First Established Pregnancies in Mediterranean Italian Buffaloes (Bubalus bubalis) Following Deposition of Sexed Spermatozoa near the Utero Tubal Junction

At the time of AI following Ovsynch protocol, a total of 51 buffaloes were randomly divided in a first group (n = 30) subjected to conventional AI into the uterine body with 20 million non-sex sorted frozen-thawed spermatozoa, while a second group (n = 21) was inseminated near the utero-tubal junction (UTJ) ipsilateral to the ovary carrying the preovulatory follicle with 2.5 million live (4 million total) sex-sorted frozen-thawed spermatozoa. The semen used for flowcytometric sorting was collected and processed on a farm in Italy, and then shipped to a laboratory in Germany. Eleven buffaloes were inseminated with X-chromosome bearing spermatozoa and 10 with Y-chromosome bearing spermatozoa. Conception rates after conventional and UTJ inseminations were 43.3% (n = 13) and 42.8% (n = 9) respectively (p = 0.97). Eight of the nine foetuses obtained after insemination with sexed spermatozoa corresponded to the sex as predicted by the cell sorting procedure (five male and four female foetuses by ultrasound vs six male and three female foetuses by cell sorting). In conclusion, for the first time buffalo semen has been successfully subjected to procedures for flowcytometric sperm sorting and freezing. Low doses of sexed spermatozoa have been deposited near the UTJ giving conception rates similar to those of conventional AI with full dose.     

Evaluation of peripheral lymphoreticular biopsy techniques and their clinical side effects in sheep

Tissue samples were collected postmortem from 126 sheep at five lymphoreticular sites by different techniques. The three most successful combinations of sites and techniques were: the third eyelids, using a forceps and scissors, which provided a mean (se) of 5.32 (0.70) lymphoid follicles per 5 microm tissue section, a mandibular lymph node, using a Biopty gun, which gave 1.19 (0.26) lymphoid follicles per 5 microm tissue section, and tonsil, using a biopsy forceps, which gave 1.14 (0.27) lymphoid follicles per 5 microm tissue section. These three techniques were repeated once a month for five months on five sheep under general anaesthesia, and their clinical effects were compared with five control sheep which were restrained and anaesthetised in the same way but from which no biopsies were taken. Most lymphoid follicles (3.47 [0.58] per 5 pm tissue section) were obtained by using the third eyelid biopsy technique. There were no clinical side effects associated with the biopsy procedure. There were increases in the plasma concentration of cortisol in all the animals, suggesting that the restraint and anaesthesia were more stressful than the biopsy procedure.     

Impact of foot-and-mouth disease on mastitis and culling on a large-scale dairy farm in Kenya

Foot and mouth disease (FMD) is a highly transmissible viral infection of cloven hooved animals associated with severe economic losses when introduced into FMD-free countries. Information on the impact of the disease in FMDV-endemic countries is poorly characterised yet essential for the prioritisation of scarce resources for disease control programmes. A FMD (virus serotype SAT2) outbreak on a large-scale dairy farm in Nakuru County, Kenya provided an opportunity to evaluate the impact of FMD on clinical mastitis and culling rate. A cohort approach followed animals over a 12-month period after the commencement of the outbreak. For culling, all animals were included; for mastitis, those over 18 months of age. FMD was recorded in 400/644 cattle over a 29-day period. During the follow-up period 76 animals were culled or died whilst in the over 18 month old cohort 63 developed clinical mastitis. Hazard ratios (HR) were generated using Cox regression accounting for non-proportional hazards by inclusion of time-varying effects. Univariable analysis showed FMD cases were culled sooner but there was no effect on clinical mastitis. After adjusting for possible confounders and inclusion of time-varying effects there was weak evidence to support an effect of FMD on culling (HR = 1.7, 95% confidence intervals [CI] 0.88-3.1, P = 0.12). For mastitis, there was stronger evidence of an increased rate in the first month after the onset of the outbreak (HR = 2.9, 95%CI 0.97-8.9, P = 0.057).

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-015-0173-4) contains supplementary material, which is available to authorized users.     

Transportation stress in young bulls alters expression of neutrophil genes important for the regulation of apoptosis, tissue remodeling, margination, and anti-bacterial function

The transportation of beef cattle results in a stress response that is associated with increased susceptibility and severity of respiratory diseases, presumably due to an alteration in immune function. Neutrophils are phagocytic immune cells important in lung defense and are also targets of the stress response. The objective of this study was to determine if a 9h transportation of young bulls by road induced changes in the expression of candidate genes known to be important in neutrophil-mediated defense and inflammation in the lung. These neutrophil genes encompassed functions of apoptosis (A1 and Fas), tissue remodeling (MMP-9), vascular margination (L-selectin), bacterial killing (BPI), and wound healing (betaglycan), as well as responsiveness of the cells to stress-induced increases in glucocorticoid hormones (GRalpha). To explore gene expression changes, blood was collected, plasma harvested, and neutrophils isolated from six Belgian Blue x Friesian bulls (231+/-7.0 kg in weight; 282+/-4 days of age) at -24, 0, 4.5, 9.75, 14.25, 24, and 48h relative to commencement of a 9h road transportation by truck. Plasma cortisol concentrations were elevated at 4.5 and 9.75h, peaking at 50.64+/-4.46 ng/mL (P<0.0001) and confirming that the animals experienced stress. Blood neutrophil count was elevated between 4.5 and 14.25h (P<0.0001), reaching a peak that was over 3-fold higher than the -24h concentration. Neutrophil Fas gene expression was acutely down-regulated (P=0.02) by transportation stress, while expressions of MMP-9, l-selectin, and BPI were profoundly up-regulated (P=0.003, 0.002, and <0.001 respectively). However, no changes in neutrophil expressions of betaglycan, GRalpha, and A1 were detected. It is concluded that a 9h transportation of young bulls induces a gene expression signature in blood neutrophils that increases their circulating numbers and may enhance their pro-inflammatory and anti-bacterial potential.     

Somatic cell mating and segregation in chimeric frogs

Both pentaploid and haploid cells were observed in a short-term culture of bone marrow of a diploid-triploid frog chimera. Apparently, diploid and triploid marrow cells fused to form "hybrid" pentaploid cells, which subsequently gave rise by somatic reduction to haploid daughter cells. The hybrid marrow cells and their haploid segregants are presumably at a selective disadvantage, as neither type of cell has been detected in the circulating blood of chimeric frogs.     

The characterisation of E. coli O157:H7 isolates from cattle faeces and feedlot environment using PFGE

The objectives of this study were to investigate the diversity of Escherichia coli O157:H7 isolates obtained over a 3-month period from a cattle feedlot in order to assess the relationship between environmental and faecal isolates and to determine the pattern of transmission of E. coli O157:H7 between groups of cattle. Faecal samples were obtained from cattle housed in four adjacent feedlot pens at monthly intervals, with environmental pen samples collected simultaneously. All E. coli O157:H7 isolates obtained were examined by pulsed field gel electrophoresis (PFGE), polymerase chain reaction (PCR) to detect eaeA, ehxA, stx1 and stx2 genes and antibiotic sensitivity profiling. Ten isolates were subjected to acid shock to imitate conditions in the acidic cattle abomasum and assess the effect on PFGE profiles. E. coli O157:H7 was isolated from 69 faecal samples and 26 environmental samples. All isolates (n=95) carried the genes for eaeA, ehxA and stx2 and were sensitive to all antibiotics tested. The PFGE profiles of all isolates differed by no more than two bands and clustered within 80% similarity following dendrogram analysis. Acid shock had no effect on the subsequent PFGE patterns. A total of 8.7% (6/69) of cattle were shedding E. coli O157:H7 in the first month with faecal shedding increasing to 52% (36/69) by the third month of the study. A single isolate of E. coli O157:H7 may be passed rapidly through cattle pens, with the environment acting as a significant reservoir for transmission. PFGE is a useful tool for tracking the direct and indirect transmission of E. coli O157:H7 isolates on the farm.     

Effect of cortisol infusion patterns and castration on metabolic and immunological indices of stress response in cattle

This study tested the hypotheses that: (1) either acute stress induced by Burdizzo castration, or cortisol infusion would modulate plasma glucose, insulin and growth hormone (GH) concentrations; and (2) immune modulation induced by cortisol would be dependent on the pattern, intensity and duration of circulating cortisol concentrations. Fifty 9.2-month-old Holstein×Friesian bulls (232±2.0 kg) were blocked by weight and randomly assigned to one of five treatments (n=10 per treatment): (1) sham handled control; (2) Burdizzo castration; (3) hydrocortisone infusion to mimic the castration-induced secretion pattern of cortisol; (4) hourly pulse infusion of hydrocortisone; and (5) sustained infusion of hydrocortisone for 8 h. Blood samples were collected intensively on day 0, and weekly from days 1 to 35. Castration acutely increased plasma cortisol, GH and haptoglobin concentrations, suppressed lymphocyte in vitro interferon-γ (IFN-γ) production, but had no effect on plasma glucose and insulin concentrations. Cortisol infusion to simulate the castration-induced secretion pattern of cortisol, and pulse infusion of cortisol did not suppress the IFN-γ production. A sustained infusion of cortisol resulted in the transient suppression of IFN-γ production. Moreover, the sustained cortisol infusion resulted in increased plasma glucose, insulin and GH concentrations. The overall 14-day feed intakes and 35-day growth rates were not affected by treatments. In conclusion, cortisol infusion to induce immune suppression in vivo occurred only at pharmacological doses. Within physiological ranges, cortisol was not associated with the suppression of immune function, indicating that during castration cortisol per se is not responsible for the suppression of in vitro IFN-γ production.     

Computed tomographic assessment of vascular invasion and resectability of mediastinal masses in dogs and a cat

AIMS: To assess the sensitivity of non-angiographic contrast-enhanced computed tomography (CT) to determine the presence of vascular invasion of cranial mediastinal masses in dogs and a cat, and to evaluate the association between vascular invasion and peri-operative mortality.

METHODS: A retrospective study was conducted on 25 dogs and one cat. CT scans were completed with slices ranging from 2 to 10 mm. CT images were evaluated by a board-certifi ed radiologist blinded to previous diagnoses and surgical fi ndings. Each CT study was evaluated for vascular invasion, defi ned as disruption of the vessel wall and extension of the mass into the vessel lumen. Data retrieved from the surgery reports included surgical approach, whether vascular invasion was present, the surgeon's decision on operability, and post-operative complications.

RESULTS: Computed tomographic evaluation revealed 25/26 masses had no evidence of vascular invasion. During surgical exploration, 10/26 masses were found to invade major regional vasculature; the cranial vena cava (CVC) was the vessel most commonly invaded (7/10 animals), and 4/7 (57%) patients with invasion of the CVC were euthanised or died in the perioperative period, from surgical or disease-related problems, which was signifi cantly higher than patients without vascular invasion (p=0.045).

CONCLUSIONS: Non-angiographic contrast-enhanced CT was signifi cantly less sensitive for detecting vascular invasion of cranial mediastinal masses when compared with surgical evaluation. If the CVC was invaded by a tumour there was a signifi cant risk of death peri-operatively when compared with non-invasive cases.

CLINICAL RELEVANCE: Due to the signifi cantly higher mortality risk associated with invasion of the CVC, a more sensitive method than CT should be investigated to determine vascular invasion of mediastinal masses pre-operatively.