The abundance and localization of heat shock proteins (HSP)-60, -70, and -90 in the oviductal ampulla of hamadryas baboon (Papio hamadryas) during the menstrual cycle.

The presence of HSPs in female reproductive and their relationship with the steroid hormone fluctuation have been reported in several mammals but not in non-human primates. The present research dealt with the oviductal expression and localization of the more studied HSPs (60, 70, and 90) as well as the morphological changes in the Hamadryas baboon (Papio hamadryas) during the follicular, preovulatory, and luteal phases of the menstrual cycle. Therefore, western blots, histomorphological, and immunohistochemical analyses were carried out. The results of western blot analysis displayed the lowest HSP expression in the luteal phase. The histomorphology showed that the mucosal epithelium consisted of undifferentiated cuboidal cells in follicular and luteal phases and well-distinguishable columnar ciliated and non-ciliated cells during the preovulatory phase. Immunohistochemistry evidenced that the mucosal epithelium contained cytoplasmic and nuclear HSP60, 70, and 90 immunostaining in the follicular and luteal phases. During the preovulatory phase, the non-ciliated cells showed: (i) cytoplasmic HSP60; (ii) nuclear and cytoplasmic HSP90. Ciliated cells showed cytoplasmic and ciliary HSP70 and ciliary HSP90. The stromal cells and myocytes of muscular layer displayed a decreased cytoplasmic HSP60 in the preovulatory phase and nuclear and low cytoplasmic HSP70 throughout the menstrual cycle. Nuclear HSP90 decreased in ampulla stromal cells and the follicular phase myocytes. These findings indicate that the expression pattern of HSP60,70, and 90 is related to the morphofunctional features of the baboon oviductal ampulla during the menstrual cycle and could represent a referent point for further studies in the oviduct of Primates.

Use of Laryngeal Mask and Anesthetic Management in Hamadryas Baboons (Papio hamadryas) Undergoing Laparoscopic Salpingectomy-A Case Series.

The study aims to describe the anesthetic and airway management of baboons (Papio hamadryas) undergoing laparoscopic salpingectomy with a laryngeal mask airway (LMA) device. Eleven baboons received tiletamine-zolazepam and medetomidine; anesthesia was induced with propofol. An LMA was positioned for oxygen and isoflurane administration in spontaneous respiration. Heart rate (HR), mean arterial pressure (MAP), respiratory rate (RR), end tidal carbon dioxide (EtCO2), minute volume (MV), and peripheral hemoglobin oxygen saturation (SpO2) were recorded before (PREPP) and immediately after abdomen insufflation (PP1), at 10 (PP2), 20 (PP3), and 30 (PP4) minutes during pneumoperitoneum, and after (POSTPP) pneumoperitoneum. The respiratory rate was significantly higher at all times compared to PREPP. The end tidal carbon dioxide concentration was significantly higher at PP2, PP3, PP4, and POSTPP, compared to the previous times. The higher values for RR and EtCO2 were registered at PP4: 22.7 (95% CI 17.6-27.8) breaths/min and 57.9 (95% CI 51.9-63.8) mmHg, respectively. The minute volume was significantly higher at PP4 and POSTPP compared to the other times. The higher value for MV was registered at POSTPP (269.1 (95% CI 206.1-331.8) mL/kg/min). This protocol is suitable for baboons undergoing laparoscopic salpingectomy. The LMA was easy to insert and allowed for good ventilation, gas exchange, and delivery of the anesthetic in spontaneous breathing baboons.

Modification of Morphology and Glycan Pattern of the Oviductal Epithelium of Baboon Papio hamadryas during the Menstrual Cycle.

The mammalian oviduct is a highly specialized structure where fertilization and early embryonic development occur. Its mucosal epithelium is involved in maintaining and modulating a dynamic intraluminal fluid. The oviductal epithelium consists of ciliated and non-ciliated (secretory) cells whose differentiation and activity are sex hormone-dependent. In this study, we investigated for the first time both the morphology and the glycan composition of baboon oviductal epithelium during the menstrual cycle. Oviducts were laparoscopically removed from 14 healthy adult female Papio hamadryas whose menstrual cycle phase was assessed based on the sex hormone levels and the vaginal cytology features. Histological investigations were carried out on fimbriae, infundibulum, ampulla, and isthmus separately fixed in 4% (v/v) paraformaldehyde, embedded in paraffin wax, and stained with hematoxylin-eosin for morphological analyses and using a panel of nine fluorescent lectins for glycoconjugate characterization. The histomorphological analysis revealed that in the entire oviduct (i) the ciliated and non-ciliated cells were indistinguishable during the follicular and luteal phases, whereas they were highly differentiated during the preovulatory phase when the non-ciliated cells exhibited apical protrusions, (ii) the epithelium height was significantly higher in the preovulatory phase compared to other menstrual phases, and (iii) the number of ciliated cells significantly (p ≤ 0.05) increased from the fimbriae to the infundibulum and progressively reduced in the other oviductal segments with the lower presence of ciliated cells in the isthmus. The glycan characterization revealed a complex and region-specific composition during the different phases of the menstrual cycle. It can be summarized as follows: (i) high-mannosylated N-linked glycans (Con A reactivity) were present throughout the oviductal epithelium during the entire menstrual cycle and characteristically in the apical protrusions of non-ciliated cells of the ampulla during the preovulatory phase; (ii) sialoglycans with α2,3-linked sialic acids (MAL II binding) were expressed along the entire oviductal surface only during the preovulatory phase, whereas α2,6-linked ones (SNA affinity) were also detected in the surface of the luteal phase, although during the preovulatory phase they were characteristically found in the glycocalyx of the isthmus cilia, and O-linked sialoglycans with sialic acids linked to Galßl,3GalNAc (T antigen) (KsPNA) and terminal N-acetylgalactosamine (Tn antigen) (KsSBA) were found in the entire oviductal surface during all phases of the menstrual cycle; (iii) GalNAc terminating O-linked glycans (HPA staining) were mainly expressed in the entire oviducts of the luteal and preovulatory phases, and characteristically in the apical protrusions of the isthmus non-ciliated cells of the preovulatory phase; and (iv) fucosylated glycans with α1,2-linked fucose (LTA reactivity) occurred in the apical surface of fimbriae during the luteal phase, whereas α1,3/4-linked fucose (UEA I binders) were present in the apical protrusions of the ampulla non-ciliated cells and in the apical surface of isthmus during the preovulatory phase as well as in the isthmus apical surface of follicular-phase oviducts. These results demonstrate for the first time that morphological and glycan changes occur in the baboon oviductal epithelium during the menstrual cycle. Particularly, the sex hormone fluctuation affects the glycan pattern in a region-specific manner, probably related to the function of the oviductal segments. The findings add new data concerning baboons which, due to their anatomical similarity to humans, make an excellent model for female reproduction studies.

Comparison of Two Radiofrequency Vessel-Sealing Device for Laparoscopic Ovariectomy in African Lionesses (Panthera Leo).

To evaluate two vessel-sealing devices with different jaw lengths for dissecting ovaries in adult obese African lionesses undergoing laparoscopic ovariectomy. Twelve lionesses (n = 12) were recruited. The surgical procedures were performed through three portals, with a retractor platform positioned at the umbilical port and cannulas placed 3-4 cm cranial and caudal to the device at the level of the midline. Ovariectomy was performed using a vessel-sealing device according to a randomization list. We evaluated the surgery time the intra- and postoperative complications. The total surgery time was 49.3 min (range 40-61 min) in the Atlas group and significantly lower in the Caiman group (mean 31.8 min, range 26-51 min). The installation phase was similar between the groups. The ovariectomy time was significantly lower in the Caiman group (mean 7.8 min, range 4-11 min) than in the Atlas group (mean 20 min, range 16-30 min). Controlled bleeding was observed at the tip of the uterine horn in two cases in the Atlas group. No other complications were noted. The results of our study confirmed the significant advantages of employing the Caiman 12 vessel-sealing device in comparison with the LigaSure Atlas in terms of the time needed to complete ovariectomy, although both instruments could be considered safe. The use of the Caiman 12 is recommended when performing laparoscopic ovariectomies in adults with obesity.

Wound Retractor Laparoscopic Port System for Laparoscopic Ovariectomy in Panthera leo.

The aim of this study was to assess the feasibility and intraoperative complications of performing ovariectomies in African lionesses (Panthera leo) using a wound retractor laparoscopic platform. Six lionesses (n = 6) were included. The surgical procedures were carried out through three portals, with a retractor platform positioned at the umbilical port and cannulas placed 3-4 cm from the cranial and caudal regions to the device at the level of the midline. An ovariectomy was performed with a vessel-sealing device. We evaluated the surgery time and the intraoperative and early postoperative complications. The mean weight was 172.83 kg. The total surgery time was 49.33 min. The installation step took a mean of 10.33 min to complete. The mean ovariectomy time was 20 min. Controlled bleeding was observed at the tip of the uterine horn in two cases due to excessive tissue thickness. The retrieval of dissected ovarian tissue and annexes was easily performed. No other complications were observed. The use of the laparoscopic platform during three-portal surgeries for laparoscopic ovariectomy in adult overweight lionesses is feasible and without intraoperative problems. The retractor meant that there were no entry-related issues due to the 25 mm mini-laparotomy. It also made it simpler to extract thick ovaries and promptly re-establish the pneumoperitoneum.

Laparoscopic salpingectomy in Papio hamadryas for birth control in captivity.

OBJECTIVE: To evaluate the feasibility of laparoscopic salpingectomy in baboons (Papio hamadryas). We hypothesized that laparoscopic salpingectomy could be performed in baboon species within a reasonable amount of surgical time, with minor complications occurring at low rates. STUDY DESIGN: Case series and technique description. ANIMALS: Sixteen baboons (n = 16). METHODS: The surgical procedures were performed using the 3-port technique, with 5 mm instruments and a telescope placed at the umbilical and hypogastric regions. A salpinx dissection was performed, using a radiofrequency bipolar vessel sealing device, from the fimbriae to the uterine attachments. We evaluated the surgical duration, learning curve, and intraoperative and early postoperative complications. RESULTS: Ten adult and 6 subadult baboons with a mean weight of 9.32 kg, a range of 4-14.2 kg, and a standard deviation (SD) of 3.09 kg were included in the study. The total duration of surgery was 28.75 min (range, 16-50 min; SD, 9.60 min). The installation phase was completed in a mean time of 7.68 min (range, 3-15 min; SD, 3.43 min), and the time to complete the salpingectomy of both salpinges was 9.68 min (range, 4-20 min; SD, 3.97 min). No complications were observed in the postoperative period. CONCLUSION: Laparoscopic salpingectomy in Papio hamadryas was feasible, with an acceptable surgical time, low invasiveness, and only minor technical perioperative complications. CLINICAL SIGNIFICANCE: Laparoscopic salpingectomy could be a viable and safe therapeutic option in nonhuman primate birth-control programs.

Clinical, haematological and biochemical findings in tigers infected by Leishmania infantum.

BACKGROUND: A large number of animal species are susceptible to Leishmania infantum (Kinetoplastida, Trypanosomatidae) in endemic areas, including domestic and wild felids such as tigers (Panthera tigris). Knowledge on the infection of this endangered species is still at its infancy, and therefore this study aims to identify clinical presentation and clinicopathological findings of tigers naturally infected by L. infantum. RESULTS: Tigers either L. infantum-positive (group A) or -negative (group B) were apparently healthy or presented visceral leishmaniasis unrelated conditions, except for one animal in which a large non-healing cutaneous lesion was observed. However, histological exam and immunohistochemistry carried out on the lesion excluded the presence of L. infantum amastigotes. Biochemical analysis showed that the average concentration of total proteins, globulins and haptoglobin were significantly higher (p < 0.01, p = 0.01 and p = 0.02, respectively), while the albumin/globulin ratio significantly lower (p = 0.05) in group A compared with group B. The biochemical alterations were partially confirmed by the serum protein electrophoresis results revealing a significant increase in the total protein value (p = 0.01) and hypergammaglobulinemia (p = 0.03) but an unmodified albumin/globulin ratio in group A. CONCLUSIONS: In this study tigers infected by L. infantum have shown to be mainly asymptomatic. The absence of clinical signs may lead veterinarians to overlook leishmaniasis in animals kept in captivity. Therefore, diagnostic and screening tests as serology should be part of routinely surveillance programs to be performed on tigers in zoological gardens located in endemic areas. Though only few protein-related laboratory abnormalities were recorded in infected animals, they could provide diagnostic clues for a first suspicion of L. infantum infection in tigers. Indeed, considering the high risk of zoonotic transmission in heavily frequented environment as zoos, a prompt diagnosis of L. infantum infection is of pivotal importance.

Leishmania infantum in Tigers and Sand Flies from a Leishmaniasis-Endemic Area, Southern Italy.

We detected Leishmania infantum infection in 45% of tigers and 5.3% of sand flies tested at a zoo in southern Italy in 2019. These infections in tigers and the abundance of Phlebotomus perniciosus sand flies represent a potential risk to other animals and humans living in or visiting the zoo.

Pharmacokinetics of injectable marbofloxacin after intravenous and intramuscular administration in red-eared sliders (Trachemys scripta elegans).

Fluoroquinolone antibacterial drugs are currently used in reptilian medicine because of their broad spectrum of activity including the most frequent pathogens of these species. The disposition kinetics of marbofloxacin (MBX) at a single dose of 2 mg/kg were determined in healthy red-eared sliders after intravenous (IV) and intramuscular (IM) administration. The influence of renal portal system on the bioavailability of the drug was investigated by using forelimb and hindlimb as IM injection sites. Apparent volume of distribution at steady-state (Vss ) and systemic clearance (Cl) of marbofloxacin after IV administration were estimated to be 48.21 ± 5.42 ml/kg and 23.38 ± 2.90 ml/hr·kg, respectively. The absolute bioavailabilities after IM route were 45.96% (forelimb) and 52.09% (hindlimb). The lack of statistically significant differences in most of the pharmacokinetic parameters after the two IM injection sites suggests a negligible influence of renal portal system in clinical use of MBX, although the Cmax after IMfore administration is advantageous, having into account the concentration-dependent action of this antibiotic. The absence of visible adverse reactions in the animals and the advantageous pharmacokinetic properties suggest the possibility of its safe and effective clinical use in red-eared sliders.

Diagnosis and treatment considerations in a case of malignant mesenchymoma in an African fur seal (Arctocephalus pusillus).

A 20-yr-old African fur seal (Arctocephalus pusillus) presented with a slowly growing mass located on the dorsum at the level of the last thoracic vertebrae. The mass was hard, 10 cm in diameter, and not adherent to the underlying tissues. Multiple biopsies were collected for histopathology and revealed extensive areas of necrosis, small nodules of malignant mesenchymal proliferation with areas of chondroid metaplasia, and atypical cells in vessel walls. The morphologic diagnosis was suggestive of malignant mesenchymal neoplasia originating from the vascular wall. The mass was removed 1 mo later due to ulceration and infection. Histologically, based on the World Health Organization's classification of neoplastic processes in domestic animals, the tumor was consistent with malignant mesenchymoma. The margins of resection revealed the presence of neoplastic cells. Based on these results, the particular species involved, the high local invasiveness, and the high metastatic index of this malignant tumor in domestic mammals and humans, the prognosis was poor. The animal died 6 mo later with metatastic disease.

Evaluation of a butorphanol, detomidine, and midazolam combination for immobilization of captive Nile lechwe antelopes (Kobus magaceros).

Field immobilization of captive antelope may be required for medical examination, blood sample collection, and animal identification. The aim of this study was to evaluate the effects of a combination of butorphanol, detomidine, and midazolam (BDM) and its partial reversibility in Nile lechwe antelope (Kobus megaceros). Nine captive lechwes, weighing 28-64 kg, were immobilized, in February 2011, with butorphanol 0.20 ± 0.05 (mean ± SD) mg/kg, detomidine 0.20 ± 0.05 mg/kg, and midazolam 0.31 ± 0.08 mg/kg administered intramuscularly (IM) with a blowpipe. Physiologic parameters and depth of anesthesia were recorded when the animals became recumbent at 19.55 ± 8.36 min after darting (T0) and after 10 (T10), 20 (T20), and 30 (T30) min. An arterial blood sample was collected at T20. At the end of the procedures, immobilization was partially reversed with atipamezole 0.25 mg/kg IM. Quality of induction, immobilization, and recovery was scored. The BDM combination induced immobilization and lateral recumbency in 13.44 ± 5.61 min. Median induction score (scored 1 [excellent] to 4 [poor]) was 1 (range 1-2). Heart rate varied 40-104 beats/min, respiratory rate 16-108 breaths/min, and rectal temperature 36.5-40.3 C. Hyperthermia was observed and rapidly treated in three animals that demonstrated insufficient immobilization after darting. Arterial blood gas analyses revealed a mean pH of 7.43 ± 0.07, partial arterial pressure of CO(2) of 44.1 ± 6.0 mmHg, partial arterial pressure of O(2) of 74.0 ± 13.5 mmHg, and an arterial O(2) saturation of 94.77 ± 3.96%. Recovery was smooth and animals were walking in 13.44 ± 7.85 min. Median recovery score (1 = excellent to 4 = poor) was 1 (range 1-2). The BDM was effective in immobilizing captive healthy lechwes with minimal cardiorespiratory changes.

Pharmacokinetics of marbofloxacin in loggerhead sea turtles (Caretta caretta) after single intravenous and intramuscular doses.

The disposition kinetics of marbofloxacin at a single dose of 2 mg/kg bodyweight were determined in a crossover trial with five clinically healthy loggerhead sea turtles (Caretta caretta) after i.v. and i.m. administration. Marbofloxacin plasma concentrations were determined by high-performance liquid chromatography (LOD/LOQ 0.05 microg/ml). Data were subjected to noncompartmental analysis. The integrated pharmacokinetic/pharmacodynamic variables showed that optimal area under the curve (AUC(0-24 h)): minimal inhibitory concentration (MIC) (>125) and Cmax: MIC (>8) ratios, as reported for concentration-dependent bactericidal antimicrobials (e.g., fluoroquinolones), were achievable with both a once daily i.v. or i.m. dose for microorganisms with MIC < or = 0.5 microg/ml, while a Cmax: MIC > 8 for MIC > or = 1 microg/ml was achievable only after the i.v. administration. The absence of adverse reactions in the animals after i.v. or i.m. administration of marbofloxacin and the favorable pharmacokinetic/pharmacodynamic properties after a single dose of 2 mg/kg suggest the possibility of its safe and effective clinical use in loggerhead sea turtles.

Pharmacokinetic behavior of enrofloxacin in estuarine crocodile (Crocodylus porosus) after single intravenous, intramuscular, and oral doses.

The disposition kinetics of enrofloxacin at a single dose of 5 mg/kg body weight were determined in clinically healthy captive-reared estuarine crocodiles (Crocodylus porosus) after intravenous (i.v.), intramuscular (i.m.), and oral (p.o.) administration. Enrofloxacin plasma concentrations were determined by diode array detection-high-performance liquid chromatography (limit of detection/limit of quantitation: 0.05 microg/ml). Data were subjected to noncompartmental analysis. The integrated pharmacokinetic-pharmacodynamic (PK-PD) variables showed that optimal area under the curve from the time of dosing to 24 hr:minimal inhibitory concentration (MIC) (>125) and peak plasma concentrations:MIC (>8) ratios, as reported for concentration-dependent bactericidal antimicrobials like fluoroquinolones, were achievable with both a single i.v. or i.m. dose for susceptible microorganisms with MIC values of < or =0.5 microg/ml, while the relatively slow onset of peak time allowed an effective plasma drug level only on day 3. The persistence of useful plasma concentrations indicated the possibility of redosing every 3 day for parenteral routes of administration, while further studies are needed for the oral route. Nevertheless, the absence of adverse reactions in the animals following i.v., i.m., or p.o. administration of enrofloxacin after a single dose of 5 mg/kg indicates the possibility of its safe and effective clinical use in captive estuarine crocodiles.

Reversible immobilization of asiatic black bear (Ursus thibetanus) with detomidine-tiletamine-zolazepam and atipamezole.

Chemical immobilization of free-ranging and captive wildlife is often required in many clinical situations. In this trial, tiletamine-zolazepam was combined with the alpha2-agonist, detomidine, in order to use the least amount of anesthetic drug possible to achieve a rapid immobilization; to ensure safety for animals and operators; and to be easily reversible with specific antagonists for a fast recovery. Twelve captive Asiatic black bears were anesthetized for clinical procedures, including clinical examination and blood sample collection, and for electrocardiographic and echocardiographic procedures. The combination detomidine-tiletamine-zolazepam, at the dosages of 0.03 mg/kg for detomidine and 1.5 mg/kg for tiletamine-zolazepam, proved to be reliable and effective in immobilizing Asiatic black bears for a 1-hr handling period for routine clinical procedures. Minimal or no respiratory and/or cardiopulmonary adverse side effects were observed, even with dosages calculated on the basis of an estimated body weight. The respiratory rate, pulse rate, and hemoglobin-oxygen saturation remained stable for the entire duration of anesthesia. Cardiac rhythm was always sinusal in all animals. Small injection volumes and darts for blowpipe use were utilized to minimize tissue damage at the site of injection. Induction and recovery were smooth and predictable, and provided for the safety of operators who could observe the bears' activities from a safe distance. Furthermore, the availability of the alpha2-antagonist atipamezole to counteract the effects of detomidine made this anesthetic regimen easily controllable and reversible. Moreover, the recovery time can be shortened by intravenous administration of this antagonist drug.

Malignant catarrhal fever in a captive American bison (Bison bison) in Italy.

Malignant catarrhal fever (MCF) is a fatal, systemic disease of cattle and other domestic and wild ruminants that, in Europe, is caused by Ovine herpesvirus 2 (OvHV-2). American bison (Bison bison) are highly susceptible to the disease. An adult American bison, housed in a zoo in southern Italy in close cohabitation with a group of domestic sheep (Ovis aries aries) displayed clinical signs that resembled the acute form of MCF. By real-time polymerase chain reaction, OvHV-2 DNA was detected intravitam in blood, in nasal and ocular swabs, and postmortem in tissue samples of the bison. By indirect fluorescent antibody test, high MCF antibody titers were found in the bison serum. Ovine herpesvirus 2 DNA and antibodies were also found in blood samples from the domestic sheep, thus suggesting a potential role of these animals as a source of the infection. To the authors' knowledge, this is the first MCF case in captive ruminants in Italy and the second confirmed case in captive bison of European zoos.

Serologic and molecular characterization of tickborne pathogens in lions (Panthera leo) from the Fasano Safari Park, Italy.

Lions (Panthera leo) are an endangered species threatened by illegal hunting, habitat loss, and infectious diseases. Little is known about the tick-borne pathogens that infect lions and could contribute to population declines. The objective of this study was to characterize Rickettsia spp., Anaplasma phagocytophilum, and Coxiella burnetii infections in 10 lions from the Fasano Safari Park in Italy by serology, polymerase chain reaction, and sequence analysis. Although animals did not show clinical signs of tick-borne diseases, evidence of infection with C. burnetii, spotted fever group Rickettsia sp., and A. phagocytophilum were found in 50%, 20%, and 10% of the lions, respectively. One of the lions tested positive for all three pathogens. This study is the first report of molecular evidence of infection with C. burnetii, Rickettsia sp., and A. phagocytophilum in lions and provides evidence that these felids become infected and serve as hosts for tick-transmitted bacteria.