Micaela Sgorbini

Cryptosporidium parvum infection in a mare and her foal with foal heat diarrhoea.

Cryptosporidium infection was molecularly investigated in mares and in their neonatal foals for which the occurrence of foal heat diarrhoea was also assessed. Thirty-seven mare/foal pairs were included in the study. All foals were born in the same stud farm during 2006-2008 breeding seasons. Two faecal samples, one prior to and one after delivery were collected from each mare, whereas three faecal samples were taken from each foal, i.e. at 8, 10 and 12 days of age. All samples (74 from mares and 111 from foals) were divided into two aliquots, one of which was examined for the presence of Cryptosporidium by a commercially available microplate ELISA kit, while the second aliquot of all ELISA-positive samples was molecularly examined. Nine out of 37 examined foals presented foal heat diarrhoea and one of them scored positive for Cryptosporidium, together with its mare. More specifically, four samples belonging to the same mare/foal pair resulted positive for Cryptosporidium upon both ELISA and PCR. The sequence analysis of the COWP gene showed the occurrence of the zoonotic species Cryptosporidium parvum. The possibility that foal heat diarrhoea-like episodes may be due to neonatal cryptosporidiosis and their relevance for the health of horses and of humans handling diarrhoeic neonatal foals and their mares are discussed.

Presence and distribution of fungi and bacteria in the reproductive tract of healthy stallions

A saprophytic bacterial flora is present on the penis and the distal part of the urethra of stallions. Little is known about the fungal flora of their reproductive tract. As micro organisms play an important role in mares fertility, the aim of the study was to describe the distribution of fungi and bacteria in the normal genital apparatus of stallions. The microbic flora of the reproductive tract of 11 healthy, fertile stallions was evaluated, collecting samples from 5 different locations: urethral fossa, penis/internal lamina of the prepuce, urethra pre- and post-ejaculation, and semen. For fungal examination samples were taken on 3 different occasions (N = 165), while for bacteriologic examination samples were taken on one occasion only (N = 55). There was a statistical difference in the presence of filamentous fungi between urethral fossa or penis/prepuce (45.4%) and urethra pre- or postejaculation or semen (15.1%, 6.0%, and 0.0%, respectively). Yeasts were isolated in 9.1% of the samples, never in semen. The most represented mycelial fungi were Penicillium spp., Aspergillus spp., Scopulariopsis spp., Trichosporon spp. and Mucoracee. The proportion of samples showing a total bacterial count ≥ 10,000 colony forming units (CFU) was higher for urethral fossa than for urethra pre- or postejaculation or for semen. Some bacterial growth was always observed in all locations, including the ejaculate. Differences between sampling locations were observed also for Staphylococci, both coagulase positive and negative. Salmonella enterica Abortus equi and sulphite reducing clostridia and other pathogens (including Klebsiella spp. and Pseudomonas spp.) were never isolated. Escherichia coli and coliforms always showed a low or absent flora. These data add information to the literature.

Occurrence of fungi from conjunctiva of healthy horses in Tuscany, Italy

The fungal flora of the normal conjunctival fornix has been described for many animal species (Urban et al., 1972; Samuelson et al., 1984; Moore et al., 1988; Davidson et al., 1994; Cooper et al., 2001) and many environmental factors, such as age, geography, habitat and husbandry are reported to influence its composition (Samuelson et al., 1984; Andrew et al., 2003). Equine keratomycosis can be caused by saprophytic fungi following an underlying corneal pathology or improper use of topical antibiotics and/or topical corticosteroids (Andrew et al., 1998; Brooks, 1999). Knowledge of differences in geographic prevalence of fungi and their frequency in the conjunctiva of the horse could allow the identification of risk factors for the development of keratomycosis. The purpose of the present paper was to culture, describe and quantify moulds and yeasts from the conjunctival fornix of healthy horses living in Tuscany (Italy), and to identify the most common fungal species.

Oral administration of tepoxalin in the horse: A PK/PD study

Tepoxalin is a non-steroidal anti-inflammatory drug with analgesic, anti-inflammatory, and antipyretic properties and has been recently introduced into veterinary medicine. The aim of this study was to evaluate the pharmacokinetic/pharmacodynamic (PK/PD) profile of tepoxalin to assess whether it would be suitable for clinical use in horses. Six female fasting/fed horses were given 10mg/kg tepoxalin orally in a cross-over study. After administration, tepoxalin underwent rapid and extensive hydrolytic conversion to its carboxylic acid metabolite RWJ-20142. In animals that had been fed, the plasma concentrations of tepoxalin were undetectable, whereas in fasting animals they were close to the limit of quantification of the method. No differences between the fasting/fed groups in RWJ-20142 plasma concentrations were shown. Tepoxalin showed a strong and long-lasting ex vivo inhibitory activity against cyclooxygenase (COX)-1, mainly due to its main metabolite RWJ-20142. Tepoxalin and RWJ-20142 do not seem to possess either COX-2 or 5-lipoxygenase inhibitory activity in the horse. These features suggest that the drug is a selective COX-1 inhibitor in horses, with no significant anti-inflammatory activity. Thus, its long term use in equine practice could be of concern.

Evaluation of pharmacokinetic and pharmacodynamic properties of cimicoxib in fasted and fed horses

Abstract AIMS: To determine the pharmacokinetics of cimicoxib and to assess the inhibition of cyclooxygenase (COX) after a 5 mg/kg, single oral administration in horses that were fasted or fed. METHODS: The study was conducted using an open, single dose (5 mg/kg), two treatment (fasted and fed), two-period, crossover design with a 2-week interval between dosages. Six healthy mares received 5 mg/kg of cimicoxib via nasogastric tube after fasting for 12 hours, or 2 hours after feeding. After administration, blood samples were collected for up to 24 hours and plasma used for pharmacokinetic analysis. Additional serum and plasma samples were used to measure concentrations of thromboxane B2 (TXB2) and prostaglandin E2 (PGE2), to assess COX-1 and -2 inhibition, respectively. RESULTS: Following cimicoxib administration, the mean maximum plasma concentration was 0.16 (SD 0.01) µg/mL and 0.14 (SD 0.03) µg/mL in fasted and fed groups, respectively. The mean time taken to reach maximum plasma concentration was longer in the fed group (5.91 (SD 3.23) hours) compared with the fasted group (3.25 (SD 1.17) hours), but this difference was not significant (p=0.12). The mean maximal inhibition of TXB2 was 62.4 (SD 13.8)% and 54.6 (SD 15.4)%, and of PGE2 was 72.1 (SD 43.3)% and 68.5 (SD 24.4)%, in fasted and fed horses, respectively. CONCLUSION: In the present study, although the COX-2 selective action of cimicoxib was not apparent, a relatively low concentration of cimicoxib resulted in both COX-1 and -2 inhibition in horses. Further investigations are required to establish an optimal dosage regimen and safety profile before clinical trials are initiated.

Plasma Procalcitonin Concentration in Healthy Horses and Horses Affected by Systemic Inflammatory Response Syndrome

Background The diseases most frequent associated with SIRS in adult horses are those involving the gastrointestinal tract. An early diagnosis should be the goal in the management of horses with SIRS. Objective The objective of this study was to evaluate the plasma procalcitonin (PCT) concentration in healthy and SIRS horses to assess differences between the two groups. Animals Seventy‐eight horses (30 healthy and 48 SIRS). Methods Prospective in vivo multicentric study. Horses were classified as SIRS if at least 2 of the following criteria were met: abnormal leukocyte count or distribution, hyperthermia or hypothermia, tachycardia, tachypnea. Healthy horses showed no clinical or laboratory signs of SIRS. Plasma PCT concentrations were measured with a commercial ELISA assay for equine species. Results were expressed as mean±standard deviation. T‐test for unpaired data was performed between healthy and SIRS group. SIRS group was divided in 4 subgroups and t‐test was performed between healthy versus each subgroup. Results PCT concentrations in healthy and SIRS horses were 18.28 ± 20.32 and 197.0 ± 117.0 pg/mL, respectively. T‐test showed statistical differences between healthy versus SIRS group and between healthy versus all subgroups. Conclusions and Clinical Importance Results showed an increase in PCT concentration in SIRS horses as previously reported in humans and dogs. PCT could be used as a single assay in equine practice for detection of SIRS.

Ocular abnormalities in healthy Standardbred foals

OBJECTIVE To determine the prevalence and describe ocular abnormalities in healthy Standardbred foals within 48 h of birth. ANIMALS One hundred and two neonatal foals. PROCEDURES All foals had an unassisted delivery. On the basis of physical examination and the results of hematological and biochemical parameters, all foals were unaffected by systemic diseases. A complete ophthalmic examination was performed within 48 h of birth. Foals with ocular hemorrhages were re-examined weekly until the abnormalities were resolved. RESULTS 65/102 (63.7%) foals did not show ocular abnormalities, while in 37/102 (36.3%) cases, ocular abnormalities were present. Retinal and subconjunctival hemorrhages were recorded in 19/102 (18.6%), and in 13/102 (12.7%), respectively. In 4/102 (3.9%) animals, an entropion of the lower eyelid was present. Only one foal (1%) showed a congenital nuclear unilateral cataract. No other ocular abnormalities were detected. However, all foals showed various degrees of remnants of hyaloid system. One week after the first ocular examination, retinal hemorrhages had resolved in 100% of the eyes, whereas subconjunctival hemorrhages had disappeared in all eyes by the second week following the first examination. CONCLUSIONS The acquired ocular lesions observed with relatively high frequency in the examined healthy Standardbred foals were ocular hemorrhages, which always showed a good outcome. Although these abnormalities were present at birth, they were not considered strictly congenital but likely acquired during parturition. Instead, congenital ocular abnormalities were rarely diagnosed, and the entropion of the lower eyelid was the most common disease in the breed.

Seasonal prevalence of fungi in the conjunctival fornix of healthy cows during a 2-year study

OBJECTIVE The aims of the present paper were to: (i) identify and quantify conjunctival fungi isolated from healthy cows; (ii) verify the influence of different methods of farm management on the prevalence (percentage of positive cultures for each fungal species per farm) of conjunctival fungi. MATERIAL AND METHODS Forty Friesian and twenty Limousin female cows aged 1-10 years stabled in three farms with different managements (farm 1: cows housed strictly indoors; farm 2: cows housed outside during the day and inside the stall during the night; farm 3: cows housed strictly outdoors) were investigated for conjunctival fungal flora. Air and food were also tested. Specimens were collected every season during a 2-year study. Identification of colonies of filamentous fungi was achieved to the genus level on the basis of macro- and microscopic features. RESULTS The total number of eyes positive for fungi ranged from 85 to 100% at farm 1, from 65 to 95% at farm 2, and from 55 to 95% at farm 3. Fungi most frequently isolated from conjunctival fornix were Cladosporium spp. and Penicillium spp. Statistical analysis did not show any differences in fungal prevalences among the three farms during the same season. Some fungal species were consistently isolated while others were intermittently isolated. CONCLUSIONS Fungi found in the conjunctival fornix of cows might represent transient seeding from the environment, as suspected in other species. The prevalence of conjunctival fungal organisms is not different in cattle housed indoors vs. outdoors.

Treatment of Chronic Atrial Fibrillation in the Horse with Flecainide: Personal Observation

Birettoni, F., Porciello, F., Rishniw, M., della Rocca, G., Di Salvo, A. and Sgorbini, M., 2007. Treatment of chronic atrial fibrillation in the horse with flecainide: Personal observation. Veterinary Research Communications, 31(Suppl. 1), 273–275

Conjunctival bacterial and fungal flora in clinically normal sheep.

Objectives The aim was to identify conjunctival bacterial and fungal flora in clinically normal sheep. Design Prospective study. Setting Tuscany. Participants 100 eyes from 50 adult Massese female sheep were examined. The sheep included in the study were considered free of anterior ophthalmic abnormalities. Primary and secondary outcome measures Bacteria were identified by morphological assessment, Gram staining, biochemical tests. Identification of filamentous fungi was achieved at the genus level, and Aspergillus species were identified based on keys provided by other authors. Yeast colonies were highlighted, but not identified. Results Positive cultures were obtained from 100/100 eyes for bacteria, and from 86/100 eyes for fungi. A total of 14 types of bacteria and 5 types of fungi were isolated. Yeasts were isolated from 13/100 eyes. The most frequent fungal isolates were saprophytic fungi. Conclusions Conjunctival bacterial and fungal flora of clinically normal eyes were reported in sheep. The positivity obtained for conjunctival bacteria was higher compared to findings in the literature by other authors in the same species (100 per cent v 40 per cent), while our results were in line with a recent work performed on mouflons (Ovis Musimon) with a 100 per cent positivity for bacterial conjunctival fornix. In our survey, Gram-positive species were prevalent, as reported by other authors in different species. Few data are available in the literature regarding conjunctival fungal flora in healthy small ruminants. The prevalence of conjunctival fungal flora in this study was higher than findings reported in mouflons (86 per cent v 45 per cent). Differences in fungal prevalence may be due to different methods of managing herds, though further studies are required to verify this hypothesis. The similarities in bacterial and fungal isolates between sheep and mouflons suggest a genera pattern of conjunctival colonisation by bacteria and fungi.