Late gestation diet supplementation of resin acid-enriched composition increases sow colostrum immunoglobulin G content, piglet colostrum intake and improve sow gut microbiota.

Resin acid-enriched composition (RAC) mainly containing tall oil fatty acid with an active component of resin acid (RA) can improve the microbial population in the digestive system, change the microbial fermentation, and improve the feed conversion ratio. We investigated the effects of dietary supplementation of RAC on sow colostrum yield (CY), colostrum composition and gut microbiota. Tall oil fatty acid and RA are commonly termed RAC and CLA, pinolenic, abietic, dehydrobiotic acids are characteristic components of RAC. The experiment was conducted in three trials in three respective herds. Sows were fed with a control diet and the same diet supplemented with 5 g RAC/day per sow during the last week of gestation. The 16S ribosomal RNA gene sequencing technique was used to assess sows' faecal microbiota populations at farrowing. Colostrum nutritional composition, acute phase proteins (APPs) and immunoglobulin (Ig) content were also assessed. Individual piglets were weighed at birth and 24 h after the birth of first piglets in order to calculate CY and later at 3 to 4 weeks to calculate average daily gain. The RAC-fed sows had significantly higher IgG levels (P0.05), but those fed RAC had higher levels of colostrum serum amyloid A. Colostrum yield was significantly higher in RAC-fed sows in herds 2 and 3 with heavier piglets between 3 and 4 weeks of age (P0.05). Resin acid-enriched composition supplementation significantly increased some beneficial and fermentative bacteria (Romboutsia and Clostridium sensu stricto) than the control diet (P<0.01) while some opportunistic pathogens (Barnesiella, Sporobacter, Intestinimonas and Campylobacter), including Proteobacteria, were suppressed. Therefore, RAC added to the sow diet at late pregnancy increases colostrum IgG, colostrum availability for neonate piglets, and seems to promote better maternal intestinal microbial sources.

Influence of the uterine inflammatory response after insemination with frozen-thawed semen on serum concentrations of acute phase proteins in mares.

The aim of this study was to investigate the clinical relevance of measuring blood concentrations of serum amyloid A (SAA), haptoglobin (Hp) and fibrinogen (Fib) in horse reproductive management, and changes in response to artificial insemination (AI) with frozen-thawed semen. Standardbred mares (n=18) with different reproductive status (eight healthy mares in first postpartum oestrus, five healthy barren mares and five mares with endometritis) were inseminated with frozen-thawed semen. Endometritis was evaluated during oestrus by bacteriological culture, cytology and presence of ultrasonically visible intrauterine fluid during oestrus. Concentrations of SAA, Hp and Fib were analysed in the blood in every 48h during oestrus and until 5, 6 or 7 days after AI. The day of sampling and number of blood samples varied between mares because of length of the oestrus and time of AI. Changes in concentrations of SAA, Hp and Fib were evaluated based on the day of sampling regard to AI and classification of the mares. There were no differences in SAA, Hp and Fib concentrations over time before or after AI or between the groups of mares. The insemination of mares with frozen-thawed semen did not increase the plasma concentrations of SAA, Hp and Fib above clinical threshold concentration and there were no differences between susceptible or healthy mares.

Efficacy of 5-day parenteral versus intramammary benzylpenicillin for treatment of clinical mastitis caused by gram-positive bacteria susceptible to penicillin in vitro.

The efficacy of parenteral (intramuscular) or intramammary (IMM) benzylpenicillin treatment for clinical mastitis caused by gram-positive bacteria susceptible to penicillin in vitro was investigated. Cows with clinical mastitis in 1 udder quarter were randomly placed into 2 treatment groups. The preliminary bacteriological diagnosis of intramammary infection (IMI) was based on on-farm culturing, and the bacteriological diagnoses were later confirmed by a quantitative PCR assay. Clinical mastitis caused by gram-positive bacteria susceptible to benzylpenicillin was treated with penicillin via either the parenteral route (20mg/kg) or IMM route (600mg) once per day for 5d. The outcome of the treatment was evaluated 3 to 4wk after the onset of the treatment. The affected quarter was examined to assess the clinical cure, and milk samples were collected from the affected quarter to determine the bacteriological cure and milk N-acetyl-ß-d-glucosaminidase activity. The survival and the composite milk somatic cell counts of the treated cows were followed up for 6 and 3mo after treatment, respectively. A total of 140 cows with clinical mastitis were included in the study, 61 being treated with benzylpenicillin parenterally and 79 via the IMM route. From all quarters treated, 108 of 140 (77.1%) were cured clinically and 77 of 140 (55.0%) were cured bacteriologically. The route of treatment did not significantly affect the outcome of the treatment; 80.3% of the quarters with parenteral treatment and 74.7% of the quarters with IMM treatment showed a clinical cure, and 54.1 and 55.7% a bacteriological cure, respectively. The milk N-acetyl-ß-d-glucosaminidase activity was significantly lower in the quarters with a clinical or bacteriological cure than in the quarters with no cure. The 6-mo survival and the proportion of cows with composite milk somatic cell counts <200,000/mL among the treated cows during the 3-mo follow-up period did not significantly differ between the treatment groups. In conclusion, the outcome of either parenteral or IMM benzylpenicillin treatment of clinical mastitis caused by penicillin-susceptible bacteria was similar.

Milk haptoglobin, milk amyloid A, and N-acetyl-ß-D-glucosaminidase activity in bovines with naturally occurring clinical mastitis diagnosed with a quantitative PCR test.

The associations between quantitative bacteriological results from a real-time PCR test and concentrations of acute-phase proteins (APP) and N-acetyl-ß-d-glucosaminidase (NAGase) activity in milk in naturally occurring clinical mastitis were investigated. Milk APP concentrations and NAGase activity in clinical mastitis caused by different udder pathogens were studied. The associations between the severity of the clinical signs and concentrations of APP and NAGase activity were estimated. Milk samples from 281 cases of clinical mastitis were collected from 3 Estonian dairy farms and analyzed by PCR to identify pathogens. Twenty-seven samples out of 281 (9.6%) were PCR negative. Milk samples containing 4 or more bacterial species (n=28) were considered possibly contaminated and excluded from all further analyses. In total, 443 bacterial identifications were made from the remaining 226 milk samples. A single bacterial species was detected in 68 samples (30.1%), 2 species were detected in 99 samples (43.8%), and 3 species were detected in 59 (26.1%) samples. To determine the inflammatory response in the udder, the concentrations of milk amyloid A (MAA) and haptoglobin (Hp) and NAGase activity in the milk were analyzed. A significant positive association was found between the severity of the clinical signs and inflammatory markers in the milk. Milk amyloid A and Hp concentrations and NAGase activity were significantly higher in samples with large quantities of bacterial DNA from Escherichia coli or Streptococcus dysgalactiae compared with milk samples not containing those species. Large quantities of bacterial DNA from Trueperella pyogenes or Streptococcus uberis in the milk were associated with elevated concentrations of Hp and high NAGase activity, but not with increased MAA concentrations. Milk samples containing Corynebacterium bovis and coagulase-negative staphylococci had significantly lower concentrations of MAA and Hp and lower NAGase activity compared with samples where these species were not detected. It can be concluded that concentrations of APP and NAGase activity in the milk were associated with the quantity of bacterial DNA in the milk samples.

Dynamics of bovine herpesvirus type 1 infection in Estonian dairy herds with and without a control programme.

Bovine herpesvirus type 1 (BHV-1) is an important bovine pathogen, exacerbating poor health and the productivity of cattle. The aims of this study were to detect the efficacy of vaccination programmes in lowering the seroprevalence of BHV-1 gE within the dairy herd and to follow the dynamics of the infection in non-vaccinated herds with uninfected heifers. A two-year longitudinal study was carried out on seven herds that were vaccinated, and in five herds with uninfected heifers without applying a control programme. After the start of the vaccination programme, calves born remained free from the virus. However, in one herd, 7 per cent (95 per cent CI 2 to 18) of these animals showed antibodies to BHV-1 two years after the first vaccination. A decline in BHV-1 antibody prevalence was found in vaccinating herds. Among the five herds not under the control programme, one experienced active virus spread, although one herd experienced self-clearance of the virus. In the herds with high BHV-1 prevalence, vaccinating all cattle from three months of age twice a year with a commercial inactivated marker vaccine efficiently protected offspring from becoming infected, and lowered the prevalence of BHV-1 within the herd. A small proportion of herds may experience self-clearance of the virus.

Association of herd BHV-1 seroprevalence with respiratory disease in youngstock in Estonian dairy cattle.

The associations between herd bovine herpesvirus 1 (BHV-1) seroprevalence, along with other infectious and farm management factors with bovine respiratory disease (BRD) in dairy calves and heifers, were investigated. Serum samples from 103 dairy cattle herds were analyzed for antibodies against BHV-1, bovine respiratory syncytial virus (BRSV), bovine viral diarrhea virus (BVDV), and Mycoplasma bovis (M. bovis). A questionnaire was used to record herd management practices. A high occurrence of respiratory disease in unweaned calves was associated with low to moderate and high prevalence of BHV-1 among cows (OR=14.8, p=0.005 and OR=19.2, p=0.002, respectively) and positive BVDV status of a herd (OR=5.1, p=0.02). The presence of BVDV in a herd was related to a high incidence of respiratory disease in heifers 3-16 months old (OR=4.3, p=0.027). Based on the results of multiple correspondence analysis, holding youngstock separately from cows until pregnancy, introducing new animals and the activities of on-farm employees may contribute to a higher incidence of BRD.

Acute phase proteins in milk in naturally acquired bovine mastitis caused by different pathogens.

The concentrations of haptoglobin (Hp) and serum amyloid A (SAA) and the activity of N-acetyl-ß-D-glucosaminidase (NAGase) in milk from 234 cows with spontaneous mastitis caused by different pathogens were measured to assess whether they corresponded with the clinical signs of mastitis and whether there were any differences between pathogens. Ninety-eight of the cows had clinical mastitis and 136 had subclinical mastitis. There were statistically significant positive correlations between the concentrations of SAA and Hp and the activity of NAGase. Significant differences in the concentrations of acute phase proteins and NAGase activity were found in milk from cows with mastitis caused by different pathogens. The highest concentrations of Hp and NAGase were found in cases of mastitis caused by Escherichia coli and Arcanobacterium pyogenes, and the lowest concentrations were from cases of mastitis caused by coagulase-negative staphylococci. Very low SAA concentrations were found in milk from the cases caused by A pyogenes, in contrast to cases caused by other major mastitis pathogens. The median concentration of SAA was over 10 times higher in cases of mastitis caused by E coli than in mastitis caused by other pathogens. There were significant differences in the mean Hp concentration and NAGase activity between clinical and subclinical mastitis. In approximately one-third of the samples, the Hp concentration was below the detection limit, potentially compromising the use of Hp as a mastitis marker.

Seroepidemiology of bovine herpesvirus 1 (BHV1) infection among Estonian dairy herds and risk factors for the spread within herds.

The objectives of this study were to reassess the herd level and within-herd prevalence of bovine herpesvirus 1 (BHV1) infection in Estonian dairy cattle, estimate the sensitivity and specificity of the enzyme-linked immunosorbent assay (ELISA) for bulk tank milk (BTM) testing and determine the risk factors related to high prevalence of the infection in herds. To estimate the herd prevalence, BTM samples from each of the 1,205 herds that sell milk to dairy companies were analysed for BHV1 antibodies. One hundred and three herds with known BHV1 infection status were selected to estimate within-herd prevalence and to calculate the sensitivity and specificity of BTM ELISA. In these herds serum samples were collected from cows and youngstock, together with BTM samples. A commercial blocking ELISA test was used to analyse samples for antibodies against BHV1. A questionnaire was completed to collect herd data. The sensitivity and specificity of the BTM ELISA were 76.5% and 97.2%, respectively, and the true herd prevalence of BHV1 was calculated to be 22.0%. The herd prevalence increased significantly with herd size, being 3.4% in the smallest category (less than 20 cows) and 85.7% in herds of size over 400. The mean within-herd prevalence was 37.8% (range 1-100, median 31.5). The mean within-herd prevalence increased with herd size. Data from 59 infected herds was used to determine the risk factors associated with high within-herd prevalence (>50%) of BHV1, using logistic regression analysis. As, in some infected herds, the youngstock were uninfected, risk factors for the presence of BHV1 among youngstock from 6 months until calving were analysed. The results indicate the importance of iatrogenic spread of the virus, since the overall within-herd prevalence was higher in those herds in which a veterinarian was an employee of the farm and an inseminator worked only for the particular farm. The presence of bovine viral diarrhoea virus (BVDV) in a herd was associated with a higher prevalence of BHV1.

Setaria tundra microfilariae in reindeer and other cervids in Finland.

Harmful parasites of the wild northern boreal mammals are still surprisingly poorly studied. In 2003-2006, a peritonitis outbreak caused by the filarioid nematode, Setaria tundra, emerged in Finland's reindeer population. In order to gain knowledge about the basic biology, epidemiology, and transmission dynamics of this parasite, samples for S. tundra were collected from reindeer and other cervids during the follow-up period 2004-2006. Using morphology and molecular biology methods, we describe here S. tundra's first larval stage, microfilaria (smf), for the first time scientifically. The prevalence and densities of smf were higher in reindeer calves than in adults, overall prevalence being 42%. The overall smf prevalences for moose, wild forest reindeer and roe deer were 1.4-1.8%, 23%, and 39%, respectively. The focus of microfilaremia moved north and settled down in the south simultaneously with the peritonitis outbreak. The peak microfilaremia occurred in the first summer after the infection, and smf disappeared from the blood after 2 years. Captive reindeer were smf positive over the year. The prepatent period of S. tundra was estimated to be about 4 months, and the life span at least 14 months. This parasite likely has an important impact on boreal ecosystems.

Host response in bovine mastitis experimentally induced with Staphylococcus chromogenes.

An experimental infection model was developed to study host response to intramammary infection in cows caused by Staphylococcus chromogenes. CNS intramammary infections have become very common in modern dairy herds, and they can remain persistent in the mammary gland. More information would be needed about the pathophysiology of CNS mastitis, and an experimental mastitis model is a means for this research. Six primiparous Holstein-Friesian cows were challenged with S. chromogenes 4 weeks after calving. One udder quarter of each cow was inoculated with 2.1 x 10(6)cfu of S. chromogenes. All cows became infected and clinical signs were mild. Milk production of the challenged quarter decreased on average by 16.3% during 7 days post-challenge. Cows eliminated bacteria in a few days, except for one cow which developed persistent mastitis. Milk indicators of inflammation, SCC and N-acetyl-beta-D-glucosaminidase (NAGase) returned to normal within a week. Milk NAGase activity increased moderately, which reflects minor tissue damage in the udder. Concentrations of serum amyloid A (SAA) and milk amyloid A (MAA) were both elevated at 12h PC. MAA was affected by the milking times, and was at its highest before the morning milking. In our experimental model, systemic acute phase protein response with SAA occurred as an on-off type reaction. In conclusion, this experimental model could be used to study host response in CNS mastitis caused by the main CNS species and also for comparison of the host response in a mild intramammary infection and in more severe mastitis models.

Association of bovine respiratory disease with clinical status and acute phase proteins in calves.

Eighty-four calves with respiratory disease from 18 herds in different parts of Finland were chosen for a study evaluating the capacity of different respiratory pathogens to cause changes in different acute phase protein concentrations, white blood cell (WBC) count and clinical signs. The selected acute phase proteins were fibrinogen, haptoglobin, serum amyloid-A, lipopolysaccharide binding protein and alpha1-acid glycoprotein. From each calf, a paired blood sample was obtained for serological studies of bovine parainfluenza virus-3, bovine respiratory syncytial virus, bovine coronavirus, bovine adenovirus-3 and bovine adenovirus-7. Tracheobronchial lavage was performed to detect bacteria and mycoplasma. Isolation of Pasteurella multocida was associated with increased concentrations of all tested acute phase proteins. For other pathogens, no significant relationships were observed. No association was present between viral or bacterial findings and WBC count.

Factors affecting fertility in loosely housed sows and gilts: vulvar discharge syndrome, environment and acute-phase proteins.

The effect of vulvar discharge syndrome (VDS) on sow and gilt fertility was studied on 26 farms. Of 824 animals inspected in 21 randomly selected and five VDS problem farms, 19 (2.3%) were afflicted with VDS. Altogether 542/799 of the examined animals (67.8%) farrowed thereafter. Nine of the 19 VDS animals (47.4%) and 533/780 non-VDS animals (68.3%) farrowed at the first chance after the examination (p = 0.05). None of the unmated gilts in this study had VDS. Environmental and individual factors likely to be associated with fertility and VDS were tested. In multivariate analysis, factors associated with farrowing were VDS, reproductive status, availability of roughage and confinement to individual stalls. None of the variables tested was associated with VDS. However, all of the VDS problem farms were overcrowded and had concrete, partly slatted floors with little or no bedding. The median value of haptoglobin (Hp) was 2.5 (range 1.3-3.1) g/l in VDS animals and 2.3 (0.5-4.3) g/l in controls (p = 0.6). The median C-reactive protein (CRP) in VDS animals was 30.3 (3.3-171.3) mg/l and in controls 25.9 (3.3-361.1) mg/l (p = 0.7). In conclusion, VDS decreased fertility of gilts and sows in the absence of a systemic acute-phase response, as indicated by stable concentrations of Hp and CRP.

Lameness and fertility of sows and gilts in randomly selected loose-housed herds in Finland.

The prevalence of lameness among 646 sows and gilts in 21 selected herds was determined; 8.8 per cent of the animals were lame and the most common clinical diagnoses were osteochondrosis, infected skin lesions and claw lesions. The lame animals had higher serum concentrations of haptoglobin and C-reactive protein than the sound animals. Animals housed on slatted floors had twice the odds of being lame and 3.7 times the odds of being severely lame than animals housed on solid floors. Yorkshire pigs had 2.7 times the odds of being lame than Landrace or crossbred animals. Higher parity and the use of roughage decreased the odds of the sows not becoming pregnant; however, lameness was not a risk factor for non-pregnancy.

Transgenic cows that produce recombinant human lactoferrin in milk are not protected from experimental Escherichia coli intramammary infection.

This is the first study describing an experimental mastitis model using transgenic cows expressing recombinant human lactoferrin (rhLf) in their milk. The aim of the study was to investigate the concentrations in milk and protective effects of bovine and recombinant human lactoferrin in experimental Escherichia coli mastitis. Experimental intramammary infection was induced in one udder quarter of seven first-lactating rhLf-transgenic cows and six normal cows, using an E. coli strain isolated from cows with clinical mastitis and known to be susceptible to Lf in vitro. Clinical signs were recorded during the experimental period, concentrations of human and bovine Lf and indicators of inflammation and bacterial counts were determined for milk, and concentrations of acute-phase proteins and tumor necrosis factor alpha were determined for sera and milk. Serum cortisol and blood hematological and biochemical parameters were also determined. Expression levels of rhLf in the milk of transgenic cows remained constant throughout the experiment (mean, 2.9 mg/ml). The high Lf concentrations in the milk of transgenic cows did not protect them from intramammary infection. All cows became infected and developed clinical mastitis. The rhLf-transgenic cows showed milder systemic signs and lower serum cortisol and haptoglobin concentrations than did controls. This may be explained by lipopolysaccharide-neutralizing and immunomodulatory effects of the high Lf concentrations in their milk. However, Lf does not seem to be a very efficient protein for genetic engineering to enhance the mastitis resistance of dairy cows.

Temporal changes in concentrations of serum amyloid-A and haptoglobin and their associations with weight gain in neonatal reindeer calves.

Age-related changes in serum concentrations of two acute phase proteins (APPs), haptoglobin (Hp) and serum amyloid-A (SAA) were investigated in newborn reindeer calves. Repeated blood samples were obtained from 51 reindeer calves at ages 0-32 days (2-4 samples from each calf). An increase of SAA concentrations was observed during the first 2 weeks of life. However, by the end of the observation period, SAA concentrations had decreased to levels below those of the first week. Serum Hp concentrations increased throughout the observation period. SAA concentrations in the second week had a negative association with weight gain during the entire study period (4 months). These time-related changes in APP concentrations suggest that these proteins have a role in the defence and adaptation mechanisms of newborn reindeer calves. Possible reasons for these changes include the presence of APP mediators in the colostrum, exposure to environmental pathogens after birth and age-related changes in hepatic synthesis of APP.

Acute phase response in reindeer after challenge with Escherichia coli endotoxin.

The serum concentrations of two acute phase proteins (APPs), haptoglobin (Hp) and serum amyloid-A (SAA), were monitored in reindeer after challenge with endotoxin. Four adult female reindeer received either 0.1 microg/kg Escherichia coli 0111:B4 lipopolysaccharide B or saline solution intravenously. At the second challenge, the treatments were reversed. In addition to the APPs, changes in blood chemistry and rectal temperature were monitored. The endotoxin challenge caused a significant increase in SAA (peak 48 h) and a sharp decrease (8-12 h) of serum iron concentrations in all animals. The mean Hp concentration increased at 8 h and remained elevated until 48 h, but no statistically significant differences were found. This investigation demonstrates that challenge with a single-bolus dose of E. coli endotoxin can activate the acute phase response (APR) and SAA appears to be a more sensitive indicator of the APR than Hp during bacterial infection in reindeer.