DNA fingerprinting of Australian isolates of Mycobacterium avium subsp paratuberculosis using IS900 RFLP

OBJECTIVES: To evaluate additional restriction enzymes for IS900 RFLP of Mycobacterium avium subsp paratuberculosis and examine the genetic diversity among Australian isolates for epidemiological studies of Johne's disease. DESIGN AND PROCEDURE: Seventy-one isolates of M paratuberculosis from cattle, sheep, goat, alpaca and rhinoceros in six Australian States and the Northern Territory, reference strains and reference DNA from previously characterised strains were tested for genetic variation. Bst EII, Pvu II and Pst I restriction enzymes were used, and four others (Bam HI, Alu I, Xho I and Dra I) were assessed for their ability to detect polymorphisms. Multiple isolates from some animals were tested. RESULTS: Bam HI, was the most effective enzyme for identifying polymorphisms (12 types), followed by Bst EII (11 types). Both Pvu II and Pst I were relatively ineffectual. Fifteen different types were identified, 12 in clinical isolates. Most isolates were cattle (C) strains and fell into the C1 (n = 28) and C3 (n = 32) groupings. All isolates from alpaca were type C1, and bovine isolates were commonly C1 (n = 15) or C3 (n = 28). All of the sheep were infected with sheep (S) strains; no S strains were identified in cattle. Two of six isolates from one animal had single band differences. CONCLUSION: The epidemiological features of M paratuberculosis in Australia are similar to those reported in New Zealand, where cattle and sheep are commonly infected with different strains. However, because of the lack of polymorphism identified within the major groups, it is unlikely that DNA fingerprinting will have a significant role in epidemiological studies of Johne's disease, unless an unusual strain in being studied.     

Bacterial infection of the lower respiratory tract in 34 horses

OBJECTIVE: To investigate associations between the bacteriology and aspects of history, clinical presentation, outcome and pathology of lower respiratory tract disease of 34 horses. PROCEDURE: Detailed aerobic and anaerobic bacteriological investigations were performed on clinical specimens from horses with pneumonia, lung abscessation and necrotic pneumonia with or without pleurisy in an attempt to identify those bacteria that might contribute to the initiation and progression of infection. RESULTS: Bacteria were cultured from 33 of the 34 horses. In ten cases, only aerobic/facultatively anaerobic isolates were cultured while aerobic/facultatively anaerobic bacteria and obligately anaerobic bacteria were isolated in the other 23 cases. Moderate to large numbers of anaerobic bacteria were isolated only when the estimated duration of illness was at least five days. Bacteria were not cultured from 12 of the pleural fluid samples but were always cultured from pulmonary samples (either transtracheal aspirates from live horses or pulmonary lesions at necropsy). Streptococcus equi subsp zooepidemicus was isolated in the three cases where only one bacterial species was cultured. In the other 30 cases, multiple species were isolated. These included most often and in greatest numbers, Streptococcus equi subsp zooepidemicus, Pasteurellaceae, Escherichia coli, anaerobic cocci, Eubacterium fossor, Bacteroides tectum, Prevotella heparinolytica, Fusobacterium spp, and pigmented members of the genera Prevotella and Porphyromonas. Aerobic/facultatively anaerobic organisms were isolated from 97% of horses, while obligately anaerobic organisms were cultured from 68% of horses. CONCLUSION: There was no association between the isolation of any specific bacterium and the outcome of disease. However, obligately anaerobic bacteria (such as anaerobic cocci, Bacteroides tectum, P heparinolytica and Fusobacterium spp) and the facultatively anaerobic species Escherichia coli, were recovered more commonly from horses that died or were euthanased than from those that survived. There was an association between failure of horses to recover from pleuropneumonia and delay in diagnosis and initiation of treatment.     

Analysis of semen quality from each testis/duct system of a turkey with a duplicated cloaca and vent

A case is described wherein a turkey male (tom) possessed two vent areas with duplicated cloacae served by one large intestine. Both cloacae were functional in that feces were excreted and semen could be collected from each. The left vas deferens and ureter emptied into the left cloaca, and the right vas deferens and ureter emptied into the right cloaca. This allowed semen from each testis/duct to be collected separately from the corresponding cloaca. Thus, a unique opportunity was presented to collect semen separately from the left and right testis/duct system for semen analysis and fertility determination. Sperm concentration and percentage of dead sperm were not significantly different when semen from left vs. right reproductive tract were compared. The concentrations of spermiophages in semen from both reproductive tracts fell into the range reported for normal semen (0-8 x 10(5)/ml); however, semen from the right side had consistently higher spermiophage concentrations than that from the left side. On the basis of observations from one male made possible by an anatomic anomaly, it appears that the fecundities of the left and right testis/duct systems of the turkey male do not significantly differ and that recruitment of spermiophages into one tract (because of immunologic challenge, etc.) does not necessarily mean that the opposite testis/ducts will respond similarly.     

Effect of short periods of high incubation temperature on hatchability and incidence of embryo pathology of turkey eggs

1. Turkey eggs were incubated at 38.0 degrees and 38.5 degrees C at different stages of embryo development and for periods of 3 to 25 d. Results were compared with control eggs incubated at 37.5 degrees C. The age of mortality, the incidence of malpositions and the incidence of morphological abnormalities were recorded from all unhatched eggs. 2. Eggs incubated at 38.5 degrees C for 5 or more days hatched significantly less well than eggs incubated at 37.5 degrees C. Eggs incubated at 38.5 degrees C for 3 d hatched worse than controls but not significantly so. Eggs incubated between 0 and 25 d and 7 and 12 d but not between 0 and 6, 13 and 18 and 19 and 25 d, at 38.0 degrees C had significantly lower hatchabilities than eggs incubated at 37.5 degrees C. 3. Lowest hatchabilities were obtained when the eggs were incubated at high temperatures between 7 to 12 d and 6 to 10 d, indicating that embryos at this stage of development are more likely to succumb to high temperature than at other ages. 4. Increases in embryo mortality due to overheating were seen in weeks 3 and 4 of incubation and at the pipping stage. This was observed even when the period of overheating occurred in weeks 1 and 2 of incubation. 5. Embryos malpositioned with their head in the small end of the egg were seen at a higher incidence when overheating in the 2nd or 3rd quarter of the incubation period.     

A case study on Salmonella enteritidis (SE) origin at three egg-laying farms and its control with an S. enteritidis bacterin

In the early 1990s, three egg-laying farms (farms S, T, and B) were thought to have the possibility of Salmonella enteritidis (SE) contamination because positive liquid egg samples originated from those farms. The present study was therefore conducted. The first clarification for SE contamination was the study on the origin of SE contamination including its vertical transmission. The results of SE contamination profiling with dust and manure, food materials, dead embryos, and residual yolks on hatch day in regular monitoring over a few years were clearly negative. Therefore, we concluded the SE transmission/infection was attributed to horizontal infection in the egg-laying farms but not vertical transmission from parental stock, hatcheries, growth, or food materials during a 7-yr experimental period. Second, we attempted to clarify if administration of an SE bacterin (Layermune SE) to growth flocks for the egg-laying farms could reduce SE incidence in liquid egg samples from each egg-laying farm. In the first experiment, we compared SE incidence in liquid egg samples from vaccinated and nonvaccinated flocks (similar age flocks). SE incidence from vaccinated and nonvaccinated flocks showed negative and <2 most probable number (MPN)/100 ml for farm B, <2 and >1600 MPN/100 ml for farm S, and negative and >1600 MPN/100 ml for farm T, respectively. In the second experiment, we compared the SE isolation incidence in the liquid egg samples from nonvaccinated and newly replaced vaccinated flocks in the same chicken houses from each of the three egg-laying farms. SE incidence in the liquid egg samples was similar to that in the first experiment. Therefore, the SE bacterin may play an important role in reducing the SE incidence of liquid egg samples.     

The effect of animal health products on the formation of injection site lesions in subprimals of experimentally injected beef calves

Two hundred and twenty beef calves were used in an experimental study to determine the occurrence of injection site lesions at slaughter (15 to 18 months of age) following subcutaneous and intramuscular injection of various products into the top hip (top butt), thigh (round), and neck or rib of calves at birth, branding, or weaning. Products tested were: 2 different preparations of selenium; a 2-way, a 7-way, and an 8-way clostridial bacterin; 2 combination 7-way clostridial and Haemophilus somnus bacterins; 2 H. somnus bacterins; 2 different 4-way modified-live viral respiratory vaccines; a 4-way killed viral and H. somnus vaccine; and penicillin, florfenicol, ceftiofur, trimethoprim-sulfa, and tilmicosin. The occurrence of lesions, number of steaks affected with lesions, the trim weight of lesions, the histological class of lesions, and the estimated economic losses are described. Generally, products administered subcutaneously in the neck produced minimal tissue damage and economic losses.     

Differential effects of dexamethasone and clenbuterol on rat growth and on beta2-adrenoceptors in lung and skeletal muscle

Beta-adrenergic agonists increase growth rate, but their efficacy is reduced over time as the number of beta2-adrenoceptors in muscle decreases. Dexamethasone increases beta2-adrenoceptor density in many tissues, but this effect has not been reported in skeletal muscle. In this study, male rats were treated daily for 10 d with either clenbuterol (4 mg/kg of feed), dexamethasone (.2 mg/kg BW, s.c.), or clenbuterol plus dexamethasone. Untreated rats served as controls. Dexamethasone caused a marked suppression of growth rate, which resulted in decreased (P < .001) body weight (-29%), carcass weight (-30%), hind-limb muscles (-22%), omental fat (-22%), and heart weight (-10%). Feed intake was reduced (-26%), but feed conversion efficiency was also impaired (P < .001). Clenbuterol caused a small increase in growth rate (+6%; P < .05), with an increase in leg muscle (+7%; P < .01) and heart mass (+8%; P < .05). Feed efficiency was improved (P < .001) by clenbuterol. Rats given the combined treatment still showed a reduction in growth rate (-81%). Clenbuterol caused only a mild attenuation of the effects of dexamethasone on feed intake, BW, and carcass weight, but reduced the catabolic effect of dexamethasone on hind-limb muscle to only -8%. Clenbuterol caused a slight increase in the affinity beta2-adrenoceptors in lung for binding to the radioligand (-)[125I]iodocyanopindolol. Relative to control values, the density of beta2-adrenoceptors in lung was +31% with dexamethasone treatment, -45% with clenbuterol, and -23% with the combined treatment. Clenbuterol also decreased beta2-adrenoceptors in skeletal muscle (-35%), but so did dexamethasone (-13%), so the effects of the beta-adrenergic agonist were not attenuated through use of the combined treatment (-40%). The results show that the inductive effect of glucocorticoids on beta2-adrenoceptors is tissue-specific and that glucocorticoid treatment is not a useful adjunct to beta-adrenergic agonist treatment in animal production.     

Net portal appearance of volatile fatty acids in sheep intraruminally infused with mixtures of acetate, propionate, isobutyrate, butyrate, and valerate

The net portal appearance of volatile fatty acids (VFA) was investigated in four ruminally fistulated and multicatheterized sheep. During the experiments, the sheep were fed once every hour for 14 h and intraruminally infused with mixtures of VFA for the 12 h commencing 2 h after the initiation of the hourly feeding protocol. Paired arterial and portal blood samples were obtained hourly during the last 6 h of the experiments. In the control treatment (1), only water was infused intraruminally. In Treatments 2 through 4, the intraruminal infusion rates of propionate (40 mmol/h), isobutyrate (5 mmol/h), and valerate (5 mmol/h) were unchanged. In Treatments 2, 3, and 4, the acetate infusion rate was 100, 60, and 20 mmol/h, respectively, and the butyrate infusion rate was 10, 30, and 50 mmol/h, respectively. Thus, the infusion rate of VFA carbon was constant across Treatments 2 through 4. Portal recovery estimated from the increased net portal appearance in Treatments 2 through 4 compared to the control treatment was 85% for propionate and 60% for isobutyrate, and these recoveries were unaffected by treatment. The portal recovery of butyrate increased (from 21 to 32%) with increasing infusion rate of butyrate and decreasing infusion rate of acetate, as did the portal recovery of valerate (from 14 to 31%). The portal recovery of acetate was 55%, when measured as net portal appearance. Thus, it seems that the capacity for beta-oxidation in ruminal epithelium is limited, which would explain the increasing portal recovery of butyrate and valerate with increasing infusion rate of butyrate, when infusion rate of VFA carbon is unchanged.     

Artificial insemination outcomes in beef females using bovine sperm with a detectable fertility-associated antigen

In this study, semen samples from 25 bulls that had passed a breeding soundness evaluation were analyzed for the presence or absence of a 31-kDa protein, known as fertility-associated antigen (FAA), on spermatozoal membranes. Eighteen bulls had FAA on sperm (FAA-positive) and seven were devoid of FAA on sperm (FAA-negative). A single ejaculate from each bull was extended and frozen with 25 to 30 x 10(6) sperm in .5-mL straws. Crossbred replacement heifers (n = 865) were estrus-synchronized and artificially inseminated either at timed AI or 12 h after they were detected in estrus. Mature cows (n = 285) were inseminated 12 h after they were detected in estrus during a 45-d AI period. Pregnancy rates (pooled) to first AI service for females (n = 764) inseminated with FAA-positive sperm were 65.6% and were 49.7% for females (n = 386) inseminated with FAA-negative sperm (P < .005). Among the estrus-synchronized replacement heifers, pregnancy rates to synchronized AI service for heifers (n = 550) inseminated with FAA-positive sperm were 62% and were 45.7% for heifers (n = 315) inseminated with FAA-negative sperm (P < .005). These data indicate that pregnancy rates to first AI service at spontaneous and synchronized estrus are higher when using semen from bulls with detectable FAA on spermatozoal membranes compared to semen from bulls devoid of FAA on membranes. Fertility-associated antigen is an important determinant for fertility potential of sperm from bulls to be used in AI breeding programs.