Prevalence and diversity of Arcobacter spp. isolated from the internal organs of spontaneous porcine abortions in Denmark

A study was conducted to determine the prevalence and possible significance of campylobacteria in pig abortions in Denmark. Surface-cauterised liver and kidney samples from 55 aborted pig fetuses submitted to the Danish Veterinary Laboratory were taken and a sensitive isolation procedure used to examine pooled tissue samples for Campylobacter, Arcobacter and Helicobacter spp. Routine microbiological, immunological, and histopathological examinations were also performed to identify concurrent infections or histopathological changes. The abortions tested negative for established abortifacient pathogens (Brucella, Leptospira, PPV, PRRSV), but Arcobacter spp. were recovered from 23/55 abortions. Co-infections with Streptococcus suis, Escherichia coli, and haemolytic streptococci were observed in 7/23 Arcobacter-positive fetuses, and in 4/32 Arcobacter-negative fetuses. Histopathological analyses identified placentitis, pneumonia, hepatitis and encephalitis among the study group. However, no obvious pathologic features were solely associated with Arcobacter-positive cases, nor were Arcobacter-like bacteria observed in tissue samples. Protein profile analyses of the 27 Arcobacter isolates identified 11 as A. cryaerophilus and 10 as A. skirrowii. Six strains could not be classified into any existing species and were phenotypically distinct, thus, potentially representing at least one new species. The identification results showed that multiple taxa could be found in a single fetus, and in distinct aborted fetuses from a single sow. The high prevalence of arcobacters in Danish pig abortions may account for at least some of the >90% of cases in which no established abortifacient agent is detected, but further studies are needed to define the role of each species, especially where co-infections with other bacteria are present.     

Influence of Boar Seminal Plasma on the Distribution of Spermatozoa in the Genital Tract of Gilts

The main purpose of the present study was to investigate whether boar seminal plasma affects the transport of spermatozoa in the genital tract of oestrous pigs or not, with special reference to the sperm transport into the oviducts. Altogether 17 gilts were used in three experiments.Experiment I. In nine gilts one uterine horn was injected surgically with 10¹⁰ spermatozoa suspended in seminal plasma and the other uterine horn with 10¹⁰ spermatozoa suspended in TESNaK-glucose buffer solution. The sperm deposition was performed under general anaesthesia. The gilts were slaughtered 1–2 or 4–6 h after insemination. The genital tract was removed and the numbers of spermatozoa determined in oviducts and in uterine horns.Experiment II. The insemination doses were prepared exactly as in Experiment I. Approx. 24 h before insemination Polyvinylchloride cannulas were inserted into the uterine lumen of the horns, drawn via the midventral incision at linea alba subcutaneously to cutaneous incisions ventral to the vulva opening. One cannula was placed in each uterine horn. At standing heat the insemination doses were slowly injected through the cannulas. The gilts were slaughtered 1 h after insemination and the numbers of spermatozoa within the genital tract were counted.Experiment III. In three gilts under general anaesthesia the uterine horns were ligated 10 cm from the uterotubal junction. The semen doses (containing 2 × 10⁹ spermatozoa), prepared as in Experiment I, were deposited into the uterine horns anterior to the ligatures through a cannula. The gilts were slaughtered 1 h after insemination, and the numbers of spermatozoa within the oviducts and ligated part of the uterine horns were counted.In all three experiments more spermatozoa were, on average, recovered in the oviducts connected to uterine horns inseminated with spermatozoa suspended in seminal plasma. In Experiments I andII this was the case for 10 of 14 gilts and in Experiment III for all the three gilts. It is therefore suggested that boar seminal plasma pro¬motes sperm transport into the oviduct of oestrous pigs. The back¬ground mechanism for this is discussed.     

In vivo confocal microscopy in the normal corneas of cats, dogs and birds

OBJECTIVE: To evaluate the applicability of in vivo confocal microscopy (IVCM) in veterinary ophthalmology and analyze the morphology of living, healthy cornea. ANIMALS EXAMINED: Thirty-seven dogs, 34 cats and five birds. PROCEDURE: Various corneal sublayers were visualized in the central region using an in vivo confocal corneal microscope (HRTII/RCM). RESULTS: An investigation method was developed and adapted for use on animals with varying skull forms and eye positions. Real-time images of the epithelial cells, the corneal stroma and the endothelial layer were obtained. The corneal stromal nerve trunks and the subepithelial and basal epithelial nerve plexus were visualized. In dogs, full corneal thickness (FCT) was 585 +/- 79 microm (mean +/- SD) and endothelial cell density (ECD) 3175 +/- 776 cells/mm(2) (mean +/- SD). In cats, FCT was 592 +/- 80 microm and ECD 2846 +/- 403 cells/mm(2). There were no significant differences between canine and feline FCT and ECD and no morphologic differences could be seen between dogs and cats. The bird images revealed a number of structural differences. CONCLUSION: Noninvasive IVCM allows accurate detection of corneal sublayers, corneal pachymetry, endothelial cell density and corneal innervation in various animal species. For clinical usage, patients must be under general anesthesia. The confocal images provided anatomic reference images of various healthy corneal structures in dogs, cats and birds.     

The humeral immune response of swine after experimental infection with Salmonella typhimurium

LPS antibody reaction of S. Typhimurium was detected in a total of 111 two- to ten-week-old pigs, which had been exposed to experimental oral infection with this serovar in 16 separate experiments. The total antibody level was determined according to the meat juice ELISA provided for the Salmonella control programme and the antibody concentrations of the isotopes IgG1, IgG2, IgA and IgM. Although there were significant differences between the individual experiments with respect to the reaction intensity and proportion of the individual immune globulin isotypes, the following general statements on the dynamics of the antibody reaction can be made: Clearly elevated antibody levels could be observed one week after the onset of infection. After one and two weeks, respectively, 22.5 and 52.9% of the infected animals showed antibody levels which were to be evaluated as positive in the sense of the Salmonella control programme (> or = 40%). While the concentration of the IgM-related antibodies decreased again during the second week already, the antibody concentration of IgG1 and IgG2 continued to increase until the end of the experiment after 3 weeks. The clear differences in the reactions between the experiments cannot be explained solely by defined conditions of infection such as strain of infection, infective dose or age of the pigs. Obviously, other non-defined or non-recognized factors may have a decisive influence on the dynamics of the antibody reaction in the animal groups.     

Herd diagnosis of low pathogen diarrhoea in growing pigs – a pilot study

Background

The major indication for antibiotic use in Danish pigs is treatment of intestinal diseases post weaning. Clinical decisions on antibiotic batch medication are often based on inspection of diarrhoeic pools on the pen floor. In some of these treated diarrhoea outbreaks, intestinal pathogens can only be demonstrated in a small number of pigs within the treated group (low pathogen diarrhoea). Termination of antibiotic batch medication in herds suffering from such diarrhoea could potentially reduce the consumption of antibiotics in the pig industry. The objective of the present pilot study was to suggest criteria for herd diagnosis of low pathogen diarrhoea in growing pigs.Data previously collected from 20 Danish herds were used to create a case series of clinical diarrhoea outbreaks normally subjected to antibiotic treatment. In the present study, these diarrhoea outbreaks were classified as low pathogen (<15% of the pigs having bacterial intestinal disease) (n =5 outbreaks) or high pathogen (≥15% of the pigs having bacterial intestinal disease) (n =15 outbreaks). Based on the case series, different diagnostic procedures were explored, and criteria for herd diagnosis of low pathogen diarrhoea were suggested. The effect of sampling variation was explored by simulation.

Results

The diagnostic procedure with the highest combined herd-level sensitivity and specificity was qPCR testing of a pooled sample containing 20 randomly selected faecal samples. The criteria for a positive test result (high pathogen diarrhoea outbreak) were an average of 1.5 diarrhoeic faecal pools on the floor of each pen in the room under investigation and a pathogenic bacterial load ≥35,000 per gram in the faecal pool tested by qPCR. The bacterial load was the sum of Lawsonia intracellularis, Brachyspira pilosicoli and Escherichia coli F4 and F18 bacteria per gram faeces. The herd-diagnostic performance was (herd-level) diagnostic sensitivity =0.99, diagnostic specificity =0.80, positive predictive value =0.94 and negative predictive value =0.96.

Conclusions

The pilot study suggests criteria for herd diagnosis of low pathogen diarrhoea in growing pigs. The suggested criteria should now be evaluated, and the effect of terminating antibiotic batch medication in herds identified as suffering from low pathogen diarrhoea should be explored.

Electronic supplementary material

The online version of this article (doi:10.1186/2046-0481-67-24) contains supplementary material, which is available to authorized users.     

Distribution of mast cell subtypes and immune cell populations in canine intestines: evidence for age-related decline in T cells and macrophages and increase of IgA-positive plasma cells

The distribution and numbers of leucocytes and mast cells (MC) in the canine gastrointestinal tract of three different age groups was investigated immunhistochemically. In all age groups, CD3+ T cells were more prominent in the villus region than in the crypt areas without differences between intestinal segments, whereas macrophages were more randomly distributed. Kresylecht-violet and tryptase-positive MC were prominent in pericrypt regions with statistic significances. Chymase-bearing mast cells, IgA-, IgG- and IgM-containing cells did not show significant differences in their distribution but, except for IgG-positive cells, subjective trends with increasing numbers towards the crypts exist. The reasons for the distribution of T cells, macrophages, immunoglobulin-containing cells and mast cells are not clear. Lamina propria CD3+ T cells and macrophages significantly decreased whilst a significant increase of IgA-containing plasma cells with increasing age was found. For mast cell subtypes, IgG- and IgM-containing cells no significant changes in numbers with increasing age exist.     

Nitrous oxide emission as affected by changes in soil water content and nitrogen fertilization

Nitrous oxide emission was measured in laboratory incubations of an alluvial soil (58% clay, pH 7.4). The soil was amended with 40 mg N kg⁻¹ as NaNO₃ or NH₄Cl, or with NaCl as a control. Each fertilization treatment was adjusted to three different water contents: constant 60% WHC (water-holding capacity), constant 120% WHC, and water content alternating between 60 and 120% WHC. During an 8-day incubation period N₂O emission rates and inorganic nitrogen concentrations in soil (NH₄⁺, NO₂⁻, NO₃⁻) were determined at regular intervals. In the control and after nitrate application small N₂O emission rates occurred with only minor variations over time, and no differences between the water treatments. In contrast, with ammonium application N₂O emission rates were much higher during the first two days of incubation, with peaks in the constant 60% WHC and 120% WHC at day 1 and in the changing-water treatment at day 2, when the first wet period (120% WHC) was completed. This N₂O peak in the changing-water treatment was 4 to 9 times higher than with constant WHC and occurred when both, NH₄⁺ and NO₂⁻ concentrations declined sharply. Thus, this N₂O emission flush can be attributed to nitrifier denitrification. After the second rewetting of the NH₄⁺-amended soil no further N₂O emission peak was observed, being in accordance with small NH₄⁺ and NO₂⁻ concentrations in soil at that time. The unexpectedly small N₂O fluxes in the constant 120% WHC treatment after nitrate application were probably caused by the reduction of N₂O to N₂ under the prevailing conditions. It can be concluded that continuous wetting or flooding of a soil is an effective measure to reduce N₂O emissions immediately after the application of NH₄⁺ fertilizers.     

Grain yield of wheat (Triticum aestivum L.) under long‐term heat stress is sink‐limited with stronger inhibition of kernel setting than grain filling

The aim of the study was to investigate source‐sink relations of wheat under continuous heat stress and to identify bottle necks of yield formation. A pot experiment was conducted in two climatic chambers exposing wheat plants (Triticum aestivum L. cv. Thasos) either to day/night temperatures of 20/20°C (control conditions) or of 30/25°C (heat stress) during the whole vegetation period in the absence of plant water deficit. Plants were harvested at four phenological stages: three‐node stage (DC 33), start of flowering (DC 61), grain filling (DC 75) and maturity (DC 94). Heat stress shortened the development phases of the plants and caused a significant decrease in total above‐ground biomass between 19% and 41%. At grain filling and at maturity, the reductions in total shoot biomass mainly resulted from grain yield depressions by 77% and 58%, respectively. The ear number per plant was significantly higher under heat stress in comparison with the control, at maturity it was more than doubled. On the contrary, under heat stress, the kernel number per ear was strongly decreased by 83% and 75% during grain filling and at maturity, respectively. The decrease in individual kernel weight was 23% at maturity. Thus, the heat‐stressed plants were able to strongly increase the number of ear‐bearing tillers which were able to set only a small number of kernels, yet these kernels showed good grain filling. The harvest index (HI) of heat‐stressed plants was significantly reduced by 36% (control: HI = 50.1% ± 0.4, heat: HI = 32.2% ± 0.9***). The plants in the stress treatment adapted to the adverse conditions by less biomass production which presumably allowed a higher transpiration without an increase in total water consumption. Nevertheless, under heat stress, the water use efficiency (WUE_grain) was strongly decreased by 62% as a result of a small grain yield. In ears and grains, the sucrose, glucose and fructose concentrations were not significantly different between control and heat stress at start of flowering and during grain filling. Thus, the supply of assimilates was not restricted (no source limitation). Sink capacity was reduced by heat stress, as lesser and smaller kernels were produced than in the control. Concerning sink activity, the sink‐limiting step during kernel set is probably the active transport of hexoses across the plasma membrane into the developing kernels, which could also affect grain filling. This needs to be investigated in more detail in further studies.