Molecular characterisation of Theileria equi and risk factors associated with the occurrence of theileriosis in horses of Punjab (Pakistan)

Theileria equi (T. equi) is an obligate intra- and extra-erythrocytic parasite that causes equine theileriosis (ET) in equids. Equine theileriosis is considered a notifiable disease of global significance, a major constraint to the international movement of horses, and endemic in many countries. This disease may be difficult to diagnose, as it can produce variable and nonspecific clinical signs. A cross-sectional study was designed for the molecular characterisation of T. equi and to investigate the associated risk factors of ET accompanied by its consequences on haematological and sero-biochemical parameters. A convenience sampling of 500 blood samples were collected from ET suspect horses from January to December 2017. PCR was performed on all blood samples targeting the 18S rRNA gene of T. equi followed by sequencing; 9% animals tested positive with confirmed sequences. The isolates of this study showed high homology with Cuban, Russian and Brazilian isolates of T. equi (accession numbers KY111762.2 , MG551915.1 and KY952237.1 , respectively). Based on multivariate analysis, the principal risk factors consisted of absence of dogs on the premises and presence of tick infestation. The haemato-biochemical parameters showed a decrease in granulocytes and erythrocytes, and an increase in lymphocytes, monocytes, mean corpuscular volume, mean corpuscular haemoglobin, mean platelet volume, glucose, phosphorus and aspartate aminotransferase in positive horses. This is the first study which identified ET in Punjab (Pakistan) using molecular techniques and risk factors together with the haemato-biochemical variations in horses.     

Use of dimethylformamide to cryopreserve alpaca semen previously incubated with collagenase

The objective of this study was to evaluate the effect of collagenase and two final dimethylformamide (DMF) concentrations (4% and 7%) on alpaca frozen-thawed sperm quality. A total of 25 ejaculates from 5 alpaca were obtained using electroejaculation. Each individual ejaculate was evaluated and then diluted 4:1 in a solution of 1 mg/ml collagenase in HEPES-TALP medium and incubated for 4 min at 37°C. Subsequently, samples were diluted in TRIS-fructose-citric acid-egg yolk and cooled to 5°C. Then, each sample was divided in two aliquots and DMF at final concentration of 4% or 7% was added, equilibrated for 1 hr at 5°C and frozen over liquid nitrogen vapours. A Kruskal–Wallis test was used to evaluate the sperm morphometry, and Completely Random Block designs were used to analyse sperm motility, viability, membrane function and acrosome status. After collagenase incubation, none of the samples showed thread formation, and sperm parameters were preserved. Non-progressive motile sperm were higher (p < .05) in equilibrated samples (4% DMF: 31.8 ± 8.3% and 7% DMF: 36.3 ± 11.8%) compared to raw (10.1 ± 4.3%) and frozen-thawed semen (4% DMF: 9.7 ± 1.8% and 7% DMF: 7.5 ± 3.2%). Sperm membrane function, membrane integrity and intact acrosomes were higher (p < .05) in raw semen (40.1 ± 12.2%, 94.6 ± 3.2% and 91.3 ± 8.1%) compared to frozen-thawed samples (4% DMF: 19.8 ± 4.7%, 53.2 ± 2.7%, 65.7 ± 8.7% and 7% DMF: 20.4 ± 4.5%, 54.1 ± 1.4%, 64.6 ± 9.1%). Length of the sperm head was lower in frozen-thawed samples, being statistically different with 4% DMF compared to pre-freezing samples. The ratio between acrosome and head areas was greater (p < .05) in frozen-thawed samples. Incubation of raw alpaca semen with collagenase decreased the thread formation without affecting sperm quality. Frozen of collagenase treated alpaca semen with 4% or 7% DMF did not preserve the sperm parameters in thawed samples.     

Fatty acid profile as affected by fat depot and the sex category of Polish Holstein-Friesian × Limousin fattening cattle fed silage ad libitum

This study was designed to compare the fatty acid profiles of four types of fat depots from bulls, steers, and heifers. All animals were Polish Holstein-Friesian and Limousin crossbreds fattened semi-intensively—silage at libitum and concentrate in the amount corresponding to 30% of their net energy requirements. The fatty acid profile in intramuscular, intermuscular, and external and internal fat was determined. The intramuscular fat of bulls was most abundant in total PUFAs and n-6 PUFAs, and functional fatty acids C 18:2, C 20:4, and C 22:5 in comparison with steers and heifers. Regardless of sex category, intramuscular and external fat were characterized by higher levels of UFAs and a more desirable MUFA/SFA ratio than the remaining fat types. It should also be noted that external fat was more abundant in CLA than other fat types, and that the highest CLA content was found in bull fat, compared with the remaining sex categories of cattle.     

Free-range system and supplementation of 25-hydroxicholecalciferol increases the performance and serum vitamin levels in mixed-parity sows

Improvements in sow productivity have raised questions regarding dietary vitamin D recommendations. The present study aimed to evaluate the effects of the housing system with access to sunlight exposure and supplementation of 25-hydroxicholecalciferol on performance and serum levels of 25(OH)D₃ in sows during gestation and lactation. Sows were distributed in an experimental design with two housing systems: gestation crates or gestation free-range system with external area for sunlight exposure; and two diets: 0 or 50 μg of 25-hydroxicholecalciferol kg⁻¹. The use of 25-hydroxicholecalciferol tended (P = 0.052) to improve total born and influenced (P = 0.046) on number of born alive. Litter weight at birth was also increased (P = 0.01) by 25-hydroxicholecalciferol supplementation; 25-hydroxicholecalciferol supplementation and housing system (free-range with sunlight exposure) tended to increase weaning weight (P = 0.07) and litter daily gain (P = 0.051) during lactation. Exposure to sunlight and 25-hydroxicholecalciferol supplementation increased 25(OH)D₃ serum levels when compared with control treatment during gestation (136.95 vs. 113.92 ng mL⁻¹; P = 0.035) and lactation (120.29 vs. 88.93 ng mL⁻¹; P = 0.026). In conclusion, the association of 25-hydroxicholecalciferol supplementation with exposure to sunlight during gestation improved significantly 25(OH)D₃ serum levels and consequently performance traits in gestation and lactation.     

Socio-spatial organization reveals paternity and low kinship in the Honduran white bat (Ectophylla alba) in Costa Rica

Ectophylla alba is a tent-making bat that roosts in mixed-sex clusters comprising adults and offspring. Our goal was to determine the genetic identity of individuals belonging to different roosting groups. We tested the hypothesis of kin selection as a major force structuring group composition. We used 9 microsatellites designed for E. alba to determine the genetic identity and probability of parentage of individuals. We analyzed parentage and kinship using the software ML-Relate, GenAIEx, and Cervus. The obtained relationship probabilities (0.5) revealed a clear maternal relationship between female adults and offspring with allele compatibility, and at least 5 relationships between male adults and pups. We found a low degree of relatedness within roosting groups. Between roosting groups at different sites, the mean probability of a half-sibling relationship ranged from 0.214 to 0.244 and, for full-sibling relationship, from 0.383 to 0.553. Genetically, adult individuals were poorly related within clusters, and kinship as an evolutionary force could not explain group membership.