Acta Veterinaria Scandinavica - Latest Articles

Comparison of Variable-Number Tandem-Repeat Markers typing and IS1245 Restriction Fragment Length Polymorphism fingerprinting of Mycobacterium avium subsp. hominissuis from human and porcine origins

Taneli Tirkkonen — 2010-03-10T00:00:00Z

Background: Animal mycobacterioses are regarded as a potential zoonotic risk and cause economical losses world wide. M. avium subsp. hominissuis is a extremely slow-growing subspecies found in mycobacterial infected humans and pigs and therefore rapid and discriminatory typing methods are needed for epidemiological studies. The genetic similarity of M. avium subsp. hominissuis from human and porcine origins using two different typing methods have not been studied earlier. The objective of this study was to compare the IS1245 RFLP pattern and MIRU-VNTR typing to study the genetic relatedness of M. avium strains isolated from slaughter pigs and humans in Finland with regard to public health aspects. Methods: A novel PCR-based genotyping method, variable number tandem repeat (VNTR) typing of eight mycobacterial interspersed repetitive units (MIRUs), was evaluated for its ability to characterize Finnish Mycobacterium avium subsp. hominissuis strains isolated from pigs (n=16) and humans (n=13) and the results were compared with those obtained by the conventional IS1245 RFLP method. Results: The MIRU-VNTR results showed a discriminatory index (DI) of 0,92 and the IS1245 RFLP resulted in DI 0,98. The combined DI for both methods was 0,98. The MIRU-VNTR test has the advantages of being simple, reproducible, non-subjective, which makes it suitable for large-scale screening of M. avium strains. Conclusions: Both typing methods demonstrated a high degree of similarity between the strains of human and porcine origin. The parallel application of the methods adds epidemiological value to the comparison of the strains and their origins. The present approach and results support the hypothesis that there is a common source of M. avium subsp. hominissuis infection for pigs and humans or alternatively one species may be the infective source to the other.

Survey radiography and computerized tomography imaging of the thorax in female dogs with mammary tumors

Carolina Otoni — 2010-03-09T00:00:00Z

Background: Accurate early diagnosis of lung metastases is important for establishing therapeutic measures. Therefore, the present study aimed to compare survey thoracic radiographs and computerized tomography (CT) scans to specifically identify lung metastases in female dogs with mammary tumors. Methods: Twenty-one female dogs, weighing 3 to 34 kg and aged from 5 years to 14 years and 10 months, with mammary tumors were studied. In all dogs before the imaging examinations, fine-needle aspiration cytology of the mammary tumors was performed to confirm the diagnosis. Three-view thoracic radiographs were accomplished: right lateral, left lateral and ventrodorsal views. Sequential transverse images of the thorax were acquired on a spiral Scanner, before and after intravenous bolus injection of nonionic iodine contrast. Soft-tissue and lung windows were applied. All the mammary tumors were surgically removed and examined histologically. Results: The correlation between the cytological and histological results regarding presence of malignancy was observed in only 17 cases. In radiographic examinations, no dog displayed signs of lung metastases or thorax chest lesions. CT detected lung metastasis in two cases, while small areas of lung atelectasis located peripherally were found in 28.57 % of the dogs. Conclusion: In this study population, spiral CT showed higher sensitivity than chest radiographies to detect lung metastasis; this indicates that CT should be performed on all female dogs with malignant mammary tumors.

The effect of the combination of acids and tannin in diet on the performance and selected biochemical, haematological and antioxidant enzyme parameters in grower pigs

Marina Stukelj — 2010-03-06T00:00:00Z

Background: The abolition of in-feed antibiotics or chemotherapeutics as growth promoters have stimulated the swine industry to look for alternatives such as organic acids, botanicals, probiotics and tannin. The objective of the present study was to compare the effects of a combination of acids and tannin with diet with organic acids and diet without growth promoters on the growth performance and selected biochemical, haematological and antioxidant enzyme parameters in grower pigs. Tannin is more natural and cheaper but possibly with the same effectiveness as organic acids with regard to growth performance. Methods: Thirty-six 7 week old grower pigs, divided into three equal groups, were used in a three week feeding trial. Group I was fed basal diet, group II basal diet with added organic acids and group III basal diet with added organic and inorganic acids and tannin. Pigs were weighed before and after feeding and observed daily. Blood was collected before and after the feeding trial for the determination of selected biochemical, haematological and antioxidant enzyme parameters. One-way ANOVA was used to assess any diet related changes of all the parameters. Paired t-test was used to evaluate changes of blood parameters individually in each group of growers before and after feeding. Results: No clinical health problems related to diet were noted during the three week feeding trial. The average daily gain (ADG) and selected blood parameters were not affected by the addition to basal diet of either acids and tannin or of organic acids alone. Selected blood parameters remained within the reference range before and after the feeding trial, with the exception of total serum proteins that were below the lower value of reference range at both times. The significant changes (paired t-test) observed in individual groups before and after the feeding trial are related to the growth of pigs. Conclusion: Diet with acids and tannin did not improve the growth performance of grower pigs but had no deleterious effects on selected blood parameters. The possibility of beneficial effects of adding acids and tannin in diets on growth performance over a longer period, however, could not be excluded.

Different DNA methylation patterns detected by the Amplified Methylation Polymorphism Polymerase Chain Reaction (AMP PCR) technique among various cell types of bulls

Nawapen Phutikanit — 2010-03-05T00:00:00Z

Background: The purpose of this study was to apply an arbitrarily primed methylation sensitive polymerase chain reaction (PCR) assay called Amplified Methylation Polymorphism Polymerase Chain Reaction (AMP PCR) to investigate the methylation profiles of somatic and germ cells obtained from Holstein bulls. Methods: Genomic DNA was extracted from sperm, leukocytes and fibroblasts obtained from three bulls and digested with a methylation sensitive endonuclease (HpaII). The native genomic and enzyme treated DNA samples were used as templates in an arbitrarily primed-PCR assay with 30 sets of single short oligonucleotide primer. The PCR products were separated on silver stained denaturing polyacrylamide gels. Three types of PCR markers; digestion resistant-, digestion sensitive-, and digestion dependent markers, were analyzed based on the presence/absence polymorphism of the markers between the two templates. Results: Approximately 1,000 PCR markers per sample were produced from 27 sets of primer and most of them (>90%) were digestion resistant markers. The highest percentage of digestion resistant markers was found in leukocytic DNA (94.8%) and the lowest in fibroblastic DNA (92.3%, P<0.05). Spermatozoa contained a higher number of digestion sensitive markers when compared with the others (3.6% vs. 2.2% and 2.6% in leukocytes and fibroblasts respectively, P<0.05). Conclusions: The powerfulness of the AMP PCR assay was the generation of methylation-associated markers without any prior knowledge of the genomic sequence. The data obtained from different primers provided an overview of genome wide DNA methylation content in different cell types. By using this technique, we found that DNA methylation profile is tissue-specific. Male germ cells were hypomethylated at the HpaII locations when compared with somatic cells, while the chromatin of the well-characterized somatic cells was heavily methylated when compared with that of the versatile somatic cells.

Application of a pig ligated intestinal loop model for early Lawsonia intracellularis infection

Torsten Boutrup — 2010-02-24T00:00:00Z

Background: Porcine proliferative enteropathy in pigs is caused by the obligate, intracellular bacterium Lawsonia intracellularis. In vitro studies have shown close bacterium-cell interaction followed by cellular uptake of the bacterium within 3 h post inoculation (PI). However, knowledge of the initial in vivo interaction between porcine intestinal epithelium and the bacterium is limited. The aims of the present study were to evaluate the usefulness of a ligated small intestinal loop model to study L. intracellularis infections and to obtain information on the very early L. intracellularis-enterocyte interactions. Methods: A ligated small intestinal loop model using three different L. intracellularis inocula was applied to 10-11-week-old pigs. The inocula were 1) wild type bacteria derived from overnight incubation of L. intracellularis bacteria from spontaneous disease, 2) crude vaccine bacteria (Enterisol(R) Ileitis Vet), and 3) vaccine bacteria propagated in cell culture. The bacteria-enterocyte interaction was visualised using immunohistochemistry on specimens derived 1, 3 and 6 h PI respectively. Results: Although at a low level, close contact between bacteria and the enterocyte brush border including intracellular uptake of bacteria in mature enterocytes was seen at 3 and 6 h PI for the vaccine and the propagated vaccine inocula. Interaction between the wild-type bacteria and villus enterocytes was scarce and only seen at 6 h PI, where a few bacteria were found in close contact with the brush border. Conclusions: The ligated intestinal loop model was useful with respect to maintaining an intact intestinal morphology for up to 6 h. Furthermore, the study demonstrated that L. intracellularis interacts with villus enterocytes within 3 to 6 h after inoculation into intestinal loops and that the bacterium, as shown for the vaccine bacteria, propagated as well as non-propagated, was able to invade mature enterocytes. Thus, the study demonstrates the early intestinal invasion of L. intracellularis in vivo.

Rumen-protected conjugated linoleic acid supplementation to dairy cows in late pregnancy and early lactation: effects on milk composition, milk yield, blood metabolites and gene expression in liver

Tanja Sigl — 2010-02-18T00:00:00Z

Background: Conjugated linoleic acid (CLA) is a collective term for isomers of octadecadienoic acid with conjugated double-bond system. Thus, it was the objective to investigate whether milk composition and metabolic key parameters are affected by adding CLA to the diet of dairy cows in the first four weeks of lactation. Methods: A study was carried out with five primiparous cows fed a CLA supplemented diet compared to five primiparous cows without CLA supplementation. CLA supplemented cows received 7.5 g CLA/day (i.e. 50% cis(c)9,trans(t)11- and 50% t10,c12-CLA) starting two weeks before expected calving and 20 g CLA/day (i.e. 50% c9,t11- and 50% t10,c12-CLA) throughout day 1 to 28 of lactation. Results: The CLA supplement was insufficiently accepted by the animals: only 61.5% of the intended amount was ingested. Fed CLA were detectable in milk fat, whereas contents of c9,t11-CLA and t10,c12-CLA in milk fat were higher for CLA supplemented cows compared to the control group. On average over the entire treatment period, there was a decrease of saturated fatty acids (FA) in milk fat of CLA supplemented cows, combined with a higher content of monounsaturated and trans FA.Our study revealed no significant effects of c9,t11- and t10,c12-CLA supplementation either on milk yield and composition or on metabolic key parameters in blood. Furthermore the experiment did not indicate significant effects of c9,t11- and t10,c12-CLA-supplementation on gene expression of peroxisome proliferator-activated receptor-alpha (PPARα), PPARγ, sterol regulatory element-binding protein-1 and tumor necrosis factor-alpha in liver tissue. Conclusions: Feeding c9,t11- and t10,c12-CLA during the first weeks after calving did not affect metabolic key parameters of blood serum or milk composition of fresh cows. Milk fatty acid composition was changed by feeding c9,t11- and t10,c12-CLA resulting in higher contents of these isomers in milk fat. High contents of long chain FA in milk fat indicate that CLA supplementation during the first four weeks of lactation did not affect massive peripheral lipomobilization.

An assessment of soybeans and other vegetable proteins as source of salmonella contamination in pig production

Martin Wierup — 2010-02-17T00:00:00Z

Background: The impact of salmonella contaminated feed ingredients on the risk for spreading salmonella to pigs was assessed in response to two incidences when salmonella was spread by feed from two feed mills to 78 swine producing herds. Methods: The assessment was based on results from the salmonella surveillance of feed ingredients before introduction to feed mills and from HACCP - based surveillance of the feed mills. Results from the mills of the Company (A) that produced the salmonella contaminated feed, were by the Chi. Square test compared to the results from all the other (B - E) feed producers registered in Sweden. Isolated serovars were compared to serovars from human cases of salmonellosis. Results: Salmonella (28 serovars) was frequently isolated from imported consignments of soybean meal (14.6%) and rape seed meal (10.0%). Company A largely imported soybean meal from crushing plants with a history of unknown or frequent salmonella contamination. The risk for consignments of vegetable proteins to be salmonella contaminated was 2.4 times (P < 0.0006) larger for A when compared to the mills of the other companies which largely were supplied by soybean meal from a crushing plant with a low risk for salmonella contamination. Also the level of feed mill contamination of salmonella was higher for feed mills belonging to Company A in comparison to the other companies before and also after heat treatment. Four (10.5%) of the 38 serovars isolated from feed ingredients (28) and feed mills (10) were on the EU 2007 top ten list of human cases of salmonellosis and all but eight (78.9%) on a 12 year list (1997-2008) of cases of human salmonellosis in Sweden. Conclusions: Salmonella contaminated feed ingredients are an important source for introducing salmonella into the feed and food chain. Effective HACCP-based control and associated corrective actions are required to prevent salmonella contamination of feed. Efforts should be taken to prevent salmonella contamination already at the crushing plants. This is challenge for the EU - feed industry due to the fact that 98% of the use of soybean/meal, an essential feed ingredient, is imported from crushing plants of third countries usually with an unknown salmonella status.

Housing system and herd size interactions in Norwegian dairy herds; associations with performance and disease incidence

Egil Simensen — 2010-02-16T00:00:00Z

Background: According to the Norwegian animal welfare regulations, it has been forbidden to build new tie-stall barns since the end of 2004. Previous studies have shown that cow performance and health differ between housing systems. The interaction between housing system and herd size with respect to performance and disease incidence has not been evaluated. Methods: Cow performance and health in 620 herds housed in free-stall barns were compared with in 192 herds housed in tie-stall barns based on a mail survey and data from the Norwegian Dairy Herd Recording and Cattle Health Systems. The housing systems herds were comparable with respect to herd size (15-55 cows). Associations between performance/disease incidence and housing system, herd size and year of building the cow barn were tested in general linear models, and values for fixed herd size of 20 and 50 cows were calculated. On the individual cow level mixed models were run to test the effect of among others housing system and herd size on test-day milk yield, and to evaluate lactation curves in different parities. All cows were of the Norwegian Red Breed. Results: Average milk production per cow-year was 134 kg lower in free-stall herd than in tie-stall herds, but in the range 27-45 cows there was no significant difference in yields between the herd categories. In herds with less than 27 cows there were increasingly lower yields in free-stalls, particularly in first parity, whereas the yields were increasingly higher in free-stalls with more than 45 cows.In free-stalls fertility was better, calving interval shorter, and the incidence rate of teat injuries, ketosis, indigestions, anoestrus and cystic ovaries was lower than in tie-stalls. All of these factors were more favourable in estimated 50-cow herds as compared to 20-cow herds. In the larger herd category, bulk milk somatic cell counts were higher, and the incidence rate of mastitis (all cases) and all diseases was lower. Conclusion: This study has shown that there is an interaction between housing system and herd size, and that performance and health is not universally better in small free-stalls than in tie-stalls.

Oestradiol-17beta plasma concentrations after intramuscular injection of oestradiol benzoate or oestradiol cypionate in llamas (Lama glama)

Maria Cavilla — 2010-02-11T00:00:00Z

Background: Llamas (Lama glama) are induced ovulators and the process of ovulation depends on dominant follicular size. In addition, a close relationship between behavioural estrus and ovulation is not registered in llamas. Therefore, the exogenous control of follicular development with hormones aims to predict the optimal time to mate. Oestradiol-17β (E2) and its esters are currently used in domestic species, including camelids, in synchronization treatments. But, in llamas, there is no reports regarding the appropriate dosages to be used and most protocols have been designed by extrapolation from those recommended for other ruminants. The aim of the present study was to characterize plasma E2 concentrations in intact female llamas following a single intramuscular (i.m.) injection of two oestradiol esters: oestradiol benzoate (EB) and oestradiol cypionate (ECP). Methods: Twelve non pregnant and non lactating sexually mature llamas were i.m. injected on day 0 with 2.5 mg of EB (EB group, n = 6) or ECP (ECP group, n = 6). Blood samples were collected immediately before injection, at 1, 6, 12, 24 h after treatment and then daily until day 14 post injection. Changes in hormone concentrations with time were analyzed in each group by analysis of variance (ANOVA) using a repeated measures (within-SS) design. Plasma E2 concentrations and area under the concentration-time curve (AUC) values were compared between groups by ANOVA. In all cases a Least-Significant Difference test (LSD) was used to determine differences between means. Hormonal and AUC data are expressed as mean ± S.E.M. Results: Peak plasma E2 concentrations were achieved earlier and were higher in EB group than in ECP group. Thereafter, E2 returned to physiological concentrations earlier in EB group (day 5) than in ECP group (day 9). Although plasma E2 profiles differed over time among groups there were no differences between them on AUC values. Conclusions: The i.m. injection of a single dose of both oestradiol esters resulted in plasma E2 concentrations exceeding physiological values for a variable period. Moreover, the plasma E2 profiles observed depended on the derivative of oestradiol administered. This basic information becomes relevant at defining treatment protocols including oestrogens in llamas.

Apoptosis of resident and inflammatory macrophages before and during the inflammatory response of the virgin bovine mammary gland

Zbysek Sladek — 2010-02-09T00:00:00Z

Background: Macrophages may play a prominent role in defense of the bovine mammary gland, and their functionality is necessary for successful eradication of bacterial pathogens. In contrast to necrosis, however, apoptosis has not yet been studied in macrophages from bovine mammary glands. Therefore, the aim of this study was to confirm the occurrence of apoptosis in macrophages from resting heifer mammary glands and during the inflammatory response. Methods: Inflammatory response was induced by phosphate buffered saline (PBS) and by lipopolysaccharide (LPS). Resident macrophages (RESMAC) were obtained before and inflammatory macrophages (INFMAC) 24, 48, 72 and 168 hours after inducing inflammatory response in mammary glands of unbred heifers. Cell samples were analyzed for differential counts, apoptosis and necrosis using flow cytometry. Results: Populations of RESMAC and INFMAC contained monocyte-like cells and vacuolized cells. Apoptosis was detected differentially in both morphologically different types of RESMAC and INFMAC and also during initiation and resolution of the inflammatory response. In the RESMAC population, approximately one-tenth of monocyte-like cells and one-third of vacuolized cells were apoptotic. In the INFMAC population obtained 24 h after PBS treatment, approximately one-tenth of monocyte-like cells and almost one-quarter of vacuolized cells were apoptotic. At the same time following LPS, however, we observed a significantly lower percentage of apoptotic cells in the population of monocyte-like INFMAC and vacuolized INFMAC. Moreover, a higher percentage of apoptotic cells in INFMAC was detected during all time points after PBS in contrast to LPS. Comparing RESMAC and INFMAC, we observed that vacuolized cells from populations of RESMAC and INFMAC underwent apoptosis more intensively than did monocyte-like cells. Conclusions: We conclude that apoptosis of virgin mammary gland macrophages is involved in regulating their lifespan, and it is involved in the resolution process of the inflammatory response.