un examen bactériologique a été réalisé sur un échantillon provenant des quatre quartiers. Les oocystes de cryptosporidies ont été mis en évidence au moyen d'une coloration de Ziehlk-Neelsen modifiée, les autres protozoaires ont été mis en évidence par la méthode SAFC et les oeufs d'helminthes ainsi que les oocystes d'Eimeria par un processus de sédimentation-flottation combiné. Un test ELISA a été utilisé pour les rota- et les coronavirus, les E. coli F5 et Cryptosporidium spp. dans les selles des bovins (Bio-X Diagnostics®, Belgique). La prévalence d'infections par des cryptosporidies était maximale entre le 7ème et le 20ème jour de vie des veaux (50.4%), 47.1% de ces veaux souffrant de diarrhée. Les stabulations libres augmentaient de façon significative le risque de diarrhée et d'excrétion de cryptosporidies entre le 1er et le 4ème jour respectivement entre le 7ème et le 20ème jour. La transmission d'IgG et l'immunité humorale des veaux n'empêchaient pas l'infection par des cryptosporidies mais la gravité de la diarrhée diminuait avec l'augmentation de la concentration des IgG totales. L'halofuginone, substance enregistrée pour la prophylaxie de la cryptosporidiose, n'a pas montré, dans cette étude, d'efficacité pour empêcher l'excrétion d'oocystes de cryptosporidies.

Morpholino-mediated SOD1 reduction ameliorates an amyotrophic lateral sclerosis disease phenotype.

Neurotoxicity due to the accumulation of mutant proteins is thought to drive pathogenesis in neurodegenerative diseases. Mutations in superoxide dismutase 1 (SOD1) are linked to familial amyotrophic lateral sclerosis (fALS); these mutations result in progressive motor neuron death through one or more acquired toxicities. Interestingly, SOD1 is not only responsible for fALS but may also play a significant role in sporadic ALS; therefore, SOD1 represents a promising therapeutic target. Here, we report slowed disease progression, improved neuromuscular function, and increased survival in an in vivo ALS model following therapeutic delivery of morpholino oligonucleotides (MOs) designed to reduce the synthesis of human SOD1. Neuropathological analysis demonstrated increased motor neuron and axon numbers and a remarkable reduction in astrogliosis and microgliosis. To test this strategy in a human model, we treated human fALS induced pluripotent stem cell (iPSC)-derived motor neurons with MOs; these cells exhibited increased survival and reduced expression of apoptotic markers. Our data demonstrated the efficacy of MO-mediated therapy in mouse and human ALS models, setting the stage for human clinical trials.

Motor neurons with differential vulnerability to degeneration show distinct protein signatures in health and ALS.

The lethal disease amyotrophic lateral sclerosis (ALS) is characterized by the loss of somatic motor neurons. However, not all motor neurons are equally vulnerable to disease; certain groups are spared, including those in the oculomotor nucleus controlling eye movement. The reasons for this differential vulnerability remain unknown. Here we have identified a protein signature for resistant oculomotor motor neurons and vulnerable hypoglossal and spinal motor neurons in mouse and man and in health and ALS with the aim of understanding motor neuron resistance. Several proteins with implications for motor neuron resistance, including GABAA receptor α1, guanylate cyclase soluble subunit alpha-3 and parvalbumin were persistently expressed in oculomotor neurons in man and mouse. Vulnerable motor neurons displayed higher protein levels of dynein, peripherin and GABAA receptor α2, which play roles in retrograde transport and excitability, respectively. These were dynamically regulated during disease and thus could place motor neurons at an increased risk. From our analysis is it evident that oculomotor motor neurons have a distinct protein signature compared to vulnerable motor neurons in brain stem and spinal cord, which could in part explain their resistance to degeneration in ALS. Our comparison of human and mouse shows the relative conservation of signals across species and infers that transgenic SOD1G93A mice could be used to predict mechanisms of neuronal vulnerability in man.

Resistance profiles and genetic diversity of Escherichia coli strains isolated from acute bovine mastitis.

Between March 2011 and February 2012 83 E. coli strains were isolated from mastitis milk samples from 83 different animals (67 farms) and tested for their sensitivity to various antibiotics by means of disk diffusion method and genotyped by determination of the phylogenetic groups as well as by pulsed field gel electrophoresis (PFGE). The antibiotics were chosen on the basis of their licenses for intramammary application in Switzerland. As many as 16.9 % of the isolates were resistant to one or more antimicrobial agents. Amoxicillin-clavulanic acid, gentamicin and third generation cephalosporins proved effective against the majority of these strains. Nevertheless, one blaCTX-M-14 harbouring extended-spectrum-beta-lactamase producing strain was found. Genetic analysis grouped most of the strains (87 %) into phylogenetic groups A and B1. PFGE genotyping demonstrated that E. coli from cows with mastitis even from the same farm were genotypically very diverse.

Entre mars 2011 et février 2012, 83 souches d'E. coli issues de 83 vaches différentes provenant de 67 exploitations ont été collectées et testées quant à leur sensibilité vis-à-vis de divers antibiotiques. Ces antibiotiques ont été choisis sur la base de leurs autorisations pour l'application intra mammaire en Suisse et le test a été effectué par diffusion sur gel d'agar. En outre toutes les souches ont été typisées quant à leur appartenance aux groupes phylogénétiques. 16.9 % des souches présentaient une résistance à un ou plusieurs antibiotiques. L'amoxicilline-acide clavulanique, la gentamicine et les céphalosporines de troisièmes générations se montraient efficaces contre la majorité des souches d'E. coli. On a toutefois trouvé une souche fabriquant un extended-spectrum-beta-lactamase qui portait le gène blaCTX-M-14. L'analyse génétique groupait la majorité des souches (87 %) dans les groupes phylogénétiques A et B1. La génotypisation par PFGE montrait une grande diversité entre les souches, même si elles provenaient de la même exploitation.

Arcanobacterium pluranimalium leading to a bovine mastitis: species identification by a newly developed pla gene based PCR.

We are describing a clinical case of bovine mastitis due to Arcanobacterium pluranimalium in a Holstein-Friesian heifer, delivering bloody milk on the left hindquarter. Moreover, we report on the development and evaluation of PCR primers based on the pluranimaliumlysin (pla) gene for the identification of this species. With the primer pair PlaF/PlaR the A. pluranimalium type strain as well as the mastitis isolate 704 revealed a correctly sized amplification product (458 bp), whereas no amplification product was obtained for all non-target strains. The established PCR provides a new and convenient tool for the mastitis diagnostic to differentiate between A. pluranimalium and Trueperella pyogenes.

Comparison of genomic and antimicrobial resistance features of latex agglutination test-positive and latex agglutination test-negative Staphylococcus aureus isolates causing bovine mastitis.

The dairy industry suffers massive economic losses due to staphylococcal mastitis in cattle. The Staphaureux latex agglutination test (Oxoid, Basel, Switzerland) was reported to lead to negative results in 54% of bovine Staphylococcus aureus strains, and latex-negative strains are thought to be less virulent than Staphaurex latex-positive strains. However, comparative information on virulence and resistance profiles of these 2 groups of Staph. aureus is scarce. Our objective was to associate the latex agglutination phenotype of Staph. aureus strains isolated from bovine mastitis milk with data on clonal complexes, virulence genes, and antibiotic resistance to (1) determine the virulence profiles of the Staphaureux test positive and Staphaurex test negative groups, and (2) provide data needed to improve treatment of bovine mastitis and to identify potential vaccine targets. Seventy-eight Staph. aureus strains isolated from 78 cows on 57 Swiss farms were characterized. Latex agglutination was tested by Staphaureux kit, and resistance profiles were generated by disk diffusion. A DNA microarray was used to assign clonal complexes (CC) and to determine virulence and resistance gene profiles. By the Staphaureux test, 49% of the isolates were latex-positive and 51% were latex-negative. All latex-negative strains were assigned to CC151, whereas latex-positive strains were assigned to various clonal complexes, including CC97 (n=16), CC8 (n=10), CC479 (n=5), CC20 (n=4), CC7 (n=1), CC9 (n=1), and CC45 (n=1). Although the latex-negative isolates were susceptible to all antimicrobial agents tested, 24% of latex-positive isolates were classified as intermediate with regard to cefalexin-kanamycin and 13% were resistant to both ampicillin and penicillin. Microarray profiles of latex-negative isolates were highly similar, but differed largely from those of latex-positive isolates. Although the latex-negative group lacked several enterotoxin genes and sak, it exhibited significantly higher prevalence rates of genes encoding enterotoxin C, toxic shock syndrome toxin, and leukocidins (lukM/lukF-P83, lukD). Our findings suggest that latex-negative isolates represent a group of closely related strains with specific resistance and virulence gene patterns.

Bacteriological survey of ready-to-eat lettuce, fresh-cut fruit, and sprouts collected from the Swiss market.

The consumption of ready-to-eat fresh vegetables has increased significantly in the recent decades. So far, no data are available on the bacteriological burden and the prevalence of foodborne pathogens in ready-to-eat lettuce, fresh-cut fruit, and sprouts on the Swiss market. This study was based on investigations carried out during 2 months of the summer season in 2011. Samples of 142 salads, 64 fresh-cut fruit, and 27 sprouts were included in this study. Escherichia coli, an indicator microorganism for fecal contamination, was only found in 5 lettuce samples, with amounts ranging between 2 and 3 log CFU/g. No Salmonella spp. were detected from any of the 233 samples analyzed in this study, and a low occurrence was found for contamination with L. monocytogenes, Shiga toxin-producing E. coli, enteropathogenic E. coli, and Cronobacter. From the results of the present study, we conclude that even in a country where the use of chlorine solutions to sanitize fruits and vegetables in the fresh-cut industry is not allowed, it is possible to produce ready-to-eat lettuce, fresh-cut fruit, and sprouts with high microbiological standards. Strict maintenance of good practices of hygiene at preharvest, harvest, and postharvest levels is of central importance to ensure both public health protection and product quality.