Detection and quantification of Anaplasma marginale DNA in blood samples of cattle by real-time PCR

A TaqMan-based real-time PCR assay was developed for the diagnosis of Anaplasma marginale infection of cattle. The established assay was proven to be highly specific, since no cross-reactions were observed with other Anaplasma species of ruminants, including the closely related Anaplasma centrale, or other haemoparasites of ruminants (Anaplasma bovis, Anaplasma ovis, Anaplasma phagocytophilum, Babesia bovis, Babesia bigemina, Theileria annulata and Theileria buffeli). The detection limit was equal to that of nested (n)PCR (10(1) copies of standard DNA and 3 x 10(1) infected erythrocytes ml(-1) of blood). The assay was also reproducible, as shown by satisfactory low intra-assay and inter-assay coefficients of variation. Fifty-four blood samples of ruminants (cattle, n = 51; sheep, n = 2; goats, n = 1), that had been tested previously by reverse line blot (RLB) hybridisation, were subjected to an nPCR assay and the newly established real-time PCR assay. By using real-time PCR, A. marginale DNA was detected in 39/51 bovine samples, with DNA titres ranging from 3.60 x 10(3) to 5.70 x 10(8) copies ml(-1) of blood, whereas sheep and goat samples tested negative. The concordance with nPCR was 100%, whereas a unique sample that had tested negative by RLB gave positive results by nPCR and real-time PCR. The established assay could overcome the limitations of existing diagnostic methods, allowing for simultaneous detection and quantification of the A. marginale DNA in bovine blood, that is essential to support the clinical diagnosis, to assess the carrier status of the animals and to evaluate the efficacy of vaccines and antirickettsial drugs.     

Thoracic aortic rupture in horses

The aorta can rupture at the aortic root or aortic arch. In most breeds, the aortic root is the likely site and rupture leads to aortocardiac fistula with communication between the aorta and the right atrium, right ventricle and/or the interventricular septum. There is a high prevalence of aortic rupture in young Friesian horses and rupture occurs at the aortic arch with pseudoaneurysm and potentially aortopulmonary fistulation. Echocardiographic and post‐mortem techniques must be adapted to identify aortic arch rupture that is not generally identified with standard approaches. Given the narrow genetic base of the Friesian breed and the significant differences found in extracellular matrix composition and metabolism between Friesians and Warmbloods, genetic factors are likely to contribute to the condition in the Friesian breed.     

In vitro control of parasitic nematodes of small ruminants using some plant species containing flavonoids

This study determined in vitro anthelmintic efficacy of three plant species: Trema orientalis, Urtica dioica and Zanthozylum capense on nematode larvae of small ruminants. Dried leaf samples (40 g) were extracted in 70% ethanol, in portions of 10 g and concentrated to 100 ml. Half and one quarter of the original crude extract were both made to 100 ml. Rectal faecal material from 10 Merino sheep and 25 Nguni goats was pooled within species and thoroughly hand-mixed. Dung samples, each of 5 g were cultured for 12 days at 27 °C. On day 13, 4 plates were watered and 4 others treated with ethanol to correct for solvent effect on mortality. The design was 2 (animal species)?×?3 (plant species)?×?3 (extract concentrations). In each of three runs, three plates were treated with each crude extract in three incremental concentrations. Surviving L3 larvae were isolated, counted and mortalities became indices of anthelmintic efficacy. Data from nematode larval mortality were analysed to determine the effect of animal species, plant species, concentration and their interactions. Efficacy was affected by concentration (P?=?0.0001), animal species (P?=?0.0046), plant species (P?=?0.0572), the interactions of animal species and concentration (P?=?0.0010), plant species and concentration (P?=?0.0123) and concentration × animal × plant species (P?=?0.0435).     

An unusual case of feline acute corneal hydrops: atypical disease presentation and possible in vivo detection of Descemet's membrane detachment in the cat's unaffected eye

A 1‐year‐old, female spayed, domestic shorthair cat presented for blepharospasm of the right eye. Slit‐lamp biomicroscopic examination showed focal corneal ulceration and presumptive keratomalacia of the right eye. Examination of the left eye was normal apart from a focal endothelial opacity. Within the first 24 h of medical management, the right eye developed marked corneal edema and globular anterior protrusion of the corneal surface consistent with feline acute corneal hydrops (FACH). Surgical management consisted of a bridge conjunctival graft, nictitating membrane flap, and temporary tarsorrhaphy. Resolution of corneal edema and pain occurred in the right eye within 24 days. Spectral domain optical coherence tomography (SD‐OCT) of the anterior segment was performed in both eyes. Conjunctival tissue from the bridge graft precluded examination of deeper corneal structures in the right eye. The left eye displayed a focal separation of the corneal endothelium and Descemet's membrane from the overlying stroma. These SD‐OCT findings are similar to the analogous syndrome found in humans and represent a potential etiology for FACH of the right eye in the case presented here. Unfortunately, the cat was lost to follow‐up and the progression of this lesion to FACH in the left eye could not be determined.     

Brucellosis in migratory sheep flock from Maharashtra,India

Brucellosis is a zoonotic disease worldwide distributed and having the economic as well as public health importance. The prevalence of brucellosis among sheep flock having history of abortions was studied. A total of 229 samples comprising of 157 blood and 72 clinical samples (vaginal swabs) were collected from 157 animals. Clinical samples were processed for the isolation of Brucella melitensis. Serum samples (n = 157) were tested by Rose Bengal plate test (RBPT) and i-ELISA. A total of 68 (43.31%) and 104 (66.24%) samples were positive by RBPT and ELISA, respectively. Brucella isolates (n = 2) were recovered from clinical samples. Both isolates demonstrated amplification for bcsp 31 and IS711 genes. On AMOS PCR, both the isolates amplified at 731 bp, i.e., belongs to B. melitensis species. The incidence of B. melitensis in a migratory flock warns the thorough testing and culling of Brucella-infected sheep from the flock on a continuous basis; otherwise, such incidence will be routine and poor farmers will be at a loss.     

Genetic evaluation of growth in farmers’ flocks of Madras Red sheep under long-term selection in a group breeding scheme

The Network Project on Sheep Improvement (NWPSI)–Madras Red field unit is a group breeding scheme involving 198 farmers’ flocks of Madras Red sheep in which selection for growth traits and rotation of rams have been practised for over two decades. Growth data collected from these flocks were used to evaluate the performance and understand the direct and expected responses to selection based on genetic parameters. The body weight at birth (BW), weaning weight (WW), 6-month weight (6W), 9-month weight (9W), 12-month weight (YW), pre-weaning average daily gain (ADG1, birth to 3 months), post-weaning ADG2 (3–6 months), ADG3 (6–9 months), ADG4 (9–12 months) and ADG5 (3–12 months) were 2.67, 10.05, 14.56, 18.36, 21.36, 80.13, 49.05, 43.00, 34.21 and 41.18 g, respectively. Univariate analyses were carried out using animal and sire models to estimate variance components. Heritability obtained from animal model for BW was 0.36 and the values for other body weight traits were almost unity. Heritability estimate for pre-weaning ADG1 was 0.31. Very high genetic variability was observed in spite of long-term selection and this sustenance of variability is one of the main advantages of a group breeding scheme, combining several flocks of smaller size. An increasing genetic and phenotypic trend was noticed for almost all the traits studied. The expected responses calculated based on genetic parameters also indicated scope for improvement.     

Viability of cattle sperm under different storage conditions in Cameroon

A study was carried out to evaluate the viability of extended cattle semen, without freezing, under different storage conditions. The semen was collected from Holstein Friesian bulls using artificial vaginas. The semen was extended and stored in a 3-by-4 factorial design (storage system × ice change). The storage media were ice boxes, buckets, and refrigerator. The ice in these media was either replaced daily, on the first and third day, first day only, or no ice at all after the semen collection. Results showed an overwhelming evidence of the effect of storage medium and ice change on sperm viability (P < 0.0001). Individual motility before processing was highest in the refrigerator with averages of 44.5%, 39.5% in ice boxes, and 10% in buckets during the 8-day experiment. There was no significant difference (P > 0.05) in progressive motility after processing in the refrigerator (34%) and in ice boxes (33%) but significantly higher (P < 0.01) to the 10% obtained in buckets. It was shown that spermatozoa in the ice box retained 45% individual motility up to the sixth day after semen was collected on the condition that the ice was changed on the third day. Progressive motility after processing in the ice box was 40% up to the sixth day with the ice changed on the third day while the spermatozoa were well preserved up to the fourth day in the same medium if the ice put on the first day was not changed. This study shows that, if farmers plan to inseminate cows within the first day after semen collection, they can use buckets with ice for the transportation of the extended semen ampoules from the artificial insemination center. Otherwise, the semen needs to be kept in ice boxes and the ice changed on the third day after collection and this semen could be used within a week.     

Incidence of swallowing during exercise in horses with dorsal displacement of the soft palate

Reasons for performing study: The relationship between dorsal displacement of the soft palate (DDSP) and swallowing is unclear. Objective: To quantify the relationship between DDSP and swallowing in horses at exercise. Hypotheses: The frequency of swallowing increases immediately prior to DDSP in horses at exercise. Methods: Videoendoscopic and upper airway pressure data were collated from horses with a definitive diagnosis of DDSP at exercise. Horses with no upper airway abnormalities were matched by age, breed and sex and used as controls. Sixty‐nine horses were identified with a definitive diagnosis of DDSP during the study interval. Airway pressure data were available for 42 horses. Results: The majority of horses displaced at high exercising speeds while accelerating; a smaller number displaced during deceleration after peak speed had been reached. Horses swallowed significantly more frequently in the 1 min immediately preceding DDSP than in the control horses at equivalent speeds. DDSP at exercise results in a significant increase in tracheal expiratory pressure, a significant decrease in pharyngeal expiratory pressure and a significantly less negative pharyngeal inspiratory pressure compared to matched controls and compared to the pressures during the 1 min interval prior to DDSP. There was no significant difference between any measure of airway pressure before or after a swallow when examined at each time interval in the DDSP population. Conclusions: The frequency of swallowing decreases with increasing speed in normal horses. In contrast, the frequency of swallowing increases immediately prior to onset of DDSP. This is not a result of pharyngeal and tracheal pressure changes. Potential relevance: The increased frequency of swallowing observed prior to DDSP may be related to the aetiology of the disease.