Assessment of platelet function in horses: ultrastructure, flow cytometry, and perfusion techniques

We studied equine platelet function and activation using ultrastructural examination, flow cytometry, and perfusion. The main aim of the study was to evaluate hemostatic mechanisms in horses using these techniques. Ultrastructural observations were done on resting and activated platelets. Flow cytometry was used to evaluate binding of antibodies to major platelet glycoproteins (GPIIb-IIIa, GPIV, and GPIb) and activation-dependent antigens (P-selectin and lysosomal integral membrane protein [LIMP]). Perfusion techniques were used to evaluate the interaction between platelets and damaged subendothelium. Aggregation experiments were done to identify the best agonists for flow cytometry. Ultrastructural observations confirmed that equine platelets lack a developed open canalicular system and that release of granule contents occurs by fusion of adjacent granule membranes that ultimately connect with external membranes. Flow cytometry identified a 2-fold increase in binding of antibodies against GPIIb-IIIa and GPIV after activation. Binding of antibodies against P-selectin and LIMP increased from 2.12 and 1.74% to 15.5 and 11.6%, respectively, in response to thrombin and to 21.86 and 10.50%, respectively, in response to collagen. Annexin V binding increased moderately after activation. Perfusion experiments with citrated blood indicated that equine platelets react more strongly to subendothelium than do human platelets. When blood was anticoagulated with low molecular weight heparin, a marked impairment of platelet interactions was observed. In conclusion, although some differences were observed between human and equine platelet function, some techniques currently used to assess human platelet function may be useful to assess equine platelets.     

Quantification of Sexual Steroid Hormones in Faeces of Iberian Wolf (Canis lupus signatus): A Non‐invasive Sex Typing Method

The determination of gender in wild animals is essential for behavioural and ecological studies, and also for conservation. The objectives of this study were (i) the determination of gender in faecal samples of Iberian wolf based on the differential concentrations of sexual steroid hormones (SSH) and (ii) to analyse the profiles of SSH in males and females (considering the gender determination carried out previously) during the non‐reproductive and reproductive periods. The quantification of androgens (testosterone, T), progestin (progesterone, P) and oestrogen (oestradiol, E) was conducted by means of enzyme immunoassay. The k‐means conglomerate analysis showed that the 59 faecal samples grouped into three different conglomerates, considering SSH levels. Groups 1 and 2 showed higher levels of T than group 3. Therefore, the faecal samples included in groups 1 and 2 (17 samples) corresponded to males and those of group 3 (42 samples) to females. The levels of T + P + E and T/P were higher in the group of males than in the group of females. The results of this study also showed that levels of T in males were higher during the reproductive period than in the non‐reproductive period. However, the concentrations of P and E turned out to be higher during the non‐reproductive season. In females, the levels of the three hormones (T, P and E) were higher during the reproductive period.     

Lymphoedema — a problem of gigantic proportions

Extract

Lymphoedema (LO) is the accumulation of protein-rich interstitial fluid caused by failure of normal lymphatic drainage. Human lymphoedema is classified as primary (congenital hereditary or idiopathic, praecox or tarda forms) or secondary (complication of surgery or radiotherapy, such as that for breast cancer, as well as bancroftian filariasis, cellulitis and lymphangitis amongst others). Primary LO may be due to lymphatic aplasia, hypoplasia, dyspla-sia, lymphangiectasia, or aplasia or hypoplasia of draining lymph nodes. Approximately one in thirty people worldwide has LO.     

Morphovariability of capulín (<Emphasis Type="Italic">Prunus serotina</Emphasis> Ehrh.) in the central-western region of Mexico from a plant genetic resources perspective

The capulín, or black cherry tree (Prunus serotina Ehrh.) is a tree with edible fruits native to North America that has been used not just as a source of food but also as a source of energy (its wood) since prehispanic times. Mexico has three of the five botanical subspecies in the P. serotina botanical complex, which have not yet been characterized for agricultural purposes. It is desirable to determine whether capulín trees comprise a homogeneous group or whether there are separate morpho-types. This study focuses on the morphological variability of seven populations from the western and central regions of Mexico. We used the Hill & Smith Analysis and graphical methods to analyze and interpret data with respect to 22 quantitative morphological characters and 17 qualitative morphological characters, all from different plant organs. Two major geographically and morphologically related groups were clearly identified, and the morphological variability in the study zone was related to taxonomic, genetic and agronomic factors. The results are discussed from a genetic resources perspective. The genetic, cultural, ecological and agronomic implications of these results are also considered, as well as the potential uses for the species. From these results, an agro-morphological characterization can be developed, in order to identify interesting types of capulín for fruit and seed production in agro-industry, or as rootstock for related species and forestry uses. Our results have implications for the conservation and sustainable use of capulín genetic resources in the central and western regions of Mexico.     

Modular hybrid total hip arthroplasty. Experimental study in dogs

Background

This prospective experimental study evaluated the surgical procedure and results of modular hybrid total hip arthroplasty in dogs.

Methods

Ten skeletally mature healthy mongrel dogs with weights varying between 19 and 27 kg were used. Cemented modular femoral stems and uncemented porous-coated acetabular cups were employed. Clinical and radiographic evaluations were performed before surgery and at 30, 60, 90, 120, 180 and 360 days post-operation.

Results

Excellent weight bearing was noticed in the operated limb in seven dogs. Dislocation followed by loosening of the prosthesis was noticed in two dogs, which were therefore properly treated with a femoral head osteotomy. Femoral fracture occurred in one dog, which was promptly treated with full implant removal and femoral osteosynthesis.

Conclusions

The canine modular hybrid total hip arthroplasty provided excellent functionality of the operated limb.     

Mastocytoma in the common carpal sheath of the digital flexor tendons of a horse

A 13-year-old Morgan gelding was examined for right forelimb lameness and tenosynovitis of the right common carpal sheath of the digital flexor tendons. The horse had moderate right forelimb lameness at the trot and marked effusion of the right common carpal sheath of the digital flexor tendons. Ultrasonographic examination revealed a soft tissue mass within the proximal pouch of the affected tendon sheath, located adjacent to the distal physis of the radius. Cytology and culture of the fluid revealed a sterile, eosinophilic tenosynovitis. Tenoscopic exploration confirmed the presence of a capsulated soft tissue mass. Thecotomy was required to fully debride the mass, which histology revealed to be a mast cell tumour. At 22 months postoperatively, the horse developed mild right forelimb lameness and eosinophilic tenosynovitis because of recurrence of the mastocytoma. Mastocytosis is a possible differential diagnosis in any horse exhibiting lameness associated with tenosynovitis. Surgical excision combined with rest and postoperative intrasynovial and systemic corticosteroids may be palliative.     

Transcutaneous ultrasound over the right flank to diagnose mid- to late-pregnancy in the dairy cow

Objective   To investigate the practical application and image quality of transcutaneous ultrasonography across the right flank for the diagnosis of mid- to late-gestation in cattle.
Procedure   Dairy cows of known pregnancy status were used to establish criteria for the identification of mid- to late-stage pregnancy by transcutaneous ultrasonography. Factors involved in the practical application of transcutaneous ultrasound for routine use in pregnancy diagnosis in dairy cattle were also examined.
Results   Transcutaneous ultrasound, using a 3.5-MHz sector transducer applied over the caudodorsal right flank fold, allowed rapid and detailed visualisation of bovine pregnancy (i.e. fetus, fetal fluids, placentomes and/or fetal membranes) and could be used to establish criteria for the diagnosis of pregnancy. The commonly available cattle holding facilities of the herringbone dairy, rotary herringbone dairy, AI race or crush, and walk-through dairy all allow safe access to the right flank of a dairy or beef cow. However, transcutaneous scanning over the right flank is difficult in modern rotary turnstile dairies.
Conclusion   A 3.5-MHz sector transducer applied over the caudodorsal right flank fold allows detailed visualisation of the bovine fetus in most types of cattle-holding facilities. Further investigation of the sensitivity and specificity of this technique over the entire gestation period and its possible use in ageing of the bovine fetus is merited.