Bronchoalveolar lavage in horses: effect of exercise and repeated sampling on cytology

SUMMARY Bronchoalveolar lavage (BAL) was performed at weekly intervals in 10 Thoroughbred horses in race training (group 1) and in 4 rested horses (group 2) for 10 weeks. Lavages were continued on a weekly basis in 4 group 1 horses for an additional 9 weeks (group 3). Cytological analysis of samples included leukocyte counts, erythrocyte counts, differential leukocyte counts, and haemosiderophage score. The mean leukocyte concentration was significantly lower in group 1(92.1 ± 4.6 cells/μL) when compared with group 2 (133.5 ± 8.2 cells/μL), P = 0.037. The differential leukocyte data were not significantly different between groups. There was a large amount of variability in the percent-age of macrophages and lymphocytes in the differential counts over time with no obvious trends. The proportion of neutrophils demonstrated a tendency to decrease over time for both groups 1 and 2. Erythrocyte counts and haemosiderin scores were significantly higher in the exercising group than the rested horses. Neither exercise nor the technique itself evoked an inflammatory response in the BAL fluid.     

Inflammation and increased numbers of bacteria in the lower respiratory tract of horses within 6 to 12 hours of confinement with the head elevated

SUMMARY: Confinement of horses with their heads elevated for periods up to 24 hours was used to evaluate the extent and the effects of bacterial contamination of the equine lower respiratory tract. Significant (P < 0.05) increases in bacterial numbers (up to 10⁹ colony forming units/mL in transtracheal aspirate derived samples) occurred within 6 or 12 hours in most horses. Pasteurella/Actinobacillus spp and Streptococcus spp were most commonly isolated. Lowering of the head for 30 minutes every 6 hours to facilitate postural drainage did not prevent multiplication of organisms to levels equivalent to those achieved by horses where the head was elevated for 24 hours. When horses were released from confinement and heads were no longer maintained in an elevated position, clearance of accumulated secretions and bacteria occurred within 8 to 12 hours. Thus, confinement with the head elevated resulted in significant bacterial contamination and multiplication within the lower respiratory tract during a period often encountered in routine management procedures, such as transportation. The clearance of accumulated secretions occurred over a prolonged period after release from such confinement.     

Survey of equine castration techniques,preferences and outcomes among Australian veterinarians

Objectives

(1) To collect the perceptions of veterinarians performing equine castrations in Australia on techniques, preferences and outcomes, (2) to investigate veterinarian use and experience with the Henderson castrating instrument and (3) to investigate potential associations between demographics, castration methods and techniques, and complications.

Design

Online survey of members of the Australian Veterinary Association’s Special Interest Group, Equine Veterinarians Australia (EVA).

Methods

A link to the survey was included in the EVA e‐newsletter and practices on the EVA website were contacted by telephone and follow‐up email. Fisher’s exact test was used to determine associations between ligation and complications. A generalised linear model with a negative binomial family was used to determine associations between count response variables and categorical independent variables.

Results

Responses were obtained from 138 veterinarians (response rate, 13.1%) who performed 5330 castrations over 12 months. Castrations were most commonly performed in the field, on anaesthetised horses, using emasculators, via an open approach and without ligation of the spermatic cord. Estimated complications after use of emasculators were swelling (25%), haemorrhage (5%) and infection (5%). The Henderson instrument was used by approximately 10% of respondents and its use for castration was associated with fewer reports of postoperative swelling compared with emasculators (P = 0.002). Rates of evisceration with the Henderson and emasculator methods were comparable (0.43% and 0.9%, respectively).

Conclusion

Castration preferences varied widely among survey participants. Reported complication types and rates were comparable to those reported previously in other countries. Perceptions that the Henderson instrument was associated with less swelling should be investigated further via a prospective controlled investigation.     

Comparison of duplex PCR and microscopic techniques for the identification of Babesia bigemina and Babesia bovis in engorged female ticks of Boophilus microplus

A single-step duplex polymerase chain reaction (PCR) technique and traditional microscopic examination of haemolymph smears were used to detect Babesia bigemina and/or Babesia bovis infection in engorged female ticks of Boophilus microplus recovered from calves raised in an endemic area of the State of Minas Gerais, Brazil. In the PCR amplification of tick-derived DNA, pairs of oligonucleotide primers specific for a 278-bp sequence from B. bigemina and for a 350-bp sequence from B. bovis were used conjointly. The microscopic examination of haemolymph revealed that 16.7% of the engorged ticks were infected with Babesia spp., although no significant differences (rho > 0.05) were found in the infection rate of ticks collected from calves of different age groups. PCR analysis showed that 77.8% of the engorged ticks whose haemolymph contained sporokinetes were infected with B. bigemina, 7.8% with B. bovis and 14.4% with both protozoan species. However, the PCR assay further revealed that, amongst the engorged female ticks whose haemolymph was apparently negative for the presence of sporokinetes, 15.6% were infected with B. bigemina, 2.2% with B. bovis and 10.0% with both species. The duplex PCR method is thus more efficient and sensitive than the microscopic assay and also permits facile identification of the protozoa species present in engorged female ticks.     

Pronuclear Embryo Yield in Canindé and Saanen Goats for DNA Microinjection

The objective of this study was to examine the effect of donor breed on pronuclear‐stage embryo yield to be used for DNA microinjection in a transgenesis goat program. Twelve Canindé and twelve Saanen goats were heat synchronized using a progestagen‐cloprostenol treatment. Forty‐eight hours before the sponge removal, superovulation was induced with a total administration of 4.4 mg/kg bodyweight NIH‐FSH‐P1, given twice daily in decreasing doses over 3 days. In addition, goats received 100 μg of GnRH and they were hand‐mated at 36 and 48 h after progestagen removal. Embryo recovery was performed by oviduct flushing at 72 h after sponge removal. Embryos were microinjected with a DNA construct and noticeable swelling of the nuclei was the criterion for successful microinjection. The total diameter, cytoplasm diameter, zona pellucida thickness and pronuclei diameter were measured for each microinjected embryo. A higher (p < 0.05) percentage of fertilized ova was observed in Canindé (89.9%) than Saanen (36.2%) goats. In addition, Canindé donors produced a higher percentage of pronuclear embryos when compared with Saanen: 72.5% vs 20.6% (p < 0.05), respectively. Successful microinjection was verified in 96.7% and 73.3% of times in Canindé and Saanen embryos, respectively (p < 0.05). Significant differences were observed for all morphometric parameters except for cytoplasm diameter. In conclusion, under our study experimental conditions, Canindé were more efficient than Saanen goats concerning the pronuclear embryo yield and manipulation. The use of Canindé goats in transgenesis programs could be increase the interest in their breeding and could be contribute to saving them from extinction.