Phenotypic and genetic characterization of Erwinia carotovora from mulberry (Morus spp.)

Phenotypic and genetic characteristics of nine bacterial strains isolated from mulberry ( Morus spp.), which were originally described as Erwinia carotovora ssp. carotovora (Ecc), were investigated. Based on the results of biochemical tests, these bacterial strains were divided into two different types, type 1 and type 2. Two strains of type 1 were similar to Ecc, whereas seven strains of type 2 were distinct from Ecc. A polyphasic study that included serological assay, specific PCR assay for E. carotovora ssp. atroseptica (Eca), PCR-RFLP of a pectate lyase ( pel ) gene and RAPD-PCR was performed on the type 2 strains, and the data were compared with those of related E. carotovora subspecies. The results of serological and specific PCR assays for Eca showed that the type 2 strains were distinct from Eca. In RFLP analysis of the pel gene using Sau 3AI, the type 2 strains showed a unique RFLP pattern. On the basis of RAPD analysis, similarity of RAPD patterns within the type 2 strains was very high. A unique RAPD fragment was isolated from the type 2 strains and used as a probe for Southern hybridization. This probe hybridized only with PCR products from the type 2 strains. Based on phenotypic, serological and genetic characteristics, the type 2 strains isolated from mulberry may belong to a distinct E. carotovora subspecies other than Eca or Ecc.     

In Vivo Embryo Recovery Rate by Laparoscopic Technique from Rabbit Does Selected for Growth Rate

Rabbit does from R line selected for growth rate present a low reproductive performance and this study aimed to evaluate both the recovery efficacy and viability of recovered embryos after vitrification and the reproductive performance of donor does subjected to in vivo recovery. Does were divided into three groups: 28 does without in vivo recovery (control), 25 does in which in vivo recovery was started in the nulliparous state (group 1) and 30 does with at least one litter before in vivo recovery (group 2). Does were superovulated with a single subcutaneous injection of 50 IU of equine chorionic gonadotropin (eCG) per female, and were then artificially inseminated 60 h later and immediately administered an intravenous dose of 75 IU of human chorionic gonadotropin (hCG) per female. Does from group 1 and 2 were recovered in vivo 76-80 h post-insemination by repeated laparoscopies at one to four times and permitted one or two parturitions between recoveries [in vivo (IV) recovery]. At the end of the experiment, about 16 does of all groups were recovered post-mortem (PM recovery). All normal embryos were vitrified, devitrified and then cultivated in vitro to evaluate the viability after thawing. A significant increase in the ovulation rate was found in does recovered PM than in those recovered IV in the nulliparous state. However, no significant differences were observed in the recovery rate, the donor rate, the number of normal embryos recovered with at least one normal embryo per doe and the viability after thawing between the PM and IV groups. A significant decrease in the fertility rate, total born, live born and weaned kids was found for does from group 1 in comparison with does from group 2. Results support the use of repeated laparoscopy to increase the number of recovered embryos per donor doe especially in such R line does, if they are permitted to produce at least one litter before the beginning of in vivo recovery.     

Use of Powdered Egg Yolk vs Fresh Egg Yolk for the Cryopreservation of Ovine Semen

Egg yolk is a common additive to sperm cryopreservation diluents. Because of its animal origin, however, it also represents a potential risk of microbiological contamination in the diluent. This potential contamination can be avoided by using powdered egg yolk, instead of fresh egg yolk, as it is pasteurized. This study was conducted to determine ram sperm cryosurvival was affected by the type of egg yolk used (powdered egg yolk or fresh egg yolk) and by yolk concentration (10, 15 or 20%) in the diluent. Microbiological analyses were also performed to quantify the microbiological contamination in the diluents containing the two types of egg yolk. Sperm cryosurvival was determined by motility and morphology analyses after thawing. Motility parameters were assessed using a computer-assisted sperm analysis (CASA) system, and the percentage of sperm with a normal apical ridge was evaluated using a differential interference contrast microscope. No significant differences were observed between diluents in the percentage of sperm with normal apical ridge. However, higher percentages of total motile cells were observed for samples containing powdered egg yolk (69%) compared to samples containing fresh egg yolk (60%). However, sperm in diluents containing fresh egg yolk, exhibited higher values for average-path velocity, straight-line velocity and beat cross frequency and lower values for amplitude of lateral head displacement (p <0.05), compared to cells in diluents containing powdered egg yolk. Microbiological contamination was similar (<200 CFU/ml) in both diluents, and no bacterial growth was observed in either, when antibiotics were added. Therefore, powdered egg yolk can be effective used in diluents for the freezing of ram semen. However, the in vivo fertility of sperm frozen in diluents containing powdered egg yolk should be tested, as some motility parameters were different for sperm treated with powdered egg yolk compared to fresh egg yolk.     

Death of a neonatal lamb due to Clostridium perfringens type B in New Zealand

ABSTRACT

Case history and clinical findings: A flock of 20 sheep was kept within three paddocks on a single property. None of the animals in the flock had been vaccinated against any disease for at least three years. Abdominal bloating and haemorrhagic diarrhoea were observed in Lamb 1 at 24 hours-of-age. The lamb subsequently died within an hour of the onset of clinical signs. Lamb 2 was 3-days-old when observed to be recumbent with opisthotonus. The lamb was treated with dextrose, vitamins B1 and B12, and penicillin G, but died 4 hours later.

Pathological findings: Examination of Lamb 1 revealed markedly increased gas within the peritoneum and within dilated loops of intestine. The intestines were dark red and contained large quantities of haemorrhagic fluid. Histology of the intestines revealed peracute mucosal necrosis with minimal accompanying inflammation. The intestinal lumen contained cell debris, haemorrhage, and myriad large Gram-positive bacilli. The intestines of Lamb 2 did not appear bloated or reddened. However, multiple fibrin clots were visible within the pericardial sac. Histopathological examination revealed small foci of necrosis within the mucosa of the distal intestine. The necrotic foci were often associated with large numbers of large Gram-positive bacilli.

Immunohistochemsitry and molecular biology: Intestinal samples from Lamb 1 were processed for Clostridium perfringens immunohistochemistry, which revealed large numbers of intralesional, positively immunostained rods. Fragments corresponding to the expected sizes for genes encoding alpha, beta, and epsilon C. perfringens typing toxins were amplified by PCR from DNA extracted from formalin-fixed sections of intestine.

Diagnosis: Lamb dysentery due to C. perfringens type B.

Clinical relevance: C. perfringens bacteria have a worldwide distribution, but disease due to C. perfringens type B has only been diagnosed in a small number of countries and has never been reported in New Zealand or Australia. C. perfringens type B produce both beta toxin and epsilon toxins, therefore both haemorrhagic enteritis and systemic vascular damage can develop. As many animals are exposed to C. perfringens without developing disease, there must be additional unknown factors that resulted in disease in these particular sheep. Vaccines that specifically protect against C. perfringens type B are available and may be recommended for use in smaller non-commercial flocks, as in the present case.     

Pharmacokinetic study of meropenem in healthy beagle dogs receiving intermittent hemodialysis

Meropenem, a second carbapenem antimicrobial agent with a broad spectrum of activity, is used to treat sepsis and resistant‐bacterial infections in veterinary medicine. The objective of this study was to identify the pharmacokinetics of meropenem in dogs receiving intermittent hemodialysis (IHD) and to determine the proper dosing in renal failure patients receiving IHD. Five healthy beagle dogs were given a single i.v. dose of 24 mg/kg of meropenem and received IHD. The blood flow rate, dialysate flow, and ultrafiltration rate were maintained at 40 mL/min, 300 mL/min, and 40 mL/h, respectively. Blood samples were collected for 24 h from the jugular vein and from the extracorporeal arterial and venous line. Urine samples and dialysate were also collected. The concentrations of meropenem were assayed using HPLC/MS/MS determination. The peak plasma concentration was 116 ± 37 μg/mL at 15 min. The systemic clearance was 347 ± 117 mL/h/kg, and the steady‐state volume of distribution was 223 ± 67 mL/kg. Dialysis clearance was 71.1 ± 34.3 mL/h/kg, and the extraction ratio by hemodialysis was 0.455 ± 0.150. The half‐life (T_1/2) in dogs with IHD decreased compared with those without IHD, and the reduction in T_1/2 was greater in renal failure patients than in normal patients. Sixty‐nine percent and 21% of the administered drug were recovered by urine and dialysate in the unchanged form, respectively. In conclusion, additional dosing of 24 mg/kg of meropenem after dialysis could be necessary according to the residual renal function of the patient based on the simulated data.     

Comparison of the rebound tonometer (TonoVet) with the applanation tonometer (TonoPen XL) in normal Eurasian Eagle owls (Bubo bubo)

OBJECTIVE: To examine the feasibility and accuracy of a handheld rebound tonometer, TonoVet, and to compare the intraocular pressure (IOP) readings of the TonoVet with those of an applanation tonometer, TonoPen XL, in normal Eurasian Eagle owls. ANIMALS STUDIED: Ten clinically normal Eurasian Eagle owls (20 eyes). PROCEDURES: Complete ocular examinations, using slit-lamp biomicroscopy and indirect ophthalmoscopy, were conducted on each raptor. The IOP was measured bilaterally using a rebound tonometer followed by a topical anesthetic agent after 1 min. The TonoPen XL tonometer was applied in both eyes 30 s following topical anesthesia. RESULTS: The mean +/- SD IOP obtained by rebound tonometer was 10.45 +/- 1.64 mmHg (range 7-14 mmHg), and by applanation tonometer was 9.35 +/- 1.81 mmHg (range 6-12 mmHg). There was a significant difference (P = 0.001) in the IOP obtained from both tonometers. The linear regression equation describing the relationship between both devices was y = 0.669x + 4.194 (x = TonoPen XL and y = TonoVet). The determination coefficient (r(2)) was r(2) = 0.550. CONCLUSIONS: The results suggest that readings from the rebound tonometer significantly overestimated those from the applanation tonometer and that the rebound tonometer was tolerated well because of the rapid and minimal stress-inducing method of tonometry in the Eurasian Eagle owls, even without topical anesthesia. Further studies comparing TonoVet with manometric measurements may be necessary to employ rebound tonometer for routine clinical use in Eurasian Eagle owls.     

Effects of electroacupuncture on intraocular pressure and hemodynamic parameters in isoflurane anesthetized dogs

The effects of electroacupuncture (EA) on intraocular pressure (IOP) and hemodynamic parameters were evaluated in isoflurane anesthetized 10 (5 males, 5 females) normal mongrel dogs (8.1-9.8 kg, 6-8 years old). After determination of baseline IOP and hemodynamic parameters (cardiac index, systolic arterial pressure, diastolic arterial pressure, heart rate and systemic vascular resistance index), EA was applied at 3 acupoints (LI-4, LIV-3 and GB-37) for 20 min. After the EA treatment, IOP was significantly decreased in the both eyes (p<0.05). However, there were not significant differences in hemodynamic parameters between those of before and after EA treatment. From these results, the EA treatment at LI-4, LIV-3 and GB-37 would be considered one of the valuable methods for the IOP treatment in dogs.     

Retrospective assessment of tolerability and efficacy of zoledronate in the palliative treatment of cancer-bearing dogs

Zoledronate is a bisphosphonate frequently used for the treatment of hypercalcaemia of malignancy and tumour-associated bone pain in dogs, however, there is a paucity of information regarding its use in veterinary medicine. The aim of this retrospective study was to report the tolerability of zoledronate in the palliative treatment of cancer-bearing dogs and secondarily to to assess the efficacy of zoledronate for the treatment of hypercalcaemia of malignancy. Thirty-seven dogs (22 with tumour-associated bone pain and 15 with hypercalcaemia of malignancy) that received 114 zoledronate infusions were included. Tolerability was assessed by the absence of post-zoledronate hypocalcaemia or other adverse events as defined by Veterinary Cooperative Oncology Group-Common Terminology Criteria for Adverse Events criteria. Efficacy was assessed by comparison of available ionized calcium levels before and after zoledronate administration in hypercalcaemic dogs. In 79% of zoledronate infusions, no adverse events were reported. The majority of adverse events which occurred in the other 21% of infusions could be attributed to concurrent chemotherapy or the underlying neoplastic disease. There was a small but significant increase in creatinine following treatment with zoledronate, however, none of the dogs developed clinically significant renal disease. In eight hypercalcaemic dogs with available ionized calcium following zoledronate administration, ionized calcium decreased rapidly within 7 days following treatment with zoledronate. Zoledronate is well-tolerated with few recorded adverse events, however, monitoring of serum creatinine is advised. Zoledronate seems to be effective in the treatment of hypercalcaemia of malignancy.