Detection of Follicular Apoptosis in Water Buffalo (Bubalus bubalis) Ovary by Histology and Nick End Labelling Technique

The objective of this experiment was to assess the features and extent of follicular apoptosis in the water buffalo (Bubalus bubalis) ovary using classical histology and nick end labelling technique. Ovaries (n = 40) procured from the slaughterhouse were used for the study. The sections (5 μm) were used for detection of terminal deoxynucleotidyl transferase‐mediated dUTP‐biotin nick end labelling (TUNEL) and classical histology (H&E). Those follicles showing ≥ 5% TUNEL positivity (TUNEL assay) and pyknotic nuclei (histology) in granulosa cells were classified as atretic. Based on histology, the atretic primary and secondary follicles (%) were 93.82 and 95.62 respectively. The histology study reveals that the rates (%) of atresia in <1, 1–3, 3–5 mm and >5 mm were 36.90, 40.50, 62.84 and 74.5 respectively. Further the atretic tertiary follicles (%) were significantly lower than the primary and secondary classes of follicles. TUNEL assay reveals that the atretic rate (%) of tertiary follicles in <1, 1–3, 3–5 and ≥ 5 mm class follicles were 50.88, 53.84, 81.81 and 36.36 respectively. The percentage of atresia in >5 mm diameter follicles were significantly lower in TUNEL than histology. Percentages of granulosa and thecal cells positive for atresia by TUNEL were 30.7 ± 0.53 and 13.82 ± 0.18 respectively per follicle. The initial structural changes in atretic follicles were seen primarily in the granulosa cells. In severely atretic follicles TUNEL positive granulosa cells along with theca cells have to be considered in assessing the rate and extent of atresia.     

Effect of feeding rumen protected rice bran on calcium homeostasis of non‐lactating multiparous cows

Milk fever in dairy cows can be prevented by activating Ca homeostasis before calving. Homeostatic adaptation can be achieved by reducing dietary Ca availability. Formaldehyde‐treated rice bran was studied to supply rumen protected phytic acid to reduce Ca availability. Twelve multiparous dry cows were used in a 3 × 3 Latin square change‐over design with 5‐day periods to test three dietary treatments. Diets consisted of a forage mix (maize silage, grass silage and hay), being 77% of ration dry matter, supplemented with three concentrates: Control (no formaldehyde‐treated rice bran), T1 (100% formaldehyde‐treated rice bran) and T2 (99.5% formaldehyde‐treated rice bran with 0.6% Ca carbonate, to equal Ca content of Control). Dietary treatments did not affect urine pH (8.14, 8.13 and 8.11 for Control, T1 and T2 respectively) or dry matter intake (13.9, 13.7 and 13.8 kg for Control, T1 and T2 respectively). Including formaldehyde‐treated rice bran in the diet resulted in lower urinary Ca/creatinine ratio (0.970, 0.457 and 0.618 for Control, T1 and T2 respectively). A sudden increase of urinary Ca excretion took place after withdrawal of T1 and T2 at introduction of Control, peaking on the first day and coming back down progressively in the second and third days. Peak was greatest after T1 and was not observed in transitions between rice bran treatments. This is understood as indirect evidence of activation of intestinal Ca absorption during formaldehyde‐treated rice bran feeding, because renal adaptations to changes in blood Ca clearance are immediate and intestinal adaptations delay 2 days. It was concluded that including formaldehyde‐treated rice bran in rations before calving may represent a dietary strategy to prevent milk fever without reducing dry matter intake.     

Effects of dietary energy concentration, nonstarch polysaccharide concentration, and particle sizes of nonstarch polysaccharides on digesta mean retention time and gut development in laying hens

1. From an experiment with 504 laying hens (ISA Brown strain, 18–40 weeks of age), 90 40-week old hens were used for determining digesta mean retention time (MRT) and gut weight development. This experiment comprised 6 dietary treatments according to a 2?×?3 factorial design. Factors were dietary apparent metabolisable energy (AME) concentration (11·8 vs 10·6?MJ/kg), insoluble nonstarch polysaccharides (NSP) concentration (65 vs 134?g/kg), and fine vs coarse particle sizes of added NSP. Titanium recovery in different gut segments was used as an indicator of MRT.

2. Increasing NSP concentration prolonged MRT in the crop (68 vs 34?min) and total foregut (91 vs 57?min) compared with control NSP. Reducing energy concentration prolonged MRT in the colon (26 vs 7?min), and total hind gut (30 vs 9?min), compared with control energy. Overall MRT was not affected by dietary treatments.

3. Increasing NSP concentration increased relative weights of the empty proventriculus-gizzard and its contents by 30% (25·2 vs 19·4?g/kg) and 18% (15·4 vs 13·0?g/kg), respectively, compared with control NSP diets.

4. MRT in the foregut was prolonged as daily insoluble NSP intake increased, and this was more pronounced in hens given coarsely ground NSP, compared with finely ground. A prolonged MRT in the foregut seemed to indicate a higher level of satiety, which may contribute to a lower feather pecking pressure in laying hens.     

Molecular Basis of Gene-for-Gene Specificity in Bacterial Speck Disease of Tomato

Transient expression of the Pseudomonas syringae avirulence gene avrPto in plant cells resulted in a Pto-dependent necrosis. The AvrPto avirulence protein was observed to interact directly with the Pto resistance protein in the yeast two-hybrid system. Mutations in the Pto and avrPto genes which reduce in vivo activity had parallel effects on association in the two-hybrid assay. These data suggest that during infection the pathogen delivers AvrPto into the plant host cell and that resistance is specified by direct interaction of Pto with AvrPto.     

Effect of Different Manganese Concentrations during in vitro Maturation of Bovine Oocytes on DNA Integrity of Cumulus Cells and Subsequent Embryo Development

Manganese (Mn) is a trace element present in forages and cereals, and its concentration depends on soil status. Manganese deficiency in cattle, goats and ewes not only impairs oestrous cycle but reduces calf birth weight. The achievement of the first oestrus is delayed, and more attempts are necessary to obtain a successful conception. This study was conducted to investigate the effect of the availability of supplemental Mn during IVM on DNA damage of cumulus cells and total glutathione (GSH) content in oocytes and cumulus cells. The effect of supplementary Mn during IVM on subsequent embryo development was also studied. The results reported here indicate (i) DNA damage in cumulus cells decreased with 0, 2, 5 and 6 ng/ml Mn supplementation during IVM (p < 0.05). (ii) Intracellular GSH‐GSSG content increased (p < 0.01) with different Mn concentrations in oocytes and cumulus cells. Also, cumulus cell number per cumulus oocyte‐complexes (COC) did not differ either before or after IVM. (iii) Addition of Mn to maturation medium resulted in similar cleavage rates (p > 0.05) at 0, 2, 5 and 6 ng/ml Mn. However, subsequent embryo development to blastocyst stage was significantly higher (p < 0.01) in oocytes matured with 5 and 6 ng/ml Mn. (iv) There was also an increase (p < 0.05) in mean cell number per blastocyst obtained from oocytes matured with 5 and 6 ng/ml respect to zero Mn (IVM alone) and 2 ng/ml Mn. This study provides evidence that optimal embryo development to the blastocyst stage was partially dependent on the presence of Mn during IVM. Moreover, the availability of Mn during oocyte maturation ensures ‘normal’ intracellular GSH content in COCs and protects DNA integrity of cumulus cells.     

Components of electroencephalographic responses to slaughter in halothane-anaesthetised calves: Effects of cutting neck tissues compared with major blood vessels

AIM: To identify whether cutting neck tissues or cutting major blood vessels initiates the mechanisms responsible for electroencephalographic (EEG) responses to slaughter by ventral-neck incision without prior stunning in halothane-anaesthetised calves.

METHODS: Calves were assigned to two groups, viz transection of neck tissues with intact blood circulation through the brain (n=10), or transection of the major blood vessels of the neck but not most other neck tissues (n=7). They were minimally anaesthetised with halothane, using an established anaesthesia protocol. The animals in the neck-tissue transection group had their carotid arteries and jugular veins exposed and cannulated proximal and distal to the proposed site of subsequent ventral-neck incision; this diverted blood fl ow through these vessels so that cerebral perfusion and drainage were preserved. In animals in the blood-vessel transection group, the carotid arteries and jugular veins were exposed bilaterally by surgical dissection. They were then transected without further damage to the remaining structures of the neck. Changes in the median frequency (F50), 95% spectral edge frequency (F95), total power of the EEG (Ptot), and arterial blood pressure were compared within each group before and after neck-tissue or blood-vessel transection, and between groups following treatments.

RESULTS: Neck-tissue transection resulted in little overall change in the F50, an increase in the F95, and an initial increase in Ptot followed by a transient decrease and eventual return to pre-treatment values. There was between-animal variation in these EEG parameters. Transection of the major blood vessels of the neck resulted in a decrease in F50 in most animals; changes in F95 were highly variable, and there was a decrease in Ptot.

CONCLUSIONS: The EEG responses seen following necktissue and blood-vessel transection were qualitatively distinct, and suggested that cutting neck tissues caused greater noxious sensory input than transection of only the major blood vessels of the neck. These observations support the conclusion that the EEG responses seen after ventral-neck incision in intact animals are primarily due to noxious stimulation as a result of incision of ventral-neck tissues and not mainly as a result of loss of blood flow through the brain.     

Retrospective study of 108 foals with septic osteomyelitis

Objective   To determine the clinical characteristics, short-term outcome and future athletic performance of foals with septic osteomyelitis.
Design   Retrospective clinical study of 108 Thoroughbred foals with radiographic evidence of bone infection that were presented at the Scone Veterinary Hospital between August 1995 and December 2001. Medical records were reviewed and information concerning signalment, the clinical, laboratory and radiographic findings, treatment and outcome was obtained. Racing records were obtained and evaluated for surviving foals that had reached racing age.
Results   Mean age of foals at initial evaluation was 39 days (range 1–180 days); 21 foals had multiple radiographic bone lesions (19.4%), and 76 had concurrent septic arthritis (70.4%). The most frequently affected bones were the femur, tibia and distal phalanx. In total, 87 foals were discharged from the hospital (80.6%), 79 survived long-term to reach racing age and 52 raced (65.8%). Overall, 48% (52/108) of the foals treated for osteomyelitis raced. Foals less than 30 days of age at the time of diagnosis, critically ill foals and those with multiple bones or joints affected were significantly less likely to be discharged from hospital. Multiple septic joints, but not multiple bone involvement, had an unfavourable prognosis for racing.
Conclusions   The prognosis for survival of foals with septic osteomyelitis or osteitis is favourable. Multiple bone or joint involvement is an important short-term prognostic indicator; however, the involvement of multiple joints, but not multiple infected bones, is associated with an unfavourable prognosis for racing.     

Inhibited development of trichostrongylid worms in grazing cattle

Inhibition of development of gastro-intestinal trichostrongylid worms was studied using successive groups of tracer calves and groups of continuously grazed calves over one year in the Tully area of North Queensland lowland wet tropics. The results, assessed by means of worms from these calves recovered at necropsy 3 weeks after their removal from pastures, showed inhibition in the development of Haemonchus placei and Cooperia punctata at the early fourth stage at the approach to and during the relatively dry period in the area. Inhibition was however minor and inhibited larvae formed but only a small percentage of worm burdens in both categories of calves, indicating that they were not in any way of major epidemiological importance. It was suggested that the minor nature of inhibition was due to the mild climatic conditions which could not produce appropriate conditioning treatment, or caused only mild selection pressure for inhibition in the area.     

Responses of broiler chickens to dietary protein: effects of early life protein nutrition on later responses

1. A study was conducted with modern broiler chicks to test the effects of early life protein nutrition and sex on responses in growth and body composition to dietary protein at a later age. Effects on the incidence of metabolic disorders were also evaluated. 2. From 11 to 26 d of age (EXP1), birds were given 8 diets varying in balanced protein to energy ratio (BPE ratio) between 0.575 and 1.100 g digestible lysine per MJ AMEn. Birds from two treatment groups in EXP1 (BPE ratio of 0.725 and 1.025 g/MJ, respectively) were subsequently used in a test from 26 to 41 d of age (EXP2). In EXP2, 8 diets were used, varying in BPE ratio between 0.500 and 1.025 g/MJ. 3. Responses in weight gain and feed conversion to BPE ratio in EXP2 changed considerably when BPE ratio in EXP1 was modified, irrespective of sex. Up to 10% improvement in both weight gain and feed conversion in EXP2 was observed if BPE ratio in EXP1 was 0.725 compared with 1.025 g/MJ. With males, however, the effect of treatment in EXP1 on weight gain in EXP2 was present only at high BPE ratios. 4. For the relative gain of breast meat and abdominal fat, but not for carcase, the responses of male broilers to BPE ratio in EXP2 were altered by the BPE ratio in EXP1. With females, responses in composition of the gain to diet in EXP2 were independent of BPE ratio in EXP1. 5. The incidence of metabolic disorders was low, irrespective of treatment in EXP1. The lower BPE ratio in EXP1 increased mortality in EXP2 from 0.8 to 3.6%. 6. Our findings show that broiler responses to dietary protein depend on previous protein nutrition and sex. Effects of early life protein nutrition on incidence of metabolic disorders were not observed. The results strongly suggest that protein levels in grower and finisher diets should not be optimised independently, but simultaneously.     

Performance, digesta characteristics, nutrient flux, plasma composition, and organ weight in pigs as affected by dietary cation anion difference and nonstarch polysaccharide

Two dietary cation anion difference (CAD) levels (-100 and 200 mEq/kg) and two dietary nonstarch polysaccharide (NSP) levels (10 and 15%) were used in a 2 x 2 factorial arrangement in two randomized blocks (trials) to evaluate performance, digesta pH and buffer capacity, apparent digestibility, plasma composition, and organ weight in pigs. Seven pigs with a mean initial weight of 7.5 kg were used in each treatment. Pigs had free access to feed and water during the 3-wk experimental period. At the end of the experiment, all pigs were killed approximately 2.5 h after addition of fresh feed. Liver, kidneys, and small and large intestine were removed and weighed immediately. Gastric and small intestinal (divided into two equal parts) digesta were collected. Dry matter, pH, buffering capacity, viscosity, acid insoluble ash, mineral content (Na+, K+, and Cl-), nitrogen, and gross energy content (only in the second part of the small intestine) of the digesta were measured. An interaction (P < 0.05) between dietary CAD and dietary NSP was observed for feed intake, growth, apparent digestibility of DM and gross energy in the distal small intestine, and chloride in the stomach and duodeno-jejunal part of the small intestine. Increasing dietary NSP content increased apparent digestibility of DM and chloride in the stomach and increased apparent digestibility of DM, chloride, sodium, and potassium in the first part of the small intestine. Lowering dietary CAD levels significantly increased apparent digestibility of DM in the first part of the small intestine and decreased chloride influx in the stomach and the small intestine. No significant differences were found for pH, buffering capacity, and viscosity of digesta, except the buffering capacity in the second part of the small intestine, which was higher (P < 0.05) in the 200 mEq/kg CAD group than in the -100 CAD mEq/ kg group. Plasma urea decreased in the low dietary CAD groups and plasma ammonia increased in the high NSP groups. Large intestine weight was significantly less for the -100 mEq/kg CAD groups than for the 200 mEq/kg CAD groups. In conclusion, the effect of CAD on feed intake and growth in pigs depends on dietary NSP levels.