Production of Buffalo (Bubalus bubalis) Embryos in vitro: Premises and Promises

Techniques for in vitro production (IVP) of buffalo embryos adopting the procedures developed in cattle have received increasing interest in the recent times. A high oocyte maturation, fertilization and cleavage rate and a low rate of blastocyst yield and calving following transfer of in vitro produced buffalo embryos have been obtained. The efficiency of IVP in buffalo is much lower than that in cattle. Several problems need to be resolved before IVP technology can be used regularly in buffalo breeding. This review attempts to present an overview of the different techniques used in buffalo to produce transferable embryos in vitro, namely in vitro maturation and fertilization of immature oocytes and in vitro development of the resulting cleaved embryos to the blastocyst stage before transfer. The problems associated with IVP, the possible solutions and the new biotechniques linked to IVP are discussed.     

The production performance of imported Friesian cattle under heavy Trypanosoma challenge in a rain forest zone of Nigeria

The productivity of 76 newly imported pregnant Friesian cattle and two bulls under heavy trypanosome challenge in the rain forest belt of Nigeria is reported. At the first visit in August 1989 and within 7 months of arrival of the heifers, the herd population had reduced by 26 (33.3%) as a result of deaths (six animals) and culls/salvages (20). The surviving 52 animals were generally in poor health with classical symptoms of trypanosomiasis. Thirty-one (40.8%) of the pregnancies were unsuccessful because of abortions (13 animals), premature births (seven), embryonic deaths (five) and death of heifers (six). Of the 45 successful calvings, 16 perinatal deaths occurred. All serum samples were negative for brucellosis. Only 41 (63.1%) of the 65 productive heifers lactated of which 24 (58.5%) yielded milk only for 6 months and less. The remaining 17 (41.5%) heifers were still at different stages of lactation ranging from 3 to 7 months within the period of analysis. Treatment with isometamidium (Samorin) at 0.5 mg/kg body weight cured the infection and prevented reinfections and/or relapses within 3 months of administration. A rise in the haematocrit and milk production after Samorin treatment was recorded. Careful analysis of the outbreak indicated that the reproductive wastages and poor lactational performance may have been induced by the severe trypanosomiasis diagnosed in the herd.     

Myofibroblastic fibrosarcoma with multifocal osseous metaplasia at the site of equine influenza vaccination

We describe a fibrosarcoma in a 12-year-old Quarterhorse × Arabian gelding as a sequela to equine influenza vaccination. Shortly after the second vaccination, swelling at the site was noticed by the owner and it continued to increase in size over the following 6 months. Biopsy of the mass indicated a fibrosarcoma had developed at the vaccination site. It was approximately 20 cm in diameter and elevated well above the level of the skin. There was no clinical evidence of metastases to the lungs or local lymph nodes. Surgical resection of the mass was performed and the wound healed by first and second intention. Histopathological examination and immunohistochemical staining confirmed a myofibroblastic fibrosarcoma with multifocal osseous metaplasia. To the authors' knowledge, this is the first equine case of a vaccine-associated fibrosarcoma.     

Indigofera spicata (creeping indigo) poisoning of three ponies

Three ponies continuously grazed a pasture containing an estimated 24% Indigofera spicata (wet weight basis) for 4–6 weeks in April and May 2004. They developed ataxia, paresis, depression, muscle fasciculations, dysphagia, ptyalism and halitosis. Two also developed corneal opacity. One pony recovered with supportive treatment, but the other two were euthanased and necropsied. Neuropathology was not present in either case, but both livers had periacinar and periportal lymphocytic infiltrations and hydropic degeneration of mid‐zonal hepatocytes, with mild to moderate periacinar necrosis also evident in one. The I. spicata contained 2.66 mg 3‐nitropropionic acid (3‐NPA)/g dry matter and 1.5 mg indospicine/g dry matter. Indospicine, but not 3‐NPA, was detected in serum from both of the euthanased ponies and indospicine was detected in heart, liver and muscle from the one pony in which this assay was performed. The clinical syndrome closely resembled ‘Birdsville horse disease’ caused by I. linnaei and was similar to that reported in horses poisoned by the closely related species I. hendecaphylla and to 3‐NPA poisoning of other animals, including humans. 3‐NPA is thought to cause this neurological syndrome. To our knowledge, this is the first authenticated report of I. spicata poisoning in grazing animals. We also report here the first published evidence that 3‐NPA and indospicine exist in naturalised I. spicata in Australia and of the formation of indospicine residues in tissues of animals grazing paddocks infested with I. spicata.     

Assessment of leaf cover and crop soil cover in weed harrowing research using digital images

Objective assessment of crop soil cover, defined as the percentage of leaf cover that has been buried in soil because of weed harrowing, is crucial to further progress in post‐emergence weed harrowing research. Up to now, crop soil cover has been assessed by visual scores, which are biased and context‐dependent. The aim of this study was to investigate whether digital image analysis is a feasible method to estimate crop soil cover in the early growth stages of cereals. Two main questions were examined: (i) how to capture suitable digital images under field conditions with a standard high‐resolution digital camera and (ii) how to analyse the images with an automated digital image analysis procedure. The importance of light conditions, camera angle, size of recorded area, growth stage and direction of harrowing were investigated, in order to establish a standard for image capture and an automated image analysis procedure based on the excess green colour index was developed. The study shows that the automated digital image analysis procedure provided reliable estimations of leaf cover, defined as the proportion of pixels in digital images determined to be green, which were used to estimate crop soil cover. A standard for image capture is suggested and it is recommended that digital image analysis be used to estimate crop soil cover in future research. The prospects of using digital image analysis in future weed harrowing research are discussed.     

Expert opinion as 'validation' of risk assessment applied to calf welfare

Background

Recently, a Risk Assessment methodology was applied to animal welfare issues in a report of the European Food Safety Authority (EFSA) on intensively housed calves.

Methods

Because this is a new and potentially influential approach to derive conclusions on animal welfare issues, a so-called semantic-modelling type ''validation'' study was conducted by asking expert scientists, who had been involved or quoted in the report, to give welfare scores for housing systems and for welfare hazards.

Results

Kendall''s coefficient of concordance among experts (n = 24) was highly significant (P < 0.001), but low (0.29 and 0.18 for housing systems and hazards respectively). Overall correlations with EFSA scores were significant only for experts with a veterinary or mixed (veterinary and applied ethological) background. Significant differences in welfare scores were found between housing systems, between hazards, and between experts with different backgrounds. For example, veterinarians gave higher overall welfare scores for housing systems than ethologists did, probably reflecting a difference in their perception of animal welfare.Systems with the lowest scores were veal calves kept individually in so-called "baby boxes" (veal crates) or in small groups, and feedlots. A suckler herd on pasture was rated as the best for calf welfare. The main hazards were related to underfeeding, inadequate colostrum intake, poor stockperson education, insufficient space, inadequate roughage, iron deficiency, inadequate ventilation, poor floor conditions and no bedding. Points for improvement of the Risk Assessment applied to animal welfare include linking information, reporting uncertainty and transparency about underlying values.

Conclusion

The study provides novel information on expert opinion in relation to calf welfare and shows that Risk Assessment applied to animal welfare can benefit from a semantic modelling approach.     

Co‐culture of Buffalo Preantral Follicles with Different Somatic Cells

The effect of co‐culture of buffalo preantral follicles (PFs) with different somatic cells, i.e, cumulus, granulosa, ovarian mesenchymal and oviductal epithelial cells was studied. Large PFs (250–450 μm) were isolated by microdissecting the trypsin (1%) digested ovarian cortical slices. Cumulus cells were isolated by repeated pipetting of oocytes, granulosa cells were isolated by aspirating from punctured PFs and ovarian mesenchymal cells were isolated from ovarian cortex by scraping the cortical slices and passing through 20 μm filter. Preantral follicles were cultured in standard culture medium without somatic cells or co‐cultured with cumulus cells, granulosa cells, ovarian mesenchymal cells and oviductal epithelial cells for 80 days. The growth rate (μm/day) of the PFs was monitored by measuring follicular diameter on day 0, 30, 60 and 80 days of culture. The viability of PFs was evaluated by trypan blue staining. The results indicated that PFs co‐cultured with cumulus, granulosa and ovarian mesenchymal cells had a better development and survivality compared with control and those co‐culture with oviductal epithelial cells. Maximum growth and survivality of PFs were achieved when cultured with cumulus cells. It is concluded that inclusion of somatic cells in PF culture media had beneficial effect on the growth of PFs and cumulus cells supported maximum growth and survivality of PFs in vitro of all somatic cells tested.