Studies about the mechanism of internalization by mammary epithelial cells of Escherichia coli isolated from persistent bovine mastitis

The purpose of this study was to investigate the interaction between Escherichia coli and primary mammary epithelial cell cultures derived from cows with persistent intramammary infection (IMI). Two strains of E. coli, isolated from the milk of two different cows suffering from persistent E. coli IMI were tested for adhesion to and invasion of three primary mammary epithelial cell cultures derived from mammary biopsies of the two infected cows. Intracellular E. coli were detected during five days post infection in vitro. Both strains of E. coli adhered to and invaded monolayers of all three primary mammary epithelial cell cultures. One strain adhered less but invaded more than the other. Comparison with other mammary pathogens indicated that E. coli invaded the cells less efficiently than Staphylococcus aureus, about as efficiently as Streptococcus dysgalactiae and more efficiently than Streptococcus uberis. The mechanism of E. coli invasion was studied using the cytoskeleton disrupting agents colchicine and cytochalasin D. These compounds inhibited the invasion of E. coli. Invasion of E. coli could also be inhibited by the phosphokinase inhibitors genistein and staurosporin in a dose-dependent fashion. Phorbol-myristyl-acetate (PMA) had no effect on the invasion of E. coli. Histology of mammary tissue revealed chronic inflammatory changes in quarters that were persistently infected by E. coli. Intracellular bacteria were not detected in mammary tissue sections. Polymerase chain reaction (PCR) analysis suggested that the two strains of E. coli lacked genes encoding for bundle-forming pili (bfpA), intimin (eae) and translocated intimin receptor (tir), which are characteristic for enteropathogenic E. coli (EPEC).     

Reticular contraction attributes and passage of digesta from the ruminoreticulum in cattle fed roughage diets.

Four nonlactating Holstein dairy cows (BW = 692 +/- 49 kg) were used in a 4 x 4 Latin square design experiment to determine how changes in attributes of reticular contractions were related to changes in passage of digesta from the ruminoreticulum in cattle fed a forage-based diet. Frequencies of reticular contraction during eating, ruminating, and resting phases of the feeding cycle were not affected (P = .09; P = .07; P = .09, respectively) by feeding level. Duration and amplitude of reticular contractions both increased linearly during eating (P = .03; P = .01), ruminating (P = .01; P = .02), and resting (P = .04; P = .02), respectively, as feed intake level increased. After removing the effects of intake, squared partial correlations computed within analyses of covariance indicated that NDF flow was not related to frequency (r2 = .15, P = .20) but was positively correlated with duration (r2 = .76, P = .0001) and amplitude (r2 = .56, P = .003) of contraction, indicating that duration and amplitude of reticular contraction were more associated with duodenal NDF flow than frequency. However, duration of reticular contraction (r2 = .47, P = .014) explained a greater variation in duodenal NDF flow than amplitude (r2 = .011, P = .74). Stepwise multiple regression also indicated that duration of reticular contraction was the attribute most associated (r2 = .76, P = .0001) with duodenal NDF flow. It was concluded that the changes in digesta passage from the ruminoreticulum were associated primarily with changes in the duration of reticular contraction.     

Immunocytochemical study of tissues from clinically normal dogs and of neoplasms, using keratin monoclonal antibodies.

Three commonly used keratin monoclonal antibodies (MAB)--AE1:AE3, CAM 5.2, and MAK-6--were compared with routinely used cytokeratin antibody. The expression of these antibodies was analyzed in several tissues obtained from clinically normal dogs and in a variety of neoplasms from dogs. Using appropriate enzymatic digestion, paraffin-embedded tissues processed in routine manner retained their typical keratin expression. Differentiated and poorly differentiated epithelial neoplasms, lymphomas, and melanomas were studied by use of the avidin-biotin-peroxidase technique. All 4 of the aforementioned antibodies had similar staining profiles. Of 3 anaplastic carcinomas, 2 had positive reaction to all 4 antibodies. All lymphomas, plasma cell tumors, and amelanotic melanomas had negative reaction to MAK-6, CAM 5.2, AE1:AE3, and cytokeratin MAB. Three basal cell epitheliomas had positive reaction to all 4 antibodies, whereas 1 basal cell tumor with a solid pattern had negative staining reaction. Two carcinoids had negative reaction to all markers and 1 of 2 malignant chemodectomas and 1 transitional cell carcinoma had staining reaction to only AE1:AE3 MAB. Comparing the 4 antibodies, use of AE1:AE3 MAB produced the strongest staining intensity followed by cytokeratin, MAK-6, and CAM 5.2 MAB. All 4 antibodies had low background staining. In conclusion, AE1:AE3 and MAK-6 MAB are as useful as cytokeratin MAB for identification of poorly differentiated epithelial neoplasms in dogs and cats.     

Resistance of Pasteurella multocida A:3,4 to phagocytosis by turkey macrophages and heterophils

A virulent field isolate and 2 vaccine strains of Pasteurella multocida A:3,4 were compared for resistance to phagocytosis by turkey macrophages and heterophils, using in vitro assays. The least virulent vaccine strain (M-9) was phagocytosed to a greater degree than was the field isolate or the other vaccine strain (Clemson University). All 3 bacteria differed significantly from each other in the amount of capsular material present as measured by polycationic ferritin labeling and electron microscopy. Removal of the capsule with hyaluronidase resulted in a significant increase in phagocytosis of the field isolate.     

Hydronephrosis and pyelonephritis associated with an anomalous vas deferens in a bull

Hydronephrosis and pyelonephritis were diagnosed in a 2-year-old Limousin bull. Salient historical and clinical observations included weight loss, anorexia, intermittent colic, polyuria, polydypsia, and an ammoniacal breath odor. Intravenous treatment with fluids and antibiotics did not induce a favorable response. An anomalous vas deferens was observed on postmortem examination. The vas deferens ended in a blind polyp that protruded into the lumen of the urethra. This polyp may have caused chronic intermittent obstruction, predisposing to the hydronephrosis and pyelonephritis.     

Effect of the synthetic androgen mesterolone upon seminal characteristics of dogs

The synthetic androgen mesterolone given at 1–5 mg/kg daily in divided doses significantly increased the total sperm output of normal dogs. A dose of 3-0 mg/kg daily in divided doses significantly improved sperm morphology of an oligoasthenoteratozoospermic dog. The effects upon plasma testosterone concentration were not conclusive.     

The key role of records in a production medicine practice

The needs of dairy farmers for veterinary assistance have changed and are still changing. It appears that many future opportunities in dairy practice will involve active consultation on herd management factors. The ability to evaluate herd records will be crucial to this effort. This article discusses organizing and then utilizing a production medicine program based on systematic records analysis.     

Contribution of propylene glycol-induced Heinz body formation to anemia in cats

Propylene glycol (PG) is a common preservative and source of synthetic carbohydrates in soft-moist pet foods. Propylene glycol was fed to cats for 5 weeks at concentrations found in commercial diets (1.6 g/kg of body weight; 12% of diet on a dry-weight basis) and for 3 weeks at concentrations exceeding usual intake (8 g/kg; 41% of diet). There was a dose-dependent increase in Heinz body percentage to 28% in cats fed the low dose of PG and to 92% in cats fed the high dose. Erythrocyte half-life, measured using [14C]-cyanate hemoglobin (Hb), decreased significantly (P less than 0.05) by 18.8% and 60% in cats fed the low and high PG doses, respectively. The PCV in cats fed the low dose was unaffected, whereas cats fed the high dose had a mean (+/- SEM) decrease in PCV from 33.5 +/- 1.05% to 26.3 +/- 1.45%, accompanied by punctate reticulocytosis and bone marrow erythroid hyperplasia. A dose-dependent increase in iron pigment was found in the liver and spleen of all cats. In cats fed the low dose of PG, erythrocyte reduced glutathione concentration actually increased from 7.02 +/- 0.56 to 9.74 +/- 0.69 mumol/g of Hb, but decreased to 2.96 +/- 0.27 mumol/g of Hb in cats fed the high dose. There was no significant increase in methemoglobin concentration. These results indicated that PG cannot be considered innocuous even at concentrations consumed by cats eating commercial diets. Heinz body-induced acceleration of RBC destruction develops in a dose-dependent manner, so that cats with greater food intake, ie, lactating queens and nursing kittens, are at greater risk for development of PG-induced Heinz body hemolytic anemia.     

Passage of chromium-mordanted and rare earth-labeled fiber: time of dosing kinetics

Coastal bermudagrass hay was labeled with Cr by the Cr-mordant procedure and with 177Lu applied to the same fiber. Neutral detergent fiber prepared from the same Coastal bermudagrass hay was labeled with Yb, 169Yb, Tb and 160Tb by soaking overnight following by thorough washing and drying. Wood chips were similarly labeled with Sm or La, and Solka Floc was labeled with 147Nd and 141Ce. The carriers, labels and times of administration to cattle were: bermudagrass fiber with both Cr and 177Lu, bermudagrass fiber with 169Yb and Solka Floc labeled with 147Nd at 0 h; bermudagrass fiber with Yb, Solka Floc with 141Ce and wood chips with Sm at 24 h; wood chips with La at 48 h; and bermudagrass fiber labeled with 160Tb at the beginning and labeled with Tb at the end of a meal. Fecal collection followed and passage characteristics were determined with a two-compartment, age-dependent model. Markers labeling the different fiber sources had different (P less than .01) passage rates (Solka Floc greater than Coastal bermudagrass greater than wood chips), but there was no difference within fiber source for rare earth passage. There also was no difference between the passage characteristics of Cr-mordant and 177Lu. However, passage rate of particles administered at the beginning of the meal (160Tb) was 42% higher than for particles at the end of the meal (Tb).(ABSTRACT TRUNCATED AT 250 WORDS)     

Serving capacity of crossbred yearling beef bulls. II. Summer grazing activity and body temperature patterns during average and heavy mating loads at pasture

Ten low (LSC) and 10 high (HSC) serving capacity yearling bulls were exposed individually to 25 naturally cyclic (N) cows for 3 d (average mating load) and subsequently to 9 estrus-synchronized (S) cows for 1 d (heavy mating load) in a randomized complete block design consisting of 10, 4-d blocks. Bulls were fitted with vibracorders and temperature acquisition modules to record grazing activity and body temperature (BT), respectively. During the N cow treatment, LSC bulls had fewer (P less than .05) services per cow and a higher mount to service ratio than HSC bulls, and LSC bulls tended (P = .12) to graze less total time than HSC bulls (7.8 vs 9.0 h/d, respectively). However, both groups of bulls exhibited similar diurnal grazing patterns with two major daily grazing periods; the first (0400 to 1300) peaked early in the morning (0600) and the second (1700 to 2200) occurred in late afternoon and evening. During the S cow treatment, LSC and HSC bulls did not differ (P = .60) in grazing time or pattern, but similar mating activity was exhibited by both SC groups while exposed to S cows. Bulls grazed little during the hottest part of the day (1300 to 1700), and LSC and HSC bulls spent only 2.2 and 2.4 h, respectively, grazing during dark, cooler periods near midnight. Body temperature did not differ between SC groups and averaged 39.3 degrees C during N cow treatment and 39.4 degrees C during S cow treatment. Body temperature had a distinct, monophasic diurnal pattern in bulls exposed to an average mating load.(ABSTRACT TRUNCATED AT 250 WORDS)