Effects of Prolonged in vitro Culture and Cryopreservation on Viability,DNA Fragmentation,Chromosome Stability and Ultrastructure of Bovine Cells from Amniotic Fluid and Umbilical Cord

The objective of this work was to study cellular types that did not participated in the gastrulation process, amniotic fluid cells (AFCs) and umbilical cord cells (UCCs), in conditions of long‐term culture and cryopreserved with different solutions. The AFCs and UCCs were used in a comparative study with ear fibroblast cells (EFCs) that were cultured in vitro until 20 cellular passages and cryopreserved in 10% dimethylsulphoxide (DMSO), 5% dimethyl formamide (DMF) and 7% glycerol (Gly) solutions. The cellular viability, ultrastructure, DNA fragmentation and chromosome stability were evaluated to determine the cellular type most resistant. In all cell types, it was possible to evaluate the AFCs until 15 passages and UCCs until 20 passages with different periods of cellular growth to reach the confluence phase. Solutions containing 10% DMSO ensured viability of 90.33 ± 5.58%, 90.56 ± 4.40% and 81.90 ± 3.31%, respectively for EFCs, AFCs and UCCs, being significantly more efficient and with less variation than other cryoprotectant solutions. The AFCs were more sensitive to cryopreservation and presented low viability rate at the passage 20 (17.2 ± 8.87%). There was no change in karyotype and nuclear fragmentation was low in all cellular passages studied. With the scanning electron analysis was possible the characterization of AFCs and UCCs in suspension. The three cellular types of cells presented different shapes and characteristics on the surface. The results demonstrate that bovine AFCs and UCCs can be isolated, cultured in vitro and cryopreserved in 10% DMSO, not causing damage to DNA and chromosomes. The UCCs were more resistant than AFCs in all aspects.     

Effects of Storing Pig Ovaries at 4 or 20°C for Different Periods of Time on the Morphology and Viability of Pre-Antral Follicles

The purpose of this study was to evaluate the effect of cooling ovarian tissue on pig pre-antral follicles. Ovaries were maintained in saline solution (0.9%) at 4 or 20 degrees C for 6, 12 or 18 h. After storage, pre-antral follicles were morphologically evaluated. While primordial follicles were not affected by the storage, the percentage of morphologically normal growing follicles was significantly reduced in ovarian tissue stored at 20 degrees C for 12 or 18 h. To test the viability of stored follicles, growing follicles isolated from ovaries stored at 4 degrees C for 18 h and at 20 degrees C for 6 h were cultured for 3 days. Follicles stored in either condition presented the same growth pattern in vitro as fresh follicles. We conclude that storage of pig ovaries at 4 degrees C for up to 18 h or at 20 degrees C for up to 6 h does not affect the morphology of growing follicles or their ability to grow in vitro.     

A Study on Postpartum Metritis in Iraqi Buffalo Cows: Bacterial Causes and Treatment

The objectives of the present study were to determine the relationship between bacteriological findings, clinical signs and histopathological changes in postpartum metritis. Evaluation of the treatment efficiency of using systemic or intra‐uterine infusion of antibiotics with some hormonal preparations for the treatment of postpartum metritis. Data were collected from 50 buffalo cows with history of calving of more than 1 month. All buffaloes were subjected to detailed clinical examination including external inspection, vaginoscopy and transrectal palpation of the cervix, uterus and ovaries. Swabs for bacteriology and biopsies for histopathology were collected from uterine lumen from each buffalo included in the present study. Bacteria identified using API systems following aerobic and anaerobic cultures. Vaginal mucus scored for character, odour and estimation of polymorphonuclear cells (PMNs). Treatment conducted using oxytetracycline in local intrauterine infusion or systemically with hormonal treatment including prostaglandinF2α (PGF2α) and oestradiol benzoate. Results revealed that the most predisposing factor for postpartum uterine infection was retained placenta and toxic puerperal metritis. The most prevalent bacteria in uterine lumen were Escherichia coli, Archanobacterium pyogenes, Bacteroides fragilis and Fusobacterium necrophorum the most prevalent bacteria in buffaloes with postpartum metritis. A. pyogenes and F. necrophorum were an important pathogens causing severe uterine inflammation as found in histopathological examinations. Buffaloes with postpartum metritis showed good clinical cure when oxytetracycline injected systemically with PGF2α. Intrauterine infusion of oxytetracycline had no advantage for the treatment of uterine infection in buffalo cows with postpartum metritis. PGF2α improved clinical cure of buffaloes with postpartum metritis.     

Effect of integrated nutrient management and crop diversification on productivity and nutrient balance sheet of rice-based cropping systems

A field study conducted for three crop years (June?–?July) from 1995?–?96 to 1997?–?98 at the Indian Agricultural Research Institute, New Delhi involving five rice-based cropping systems and six nutrient combinations indicated that rice?–?wheat?–?mungbean (RWM), rice?–?potato?–?mungbean (RPM), rice?–?rapeseed?–?mungbean (RRsM) and rice?–?clover (RC) cropping systems gave 0.7?–?1.0, 3.2?–?5.9, 0.2?–?2.2 and 1.5?–?3.6 t ha^?1 yr^?1, respectively, higher productivity and removed 7.9?–?22.6, 38.0?–?64.5, 53.0?–?61.8 and 51.4?–?66.1?kg ha^?1 yr^?1, respectively, more nitrogen, 2.3?–?7.1, 14.5?–?22.8, 3.8?–?7.2 and 17.3?–?21.7?kg ha^?1 yr^?1, respectively, more phosphorus and 1.6?–?11.4, 15.3?–?42.3, 8.2?–?22.7 and 40.8?–?57.8?kg ha^?1 yr^?1, respectively, more potassium than the rice?–?wheat (RW) cropping system which led to a net balance of +?151, +?58, ??153 and ??167?kg ha^?1 of nitrogen, +?13, ??27, ??8 and ??59?kg ha^?1 of phosphorus and ??549, ??551, ??558 and ??691?kg ha^?1 of potassium after three cycles of RWM, RPM, RRsM and RC cropping systems, respectively against a net balance of +?201, +?26 and ??533?kg ha^?1 of N, P and K, respectively in the RW cropping system. Application of FYM along with NPK in these cropping systems changed the negative balance of nitrogen and phosphorus into positive balance and reduced the negative balance of potassium by 151?–?378?kg ha^?1. Application of P and K fertilizers along with nitrogen also helped in arresting the negative balance of P and K under different rice based cropping systems. These results thus indicate that balanced use of NPK and FYM plays a major role for sustaining the productivity of a cropping system.     

Direct demonstration of milk as an element of archaeological economies

The stable carbon isotope (delta13C) compositions of individual fatty acid components of remnant fats preserved in archaeological pottery vessels show that dairying was a component of archaeological economies. Characteristic delta13C values arise from biases in the biosynthetic origins of the C18:0 fatty acids in milk and adipose fat. Milk and adipose fat from animals raised on similar pastures and fodders have distinct isotopic signatures.     

Buffalo (Bubalus bubalis) Pre-antral Follicle Population and Ultrastructural Characterization of Antral Follicle Oocyte

The main objectives of the present study were to determine the ultrastructural modifications occurring in the oocyte during late folliculogenesis and to estimate pre-antral follicle population in buffalo. Half the collected ovaries were fixed and prepared for optic microscopy; the antral follicles from the other ovaries were measured and individually punctured. The cumulus–oocyte complexes (COCs) were processed for transmission electron microscopy. The number of pre-antral follicles in buffalo ovaries was estimated at 19 819 structures. Cumulus–oocyte complexes derived from 1-mm antral follicle had an eccentrical nucleus and compact corona radiata , ooplasm vilosities were fully embedded in zona pellucida (ZP) and a well-defined junction could be observed. Mitochondria were predominantly round and well distributed in ooplasm, as were small lipid vacuoles. In COCs derived from 2-mm antral follicles, the initial formation of perivitelline space was observed. The nucleus was peripherally located and the number of pleomorphic mitochondria increased. Cortical granules were clustered at oocyte periphery and lipid vacuoles increased in number and size. In COCs derived from 6-mm antral follicles, the organelles were located mainly in the perinuclear region. Golgi complexes and smooth endoplasmic reticulum (SER) were more developed. Mitochondria migrated to the cortical region and lipid vacuoles migrated to the medullar region. In COCs derived from 10-mm antral follicles, the lipid vacuoles coalesced and occupied the medullar region of the oocyte, together with a well-developed SER. Mitochondria were pleomorphic and located at the oocyte periphery. In conclusion, the morphological differences described in this paper could be responsible for some functional differences observed in in vitro embryo production and follicular dynamics for buffalo, when compared with cattle.