Clinical, biochemical, and molecular aspects of Glanzmann's thrombasthenia in humans and dogs

Glanzmann's thrombasthenia (GT) is an inherited, intrinsic platelet function defect that involves the platelet glycoprotein complex IIb-IIIa, also known as the fibrinogen receptor and the integrin alphaIIbbeta3. The defect was originally described by Dr. Glanzmann in humans in 1918 as a bleeding disorder that differed clinically from other known coagulopathies. Over the decades that followed, researchers determined the biochemical and molecular basis for the disease in humans. Otterhounds with thrombasthenic thrombopathia, described in the 1960s, were the only animal model that closely resembled the disease described in humans until 1996. At that time, a Great Pyrenees dog was identified with unequivocal clinical and biochemical features of Type I GT The cDNA encoding for glycoproteins IIb and IIIa were sequenced in normal dogs in 1999, allowing for identification of specific mutations causing Type I GT in both Otterhounds and Great Pyrenees dogs. Knowing the molecular basis for Type I GT in dogs as well as the cDNA sequences in normal dogs should enhance the understanding of structure/function relationships of the alphaIIbbeta3 integrin and provide an excellent animal model for studies aimed at correction of GT in humans. The following review focuses on the structure and function of this platelet receptor and reviews the molecular, biochemical, and clinical aspects of Glanzmann's thrombasthenia in humans and dogs.     

Comparison of the Value of Pulsed-field Gel Electrophoresis,Random Amplified Polymorphic DNA and Amplified rDNA Restriction Analysis for Subtyping Taylorella equigenitalis

Eight strains of Taylorella equigenitalis were identified by a polymerase chain reaction using a primer pair specific to the 16S rDNA of T equigenitalis. These eight strains were chosen because they had previously been shown to represent eight distinct genotypes by pulsed-field gel electrophoresis analysis after separate digestion of the genomic DNA with ApaI or NotI. The eight strains could be classified into six or seven types by random amplified polymorphic DNA analysis using different kinds of primers. Amplified rDNA restriction analysis after separate digestion with five restriction enzymes, including AluI and MboI, of the 1,500 bp fragments of rDNA amplified by polymerase chain reaction did not discriminate the genomic variations among the eight strains of T equigenitalis. Thus, pulsed-field gel electrophoresis was shown to discriminate these eight organisms better than random amplified polymorphic DNA analysis, while amplified rDNA restriction analysis was found to be unsuitable for subtyping T equigenitalis.     

Laparoscopic surgical technique for repair of rectal and colonic tears in horses: an experimental study

OBJECTIVE: To determine the efficacy of laparoscopic surgical techniques for repair of rectal lacerations in horses. STUDY DESIGN: Experimental study. ANIMAL OR SAMPLE POPULATION: Thirty-two segments of equine bowel placed in an equine pelvitrainer, 8 equine cadavers, and 3 normal horses. METHODS: In experiment 1, 3 laparoscopic intestinal-repair techniques were evaluated in an equine pelvitrainer: suturing with needle holders, with an automatic suture device, and stapling with a hernia stapler. In experiment 2, descending colon lacerations were sutured laparoscopically using needle holders in a pelvitrainer and in equine cadavers. In experiment 3, iatrogenic rectal lacerations were sutured laparoscopically with needle holders in horses under general anesthesia. These horses were evaluated for 7 days' postoperatively by clinical examination and blood and peritoneal fluid analysis. The horses were euthanatized 7 days' postoperatively and necropsied. The repaired colonic segments were collected for determination of bursting pressures, degree of luminal narrowing, and microscopic examination of the suture line. RESULTS: For the 3 techniques, there was no significant difference in repair time or degree of luminal narrowing, but bursting pressure was higher in segments repaired by use of needle holders or with the suture device. Colonic and rectal lacerations were sutured successfully with laparoscopic needle holders. No major complications were recorded. At necropsy, adequate healing was confirmed by a high bursting strength (>140 mm Hg) and by histologic examination. CLINICAL RELEVANCE: A novel laparoscopic method can be used to repair iatrogenic rectal lacerations in horses. Whether this method may be used for clinical cases of rectal tears must be investigated.     

Endotoxin activity in whole blood measured by neutrophil chemiluminescence is applicable to canine whole blood

The dog is widely used as a translational experimental model studying the host response and new treatments for human endotoxemia. The present study evaluated the applicability of a novel patient-near neutrophil chemiluminescence assay for the measurement of endotoxin activity in human blood when applied to canine blood samples. The assay was observed to be analytically sensitive and specific to endotoxin when tested in vitro, spiked with purified Escherichia coli lipopolysaccharide and live E. coli. The diagnostic sensitivity was sustained during Gram-positive contamination. Finally, it also demonstrated diagnostic potential when able to discriminate dogs with spontaneously occurring endotoxemia from both healthy dogs and diseased dogs without endotoxemia. The rapid patient-near assessment of endotoxin activity in canine blood should facilitate future studies on endotoxemia in both spontaneous disease and in experimental settings.     

Mineral content of hay harvested in Bavarian and Swiss horse farms. Predictive value of cutting time,number of cut,botanical composition,origin and fertilization

Hay samples from 29 horse farms in Southern Upper Bavaria and 31 horse farms in Switzerland were taken and analysed for minerals such as calcium, magnesium, phosphorus and potassium. The content of herbs and legumes in the hay was determined and the Weende analysis was performed. Comparison between Bavaria and Switzerland resulted in comparable calcium contents of approximately 4 g Ca/kg dry matter (DM) in grass hay. Hay with more than 10% of herbs and legumes found only in Switzerland showed higher Ca contents of >7 g Ca/kg DM in hay. The mineral contents of phosphorus and potassium were lower in hay from Switzerland (P: 1.8 ± 0.7 g/kg DM, K: 15.4 ± 5.1 g/kg DM) than from Bavaria (P: 3.8 ± 0.6 g/kg DM, K: 20.0 ± 6.0 DM), whereas the magnesium content of the hay showed no difference between the regions (∼1.5 g Mg/kg DM). Very late first grass hay cuts showed low magnesium and calcium (<4 g/kg DM) contents. Further minerals in the hay for horses differed from those in dairy cattle in the same region. Fertilization showed only small effects on the mineral contents of the hay produced for horses. Therefore, in calculation of ration and production of supplements for horses, these differences should be taken into account.     

Stress response of extensively reared young bulls being transported to growing-finishing farms under Spanish summer commercial conditions

To evaluate the effect of Spanish summer commercial journeys on the stress response of young bulls born and reared under extensive conditions, 2 replicates of a transport from an assembly centre to a growing-finishing farm were studied. Journeys lasted 27 h, involving a total of 62 young bulls. Variables under study included haematocrit, red blood cell count (RBC), total white blood cell count (WBC), differential WBC counts, serum haptoglobin (Hp), cortisol, glucose, creatine phosphokinase (CPK), lactate dehydrogenase (LDH), total protein, and albumin at loading, at the end of an intermediate market stop, and at the unloading. Before the beginning of the journey elevated WBC and neutrophil counts, and high Hp values were detected, reflecting high stress levels probably as a consequence of previous procedures associated with the grouping at the assembly centre. Some stress was also detected at the end of the market stop, with cortisol increasing from 6.5 to 12.6 ± 2.0 ng/mL (P < 0.001), although a change in Hp concentration was not observed. Neither CPK and LDH activities, related to muscular tissue damage, nor haematocrit and RBC count, related to dehydration revealed a significant effect of this first stage of the journey on the physical stress of the young bulls. Subsequent 13 h transport to the growing-finishing farm induced an increase in Hp levels from 0.48 to 0.78 ± 0.16 mg/mL (P < 0.001), reflecting an onset of the acute stress response, although cortisol levels immediately after the unloading were similar to those found before loading at the market, suggesting that calves got accustomed to transport. At the end of the journey some dehydration and physical stress were also detected. Overall, our study provides new information to the discussion of the effect of temperatures during cattle transport. Although an improvement in pre-transport conditions is essential if the welfare of assembled and transported cattle is to be improved, the stress-related alteration of cattle physiology under Spanish summer commercial transport conditions is similar to that observed under colder conditions.     

The impact of the phyto‐oestrogen genistein on swine granulosa cell function

Soya and soybean products used in swine feeding contain genistein, a non‐steroidal phyto‐oestrogen which has been demonstrated to influence endocrine functions. This observation leads us to design this study to evaluate the effect of genistein on swine granulosa cell steroidogenesis and proliferation. In the attempt to unravel the genistein signal transduction mechanisms, we verified the effect of lavendustin, a Tyrosine Kinase (TK) inhibitor, and the potential involvement of NO/cGMP pathway. Finally, as angiogenesis is essential for follicle development, we tested the effect of the phyto‐oestrogen on vascular endothelial growth factor production and on granulosa cell redox status, because free‐radical species modulate neovascularization. Our data provide evidence that genistein interferes with granulosa cell steroidogenesis while it does not modulate cell growth: this effect could be at least partially produced by inhibiting TK‐dependent signalling systems. On the contrary, NO/cGMP pathway or vascular endothelial growth factor production can be excluded as signalling mechanism involved in phyto‐oestrogen effects. Remarkably, genistein stimulates hydrogen peroxide production thus potentially inhibiting follicular angiogenesis. Collectively, these results suggest that genistein consumption could potentially negatively impact swine reproductive function.     

Repeat tuberculin skin testing leads to desensitisation in naturally infected tuberculous cattle which is associated with elevated interleukin-10 and decreased interleukin-1 beta responses

The principal surveillance tool used to control bovine tuberculosis in cattle is the removal of animals that provide a positive response to the tuberculin skin-test. In this study we performed a longitudinal investigation of the immunological and diagnostic consequences of repeated short-interval skin-tests in cattle naturally infected with Mycobacterium bovis. Tuberculin skin-test positive cattle were subjected to up to four further intradermal comparative cervical skin-tests at approximately 60-day intervals. A significant progressive reduction in the strength of the skin-test was observed after successive tests. In contrast, the magnitude of interferon-γ (IFN-γ) responses was not influenced by repeat skin-testing either transiently around the time of each skin-test or longitudinally following repeated tests. A significant boost in blood interleukin-10 (IL-10) production was observed within 3 days following each skin-test although the magnitude of this boosted response returned to lower levels by day 10 post-test. The application of a novel multiplex assay to simultaneously measure seven cytokines and chemokines also identified that skin-testing resulted in a significant and progressive reduction in antigen specific interleukin-1β (IL-1β) whilst confirming stable IFN-γ and elevated IL-10 responses in the blood. Therefore, we have demonstrated that in cattle naturally infected with M. bovis, repeat short-interval skin-testing can lead to a progressive reduction in skin-test responsiveness which has potential negative consequences for the detection of infected animals with marginal or inconclusive skin-test responses. The desensitising effect is associated with decreased IL-1β and elevated IL-10 responses, but importantly, does not influence antigen specific IFN-γ responses.     

Analysis of Amblyomma surveillance data in the Caribbean: Lessons for future control programmes

Amblyomma variegatum, the Tropical Bont Tick (TBT), is the principal vector of heartwater and is associated with dermatophilosis, major causes of losses in animal production and mortality in Caribbean livestock. From 1995 to 2007, the Caribbean Amblyomma Programme (CAP) supported treatment and surveillance activities in 11 islands of the Eastern Caribbean with an initial objective of eradicating TBT. In addition to control activities, surveillance data were collected between 1997 and 2006 in a unique regional database. We report the analysis of the surveillance data from four islands (Nevis, St Kitts, St Lucia, and Barbados) where control and surveillance followed the initial protocol and where enough data were collected. We describe the evolution of TBT infestation levels and the efforts carried out throughout the surveillance period. Logistic regression identified factors associated with herds found infested with TBT. Overall, treatment programmes were associated with a decrease in proportion of TBT-infested farms. High surveillance efforts were carried out throughout the 1997–2007 period for all island of interest, but inadequate level of surveillance was observed in several quarters especially for St Kitts. Third quarter of the year, as indication of adult TBT seasonality on livestock, was significantly associated with the risk of detecting TBT in Nevis and St Kitts livestock farms. Also, presence of cattle in Nevis farms was shown associated with an increasing probability of farms being declared TBT-infested. Outcomes of these analyses provide basis for recommendations to improve future national and regional control and surveillance activities. This analysis demonstrates the usefulness of long term and adequate surveillance data for control programmes and identification of factors associated with risk of having infested herds.