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Biomedical subjects

T A Niewold

Publications and source records attributed to T A Niewold.

At least 19 recordsLinked to original sources

Kinetics of local and systemic isoforms of serum amyloid A in bovine mastitic milk.

The aim of the present study was to characterise the serum amyloid A (SAA) response to intramammary inoculation of Escherichia coli and to examine the distribution of hepatically and extrahepatically produced SAA isoforms in plasma and milk from cows with mastitis. Milk and plasma SAA concentrations were determined before and after experimental induction of E. coli mastitis in six dairy cows. The milk SAA response was characterised by low or undetectable levels before inoculation, very rapid and large increases in concentration after inoculation, and rapid decline towards baseline levels after resolution of disease. In plasma from cows with experimentally induced E. coli mastitis, four hepatically derived SAA isoforms with apparent isoelectric point (pI) values of 5.8, 6.2, 6.8 and 7.4 were demonstrated by denaturing isoelectric focusing. In milk three highly alkaline isoforms with apparent pI values above 9.3 appeared 12 h post-inoculation. These isoforms were not present in any of the plasma samples, and it therefore seems likely that they were locally produced, tissue-specific isoforms. At 24-36 h post-inoculation one or more acidic isoforms corresponding to those found in plasma appeared in the milk samples. The isoforms demonstrated in plasma from cows with E. coli mastitis were also present in serum obtained from three cows with clinical Streptococcus uberis mastitis. In conclusion, experimentally induced E. coli mastitis is accompanied by a prominent SAA response. The results of the present study indicate that SAA accumulation in mastitic milk is the result of both local synthesis of SAA and of hepatically derived SAA gaining access to the milk due to increased permeability of the blood-milk barrier.

Animals↗

Bacterial growth during the early phase of infection determines the severity of experimental Escherichia coli mastitis in dairy cows.

The aim of this study was to investigate the importance of bacterial growth for the severity of experimental Escherichia coli mastitis, indirectly expressed as the area under the curve of bacterial counts in milk over time. The association of pre-infusion somatic cell count and post-infusion influx of inflammatory cells in milk with severity of infection was also examined. Bacterial growth was studied through culture in milk samples (in vitro) and through monitoring of bacterial counts in milk during the early phase of infection (in vivo) in 36 cows. Individual variation in bacterial counts was more than 2 x 10(2)-fold after 6 h of in vitro incubation, and more than 8 x 10(2)-fold 6 h after intramammary infusion. In vitro growth in milk was not associated with in vivo growth during the early phase of infection, nor with severity of E. coli mastitis. Somatic cell count before experimental E. coli mastitis was negatively associated with in vivo bacterial growth during the early phase of infection (R2 = 0.28), but was not associated with severity of E. coli mastitis (R2 = 0.06). In vivo bacterial growth during the early phase of infection (positive association; R2 = 0.41), together with influx of inflammatory cells in milk, expressed as mean hourly increase of somatic cell count between 6 and 12 h post-infusion (negative association; R2 = 0.11), are major determinants for the severity of experimental E. coli mastitis (R2 = 0.56).

Animals↗

Acute phase protein concentrations in serum and milk from healthy cows, cows with clinical mastitis and cows with extramammary inflammatory conditions.

The concentrations of the two acute phase proteins, serum amyloid A and haptoglobin, in serum and milk were compared in 10 cows with clinical mastitis, 11 cows with extramammary inflammatory conditions and 10 clinically healthy control cows. The concentrations of both acute phase proteins were higher in the serum and milk of the cows with mastitis than in the cows in the other two groups. Four of the cows with extramammary inflammatory conditions had serum amyloid A concentrations in serum above 100 microg/ml, but negligible concentrations in milk, indicating that a pathogen must be present in the mammary gland for serum amyloid A to accumulate in milk. The acute phase protein concentrations in milk increased significantly with increasing somatic cell count, suggesting that they may be indicators of the severity of an infection.

Animals↗

Plasma intestinal fatty acid binding protein (I-FABP) concentrations increase following intestinal ischemia in pigs.

Intestinal fatty acid binding protein (I-FABP) is an intracellular epithelial protein in the intestinal mucosa of many animals. I-FABP appears in the circulation following epithelial damage, and in humans, is proven to be a parameter for damage to the mucosa. In this paper, an ELISA test designed for human I-FABP analysis was used to assay pig blood samples. The test recognized I-FABP cloned from pig small intestine and expressed in Escherichia coli. Furthermore, in our experimental model of (low flow) intestinal ischemia and reperfusion a significant rise in plasma I-FABP concentrations 15-30 min after clamping of the mesenteric artery was demonstrated. This is the first report that in pigs circulating I-FABP is a useful marker for (mild) intestinal injury, and could possibly be used to monitor (intestinal) health in clinical practice.

Animals↗

In vitro growth of mastitis-inducing Escherichia coli in milk and milk fractions of dairy cows.

The outcome of E. coli mastitis in cows ranges from mild to severe in individual animals. This study explored the hypothesis that milk from individual cows differs in its growth medium properties for E. coli, and whether possible variation could be related to specific milk constituents. To mimic the early phase of intramammary E. coli infection, a low inoculum size and a short incubation period were used. Cell-reduced, cell- and fat-free (skim) and cell- and fat-free and protein-reduced (whey) fractions were prepared from whole milk samples (n=18). Ten ml of whole milk, milk fractions and brain heart infusion broth (BHI) were inoculated with approximately 100cfu E. coli. After 6h of incubation, bacterial counts were assessed by dilution plating in triplicate. Bacterial counts in whole milk differed up to a 100-fold between cows, which was not associated with SCC. Bacterial counts were significantly higher in whey fractions than in whole milk, cell-reduced and skim fractions and variation in whey was smaller, indicating that the acid-precipitable protein fraction contains the milk constituents of major relevance for inhibition of and variation in bacterial growth. The presence of fat and cells added to bacterial growth inhibition to a lesser extent. In conclusion, in vitro growth of E. coli in milk differs substantially between individual cows within an incubation period comparable with the early phase of intramammary infection. This suggests that the growth medium properties of milk could be of importance in the pathogenesis of E. coli mastitis and subsequent outcome of disease.

Animals↗

Differences in polymorphonucleocyte function and local inflammatory response between horses and ponies.

REASONS FOR PERFORMING STUDY: Wound healing proceeds faster in ponies than in horses and complications during healing, such as wound infection, occur less frequently in ponies. Earlier studies suggested that this difference might be related to differences in the initial post traumatic inflammatory response. HYPOTHESIS: That polymorphonuclear leucocyte (PMN) function and profiles of humoral factors in local inflammatory processes are different in horses and ponies. METHODS: PMNs were isolated from venous blood of horses and ponies. Chemotaxis and reactive oxygen species (ROS) production was determined. Tissue cages were implanted in limbs and necks of horses and ponies and injected with carrageenan and, 3 weeks later, with LPS. In sequential samples of inflammatory exudate, the numbers of macrophages and PMNs and the production of PGE2, TNFalpha, IL-1, IL-6 and chemoattractants were determined. RESULTS: In vitro ROS production of PMNs was significantly higher in ponies than in horses, whereas in vitro PMN chemotaxis was significantly lower in ponies. In the tissue cages for both stimuli, the production of IL-1 and chemoattractants was significantly higher in ponies than in horses and remained so towards the end of the observation period in ponies. CONCLUSIONS: This study demonstrated a higher production of various inflammatory mediators by pony leucocytes. Despite the lower in vitro chemotaxis of pony PMNs, this higher in vivo production resulted in a stronger initial inflammatory response in ponies, as has been reported in studies on wound healing, through the attraction of leucocytes and triggering of the production of other cytokines. A stronger initial inflammation may promote healing by more rapid elemination of contaminants and earlier transition to repair. POTENTIAL RELEVANCE: Modulation of the initial inflammatory response might therefore be a valid option for therapeutic intervention in cases of problematic wound healing. Further, the intraspecies differences in leucocyte function may have an impact on many fields in equine medicine.

Animals↗

Peritoneal, systemic, and distant organ inflammatory responses are reduced by a laparoscopic approach and carbon dioxide versus air.

BACKGROUND: Advantages of laparoscopic surgery have, among other factors, been attributed to a shorter length of abdominal incision and the use of CO2 versus air. An analysis of these factors taking pressure-induced alterations into account is lacking. The objective of the study was to determine the impact of laparoscopy and laparotomy with exposure to CO2 and room air under a similar pressure on local, systemic, and distant organ immune responses. METHODS: Twenty piglets were randomized into four groups: CO2 laparoscopy, air laparoscopy, CO2 laparotomy, and air laparotomy. Laparotomy was performed in a sterile balloon pressurized similar to laparoscopy. Peritoneal interleukin-1, interleukin-6, tumor necrosis factor-a, and counts of polymorphnuclear cells (PMNs), and macrophages (MFs) were determined in abdominal lavage fluids at 0, 2, and 48 h. Macrophages were assessed for reactive oxygen species (ROS) production. Systemic responses were gauged by white blood cell count (WBC) and cytokines. Alveolar lavage was performed at 48 h to determine cytokine levels, cell counts, and MF ROS production. Blood, lavage fluids, and mesenteric lymph nodes were tested for bacterial translocation. RESULTS: Regarding the peritoneal response, laparotomy versus laparoscopy when performed with CO2 significantly increased PMN and decreased the percentage of macrophages (%MF) up to 48 h. There was a significant increase in interleukin-6, and there was a fourfold increase in MF ROS production. Similar differences between the procedures were found with exposure to air. The use of air versus CO2 in laparoscopy, but not in laparotomy, resulted in an increase of peritoneal PMN and a decrease of the %MF up to 48 h. Air increased the local interleukin-6 release in both procedures and increased MF ROS production fourfold. Regarding the systemic response, laparotomy produced a significant increase in WBC, which was significantly more pronounced with exposure to air. No alteration of other systemic cytokines was seen. Regarding the pulmonary response, the number of MFs and MF ROS production were significantly increased after air versus CO2 laparoscopy. There were no such differences between the laparotomy groups. Regarding bacterial translocation, no bacteria were cultured from peritoneal fluids, lymph nodes, or blood. CONCLUSIONS: Inflammatory responses were reduced by a laparoscopic approach and by exposure to CO2 versus air. Peritoneal responses were affected to a larger degree than systemic parameters. Laparotomy overruled the effects of CO2 on chemotaxis and distant organ injury but not on peritoneal cytokine release.

Air↗

The effect of dietary spray-dried porcine plasma on clinical response in weaned piglets challenged with a pathogenic Escherichia coli.

Weaned piglets were used to determine the effect of dietary spray-dried porcine plasma (SDPP) on the clinical response to an infection with a pathogenic Escherichia coli (E. coli) O139:K82 LT(-). The piglets were divided into two groups of 10 animals each. One group was fed the control diet containing soybean(meal) plus whey powder. The test piglets were fed a diet with 8% SDPP. Piglets were orally infected with the challenge strain on days 6 and 7 after weaning. The experimental period lasted 14 days after which the piglets were euthanised and necropsied. Faecal samples were collected daily for bacteriological analysis. Segments of jejunum, caecum and rectum were removed for bacteriological analysis post mortem. Feed intake and weight gain, faecal and condition scores and body temperature were measured daily. In the control and SDPP groups, 6 and 7 piglets died from diarrhoea. The average daily feed intake (ADFI) and average daily gain (ADG) were substantially higher in the SDPP group than in the control group. SDPP-fed piglets generally had a more favourable faecal score and a healthier appearance than did the control piglets. The faecal excretion of E. coli O139:K82 was similar for control and test piglets. There were no diet effects on the E. coli O139:K82 counts at different sites of the intestine. In this experiment, the inclusion of SDPP at an economically acceptable percentage in the diet could not prevent piglet losses due to challenge with a pathogenic E. coli, but improvements of ADG, ADFI and faecal and condition scores were achieved.

Animal Feed↗

Small intestinal morphology and disaccharidase activities in early-weaned piglets fed a diet containing spray-dried porcine plasma.

The purpose of this study was to test whether dietary spray-dried porcine plasma (SDPP) in early-weaned piglets prevents small intestinal villus atrophy by trophic or protective activity. Fifty-four weaned, 18-day-old piglets were used to determine the effect of dietary SDPP on small intestinal villus length, crypt depth, enterocyt mitotic activity and brush border enzyme activities during the first week after weaning. The piglets were offered a diet containing either 8% SDPP or 8% casein. At 2 and 7 days after weaning, piglets were anaesthetized to provide samples of the small intestinal wall and killed immediately afterwards. There were no differences in daily gain and daily feed intake between the two dietary treatments. At day 2 after weaning, all piglets showed a marked reduction in villus height when compared with baseline values. In all piglets, small intestinal enterocyte mitotic activity had decreased by day 2 and was increased again on day 7. There were no significant effects of dietary SDPP on small intestinal villus length, crypt depth and enterocyt mitotic activity. This indicates that SDPP has no trophic effect on the small intestinal mucosa and that it does not protect against the damaging effect on the small intestinal villi that is associated with the process of weaning. There was no effect of SDPP on lactase-, sucrase- or maltase-specific activities that are a measure of the digestive function of the small intestine. It can be concluded that SDPP versus casein has no effect on small intestinal morphology and disaccharidase activities in early weaned piglets kept under low infection pressure.

Animal Feed↗

Assessment of respiratory herd health in weaner pigs by measuring cellular composition of bronchoalveolar lavage fluid.

Cell differentiation of bronchoalveolar lavage fluid (BALF) was compared among 11 herds having a history of recurrent respiratory disease in weaner pigs and nine herds lacking such a history. In every herd, 20 pigs aged 8-10 weeks were lavaged. The two groups differed significantly on median percentage of macrophages, neutrophils and lymphocytes, but not on white blood cell count of the BALF. Logistic regression showed the percentage of samples per herd exceeding the reference value for neutrophils of 0-8% to be the most promising parameter to assess the health status in weaner pigs.

Animals↗

Disease incidence and immunological traits for the selection of healthy pigs. A review.

Disease is a major issue in animal production systems and society demands that the use of medicines and vaccines be reduced. This review describes the breeding approaches that could be used to improve disease resistance and focuses especially on their application to pigs. Disease reduction by genetic means has certain advantages through cumulative and permanent effects, and direct and indirect selection methods are available. Direct selection for disease incidence requires, besides a unique pig identification and disease registration system, challenge routines that are inconvenient in intensive pig production. Indirect selection for the expression of immune capacity may be an alternative but requires detailed knowledge of the different components of the immune system. There is ample opportunity for genetic improvement of the immune capacity because immune traits show substantial genetic variation between pigs. We therefore conclude that indirect selection via immune traits is very interesting, also for practical implementation, and that there is an urgent need for knowledge, within lines, about the genetic relationships between immune capacity traits and resistance to specific diseases or to disease incidence in general. Furthermore, knowledge about the relationship between immune system traits and production traits is needed as well as knowledge about the effect of selection on the epidemiology of disease at a farm/population level and on the host-pathogen interaction and coevolution.

Animals↗

Overview of the Third International Workshop on Swine Leukocyte Differentiation Antigens.

The aim of the Third International Workshop on Swine Leukocyte Differentiation Antigens (CD workshop), supported by the Veterinary Immunology Committee (VIC) of the International Union of Immunological Societies (IUIS), was to standardize the assignment of monoclonal antibodies (mAb) reactive with porcine leukocyte differentiation antigens and to define new antibody clusters, using nomenclature in accordance with human and ruminant CD nomenclature, as agreed at the summary meeting of the Second International Swine CD Workshop in Davis, 1995: only mAb with proven reactivity for the orthologous porcine gene product or cross-reactivity for the human gene products, were given the full CD nomenclature, all other allocations were prefixed with "w". As in previous workshops, the overall organization was entrusted to the chair and first author, with support by the chair of the previous workshop and second author. In addition to the existing 26 pig leukocyte CD/SWC determinants established in previous workshops, this workshop established/confirmed another 11 CDs for pig leukocytes, identified by a total of 21 mAb: CD11R1 (2 mAb), CD11R2 (1 mAb), CD11R3 (4 mAb), wCD40 (1 mAb), wCD46 (4 mAb), wCD47 (3 mAb), wCD49d (1 mAb), CD61 (1 mAb), wCD92 (1 mAb), wCD93 (1 mAb) and CD163 (2 mAb).

Animals↗

Summary of the first round analyses of the Third International Workshop on Swine Leukocyte Differentiation Antigens.

The reactivity of 155 monoclonal antibodies submitted to the Third International Workshop on Swine Leukocyte Differentiation Antigens, together with 41 internal standards, was analysed by flow cytometry on 29 different pig cell targets as well as two human cell targets as a means of establishing suitable panels of monoclonal antibodies for more detailed clustering analyses by the various subsections of the workshop. Results were collected either without further gating, with gating based on FS/SS characteristics or with gating based on the co-expression of a reference antibody in two-colour flow cytometry. The CD or SWC reactivity of the internal standards had been established in previous workshops. Data sets were subsequently analysed by statistical clustering using the Leucocyte Typing Database IV software. The resulting 18 cluster groups were allocated to the appropriate second round sections of the workshop, after reviewing the overall cellular reactivity of each cluster as well as the specificity of known standards which clustered in a group.

Animals↗

Thirty minutes transport causes small intestinal acidosis in pigs.

Long duration (>10 hours) transport has been described as having either adverse or no effects on porcine health. However, the effect of short duration transport on porcine health is unknown. In the present paper, pigs fed ad libitum (n = 6) were transported for 30 minutes, anaesthetised, instrumented, and cardiovascular and gastrointestinal parameters were measured. Non-transported pigs (n = 6) served as controls. No significant differences between groups were found concerning blood flow in the arteria mesenterica cranialis, heart rate, cardiac output, pulmonal blood pressure, haemoglobin content and haematocrit value. Systolic systemic blood pressure was higher (though not significant), and diastolic systemic blood pressure was significantly higher in the transported pigs than in the controls. Small intestinal pH was significantly lower in transported pigs than in control pigs. We conclude that a 30-minute transport of fed pigs results in small intestinal acidosis. As small intestinal acidosis predisposes to bacterial translocation, even short duration transport should be avoided when possible.

Acidosis↗

Oedema disease is associated with metabolic acidosis and small intestinal acidosis.

Limited information is available about the pathogenesis and pathophysiology of oedema disease (OD). Oedema disease is caused by specific enterotoxemic Escherichia coli (SLTIIv-toxin producing) strains; however, the same strains are also found in non-afflicted pigs. Furthermore, it is unclear how the 80 kDa SLTIIv-toxin can pass the intestinal barrier. In the present paper, piglets showing signs of acute OD were anaesthetised, instrumented and cardiovascular and intestinal parameters were determined at 0, 1, 2 and 3 hours. Healthy piglets from the same herd were used as a control. Cardiac output, blood pH and bicarbonate, small intestinal intramucosal pH, and (pulmonary) blood pressure were significantly lower in OD-pigs than in control pigs. It is concluded that OD is associated with metabolic and intestinal acidosis. Intestinal acidosis is known to increase macromolecular permeability. This suggests that once OD has developed, influx of SLTIIv-toxin into the blood stream is facilitated, thus perpetuating the disease. Since intestinal permeability appears to be central in OD, it is argued that post-weaning events increase intestinal permeability and predispose individuals to OD.

Acidosis↗

Small intestinal morphology in weaned piglets fed a diet containing spray-dried porcine plasma.

The hypothesis tested in this study was that the reported beneficial effects of spray-dried porcine plasma (SDPP) on piglet post-weaning performance and health are associated with a trophic effect on small intestinal mucosa. At 24 days of age, the piglets of seven sows were assigned to one of three treatment groups. One group continued to be suckled. The other two groups were weaned and offered a diet containing either 15 per cent SDPP or casein. From each treatment group, one piglet was anaesthetised and samples were taken from the small intestinal wall at 26, 28 and 31 days of age. There were no significant effects of SDPP versus casein on villus length. On average, there was less mitotic activity in the SDPP -fed piglets than in those fed casein on days 4 and 7 after weaning. As less mitotic activity leads to less immature enterocytes, this may provide a mechanism for the reported beneficial effects of SDPP on performance and health.

Animal Feed↗

Llama heavy-chain V regions consist of at least four distinct subfamilies revealing novel sequence features.

In addition to conventional antibodies (Abs), camelids possess Abs consisting of only heavy chains. The variable domain of such a heavy-chain Ab (VHH) is fully capable of antigen (Ag) binding. Earlier analysis of 47 VHHs showed sequence features unique to VHH domains. These include the presence of characteristic amino acid substitutions in positions which, in conventional VH domains are involved in interdomain interactions, and the presence of a long third complementarity-determining region (CDR3) which is frequently constrained by an interloop disulphide bond. Here, we describe a large (152) set of Lama glama VHH cDNAs. Based on amino acid sequence similarity, these and other published camelid VHHs were classified into four subfamilies. Three subfamilies are absent in dromedaries, which have been the primary source of VHHs thus far. Comparison of these subfamilies to conventional VH regions reveals new features characteristic of VHHs and shows that many features earlier regarded as characteristic of VHHs in general are actually subfamily specific. A long CDR3 with a concomitant putative additional disulphide bond is only observed in two VHH subfamilies. Furthermore, we identified new VHH-characteristic residues at positions forming interdomain sites in conventional VH domains. The VHH subfamilies also differ from each other and conventional VH domains in the canonical structure of CDR1 and CDR2, mean CDR3 length, and amino acid residue variability. Since different VHH-characteristic residues are observed in all four subfamilies, these subfamilies must have evolved independently from classical VH domains.

Amino Acid Sequence↗

A review of porcine pathophysiology: a different approach to disease.

Diseases are often thought to result from a single cause. Although this is sometimes the case, e.g. with a highly virulent infection such as Classical Swine Fever (CSF), more often clinical disease in swine herds results from multiple predisposing factors. This is especially true in modern intensive pig husbandry, in which the role of highly infectious diseases is limited to (nonetheless devastating) outbreaks. More important nowadays are diseases, although associated with an agent, without a clear pathogenesis. The emphasis in disease control thus far has been on treatment, eradication and prevention. This has been achieved by focusing attention on husbandry factors, such as climate, housing, hygiene, management, and nutrition. Although this approach has been successful for a number of diseases, several health problems are persistent. There are strong indications that in the latter, intrinsic animal factors are important. Successful handling of these problems requires knowledge of the (patho)physiology of the pig. In this article, several characteristics of pig physiology associated with the occurrence of disease are described. It appears that the modern (fattening) pig is exceptional among other animal species in that its cardiovascular system is mismatched to its body weight. It is argued that this particular disposition causes relatively minor disturbances to have major consequences in the pig. This concept of pig physiology is central to the understanding of the hitherto poorly understood pathogenesis of several diseases, such as oedema disease.

Animal Husbandry↗