Francesco Di Ianni

Conjunctival flora of clinically normal and diseased turtles and tortoises

BackgroundIn captive breed turtles and tortoises conjunctival disease is common. Our aim was to investigate the bacterial and fungal flora present in the eyes of healthy and pathological chelonians and to compare findings in turtles with those in tortoises.ResultsSamples were taken from the conjunctival sacs of 34, diseased and healthy, chelonians (18 tortoises and 16 turtles) and submitted to bacterial and fungal investigation. All samples showed bacterial growth. Thirteen animals (38%), harboured a single bacterial species as sole isolate and twenty-one animals (62%) harboured more than one species. Detection of multiple bacterial infection was clearly greater in tortoises compared to turtles. Most frequently isolated bacterial species were Bacillus spp. (13 isolates), Staphylococcus xylosus (10 isolates), Sphingomonas paucimobilis (6 isolates), Staphylococcus sciuri and Aeromonas hydrophila/caviae (each 5 isolates), Ochrobactrum anthropi (3 isolates), Citrobacter freundii, Enterobacter cloacae and Pseudomonas luteola (each 2 isolates). Only one isolate of Kocuria varians/rosea, Staphylococcus aureus, Staphylococcus auricularis, Staphylococcus haemolyticus, Staphylococcus lentus, Morganella morganii, Pasteurella multocida, Pasteurella pneumotropica/haemolytica, Proteus spp., Pseudomonas putida, Salmonella enterica ssp. arizonae, Stenotrophomonas maltophilia and Vibrio parahaemolyticus was evidenced. The presence in 8 animals of Mycoplasma spp. and in 1 animal with severe conjunctivitis of Chlamydia spp. was detected by PCR. Candida spp. was also isolated from two healthy animals.ConclusionsA clear predominance of Gram positive isolates in tortoises and Gram negative isolates in turtles was found. However, we cannot ascribe the observed difference to the diversity of animal species, as other factors, including especially different characteristics of the living environments, may play a role. Almost all bacterial species isolated may have clinical significance, mostly as opportunistic pathogens, both for humans and animals. That chelonians are often carrier of bacteria with zoonotic potential is a well-known fact, in particular with regard to Salmonella spp. Therefore, it is not surprising the detection of a strain of Salmonella enterica ssp. arizonae in the eye of one of the animals tested. Worthy of note is the finding of Chlamydia spp. in a severe case of conjunctivitis, though we cannot epidemiologically assess a cause-effect relationship between presence of chlamydia and disease.

Boar semen bacterial contamination in Italy and antibiotic efficacy in a modified extender

The aims of the study were to identify microbial flora in boar semen under field conditions in northern Italy, to investigate antibiotic resistance and sensitivity of isolated bacteria, and to evaluate elimination of bacteria after storage in two types of extenders added with different antibiotics (amikacin vs gentamicin). A total of 60 boars were collected in 13 pig farms. Bacteriological and mycological investigations were performed immediately on raw semen samples, then at 48 and 120 h of storage on semen diluted randomly in a new short-term modified extender (ME-S) or in a commercial one (CRONOSTM). Bacterial contamination was found in 63% of raw semen samples and different bacterial species were isolated: E.coli, Serratia marcescens, Staphylococcus epidermidis and aureus, Proteus spp., Streptococcus spp. and Pseudomonas aeruginosa. E. coli was the most isolated contaminant (53%); Pseudomonas aeruginosa was found only in one semen sample. The analysis of variance of factors affecting contamination levels was significant for the farm of origin (P<0.05) and not significant for the breed. Antibiotic resistance of these bacteria was assessed using different antibiotics. Significant differences (P<0.05) between observed and expected frequencies of bacterial isolates resistant or not to the antibiotics contained in the extenders were found. At 48 h of storage a reduction of aerobic contamination was found after ME-S dilution by 85.3% and after CRONOSTM by 63.8%. This paper proved the presence of pathogenic bacteria in semen. We thus believe it is highly advisable to perform periodic microbiological screening of boar semen in the swine industry to avoid the use of low sperm quality.

Use of aglepristone for the treatment of P4 induced insulin resistance in dogs.

Insulin resistance (IR) in dogs is suspected when hyperglycemia is present despite administration of insulin doses greater than 1.0 to 1.5 UI/kg. IR is caused by increases in counter regulatory hormones concentrations (glucagon, glucocorticoids, catecholamines and growth hormone). This study was conducted to investigate the use of aglepristone (RU 46534), a P4 receptor antagonist, for the treatment of IR diabetes mellitus in bitches during the luteal phase. All animals were treated with porcine insulin zinc suspension (Caninsulin) and aglepristone (Alizin) 10 mg/kg subcutaneously at day 1, 2, 9 and 17 from diagnosis. At day 5, no significant variation in glycemia was shown. At day 12 and 20, serum glucose concentrations were significant lower (p < 0.05). From day 12 the insulin dose was reduced to 0.8 IU BID. Insulin was reduced in the following weeks and glycemia was controlled.

Diagnostic sensitivity of ultrasound, radiography and computed tomography for gender determination in four species of lizards.

Gender determination is frequently requested by reptile breeders, especially for species with poor or absent sexual dimorphism. The aims of the current study were to describe techniques and diagnostic sensitivities of ultrasound, radiography, and computed tomography for gender determination (identification of hemipenes) in four species of lizards. Nineteen lizards of known sex, belonging to four different species (Pogona vitticeps, Uromastyx aegyptia, Tiliqua scincoides, Gerrhosaurus major) were prospectively enrolled. With informed owner consent, ultrasound, noncontrast CT, contrast radiography, and contrast CT (with contrast medium administered into the cloaca) were performed in conscious animals. Imaging studies were reviewed by three different operators, each unaware of the gender of the animals and of the results of the other techniques. The lizard was classified as a male when hemipenes were identified. Nineteen lizards were included in the study, 10 females and nine males. The hemipenes were seen on ultrasound in only two male lizards, and appeared as oval hypoechoic structures. Radiographically, hemipenes filled with contrast medium appeared as spindle-shaped opacities. Noncontrast CT identified hemipenes in only two lizards, and these appeared as spindle-shaped kinked structures with hyperattenuating content consistent with smegma. Hemipenes were correctly identified in all nine males using contrast CT (accuracy of 100%). Accuracy of contrast radiography was excellent (94.7%). Accuracy of ultrasound and of noncontrast CT was poor (64.3% and 63.1%, respectively). Findings from the current study supported the use of contrast CT or contrast radiography for gender determination in lizards.

Preparation and application of an innovative thrombocyte/leukocyte-enriched plasma to promote tissue repair in chelonians

Platelet concentrates are widely used in mammalian regenerative medicine to improve tissue healing. Chelonians (Testudines) would benefit from the application of thrombocyte preparations to regenerate damaged tissues, since traumatic injuries are leading causes of morbidity and mortality for both wild-living and domesticated animals. The aim of this study was to establish a protocol that optimized the recovery of the thrombocytes from blood samples and to show the efficacy of thrombocyte-enriched plasma in chelonians. Peripheral blood samples were obtained from Testudo spp. (n = 12) and Trachemys scripta elegans (n = 10). Blood cells were fractionated by sodium diatrizoate-sodium polysucrose density gradient using a two-step centrifugation protocol. Thrombocytes and leukocytes were isolated and resuspended to obtain thrombocyte-leucocyte rich plasma (TLRP). The mean recovery of leukocytes and thrombocytes was 48.9% (±4.0 SEM, n = 22) of the whole blood cell content. No statistically significant difference was observed between blood samples collected from different turtle species. The ability of TLRP to form a gel was evaluated by adding variable concentrations of calcium gluconate at room temperature and at 37°C. A reliable and consistent clotting of the TLRP was obtained in glass tubes and dishes by adding 5-20% v/v of a 100 mg/ml solution of calcium gluconate. Furthermore, in order to test the clinical efficacy of TLRP, a preliminary evaluation was performed on four turtles (Testudo spp.) with traumatic injuries. In all the four animals, a successful clinical outcome was observed. The results demonstrated that a thrombocyte-enriched plasma, comparable to mammalian platelet rich plasma, can be prepared from chelonian blood samples. Furthermore, although the low number of cases presented does not allow definitive conclusions from a clinical point of view, their outcome suggests that TLRP application could be further investigated to improve the healing process of both soft and hard tissue injuries in chelonians.

Monitoring the reproductive activity in captive bred female ball pythons (P. regius) by ultrasound evaluation and noninvasive analysis of faecal reproductive hormone (progesterone and 17β-estradiol) metabolites trends

The royal python (Python regius) is commonly bred in captivity. To have a successful breeding season, accurate monitoring of the reproductive activity is necessary. The use of non-invasive monitoring methods in exotics is important in order to minimize stress. For this purpose ultrasound has been anecdotally used to monitor royal python reproductive activity. However, there is limited information regarding the reproductive cycle of this species. The aim of the present study is to monitor the female reproductive cycle of the royal python using ultrasonography and gonadal steroid metabolite measurements in the faeces. The reproductive activity of one hundred twenty-nine adult female P. regius was examined during two consecutive years. We performed brief scans on non-anaesthetized snakes using a portable ultrasound system and a 10–12 MHz linear array transducer (MyLab™ 30 Gold, Esaote). Ultrasound features, dimension and echogenicity of the reproductive structures were determined. During the second reproductive cycle, the hormonal profiles of 30 animals were also evaluated, with a monthly collection of faecal samples. These samples were classified according to reproductive stage, as identified by ultrasonographic examination, and the mean faecal progesterone and 17β-estradiol levels were calculated using the results from an enzyme-linked immunosorbent assay (ELISA). Progesterone levels increased during the reproductive cycle. Estradiol levels showed greater variability, although they appeared to increase before coupling when compared to the levels between coupling and egg laying. The present study suggests that it is possible to identify different phases in the female royal python reproductive cycle: anovulatory phase, transition, folliculogenesis and embryogenesis. Ultrasound is also useful for identifying follicular regression or slugs. Gonadal steroid metabolite measurements from the faeces could help integrate reproductive information. The use of ultrasonography in addition to the steroid metabolite measurement in the faeces gives an accurate picture of ovarian activity in captive adult female royal pythons.

Evaluation of effectiveness of an innovative semen extender (Formula®) comparing with a traditional extender (Lepus®) for artificial insemination in rabbits does

Abstract This study aimed to investigate the preservability and viability of the rabbit spermatozoa diluted in a new semen extender Formula® in comparison with Lepus® at 17 °C of storage. The main characteristic of the new extender formulation is the use of an enzymatic agent associated to a polysaccharide as energy source precursor, added with gentamycin. During eight trials, ejaculates from 70 bucks were collected and diluted at 1:10 ratio with both the extenders, after 24 h of storage the semen doses were used for the artificial insemination (AI). Aliquots of the semen doses for each trial were stored at 17 °C, the total and progressive motility were checked at 0, 4, 12, 18, 24, 36, 48, 60, 72, 84, 96, 108 h of storage. A total of 1267 and 1525 does were inseminated, respectively with Formula® and Lepus®. During storage the mean total and progressive motility (77.23% and 72.854%, respectively) were significantly higher for Formula® (p < .01) and the progressive motility at almost 70% was maintained for at least 60 h vs the 24 h of storage for Lepus® with significant differences after 12 h of storage (p < .05). The new extender reported a higher pregnancy rate (p < .05) and an average of 9.25 rabbits born per litter vs 8.83 for the traditional extender (p < .05), while the mean of the newborn alive was 9.08 using Formula® vs 8.51 with Lepus® (p < .05). In conclusion, the use of Formula® is recommended for rabbit semen AI programmes.

Modulation of the Bifidobacterial Communities of the Dog Microbiota by Zeolite.

During last decades canine health and well being is becoming an important issue for human owners. In dogs, several factors including diet, pathogenic bacterial and stress conditions can affect the composition of the gut microbiota. In this study, we evaluated the effect of dietary chabazitic zeolitite (CZ) supplementation on the contribution of bifidobacteria to the fecal microbiota in training hunting dogs. Fecal microbiota cataloging based on 16S rRNA microbial profiling analyses highlighted an increase of Lactobacillus and Bifidobacterium in animals treated with CZ, with a simultaneous decrease of pathogens associated with dog gastrointestinal infections, such as Klebsiella and Enterobacter. A detailed profiling of the bifidobacterial population of dogs receiving CZ based on the ITS-based sequencing approach, revealed an enhancement bifidobacterial of species typical of animals such as Bifidobacterium animalis and B. pseudolongum. Moreover, these analyses identified the occurrence of putative new bifidobacterial taxa in both treated and untreated samples.

COMPUTED TOMOGRAPHIC FEATURES OF INCISOR PSEUDO-ODONTOMAS IN PRAIRIE DOGS (CYNOMYS LUDOVICIANUS)

Maxillary incisor pseudo-odontomas are common in pet prairie dogs and can cause progressive respiratory obstruction, while mandibular pseudo-odontomas are rarely clinically significant. The aim of this retrospective cross-sectional study was to describe CT features of maxillary and mandibular incisor pseudo-odontomas vs. normal incisors in a group of pet prairie dogs. All pet prairie dogs with head CT scans acquired during the period of 2013-2015 were included. A veterinary radiologist who was aware of final diagnosis reviewed CT scans and recorded qualitative features of affected and normal incisors. Mean density values for the pulp cavity and palatal and buccal dentin were also recorded. A total of 16 prairie dogs were sampled (12 normal maxillary incisors, 20 confirmed maxillary incisor pseudo-odontomas, 20 normal mandibular incisors, 12 presumed mandibular incisor pseudo-odontomas). Maxillary incisors with confirmed pseudo-odontomas had a significantly hyperattenuating pulp and dentin in the reserve crown and apical zone, when compared to normal maxillary incisors. Pseudo-odontomas appeared as enlargements of the apical zone with a globular/multilobular hyperattenuating mass formation haphazardly arranged, encroaching on midline and growing caudally and ventrally. Presumed mandibular incisor pseudo-odontomas had similar CT characteristics. In 60% of prairie dogs with maxillary incisor pseudo-odontomas, the hard palate was deformed and the mass bulged into the oral cavity causing loss of the palatine bone. The common nasal meatus was partially or totally obliterated in 81.8% of prairie dogs with maxillary pseudo-odontomas. Findings supported the use of CT for characterizing extent of involvement and surgical planning in prairie dogs with pseudo-odontomas.

Effects of three different designed farrowing crates on neonatal piglets crushing: preliminary study

Abstract In swine production, the economic gain in the farrowing room corresponds to the number of weaned piglets/sow per year. Pre-weaning mortality is at least 11–13% of total losses within one herd, considering a previous 7–8% of the stillbirths. Piglet crushing by the sow contributes significantly to the overall piglet mortality. It is caused by multiple factors related to the sow, the piglet and due to modern swine husbandry along the last 50 years. This study aimed to compare three different designs of farrowing crates during the first three days of life in relation to piglet-crushing mortality in intensive herd. One hundred fifty-eight sows with spontaneous deliveries were housed in the following farrowing crates groups: group A (n = 51) farrowing within conventional crates, group B (n = 47) farrowing within the slide cage and group C (n = 60) in the up and down designed, for a total of 2487 live born piglets. In the first three days of life, group C reported the lower crushing mortality rate (0.54%), while groups B and A reported 2.37% and 5.46%, respectively. The comparison between the group C value compared with group A values (p<.001; Chi-square = 36.90) and B (p = .003; Chi-square = 8.81) were statistically significant. In conclusion, the approach to create more space for both mother and litter within an acceptable size of farrowing crates (slide cage and up & down designed) allowed a significant reduction in mortality crushing rate of the piglets during the first three days of life.