Acta Veterinaria Scandinavica - Latest articles

Tumor slices as a model to evaluate doxorubicin in vitro treatment and expression of trios of genes PRSS11, MTSS1, CLPTM1 and PRSS11, MTSS1, SMYD2 in canine mammary gland cancer

2008-07-04

Background: In women with breast cancer submitted to neoadjuvant chemotherapy based on doxorubicin, tumor expression of groups of three genes (PRSS11, MTSS1, CLPTM1 and PRSS11, MTSS1, SMYD2) have classified them as responsive or resistant. We have investigated whether expression of these trios of genes could predict mammary carcinoma response in dogs and whether tumor slices, which maintain epithelial-mesenchymal interactions, could be used to evaluate drug response in vitro. Methods: Tumors from 38 dogs were sliced and cultured with or without doxorubicin 1uM for 24 h. Tumor cells were counted by two observers to establish a percentage variation in cell number, between slices. Based on these results, a reduction in cell number between treated and control samples [greater than or equal to] 21.7%, arbitrarily classified samples, as drug responsive. Tumor expression of PRSS11, MTSS1, CLPTM1 and SMYD2, was evaluated by real time PCR. Relative expression results were then transformed to their natural logarithm values, which were spatially disposed according to the expression of trios of genes, comprising PRSS11, MTSS1, CLPTM1 and PRSS11, MTSS1, SMYD2. Fisher linear discrimination test was used to generate a separation plane between responsive and non-responsive tumors. Results: Culture of tumor slices for 24 h was feasible. Nine samples were considered responsive and 29 non-responsive to doxorubicin, considering the pre-established cut-off value of cell number reduction [greater than or equal to] 21.7%, between doxorubicin treated and control samples. Relative gene expression was evaluated and tumor samples were then spatially distributed according to the expression of the trios of genes: PRSS11, MTSS1, CLPTM1 and PRSS11, MTSS1, SMYD2. A separation plane was generated. However, no clear separation between responsive and non-responsive samples could be observed. Conclusion: Three-dimensional distribution of samples according to the expression of the trios of genes PRSS11, MTSS1, CLPTM1 and PRSS11, MTSS1, SMYD2 could not predict doxorubicin in vitro responsiveness. Short term culture of mammary gland cancer slices may be an interesting model to evaluate chemotherapy activity.

Transfer of immunoglobulins through the mammary endothelium and epithelium and in the local lymph node of cows during the initial response after intramammary challenge with E. coli endotoxin

Karin Ostensson and Shichun Lun — 2008-07-02

Background: The first hours after antigen stimulation, interactions occur influencing the outcome of the immunological reaction. Immunoglobulins originate in blood and/or are locally synthesized. The transfer of Ig isotypes (Igs) in the udder has been studied previously but without the possibility to distinguish between the endothelium and the epithelium. The purpose of this study was to map the Ig transfer through each barrier, separately, and Ig transfer in the local lymph nodes of the bovine udder during the initial innate immune response. Methods: The content of IgG1, IgG2, IgM, IgA and albumin (BSA) was examined in peripheral/afferent mammary lymph and lymph leaving the supramammary lymph nodes, and in blood and milk before (0 h) and during 4 hours after intramammary challenge with Esherichia coli endotoxin in 5 cows. Results: Igs increased most rapidly in afferent lymph resulting in higher concentrations than in efferent lymph at postinfusion hour (PIH) 2, contrary to before challenge. Ig concentrations in milk were lower than in lymph; except for IgA at 0 h; and they increased more slowly. Afferent lymph:serum and efferent lymph:serum concentration ratios (CR) of Igs were similar to those of BSA but slightly lower. Milk:afferent lymph (M:A) CRs of each Ig, except for IgG2, showed strikingly different pattern than those of BSA. The M:A CR of IgG1, IgM and IgA were higher than that of BSA before challenge and the CR of IgA and IgG1 remained higher also thereafter. At PIH 2 there was a drop in Ig CRs, except for IgG2, in contrast to the BSA CR which gradually increased. The M:A CR of IgM and Ig A decreased from 0 h to PIH 4, in spite of increasing permeability. Conclusions: The transfer of Igs through the endothelium appeared to be merely a result of diffusion although their large molecular size may hamper the diffusion. The transfer through the epithelium and the Ig concentrations in milk seemed more influenced by selective mechanisms and local sources, respectively. Our observations indicate a selective mechanism in the transfer of IgG1 through the epithelium also in lactating glands, to our knowledge not previously shown; a local synthesis of IgA and possibly of IgM, released primarily into milk, not into tissue fluid; that IgG2 transfer through both barriers is a result of passive diffusion only and that the content of efferent lymph is strongly influenced by IgG1, IgM and IgA in the mammary tissue, brought to the lymph node by afferent lymph.

Post mortem findings in sows and gilts euthanised or found dead in a large Swedish herd

2008-07-01

Background: The aim of this study was to get information on post mortem diagnoses of sows found dead or euthanised and to understand the diagnoses aetiology (causative background). Moreover, the study was to evaluate the association between the clinical symptoms observed on farm and post mortem findings. Methods: A large Swedish herd was studied from January to September 2006. During the 32-week period 3.9% of the removed sows and gilts (old enough to be mated) were found dead, 12.0% were euthanised and the rest were sent to slaughter. Of 32 sows/gilts found dead 17 (53%) were post mortem examined, and of 98 sows euthanised 79 (81%) were examined. The 96 examined carcasses were after 70 sows and 26 gilts. The findings at examination were together with data from the herd monitoring program PigWin Sugg the base for the descriptive statistics presented. Results: The average parity number at removal was 2.8 for those found dead and 2.1 for those euthanised. The highest number euthanised and found dead was in parity 0 (gilts). The main proportion of post mortem examinations was made on sows being in the period [greater than or equal to] 28 d of gestation at death (37.5%), followed by weaning to next service period (24.0%). Arthritis, with an incidence of 36.4% was the most common main finding of pathological-anatomical diagnosis (PAD). Of sows/gilts found dead were circulatory/cardiac failure (23.5%) and trauma related injuries (23.5%) most common PAD. The most commonly observed clinical symptom and reason for euthanasia of the sows/gilts was lameness. Notably, in 43% of the cases with PAD arthritis, the clinical symptoms suggested it being a fracture. Further one or more abscesses (38.5%) and teeth injuries (31.0%) were common findings when also incidental findings were included. Conclusions: This post mortem study based on carcasses from sows/gilts found dead or euthanised showed that arthritis was a significant problem in the studied herd and that post mortem examination was important to get proper diagnosis.

Increasing daily feeding occasions in restricted feeding strategies does not improve performance or well being of fattening pigs

Eva Persson, Margret Wülbers-Mindermann, Charlotte Berg and Bo Algers — 2008-06-24

Background: The natural feeding behaviour of the pig is searching for feed by rooting activities throughout the day; self-feeding pigs randomly space their eating and drinking periods throughout the day consuming ten to twelve meals per day. Pigs in conventional fattening pig production are normally fed 2–3 times daily with the feed consumed within 15 minutes. The aim of this study was to determine if more frequent feedings could improve the performance of conventionally kept fattening pigs. Methods: The experiment was carried out on 360 fattening pigs (27–112 kg live weight), weighed and assigned to pens stratified by weight and sex. Each treatment group consisted of 180 pigs, allocated to 20 pens with nine pigs in each pen. To evaluate how more feeding occasions affects performance and well-being the pigs were divided into two groups and fed three (control group) or nine (treatment group) times daily. The same total amount of liquid feed was fed to each group and the feed ration was correlated to the live weight of the pigs. All weight and slaughter recordings were made individually and recordings of feed consumption were made pen-wise. At slaughter the stomach of each pig was examined for lesions in the pars oesophagea and scored on a scale from 1–6. Results: Frequent feeding occasions influenced both performance and status of gastric lesions of the pigs adversely. Pigs in the treatment group grew slower compared to pigs in the control group; 697 g/day (± 6.76) versus 804 g/day (± 6.78) (P < 0.001) with no difference in within-pen variation. There was also a lower prevalence of gastric lesions within pigs in the control group (2.4 (± 0.12) compared to 3.0 (± 0.12) (P < 0.01)). There was a positive correlation between gastric lesions in the treatment group and daily weight gain (r = 0.19; P < 0.01). Conclusion: Increased daily feeding occasions among group housed pigs resulted in a poorer daily weight gain and increased mean gastric lesion score as compared with pigs fed three times daily. This may be a consequence of more frequently occurring competition for feed in the treatment group. The present study does not support increased daily feeding occasions in fattening pigs.

Glutathione peroxidase (GPX) activity in blood of ewes on farms in different scrapie categories in Iceland

2008-06-23

Background: Preliminary studies indicated decreased glutathione peroxidase (GPX) activity in blood of ewes on scrapie-afflicted farms. Other studies have shown decreased GPX activity in brain of prion-infected mice and in prion-infected cells in vitro. The aim of this study was to examine the GPX activity in blood as well as the distribution of GPX-activity levels from ewes on farms in scrapie-afflicted areas in Iceland. Methods: Blood samples were collected from 635 ewes (non-pregnant [n = 297] and pregnant [n = 338]) on 40 farms in scrapie-afflicted areas during the years 2001–2005, for analysis of GPX activity. The farms were divided into three categories: 1. Scrapie-free farms (n = 14); 2. Scrapie-prone farms (earlier scrapie-afflicted, restocked farms) (n = 12); 3. Scrapie-afflicted farms (n = 14). For comparison, 121 blood samples were also collected from non-pregnant ewes on one farm (farm A) in a scrapie-free area (scrapie never registered). Chi-square test was used to test for normal distribution of GPX-results, and Kruskal-Wallis test to compare GPX-results between categories. Results: The GPX-results appeared to be biphasically distributed in ewes in all three scrapie categories and on farm A. The presumptive breaking point was about 300 units g Hb-1. About 30–50% of the GPX-results from ewes in all three scrapie categories were below 300 units g Hb-1 but only about 13% of the GPX-results from ewes on farm A. The mean GPX activity was highest on farm A, and was significantly lower on scrapie-prone farms than on scrapie-free or scrapie-afflicted farms (non-pregnant and pregnant ewes: P < 0.005, respectively; non-pregnant and pregnant ewes combined: P < 0.0005). Conclusions: 1) the distribution of GPX-results in blood of Icelandic ewes apparently has a biphasic character; 2) the GPX-results were higher in ewes on one farm in a scrapie-free area than in ewes on farms in the scrapie-afflicted areas; 3) GPX-activity levels were significantly lowest on earlier scrapie-afflicted, restocked farms, which might have a bearing on the recurrence of sporadic scrapie on these farms; 4) further study on the possible role of GPX activity in the occurrence of scrapie in Iceland is warranted.

Prevalence of myocardial hypertrophy in a population of asymptomatic Swedish Maine coon cats

Suzanne Gundler, Anna Tidholm and Jens Häggström — 2008-06-18

Background: Maine coon cats have a familial disposition for developing hypertrophic cardiomyopathy (HCM) with evidence of an autosomal dominant mode of inheritance 1. The current mode to diagnose HCM is by use of echocardiography. However, definite reference criteria have not been established. The objective of the study was to determine the prevalence of echocardigraphic changes consistent with hypertrophic cardiomyopathy in Swedish Maine coon cats, and to compare echocardiographic measurements with previously published reference values. Methods: All cats over the age of 8 months owned by breeders living in Stockholm, listed on the website of the Maine Coon breeders in Sweden by February 2001, were invited to participate in the study. Physical examination and M-mode and 2D echocardiographic examinations were performed in all cats. Results: Examinations of 42 asymptomatic Maine coon cats (10 males and 32 females) were performed. The age of the cats ranged from 0,7 to 9,3 years with a mean of 4,8 ± 2,3 years. Four cats (9,5%) had a diastolic interventricular septal (IVSd) or left ventricular free wall (LVPWd) thickness exceeding 6,0 mm. In 3 of these cats the hypertrophy was segmental. Two cats (4,8%) had systolic anterior motion (SAM) of the mitral valve without concomitant hypertrophy. Five cats (11,9%) had IVSd or LVPWd exceeding 5,0 mm but less than 6,0 mm. Conclusion: Depending on the reference values used, the prevalence of HCM in this study varied from 9,5% to 26,2%. Our study suggests that the left ventricular wall thickness of a normal cat is 5,0 mm or less, rather than 6,0 mm, previously used by most cardiologists. Appropriate echocardiographic reference values for Maine coon cats, and diagnostic criteria for HCM need to be further investigated.

Risk factors for post-weaning diarrhoea on piglet producing farms in Finland

Taina M. Laine, Tapani Lyytikainen, Maija Yliaho and Marjukka Anttila — 2008-06-18

Background: Post-weaning diarrhoea (PWD) is a significant gastrointestinal disease in pigs. It is considered a multifactorial disease associated with proliferation of enterotoxigenic Escherichia coli in the intestinal tract of affected pigs. The aim of this study was to analyse risk factors related to the occurrence of PWD on Finnish piglet producing farms. Methods: The data of a follow-up study of 73 conventional piglet producing farms was used in the case-control study. The selection of the 41 PWD case and 28 control farms was based on the use of antimicrobials for treating diarrhoea in weaned pigs and the answers related to the occurrence of diarrhoea after weaning in the questionnaire. Four intermediate farms were excluded from the statistical analysis. Altogether 39 factors related to herd characteristics, weaner pig management and pig health were studied. The median number of sows was 59.0 (IQR = 44.0; 74.5) and 52.5 (IQR = 36.8; 61.5) on the case and the control farms, respectively. The significances of the univariable associations between the explanatory variables and the outcome variable were tested, and in the multivariate analysis quasibinomial generalized linear models were applied. Results: An increased risk of PWD was associated with the regimen of twice a day feeding and feed restriction after weaning (P = 0.02; compared to feeding three or more meals a day or the use of ad libitum feeding) and with a higher number of sows on the farm (P = 0.02; risk increasing with increasing number of sows). Automatic temperature control was associated with a decreased risk of PWD (P = 0.03; compared to manual temperature control). Conclusions: Twice a day feeding of newly-weaned pigs should be avoided if the amount of feed given is restricted. Variation in ambient temperature should be minimized in housing of newly-weaned pigs and this can be achieved by using automatic temperature control. With increasing number of sows in the herds the risk of PWD increases and more attention should be paid to prevention of post-weaning diarrhoea.

Filarioid Nematodes in Cattle, Sheep and Horses in Finland

2008-06-16

Background: In autumn 2006, Finnish meat inspection data revealed lesions in tendons, muscles and ligaments of bovine hind legs leading to partial condemnation of carcasses. In gross pathological examination at Finnish Food Safety Authority Evira, Oulu (now Fish and Wildlife Health) Research Unit, Onchocerca sp. (Filarioidea; Onchocercidae) nematodes were detected in lesions. Due to this, a pilot study was made in order to find out what filarioid nematodes do occur in cattle, horses and sheep in Finland. Methods: Ventral skin biopsies from 209 dairy cattle and 42 horses, as well as blood samples from 209 cattle, 146 horses and 193 sheep, were collected from different parts of Finland and examined for microfilariae. Visceral organs and other tissues from 33 cattle with parasitic lesions were studied histopathologically. Results: Onchocerca sp. microfilariae (mf), 240 mum long, range 225-260 mum, 5.4 mum thick, were found in 37 % of the skin biopsies of cattle. All blood samples from cattle, horses and sheep and skin biopsies from horses were negative for mf. Ventral skin microfilaria prevalence in cattle was higher in southern Finland than in the North (p=0.001). Animal age and sampling time was not associated with mf prevalence. The infection was evenly distributed among young and older animals. Macroscopic lesions on tissues included greenish-grey discolouration and often oedema. In most of the lesions, small pale nodules were seen on the fasciae. Histopathologic examination of the samples revealed mild to intense infiltration with eosinophilic granulocytes and multifocal nodular lymphoplasmacytic aggregations were seen. In some samples, there were granulomatotic lesions with central necrotic tissue and cell detritus, surrounded by eosinophilic granulocytes, lympho-, plasma- and histiocytes and some multinucleated giant cells. Around living nematodes no or only weak inflammatory changes were observed. Conclusions: Onchocerca sp. infection in cattle was found to be common in Finland, but the amount of pathological changes leading to condemnation of infected parts is low compared to the mf prevalence. Pronounced pathological changes are distinct but rare and mild changes are difficult to distinguish. No other filarioid nematodes were observed from the animals and it appears that horses and sheep may be free from filarioid nematodes in Finland.

Occurrence of antimicrobial resistance among bacterial pathogens and indicator bacteria in pigs in different European countries from year 2002 - 2004; the ARBAO-II study

2008-06-13

Background: The project "Antibiotic resistance in bacteria of animal origin - II" (ARBAO-II) was funded by the European Union (FAIR5-QLK2-2002-01146) for the period 2003-05. The aim of this project was to establish a program for the continuous monitoring of antimicrobial susceptibility of pathogenic and indicator bacteria from food animals using validated and harmonised methodologies. In this report the first data on the occurrence of antimicrobial resistance among bacteria causing infections in pigs are reported. Methods: Susceptibility data from 17,642 isolates of pathogens and indicator bacteria including Actinobacillus pleuropneumoniae, Streptococcus suis and Escherichia coli isolated from pigs were collected from fifteen European countries in 2002-2004. Results: Data for A. pleuropneumoniae from infected pigs were submitted from five countries. Most of the isolates from Denmark were susceptible to all agents tested with the exceptions of a low frequency of resistance to tetracycline and trimethoprim - sulphonamide. Data for Streptococcus suis were obtained from six countries. In general, a high level of resistance to tetracycline (48.0 - 92.0 %) and erythromycin (29.1 - 75.0 %) was observed in all countries whereas the level of resistance to ciprofloxacin and penicillin differed between the reporting countries. Isolates from England (and Wales), France and The Netherlands were all susceptible to penicillin. In contrasted the proportion of strains resistant to ciprofloxacin ranged from 12.6 to 79.0 % (2004) and to penicillin from 8.1 - 13.0 % (2004) in Poland and Portugal. Data for Escherichia coli from infected and healthy pigs were obtained from eleven countries. The data reveal a high level of resistance to tetracyclines, streptomycin and ampicillin among infected pigs whereas in healthy pigs the frequency of resistance was lower. Conclusion: Bacterial resistance to some antimicrobials was frequent with different levels of resistance being observed to several antimicrobial agents in different countries. The occurrence of resistance varied distinctly between isolates from healthy and diseased pigs, with the isolates from healthy pigs generally showing a lower level of resistance than those from diseased pigs. The study suggests that the choice of antimicrobials used for the treatment of diseased animals should preferably be based on knowledge of the local pattern of resistance.

Acute phase response in two consecutive experimentally induced E. coli intramammary infections in dairy cows

2008-06-13

Background: Acute phase proteins haptoglobin (Hp), serum amyloid A (SAA) and lipopolysaccharide binding protein (LBP) have suggested to be suitable inflammatory markers for bovine mastitis. The aim of the study was to investigate acute phase markers along with clinical parameters in two consecutive intramammary challenges with Escherichia coli and to evaluate the possible carry-over effect when same animals are used in an experimental model. Methods: Mastitis was induced with a dose of 1500 cfu of E. coli in one quarter of six cows and inoculation repeated in another quarter after an interval of 14 days. Concentrations of acute phase proteins haptoglobin (Hp), serum amyloid A (SAA) and lipopolysaccharide binding protein (LBP) were determined in serum and milk. Results: In both challenges all cows became infected and developed clinical mastitis within 12 hours of inoculation. Clinical disease and acute phase response was generally milder in the second challenge. Concentrations of SAA in milk started to increase 12 hours after inoculation and peaked at 60 hours after the first challenge and at 44 hours after the second challenge. Concentrations of SAA in serum increased more slowly and peaked at the same times as in milk; concentrations in serum were about one third of those in milk. Hp started to increase in milk similarly and peaked at 36–44 hours. In serum, the concentration of Hp peaked at 60–68 hours and was twice as high as in milk. LBP concentrations in milk and serum started to increase after 12 hours and peaked at 36 hours, being higher in milk. The concentrations of acute phase proteins in serum and milk in the E. coli infection model were much higher than those recorded in experiments using Gram-positive pathogens, indicating the severe inflammation induced by E. coli. Conclusion: Acute phase proteins would be useful parameters as mastitis indicators and to assess the severity of mastitis. If repeated experimental intramammary induction of the same animals with E. coli is used in cross-over studies, the interval between challenges should be longer than 2 weeks, due to the carry-over effect from the first infection.

Radiographic parameters for diagnosing sand colic in horses

Anna Kendall, Charles Ley, Agneta Egenvall and Johan Brojer — 2008-06-13

Background: Ingestion of sand can cause colic, diarrhoea and weight loss in horses, but these signs are unspecific and can have many other causes. The amount of sand that induces disease may vary between individuals. To avoid over-diagnosing, it is important to determine the amount of sand that can be found in horses without clinical signs of gastrointestinal disease. The aim of this study was to use previously suggested parameters for establishing a radiographic diagnosis of sand colic, and compare these findings between a sand colic group and a control group. Methods: Abdominal radiographs were obtained in 30 horses with a complaint unrelated to the gastrointestinal tract. In addition, archived abdominal radiographs of 37 clinical cases diagnosed with sand impaction were investigated. The size of the mineral opacity indicative of sand in the abdomen was measured and graded according to a previously published protocol based on height and length. Location, homogeneity, opacity and number of sand accumulations were also recorded. Results: Twenty out of 30 control horses (66%) had one or more sand accumulations. In the present study; height, length and homogeneity of the accumulations were useful parameters for establishing a diagnosis of sand colic. Radiographically defined intestinal sand accumulation grades of up to 2 was a common finding in horses with no clinical signs from the gastrointestinal tract whereas most of the clinical cases had much larger grades, indicating larger sand accumulations. Conclusions: Further work to establish a reliable grading system for intestinal sand content is warranted, but a previously proposed grading system based on measurements of height and length may be an alternative for easy assessment of sand accumulations in the meantime. The present study indicates that a grade 1 - 2 sand accumulation in the intestine is a frequent finding in horses. When working up a case with clinical signs from the gastrointestinal tract, one or more accumulations of this grade should not be considered the cause until other possibilities have been ruled out.

Reproductive biology of the pampas deer (Ozotoceros bezoarticus): a review

2008-06-05

The pampas deer (Ozotoceros bezoarticus) is a South American grazing deer which is in extreme danger of extinction. Very little is known about the biology of the pampas deer. Moreover, most information has not been published in peer-reviewed scientific journals, and is only available in local publications, theses, etc. Therefore, our aim was to update and summarize the available information regarding the reproductive biology of the pampas deer. Moreover, in most sections, we have also included new, unpublished information. Detailed descriptions are provided of the anatomy of both the female and the male reproductive tract, puberty onset, the oestrous cycle and gestational length. Birthing and the early postpartum period are described, as are maternal behaviour and early fawn development, seasonal distribution of births, seasonal changes in male reproduction and antler cycle, reproductive behaviour, semen collection, and cryopreservation. Finally, an overview is given and future directions of research are proposed.