Stress alters the cellular and proteomic compartments of bovine bronchoalveolar lavage fluid

The stresses of transportation, weaning and commingling are associated with an increased incidence of bacterial and viral pneumonia in cattle. Proteins expressed in the epithelial lining fluid (ELF) of the lungs, in conjunction with resident leukocytes, represent the first line of defence against opportunistic pathogens, and stress-induced alterations in their expression may reveal markers of disease susceptibility. Bronchoalveolar lavage fluid was sampled in weaned and transported calves and ELF protein expression was compared to a control group of calves using two-dimensional electrophoresis (2DE). Serum and pulmonary haptoglobin were increased following stress concurrent with the number of blood neutrophils. Using 2DE, significant changes in expression were observed in spots identified by mass spectrometry as annexin A1 and A5, odorant-binding protein (OBP), isocitrate dehydrogenase, fibrinogen, heme-binding protein, alpha-2-HS-glycoprotein, alpha-1-antichymotrypsin and albumin. Quantification of OBP mRNA by real-time RT-PCR and OBP protein by western blot revealed gender-dependent differences in relative OBP expression in response to stress. These findings reveal stress-associated protein changes in pulmonary ELF and suggest a mechanism through which stress alters respiratory disease susceptibility.     

Sub-cellular localisation of alkaline phosphatase activity in the cytoplasm of tammar wallaby (Macropus eugenii) neutrophils and eosinophils

Alkaline phosphatase (ALP) has been used in studies of neutrophil morphology and function as a marker for identifying different granule populations. In human neutrophils, ALP is found within secretory vesicles, a rapidly mobilisable vesicle population important for upregulating membrane receptors during early activation. Intra-cellular ALP activity in the heterophils of rabbits and guinea pigs, in contrast, is found only in secondary granules. The neutrophils and eosinophils of tammar wallabies (Macropus eugenii) have previously been reported to contain large amounts of ALP activity when stained using routine cytochemical techniques. To define the subcellular location of ALP in this species, cell suspensions were examined using cerium chloride cytochemistry and transmission electron microscopy (TEM). ALP was found in 2 distinct cytoplasmic compartments. One compartment displayed morphology consistent with a subpopulation of secondary granules while a second tubulo-vesicular population appeared similar to the secretory vesicles of human neutrophils. Thin tubular vesicles containing ALP were also identified within the cytoplasm of tammar wallaby eosinophils. Large numbers of ALP-containing vesicles have not been recognised previously in eosinophils and this may represent a novel cytoplasmic compartment. In both cell types, ALP-containing structures showed alteration in morphology following stimulation with N-formyl-Met-Leu-Phe (fMLP) and PMA.     

Affinity of detomidine, medetomidine and xylazine for alpha-2 adrenergic receptor subtypes

α₂-Adrenergic receptor agonists are widely used in veterinary medicine as sedative/hypnotic agents. Four pharmacological subtypes of the α₂-adrenergic receptor (A, B, C and D) have been identified based primarily on differences in affinity for several drugs. The purpose of this study was to examine the affinities of the sedative agents, xylazine, detomidine and medetomidine at the four α₂-adrenergic receptor subtypes. Saturation and inhibition binding curves were performed in membranes of tissues containing only one subtype of a₂-adrenergic receptor. The K_D for the α₂-adrenergic receptor radioligand, [³H]-MK-912, in HT29 cells (α_2A-), neonatal rat lung (α_2B-), OK cells (α_2C-) and PC12 cells transfected with RG20 (α_2D-) were 0.38 ± 0.08 n m , 0.70 ± 0.5 n m , 0.07 ± 0.02 n m and 0.87 ± 0.03 n m , respectively. Detomidine and medetomidine had approximately a 100 fold higher affinity for all the α₂-adrenergic receptors compared to xylazine but neither agonist displayed selectivity for the α₂-adrenergic receptor subtypes. These data suggest that available sedative/hypnotic α₂-adrenergic receptor agonists can not discriminate between the four known α₂-adrenergic receptor subtypes.     

Growth response of broilers to spray-dried plasma in pelleted or expanded feed processed at high temperature

A series of 4 experiments evaluated effects of mash conditioning temperature from a pellet mill or expander on performance of broilers fed pelleted diets containing spray-dried plasma (SDP). All experiments utilized Ross x Ross 308 male broilers randomly assigned to their respective treatments (6 or 10 broilers/pen and 8 or 10 pens/treatment). Treatments in Exp. 1 consisted of a control (0% SDP), SDP coated postpelleting, or SDP blended into the meal prepelleting. Experiment 2 and 3 included the same 3 treatments as in Exp. 1 but with additional treatments of SDP blended into the meal and conditioned at 90 or 95 degrees C before pelleting. In Exp. 4, treatments consisted of a control (0% SDP) or SDP blended into the meal and pelleted (85 degrees C conditioning temperature) or expanded (149 degrees C final effective temperature) and then pelleted. Corn-soybean meal-based diets were formulated to be equal in lysine and ME in all experiments. Pelleted diets were conditioned for 15 s at 85 degrees C, and expanded diets were conditioned at 95 degrees C, 29.7 MJ/t, 13.95 kg/cm2 cone pressure, exit temperature of 149 degrees C, and then pelleted through a 4 x 32-mm die. In Exp. 1, ADG and feed intake were improved (P < 0.05) for broilers fed SDP from d 1 to 28 of age, with greater BW at d 42. In Exp. 2, both in early (d 1 to 28 of age) phases, and overall (d 1 to 42 of age), broilers fed SDP had improved (P < 0.05) gain and efficiency. In Exp. 3, ADG, feed intake, efficiency of gain, and BW were improved (P < 0.01) for broilers fed SDP from d 1 to 21 of age, regardless of conditioning temperature. In Exp. 4, broilers fed SDP had improved (P < 0.05) gain, BW, and feed intake regardless of processing method. Overall, the results of all of the experiments demonstrated that pellet conditioning temperature from 85 to 95 degrees C and expander temperatures to 149 degrees C did not impair the positive growth effects of SDP in pelleted or expanded broiler feed.     

Relationship between accumulated heat stress during the dry period, body condition score, and reproduction parameters of Holstein cows in tropical conditions

To estimate the relationship between heat stress during the last 60 days prepartum, body condition score and certain reproductive traits in the subsequent lactation of Holstein cows, 564 multiparous cows and 290 primiparous cows from four dairy herds were used in a hot, humid region. Maximum prepartum degree days were estimated to quantify the degree of heat stress. Multiple regressions analyses and logistic regression analysis were performed to determine the effect of prepartum heat stress and body condition change on reproductive parameters, which were obtained from DHIA forms at the end of the lactation. Multiparous and primiparous cows which gained body condition score from calving to 60 d postpartum exhibited 28 and 27 fewer days open (P?<?0.05), respectively, than cows not gaining. There was no effect (P?>?0.05) of heat stress measurement on days open or services per conception in either multiparous or primiparous cows. During hotter months of calving, multiparous cows showed higher services per conception and primiparous cows showed higher days open and services per conception (P?<?0.05). Maximum prepartum degree-days were positively associated (P?<?0.05) with calving difficulty score. Multiparous cows with high body condition score at calving were 1.47 times more likely to present a very difficult calving than cows that calved in October (P?<?0.05). Collectively, these results suggest that reproductive performance was not affected by cumulative prepartum heat stress although it was associated with very difficult calving score.     

Obituary: James Thomas Kelly,BVSc

Extract

On 28 August 1987 James Thomas Kelly, veterinary surgeon, of Milson Line, Palmerston North, died at 68 years of age.