Quantification of the DNA Difference,and Separation of X‐ and Y‐Bearing Sperm in Alpacas (Vicugna pacos)

Sperm sexing is an emerging reproductive technology which has been successfully used to produce offspring of a pre‐determined sex in domestic and wildlife species but has yet to be applied to New World camelids. The aims of the present study were to (i) optimize the Hoescht 33342 (H33342) staining concentration for the flow cytometric separation of X and Y chromosome‐bearing alpaca (Vicugna pacos) sperm nuclei, (ii) separate alpaca sperm nuclei into high purity (>90%) populations bearing the X‐ and Y‐chromosome and (iii) determine the DNA difference between X‐ and Y‐bearing sperm in alpacas. Semen was collected from alpacas and sperm nuclei stained with H33342, incubated and analysed using a high‐speed cell sorter (SX‐MoFlo^®). H33342 staining concentrations of 36, 54, 72 or 90 μm did not affect the proportion of correctly oriented sperm nuclei (43.3 ± 3.9, 46.4 ± 3.7, 44.5 ± 4.0 and 51.1 ± 2.5% respectively) nor the speed of sorting (1381 ± 160, 1386 ± 123, 1371 ± 133 and 1379 ± 127 sperm nuclei/s). Sort reanalysis determined high levels of purity for X‐ and Y‐enriched populations (96.6 ± 0.7% and 96.1 ± 1.1% respectively). The DNA difference, based on fluorescence intensity (determined by the SX‐MoFlo^®), was 3.8 ± 0.06%. These data demonstrate for the first time that alpaca sperm nuclei can be separated into high purity populations and the potential for applying sperm sexing technology to New World camelids.     

Cover Image,Volume 14, Issue 2

The cover image, by Riccardo Finotello et al., is based on the Original Article Canine pancreatic islet cell tumours secreting insulin‐like growth factor type 2 (IGF‐II): a rare entity, DOI: 10.1111/vco.12085 .

     

Trace-element studies in a remote scottish upland catchment

Fresh and aged within-pack snow samples were collected and analyzed for 6 trace-elements during the winters of 1984 and 1985 at a remote site located within the Cairngorm Mountains, Scotland. All of the fresh snow samples were acidic and highly variable in composition. The variability of pH and trace-element concentrations of snowfall are demonstrated to be associated with different air trajectory categories, and the study catchment is susceptible to episodic pollution events due to long range atmospheric transport. Partial melt experiments have shown that both fractionation and preferential elution of trace-elements occur during melting, the concentrations being 1.3 to 5.4 times higher in the first 10% meltwater fraction than in the bulk snow. Cadmium and Mn appear to be preferentially eluted from the melting snow with respect to the remaining elements. Upon snowpack melting, the trace-elements may be mobilized and redistributed within the snow profile. The ions concentrate at depth from where they can be quickly removed with the early water runoff during the spring.     

Investigation of thyroid function in dogs treated with the tyrosine kinase inhibitor toceranib

Tyrosine kinase inhibitors are widely utilized in veterinary oncology for the treatment of mast cell and solid tumours. In man, these drugs are associated with thyroid dysfunction: however, to date only one study has investigated this in dogs. The aim of this study was to prospectively assess thyroid function in a group of dogs with cancer receiving toceranib. Thirty‐four dogs were prospectively enrolled at two referral hospitals into two groups; those receiving toceranib with prednisolone and those receiving toceranib alone. Total thyroxine (TT4) and thyroid stimulating hormone (TSH) was monitored at regular time points during treatment. Follow‐up data was available for 19 dogs. Overall, 12 incidences of elevated TSH occurred but none of these dogs had concurrent low TT4 concentrations. There was a significant difference in median TSH at week six compared with baseline. Hypothyroidism was not diagnosed in any patient during the study period. Patient drop‐out was higher than anticipated which prevented the assessment of longer term toceranib administration on thyroid function. Toceranib therapy was not associated with hypothyroidism in this study but did result in elevations in TSH which confirms what has been previously reported. Toceranib should be considered to cause thyroid dysfunction in dogs and monitoring is advised.     

Anterior uveitis as an atypical presentation of large granular lymphoma in a caracal (Caracal caracal)

An 11-year-old, female, spayed caracal (Caracal caracal) presented with a 3-month history of intermittent anorexia, vomiting, and weight loss. At examination, bilateral anterior uveitis with anisocoria was present. Further examination under general anesthesia revealed the anterior chamber of the right eye contained copious amounts of thick fibrin, hypopyon, and hyphema obscuring the ventral pupil margin and ventral iris. Aqueous paracentesis revealed a cytologic diagnosis of large granular lymphoma. Additional antemortem diagnostics (computed tomography, ultrasound with fine needle aspiration, and cytology) confirmed the diagnosis of multicentric large granular lymphoma with associated intestinal perforation. Necropsy and histopathology confirmed multicentric large granular lymphoma in the right eye, jejunum, mesenteric lymph nodes, and right kidney. Large granular lymphoma in nondomestic felids is likely rare since it has not been previously reported, but should be considered as a differential in any felid species with anterior uveitis or when lymphoma is considered. In the case of this caracal, the use of aqueous paracentesis with cytology was a powerful diagnostic.     

Mycotic dermatitis with digital gangrene and osteomyelitis,and protozoal intestinal parasitism in Marlborough green geckos (Naultinus manukanus)

CASE HISTORY: Thirty adult Marlborough green geckos (Naultinus manukanus) were collected from Stephens Island and held over winter, prior to their translocation. Five adult geckos developed skin lesions after husbandry changes affected the humidity of their enclosures. Two geckos underwent ecdysis and recovered. One animal died and two others progressively worsened and were presented for treatment.

CLINICAL AND PATHOLOGICAL FINDINGS: The geckos were in poor body condition and had multiple black powdery lesions and solitary raised white nodules on their skin. Both geckos died despite topical and supportive treatment. Histopathology showed the skin nodules contained branching non-septate hyphae infiltrating necrotic epidermal tissue, and associated dermal inflammation. There was necrosis of several digits and mycotic osteomyelitis. Mucor ramosissimus was cultured from skin biopsies from each animal. Large numbers of motile protozoa, resembling Trichomonas, and another unidentifiable, were recovered from fresh faecal smears, and Nyctotherus sp protozoa were present in the lumen of the intestine of one animal post mortem.

DIAGNOSIS: Mycotic dermatitis with digital gangrene and osteomyelitis due to Mucor ramosissimus, and enteric protozoal parasitism with Trichomonas sp and Nyctotherus sp.

CLINICAL RELEVANCE: The clinical course and pathological findings of mycotic dermatitis in two Marlborough green geckos involved in a wildlife translocation in New Zealand are reported, and also the first record of the Marlborough green gecko as a host for the enteric protozoa Trichomonas sp and Nyctotherus sp.