First evidence of Schmallenberg virus introduction in Portugal, 2014

Objectives: Serra da Estrela native (SEN) breed of sheep is one of the most important in Portugal, standing responsible for the production of one of the most internationally recognized cheeses in the world, "Queijo Serra da Estrela” (protected designation of origin). One of the major risks to the sustainability of SEN dairy flocks if footrot, an infectious diseases that causes lameness, decrease in milk production, weight loss and decrease in fertility. The aim of this work was to determine which parameters become decisive for the appearance of footrot in SEN dairy flocks, while establishing associations with environmental and nutritional variables. Materials and Methods: The present study was carried out by performing an especially designed 27 multiple choice questionnaire, based on the underlying causes of lameness in livestock and in the clinical diagnosis performed by the vet techinichian of the clinical cases evaluated at the moment of the inquiry. The survey was performed during the execution of the oficial veterinary health program between February and September of 2014 by a veterinarian team from the Association of SEN Sheep Breeders (ANCOSE). The ovine producers (N=30, with a total of 1270 animals) were randomly selected from the extense area of production of “Queijo Serra da Estrela”. The parameters evaluated in the study were: season and consequent weather changes during the period of the study, floor types, hygiene conditions, bed types, the existence and use of footbaths, location of footbaths, foot trimming and foot hygiene procedures. After the construction of the database and using the Statistical Package for Social Sciences version 16.0 the frequency response for each item was calculated. Results: All SEN livestock producers are proactive in the treatment of lameness (70%). About 99.7% of lameness was related with footroot and most appeared in winter (36.7%). In some occasions there was not a specific season distribution (56.7% rainiest years). From the analyzed farms, 70% use straw as a floor bed, followed by bush (21%). 45.6% of animals were clinically diagnosed with footrot and were sheltered in straw floor. Bed quality is good in 40%, however frequent changes in the floor bed in the preceding 15 days had a higher rate of footroot diagnosed cases (33,23%) compared to monthly changes. Regarding sheepfold animal number, the distribution is proper in 36.7%, elevated in 30%, overcrowded in 6.7% and low in 23.3%. Clinically ill diagnosed animals in last category were the lowest observed (3%). Concerning hoof trimming, 76.7% trims while 23.3% reported not to perform that task. From those that perform trimming, 73.9% do it only when necessary, 21.7% at the time of clipping and 4.4% when the animal is affected. One curious result was that animals who perform trimming at clipping had higher footrot cases (52.6%), unlike those who trim only when necessary (40.2%) or even when animals are clinically ill (0.91%). Mostly all in the presence of footrot choose a local treatment (95.2%) using antibiotic sprays, instead of parenteral antibiotic treatment. Footbath is rarely used in the prevention (13.3%) of this disease and when it is misconceived (25%) and incorrectly formulated (100%). Conclusions: This study was the first performed in Portugal focusing footrot caracterization in native SEN flocks.Economic impact in SEN dairy flocks is atributed to factors such as direct decrease in milk and meat production,early slaughter of affected animals due to non responsive treatment and medical expenses.The most effective eradication method combines first of all the ability to understand the problems of SEN producers and implementation of preventive measures and treatment of footrot.Often linked to lack of formation, the advance age of portuguese producers the mentality and social factors block the advance of veterinarian intervention itself.

Footrot caracterization in serra da estrela native flocks

Objectives: Serra da Estrela native (SEN) breed of sheep is one of the most important in Portugal, standing responsible for the production of one of the most internationally recognized cheeses in the world, "Queijo Serra da Estrela” (protected designation of origin). One of the major risks to the sustainability of SEN dairy flocks if footrot, an infectious diseases that causes lameness, decrease in milk production, weight loss and decrease in fertility.The aim of this work was to determine which parameters become decisive for the appearance of footrot in SEN dairy flocks, while establishing associations with environmental and nutritional variables. Materials and methods: The present study was carried out by performing an especially designed 27 multiple choice questionnaire, based on the underlying causes of lameness in livestock and in the clinical diagnosis performed by the vet techinichian of the clinical cases evaluated at the moment of the inquiry. The survey was performed during the execution of the oficial veterinary health program between February and September of 2014 by a veterinarian team from the Association of SEN Sheep Breeders (ANCOSE). The ovine producers (N=30, with a total of 1270 animals) were randomly selected from the extense area of production of “Queijo Serra da Estrela”. The parameters evaluated in the study were: season and consequent weather changes during the period of the study, floor types, hygiene conditions, bed types, the existence and use of footbaths, location of footbaths, foot trimming and foot hygiene procedures. After the construction of the database and using the Statistical Package for Social Sciences version 16.0 the frequency response for each item was calculated. Results: All SEN livestock producers are proactive in the treatment of lameness (70%). About 99.7% of lameness was related with footroot and most appeared in winter (36.7%). In some occasions there was not a specific season distribution (56.7% rainiest years). From the analyzed farms, 70% use straw as a floor bed, followed by bush (21%). 45.6% of animals were clinically diagnosed with footrot and were sheltered in straw floor. Bed quality is good in 40%, however frequent changes in the floor bed in the preceding 15 days had a higher rate of footroot diagnosed cases (33,23%) compared to monthly changes. Regarding sheepfold animal number, the distribution is proper in 36.7%, elevated in 30%, overcrowded in 6.7% and low in 23.3%. Clinically ill diagnosed animals in last category were the lowest observed (3%). Concerning hoof trimming, 76.7% trims while 23.3% reported not to perform that task. From those that perform trimming, 73.9% do it only when necessary, 21.7% at the time of clipping and 4.4% when the animal is affected. One curious result was that animals who perform trimming at clipping had higher footrot cases (52.6%), unlike those who trim only when necessary (40.2%) or even when animals are clinically ill (0.91%). Mostly all in the presence of footrot choose a local treatment (95.2%) using antibiotic sprays, instead of parenteral antibiotic treatment. Footbath is rarely used in the prevention (13.3%) of this disease and when it is misconceived (25%) and incorrectly formulated (100%). Conclusion: This study is the first performed in Portugal focusing on footrot caracterization in native SEN flocks. The economic impact in SEN dairy flocks is atributed to factors such as the direct decrease in milk and meat production, the early slaughter of affected animals due to a non responsive treatment, and medical expenses. The most effective eradication method combines first of all the ability to understand the problems of SEN producers and then the implementation of preventive measures and treatment of footrot. Often linked to the lack of formation, and the advanced age of portuguese producers, the mentality and social factors block the advance of veterinarian intervention itself.

Percutaneous vertebroplasty: a new animal model.

Background Context Percutaneous vertebroplasty (PVP) is a minimally invasive surgical procedure and is frequently performed in humans who need surgical treatment of vertebral fractures. PVP involves cement injection into the vertebral body, thereby providing rapid and significant pain relief. Purpose The testing of novel biomaterials depends on suitable animal models. The aim of this study was to develop a reproducible and safe model of PVP in sheep. Study Design This study used ex vivo and in vivo large animal model study (Merino sheep). Methods Ex vivo vertebroplasty was performed through a bilateral modified parapedicular access in 24 ovine lumbar hemivertebrae, divided into four groups (n=6). Cerament (Bone Support, Lund, Sweden) was the control material. In the experimental group, a novel composite was tested—Spine-Ghost—which consisted of an alpha-calcium sulfate matrix enriched with micrometric particles of mesoporous bioactive glass. All vertebrae were assessed by micro-computed tomography (micro-CT) and underwent mechanical testing. For the in vivo study, 16 sheep were randomly allocated into control and experimental groups (n=8), and underwent PVP using the same bone cements. All vertebrae were assessed postmortem by micro-CT, histology, and reverse transcription-polymerase chain reaction (rt-PCR). This work has been supported by the European Commission under the 7th Framework Programme for collaborative projects (600,000–650,000 USD). Results In the ex vivo model, the average defect volume was 1,275.46±219.29 mm3. Adequate defect filling with cement was observed. No mechanical failure was observed under loads which were higher than physiological. In the in vivo study, cardiorespiratory distress was observed in two animals, and one sheep presented mild neurologic deficits in the hind limbs before recovering. Conclusions The model of PVP is considered suitable for preclinical in vivo studies, mimicking clinical application. All sheep recovered and completed a 6-month implantation period. There was no evidence of cement leakage into the vertebral foramen in the postmortem examination.

Antibiofilm activity of Brasilian propolis extracts against Staphylococcus aureus producers isolated from goat and sheep mastitic milk

The aim of this study was to investigate the action of inhibiting S. aureus biofilm formation, and the ability to eliminate formed biofilm, by alcoholic extracts of green, red and brown propolis from Brazil. Ten isolates of S. aureus have been tested, 8 field isolates, 1 MRSA and 1 ATCC 25923, by microplate quantitative method. For the evaluation of inhibitory action, the isolates were inoculated, in triplicate, in TSB 1% glucose in the presence of green (1), red (2) and brown (4) propolis extracts. Biofilm formation was evaluated by optical reading, compared to a negative control consisting of a mixture of TSB and extract. For biofilm elimination assay, extracts were added to plates with 24h cultures of the same isolates. Assays were repeated three times on three different days. Eight out of the 10 isolates produced less biofilm in the presence of the green propolis extracts, so the inhibitory effect is 80%. Brown propolis extracts inhibited the formation of biofilm in 10% to 70% of the isolates and the red extracts in 30% to 80%. Regarding the biofilm elimination activity, green propolis extract was positive for 9 out of the 10 isolates (90%), the brown propolis extracts were positive for 0% to 100% isolates and red extracts for 0% to 10% isolates.

Propolis components accountable for bactericidal accomplishment and antibiofilm activity

This study aimed at evaluating antimicrobial and antibiofilm activity of phenolic compounds present in propolis ethanol extracts (PEE). Seventy per cent ethanol extracts from seven types of propolis, one Green, two Red and four Brown collected in four Brazilian States were prepared and total phenolics, flavonoids, tannins and anthocyanins were assessed by high-performance liquid chromatography (HPLC). Minimum bactericidal concentration (MBC) and inhibitor effect on Staphylococcus aureus biofilm formation and capacity to disrupt established biofilms were assessed towards eight S. aureus isolates from milk of small ruminants with mastitis, one methicillin-resistant S. aureus (MRSA) and S. aureus ATCC 25923. To evaluate different propolis components accountability for bactericidal accomplishment and antibiofilm activity, the results were analysed by the non-parametric Spearman coefficient. Results of phenolic compounds were 216,21 to 312,08 gallic acid milligram equivalent per extract gram (mg EGA/g) of total phenolics, 55,08 to 140,6 quercetin milligram equivalent per extract gram (mg EQ/g) of flavonoids, 118,51 to 3766,16 catechin milligram equivalent per extract gram (mg EC/g) of tannins and 1,03 to 8,39 milligram per extract gram (mg/g) of anthocyanins. Red1 and Red2 showed higher tannin contents, while Red2 exhibited superior amount of anthocyanins and total phenolics. Brown3 presented higher flavonoid quantity. Green, Red1 and Red2 PEE showed the lowest levels of flavonoids, but the higher antimicrobial activity. Most PEE exhibit bactericidal activity at a concentration of 1.6 mg/mL. Brown4 PEE showed the worst capacity to inhibit S. aureus. Green PEE showed to be the most efficient in both preventing and disrupting biofilm. All PEE studied exhibited a better inhibitory activity prior-to than post-biofilm formation. According to non-parametric Spearman correlation analysis, there seems to be a significant negative correlation between the ability to disrupt biofilm and both tannins and anthocyanins contents.

“Biomarkers in a One Health perspective: current knowledge of their ability to merge human and animal health

Biomarkers are nowadays essential tools to be one step ahead for fighting disease, enabling an enhanced focus on disease prevention and on the probability of its occurrence. Research in a multidisciplinary approach has been an important step towards the repeated discovery of new biomarkers. Biomarkers are defined as biochemical measurable indicators of the presence of disease or as indicators for monitoring disease progression. Currently, biomarkers have been used in several domains such as oncology, neurology, cardiovascular, inflammatory and respiratory disease, and several endocrinopathies. Bridging biomarkers in a One Health perspective has been proven useful in almost all of these domains. In oncology, humans and animals are found to be subject to the same environmental and genetic predisposing factors: examples include the existence of mutations in BR-CA1 gene predisposing to breast cancer, both in human and dogs, with increased prevalence in certain dog breeds and human ethnic groups. Also, breast feeding frequency and duration has been related to a decreased risk of breast cancer in women and bitches. When it comes to infectious diseases, this parallelism is prone to be even more important, for as much as 75% of all emerging diseases are believed to be zoonotic. Examples of successful use of biomarkers have been found in several zoonotic diseases such as Ebola, dengue, leptospirosis or West Nile virus infections. Acute Phase Proteins (APPs) have been used for quite some time as biomarkers of inflammatory conditions. These have been used in human health but also in the veterinary field such as in mastitis evaluation and PRRS (porcine respiratory and reproductive syndrome) diagnosis. Advantages rely on the fact that these biomarkers can be much easier to assess than other conventional disease diagnostic approaches (example: measured in easy to collect saliva samples). Another domain in which biomarkers have been essential is food safety: the possibility to measure exposure to chemical contaminants or other biohazards present in the food chain, which are sometimes analytical challenges due to their low bioavailability in body fluids, is nowadays a major breakthrough. Finally, biomarkers are considered the key to provide more personalized therapies, with more efficient outcomes and fewer side effects. This approach is expected to be the correct path to follow also in veterinary medicine, in the near future.

Identificação molecular de estirpes de Staphylococcus aureus envolvidas em casos de mastite caprina subclínica.

2015

Uso de antimicrobiano nanoparticulado para o tratamento da mastite subclínica de ovelhas de corte no período seco.

A inflamação da glândula mamária é uma das principais causas de prejuízo na ovinocultura. Este estudo teve como objetivo investigar as taxas de cura do tratamento da mastite subclínica após infusão intramamária de princípio ativo antimicrobiano no momento da secagem, em formulações convencional e nanoparticulada. Os rebanhos estavam localizados em São Carlos, São Paulo, Brasil. Analisou-se um total de 584 glândulas mamárias de 307 ovelhas de aptidão para produção de carne. Triagem prévia dos casos subclínicos de mastite foi efetuada por meio do California Mastitis Test (CMT) e/ou da contagem de células somáticas (CCS). Análises microbiológicas foram realizadas para confirmação da etiologia infecciosa. As glândulas mamárias com mastite subclínica foram distribuídas em três grupos: G1 (Controle; glândulas mamárias que não receberam tratamento antimicrobiano); G2 (glândulas mamárias em que foi administrado 100 mg de cloxacilina benzatina em estrutura convencional) e G3 (glândulas mamárias em que foi administrado 50 mg de cloxacilina benzatina em estrutura nanoencapsulada). O tratamento aplicado ao G3 mostrou-se mais eficiente (P=0,047) na cura de glândulas mamárias com mastite subclínica. O uso da cloxacilina nanoencapsulada no momento da secagem de ovelhas de corte auxilia no controle da mastite subclínica infecciosa e reduz os prejuízos consequentes.