Genetic diversity and phylogenetic relationships between Leishmania infantum from dogs,humans and wildlife in south‐east Spain

Leishmania infantum causes human and canine leishmaniosis. The parasite, transmitted by phlebotomine sand flies, infects species other than dogs and people, including wildlife, although their role as reservoirs of infection remains unknown for most species. Molecular typing of parasites to investigate genetic variability and evolutionary proximity can help understand transmission cycles and designing control strategies. We investigated Leishmania DNA variability in kinetoplast (kDNA) and internal transcribed spacer 2 (ITS2) sequences in asymptomatically infected wildlife (n = 58) and symptomatically and asymptomatically infected humans (n = 38) and dogs (n = 15) from south‐east Spain, using single nucleotide polymorphisms (SNPs) and in silico restriction fragment length polymorphism (RFLP) analyses. All ITS2 sequences (n = 76) displayed a 99%–100% nucleotide identity with a L. infantum reference sequence, except one with a 98% identity to a reference Leishmania panamensis sequence, from an Ecuadorian patient. No heterogeneity was recorded in the 73 L. infantum ITS2 sequences except for one SNP in a human parasite sequence. In contrast, kDNA analysis of 44 L. infantum sequences revealed 11 SNP genotypes (nucleotide variability up to 4.3%) and four RFLP genotypes including B, F and newly described S and T genotypes. Genotype frequency was significantly greater in symptomatic compared to asymptomatic individuals. Both methods similarly grouped parasites as predominantly or exclusively found in humans, in dogs, in wildlife or in all three of them. Accordingly, the phylogenetic analysis of kDNA sequences revealed three main clusters, two as a paraphyletic human parasites clade and a third including dogs, people and wildlife parasites. Results suggest that Leishmania infantum genetics is complex even in small geographical areas and that, probably, several independent transmission cycles take place simultaneously including some connecting animals and humans. Investigating these transmission networks may be useful in understanding the transmission dynamics, infection risk and therefore in planning L. infantum control strategies.     

Transvenous coil embolisation for the treatment of single congenital portosystemic shunts in six dogs

This article describes the treatment of single congenital portosystemic shunts (CPSs) (intrahepatic and extrahepatic) using an interventional radiology technique involving embolisation of anomalous vessels with percutaneous coils. Briefly, a multipurpose catheter was introduced into the caudal vena cava and then into the portosystemic shunt. An autoexpandable stent was placed in the caudal vena cava, next to the shunt, in order to avoid coil migrations, and a cobra-like vascular catheter was used to pass through the stent and to place the coils in the shunt. This technique was used for treatment of CPS in six dogs. The results indicate that percutaneous embolisation of a CPS using coils, a less invasive technique than the traditional surgical technique, may result in complete closure of the anomalous vessel without development of portal hypertension.     

Endotracheal stenting therapy in dogs with tracheal collapse

Tracheal collapse in dogs is a common respiratory disorder, typically presenting with a history of chronic cough, increasing respiratory difficulties, and episodes of dyspnoea. Medical treatment is the therapy of choice and surgical repair is considered when patients do not respond well. Minimally invasive endotracheal stenting is a promising new therapy under investigation, but there remain significant challenges to overcome potential complications. The purpose of this article is to provide a comparative overview of intra-luminal stenting of the trachea in human and veterinary medicine. The currently available stents and their potential clinical application to the veterinary patient will be discussed.     

Low‐density lipoproteins and milk serum proteins improve the quality of stallion sperm after vitrification in straws

Lipids and proteins can be used for sperm vitrification to preserve the integrity of sperm membranes or to increase the viscosity of the medium. This study evaluated the effect of low‐density lipoproteins (LDL) and milk serum proteins (Pronexcell) for stallion sperm vitrification. Hippex extender (Barex Biochemical Products, The Netherlands), plus 1% of bovine serum albumin and 100 mM of trehalose, was used as control for sperm vitrification. In experiment 1, different concentrations of LDL (L1 = 0.25, L2 = 0.5, L3 = 1%) and in experiment 2 of Pronexcell (P1 = 1, P2 = 5, P3 = 10%) were added to control extender. Vitrification was performed in 0.25‐ml straws directly plunged into liquid nitrogen. Total motility (TM, %) and progressive motility (PM, %) were analysed by CASA, and plasma membrane (IMS, %) and acrosome membrane integrity (AIS, %) were assessed under epifluorescence microscopy. Post‐warmed sperm parameters were compared between treatments by ANOVA. Results were expressed as mean ± SEM. In both experiments, the minimum concentration of LDL and Pronexcell obtained significantly higher values (p < 0.01) than the control extender for TM (L1 = 52.95 ± 4.4; P1 = 58.99 ± 4.6; C = 30.88 ± 3.0), PM (L1 = 36.79 ± 5.5; P1 = 47.25 ± 4.3; C = 19.20 ± 2.4), IMS (L1 = 68.88 ± 3.6; P1 = 47.25 ± 4.3; C = 52.81 ± 2.6) and AIS (L1 = 45.88 ± 3.6; P1 = 47.25 ± 4.3; C = 26.00 ± 2.1). No differences in sperm parameters were found among different concentrations of LDL or Pronexcell. In conclusion, the addition of 0.25% LDL and 1% Pronexcell to the vitrification extender is recommended to improve the quality of stallion sperm after vitrification.     

Is sperm cryopreservation in absence of permeable cryoprotectants suitable for subfertile donkeys?

Sperm from fertile donkeys have been successfully frozen in absence of permeable cryoprotectants. The aim of this study was to determine whether this cryopreservation method is suitable for subfertile donkeys in comparison to conventional sperm freezing with glycerol. Ejaculates were collected from four Andalusian Donkeys: three fertile and one subfertile. Semen was frozen with an extender containing glycerol (GLY), or adding instead sucrose 0.25 molar and 1% bovine serum albumin (SUC) as non‐permeable cryoprotectants. After thawing, samples were assessed for total (TM, %) and progressive (PM, %) sperm motility by CASA, plasma membrane integrity (PMI, %) by epifluorescence microscopy and DNA integrity (DFI, %) by SCSA. Results (mean ± SD) were compared between extenders in fertile and subfertile donkeys using the Student's t test. No differences between GLY and SUC treatments were found in the fertile group for the sperm parameters assessed. In subfertile donkey ejaculates, GLY resulted in significantly higher values than SUC for TM (25.5 ± 3.1 vs. 19.6 ± 1.9) and PM (13.3 ± 5.1 vs. 4.0 ± 1.2), respectively. In conclusion, considering all the sperm parameters assessed, sperm freezing in absence of permeable cryoprotectants may not be still an option for cryopreservation of subfertile donkey sperm.     

Hematology, blood chemistry, and bacteriology of the free-ranging Mexican beaded lizard (Heloderma horridum).

Blood samples from 16 adult, wild Mexican beaded lizards were obtained for complete blood count and serum biochemical analysis. In addition, both oral and cloacal swab samples were collected and submitted for bacteriologic culture. This is the first report of hematologic and bacterial data from free-living beaded lizards. This information will serve as baseline reference values for future health assessment studies in conservation, captive breeding, and research programs.     

INTRAVENOUS ALFAXALONE ANAESTHESIA IN LEOPARD GECKOS (EUBLEPHARIS MACULARIUS)

Intravenous alfaxalone, administered at a dose of 5 mg/kg in the jugular vein, was evaluated in 20 leopard geckos (Eublepharis macularius) to ascertain its ability to provide anesthesia. The induction time, time to loss of mandibular tone, interval of deep anesthesia, and full recovery time were 27.5 ± 30.7 seconds (10 to 56 seconds), 1.3 ± 1.4 minutes (11 seconds to 4 minutes), 12.5 ± 2.2 minutes (11.11 to 15.39 minutes), and 18.8 ± 12.1 minutes (10.4 to 52.31 minutes), respectively. A significant reduction in heart rate (74 ± 12.9 beats/minute) was recorded between 2 and 24 minutes after alfaxalone administration. A significant decrease in respiratory rate (26.8 ± 10.1 breaths/minute) was recorded 2 minutes after alfaxalone administration, and respiratory rate remained lower than the basal rate (31.4 ± 3.1 breaths/minute) for 24 minutes but without statistical significance. The intravenous administration of alfaxalone in leopard geckos achieved a rapid onset of anesthesia and a suitable recovery time. Based on this investigation, an afaxalone dose of 5 mg/kg intravenously proved to be suitable for sedation before tracheal intubation. Moreover, the administration route via the jugular vein, was acceptable in leopard geckos; a species in which other venipuncture sites can be challenging or inaccessible.     

COMPUTED TOMOGRAPHIC FINDINGS IN OVINE COENUROSIS

Computed tomographic imaging was conducted in twenty ewes with cerebral coenurosis. CT imaging allowed precise evaluation of the size and location of the cyst, which appeared as a hypoattenuating structure with a mass effect. No meaningful correlation between clinical signs and the location of parasitic cyst was detected.