Effects of different media on proliferation and differentiation capacity of canine, equine and porcine adipose derived stem cells

Adult stem cells are of particular interest for therapeutic use in the field of regenerative medicine. Adipose-derived mesenchymal stem cells (ASCs) are an attractive stem cell source for all fields of regenerative medicine because adipose tissue - and therewith cells - can easily be harvested from each donor. However, common expansion using fetal bovine serum (FBS) can not be used for clinical applications as xenogenic proteins must be avoided. Adipose tissue from equine, canine and porcine donors was digested with collagenase to isolate ASCs. ASCs were either expanded in a cell culture medium supplemented with FBS or in a serum-free medium (UltraCulture; UC) supplemented with a serum substitute (UltroserG). From all three animal species, the adipogenic and osteogenic differentiation potential of ASCs cultured with different media was analyzed in vitro. Cell proliferation analysis showed a population doubling time of 48-68 h for canine cells, 54-65 h for porcine cells and 54-70 h for equine cells, expanded in different media. Except for porcine ASCs, cells cultured in media supplemented with FBS grew faster than cells expanded in UC medium with UltroserG. Yet, all cells maintained their potential to differentiate into adipocytes and osteoblasts. UltraCulture medium containing UltroserG can for all examined species be recommended if FBS needs to be avoided in the expansion of donor-derived (stem) cells.     

Cardiac troponin I and the occurrence of cardiac arrhythmias in horses with experimentally induced endotoxaemia

Branding is the traditional and well-established method used to mark horses, but recently microchip transponders for implantation have become available. In this study, behaviour, physiological stress variables and skin temperature in foals were determined in response to hot-iron branding (n=7) and microchip implantation (n=7). Salivary cortisol concentrations increased in response to branding (1.8 ± 0.2 ng/mL) and microchip implantation (1.4 ± 0.1ng/mL), but cortisol release over time did not differ. In response to both manipulations there was a transient increase in heart rate (P<0.001) and heart rate variability (P<0.01). Branding and microchip implantation induced a comparable aversive behaviour (branding, score 3.86 ± 0.85; microchip, score 4.00 ± 0.82). Both techniques thus caused similar physiological and behavioural changes indicative of stress. Acutely, implantation of a microchip was as stressful as branding in foals. Branding caused a necrotising skin burn lasting at least 7 days. Moreover branding, but not microchip implantation (P<0.001), was accompanied by a generalized increase in skin temperature which was comparable to low degree post-burn hypermetabolism in humans.     

Biosecurity and arable use of manure and biowaste — Treatment alternatives

The potential negative environmental impact of manure and biological waste (BW) can be minimised at a profit by recycling plant nutrients in the food chain. Current large-scale livestock production, epizootic diseases and increasing globalisation increase the need for biosecurity, to minimise the risk of disease transmission to the food chain. Arable use of manure and BW can inadvertently spread infectious diseases; opinion differs concerning the risk levels. To obtain general acceptance for arable use, a hygienically safe end-product is needed. This paper provides a detailed discussion of treatment alternatives for co-treatment, i.e., mixture before treatment of manure and BW. Composting, anaerobic digestion and ammonia treatment are the three options given. A decision support tool is also presented and discussed. Suitable treatment methods must combine biosecurity aspects with environmental, economic and nutrient recycling aspects to create a beneficial whole-farm approach.     

Dairy goat production systems

Goat production concentrated in developing countries (tropics, dry areas), contributes largely to the livelihoods of low and medium income farmers. Farming systems in these areas have evolved to cope with the formidable constraints imposed by harsh natural and economic conditions by adapting integrated crop/livestock production strategies. In Asia, Africa and Latin America, due to its almost exclusive extensive nature, goat production relies mainly on grazing on communal lands that hardly provide the minimum nutrient requirements due to overstocking and degradation. While some of these production systems are becoming semi-intensive, appropriate breeding strategies should be designed to promote conservation and improvement of their unique attributes, such as adaptability, water use efficiency and suitability under harsh climatic conditions. In Europe, dairy goat production is more common around the Mediterranean basin, where it is important from an economic, environmental and sociological perspective to the Mediterranean countries: Spain, France, Italy and Greece. Europe owns only 5.1 % of the world’s dairy goat herds, but produces 15.6 % of the world’s goat milk; this is the only continent where goat milk has such an economic importance and organization. In developing countries the dairy goat sector requires a systemic approach, whereby nutrition, animal health, breeding, know-how, inputs and technologies must be assembled. This would allow the optimization of natural and local resources and would promote the transition from a risk reduction strategy towards an increased productivity strategy. Such an increase would privilege production efficiency based on clean, green and ethical practices for responsible innovation.     

Outbreak of Mycobacterium tuberculosis infection among captive Asian elephants in a Swedish zoo

Between 2001 and 2003, there was an outbreak of tuberculosis in a Swedish zoo which involved elephants, giraffes, rhinoceroses and buffaloes. Cultures of trunk lavages were used to detect infected elephants, tuberculin testing was used in the giraffes and buffaloes, and tracheal lavage and tuberculin testing were used in the rhinoceroses. The bacteria isolated were investigated by spoligotyping and restriction fragment length polymorphism. Five elephants and one giraffe were found to have been infected by four different strains of Mycobacterium tuberculosis.     

Genetic diversity of Staphylococcus aureus isolated from ovine intramammary infections in Norway

Three hundred and eighty-four Staphylococcus aureus isolates obtained from mammary secretions from 332 ewes kept for meat production were typed by pulsed-field gel electrophoresis (PFGE). The ewes were from 242 flocks located in 13 counties distributed in four regions of Norway. In total, 64 different pulsotypes were identified, 31 of these were represented by a single isolate. Fifty-nine percent of the isolates belonged to one of five closely related pulsotypes. This group of pulsotypes occurred in all the counties. Although widely disseminated, the proportions of the prevalent and closely related pulsotypes differed between the regions. Nine pulsotypes were unique to single regions but the number of isolates belonging to each of these pulsotypes was low. Resistance to penicillin was found in only 3 of the 384 S. aureus isolates. These represented three different single banding patterns, not related to any of the prevalent pulsotypes found.     

Mycobacterium avium subsp. paratuberculosis enters the small intestinal mucosa of goat kids in areas with and without Peyer's patches as demonstrated with the everted sleeve method

The main lesions of paratuberculosis in ruminants are in the small intestine. Previous studies have shown that the bacterium enters the small intestine through M cells found in the follicle-associated epithelium lining the domes of the Peyer's patches. The everted sleeve method, devised for the in vitro study of intestinal absorption, was used in this study to investigate the uptake of Mycobacterium avium subsp. paratuberculosis in goat intestine. Everted small intestinal sleeves of goat kids, prepared from areas with and without Peyer's patches, were incubated for 60 min in 3H-labeled bacterial solution. The results of this study imply that the bacteria can enter the intestinal mucosa of the jejunum, both in areas with and without Peyer's patches. These findings indicate, therefore, that M. avium subsp. paratuberculosis bacteria not only enter through M cells but also through enterocytes.     

Description of the infection status in a Norwegian cattle herd naturally infected by Mycobacterium avium subsp. paratuberculosis

The Norwegian surveillance and control programme for paratuberculosis revealed 8 seroreactors in a single dairy cattle herd that had no clinical signs of Mycobacterium avium subsp. paratuberculosis (M. a. paratuberculosis) infection. Paratuberculosis had been a clinical problem in goats several years previously in this herd. All 45 cattle were culled and a thorough investigation of the infection status was conducted by the use of interferon-gamma (IFN-gamma) immunoassay, measurement of antibodies, and pathological and bacteriological examination. In the IFN-gamma immunoassay, 9 animals gave positive results, and 13 were weakly positive, while 19 animals were negative. In the serological test, 10 animals showed positive reactions, and 5 were doubtful, while 30 animals gave negative reactions. There appeared to be a weak trend toward younger animals having raised IFN-gamma and older animals having raised serological tests. Histopathological lesions compatible with paratuberculosis were diagnosed in 4 animals aged between 4 and 9 years. Three of these animals had positive serological reaction and one animal gave also positive results in the IFN-gamma immunoassay. Infection was confirmed by isolation of M. a. paratuberculosis from 2 of these 4 animals. One single bacterial isolate examined by restriction fragment length polymorphism (RFLP) had the same profile, B-C1, as a strain that had been isolated from a goat at the same farm several years previously. Despite many animals being positive in one or both of the immunological tests, indicative of a heavily infected herd, none of the animals showed clinical signs and only one cow was shown to be shedding bacteria. A cross-reaction with other mycobacteria might have caused some of the immunoreactions in these animals. It is also possible that the Norwegian red cattle breed is resistant to clinical infection with M. a. paratuberculosis.     

Seroprevalence of Q fever (coxiellosis) in sheep from the Southern Marmara Region, Turkey

Little information is available in Turkey on Q fever, a zoonose caused by Coxiella burnetii and transmitted from domestic ruminants. This study aimed at investigating the seroprevalence in sheep flocks from three provinces (Bursa, Balikesir and Canakkale). Serosurvey was undertaken on 42 flocks, which were categorised by sizes. Sera were collected randomly from specific age groups within the young population. CHEKIT Q-fever ELISA kit was used to identify the infection in sheep. The results showed that 20% (n=151) of sheep were seropositive. A total of 34 flocks (81%) revealed at least one seropositive animal. Higher seroprevalence was observed in Balikesir region. Larger flocks resulted more infected than medium and small flocks. An association was found between seropositivity and age, when the primiparous ewes (1-year old) had higher antibodies rates than newborn sheep (aged less than 10 months) or biparous ewes (2 years old). These results showed that Q fever infection was common and circulating in the studied region, hence encourage efforts to propose measures that could reduce the spread and the zoonotic risk.