Automated hematological cell count using sysmex XN-1000V in Testudo hermanni: Agreement with manual count.

Mediterranean area represents the main habitat of Testudo hermanni. Clinical signs of disease of these tortoises are non-specific, making the hematology results crucial in revealing underlying pathological conditions. However, accurate automated identification of blood cell populations is hampered by the presence of nucleated erythrocytes (NRBC) and thrombocytes (Thr), necessitating manual methods such as counting chambers. The aim of the study was to assess the performance of the novel automated hematology analyzer Sysmex XN-1000 V, which includes a a specific channel (WNR) for counting NRBC, in accurately identify and quantify the different blood cell populations of Testudo hermanni. Additionally, its agreement with manual counts was evaluated. Fifty heparinized blood samples were initially counted using the Neubauer improved chamber and then analysed twice with Sysmex XN-1000 V. Thirteen out of 50 samples were instrumentally counted again after 48 h to assess the inter-assay precision. All WNR scattergrams were re-analysed using an ad hoc gate panel to differentiate two populations: NRBCs (weak fluorescence signal) and WBC + Thr (high fluorescence signal). Sysmex XN-1000 V demonstrated optimal intra- and inter-assay precision for NRBCs (CV 0.98% ± 1.96; 1.31% ± 2.98) and moderate precision for WBC + Thr (CV 9.24% ± 16.61; 12.69% ± 10.35). No proportional nor constant errors were observed between the methods for both the populations. The instrumental NRBC counts were consistently slightly lower, while WBC + Thr counts were slightly higher compared to manual counts. These findings suggest that Sysmex XN-1000 V can be used for analyzing cell populations in heparinized blood of Testudo hermanni. However, specific instrumental reference intervals are suggested.

Determination of Efficacy of Single and Double 4.7 mg Deslorelin Acetate Implant on the Reproductive Activity of Female Pond Sliders (Trachemys scripta).

The use of long-acting gonadotropin-releasing hormone (GnRH) agonists to suppress fertility has been poorly investigated in reptiles, and the few available studies show inconsistent results. The efficacy of single and double intramuscular 4.7 mg deslorelin acetate implants in captive pond sliders (Trachemys scripta) was investigated, with 20 animals divided into three groups: a single-implant group (6 animals), a double-implant group (6 animals), and a control group (no implant). During one reproductive season (March to October), plasmatic concentration of sexual hormones (estradiol, progesterone, and testosterone) and ovarian morphometric activity via computed tomography were monitored about every 30 days. A significative decrease in the number of phase II ovarian follicles was detected in the double-implant group compared with the control group, but no significant difference was noted in the number of phase III and phase IV follicles, egg production, and plasmatic concentration of sexual hormones. Results show that neither a single nor a double deslorelin acetate implant can successfully inhibit reproduction in female pond sliders during the ongoing season, but the lower number of phase II follicles in the double-implant group can possibly be associated with reduced fertility in the following seasons.

Molecular Survey on Toxoplasma gondii and Neospora caninum Infection in Wild Birds of Prey Admitted to Recovery Centers in Northern Italy.

Toxoplasma gondii and Neospora caninum (Apicomplexa, Sarcocystidae) are protozoan parasites infecting a wide range of intermediate hosts worldwide, including birds. Raptors acquire the infections through the ingestion of both infected preys and oocysts in the environment suggesting they might be used as indicators of the spread of these pathogens. Here, we report an epidemiological survey with the aim of determining the prevalence of T. gondii and N. caninum infections in wild birds of prey, hospitalized in two Wildlife Recovery Centres (WRCs) in Northern Italy. Genomic DNA extracted from brain tissue samples was submitted to Real Time PCR targeting T. gondii B1 and N. caninum Nc5 genes. T. gondii genotyping was then performed by multilocus sequence typing (MLST) analysis, targeting three polymorphic genes (GRA6, BTUB, and altSAG2). T. gondii DNA was found in 35 (62.5%) out of 56 examined samples; concerning genotyping, it was possible to amplify at least one gene for 26 animals, and obtained sequences belonged to Type II. N. caninum DNA was only detected in two (3.6%) common kestrels (Falco tinnunculus), adding a new species to the list of suitable intermediate hosts for this pathogen. Data obtained in the present study thus confirmed the spread of both T. gondiiand N. caninum in wild bird of prey, endorsing the role of WRCs in the epidemiological surveillance of wildlife.

Comparison of Lateral and Dorsal Recumbency during Endoscope-Assisted Oophorectomy in Mature Pond Sliders (Trachemys scripta).

Ovariectomy via the prefemoral fossa is a well-described technique for sterilization in chelonians. The choice between lateral and dorsal recumbency is generally left to the surgeon's preference, with no data supporting an objective superiority of one over the other. Twenty-four sexually mature female pond sliders (Trachemys scripta) were enrolled for elective prefemoral endoscope-assisted oophorectomy, and were randomly divided in two groups: 12 animals were placed in right lateral recumbency with a left fossa approach (Group A), and 12 in dorsal recumbency with a right fossa approach (Group B). Scoring systems were applied to assess the ease of access to the coelomic cavity, and the ease of identification of the ovary opposite to the surgical incision. A negative correlation was found between the body weight of the animals and the ease of access to the coelom (p = 0.013), making the access easier in smaller animals. No significant difference was found between the two groups in terms of ease of access to the coelomic cavity, first ovary removal time, ease of identification of the second ovary, second ovary removal time, or total surgery time.

Toxoplasma gondii infection in raptors from Italy: Seroepidemiology and risk factors analysis.

Toxoplasma gondii is a zoonotic parasite infecting a wide range of intermediate hosts, including birds. Nevertheless, scant information on the spread of infection in wild bird populations is available to date. With the aim of updating information on T. gondii infection in birds of prey and possible risk factors associated with the infection, a serosurvey was planned on both wild and captive raptors. An overall of 93 raptors from Northern Italy were tested for the presence of anti-T. gondii antibodies with a commercial modified agglutination test (MAT). A T. gondii prevalence of 10.7% was recorded; the highest seroprevalence was observed within the Family Strigidae (12.5%). Only wild animals tested positive; any statistical difference among species, taxonomic family, age, origin, use, migratory behavior and diet composition was not highlighted. Toxoplasmosis in birds of prey, sentinel species for the environmental spread of T. gondii, should always be considered within sanitary programs devoted to avian species protection.