Influence of bacteriocin-like substance, generation times, and genetic profiles of Listeria innocua on the isolation of Listeria monocytogenes

Inhibition of isolation of Listeria monocytogenes by bacteriocin-like substance (BLS)-producing Listeria innocua after enrichment culture was investigated. When 26 L. monocytogenes strains were examined in combination with eight L. innocua strains using the spot on lawn method, 52/208 (25.0%) combinations showed the growth inhibition of L. monocytogenes. When two Listeria species were cultured simultaneously in selective enrichment broth, inhibition of isolation of L. monocytogenes was observed in 12/52 of the combinations at 24h (23.1%), in 24/52 at 48h (46.2%) and in 30/52 (57.7%) after 7 days of incubation. The randomly amplified polymorphic DNA profiles showed no interstrain similarities between either strains of the BLS-producing L. innocua or the BLS-sensitive L. monocytogenes strains. Therefore inhibition by BLS-producing L. innocua of isolation of L. monocytogenes after enrichment culture is unlikely to be dependent upon a particular genetic profile.     

The marked increase of Listeria monocytogenes isolation from contents of swine cecum

The actual prevalence of Listeria monocytogenes from contents of swine cecum was investigated. The efficiency of Listeria enrichment broth (LEB) for isolation was examined by the recovery of artificially inoculated L. monocytogenes in contents of swine cecum. The numbers of organisms did not increase after 48 h incubation, but increased when the rapid decrease in pH of the LEB was adjusted. Between 1991 and 1993, 250 contents of swine cecum were examined for the prevalence of L. monocytogenes using LEB enrichment, either with or without pH adjustment. L. monocytogenes was isolated from 74 samples in 1993 with pH adjustment, however, no organisms were isolated in 1991 and 1992. It was suggested that the marked rise of the L. monocytogenes isolation was due to the spread of the organism among swine. Furthermore, 67 out of the 74 isolates were identified as 1/2c by serotyping. The serovar 1/2c strains showed genetic diversity by random amplified polymorphic DNA.     

Relationship between growth and plasma concentrations of ghrelin and growth hormone in juvenile beagle dogs

Although the release of growth hormone (GH) is known to be regulated mainly by GH-releasing hormone (GHRH) and somatostatin (SRIF) secreted from the hypothalamus, ghrelin also may be involved in GH release during juvenile period. We have examined plasma concentrations of acylated ghrelin, desacyl ghrelin, and GH in juvenile beagle dogs. Plasma acylated and desacyl ghrelin levels changed through aging; however, there was no closely correlation between ghrelin, body weight and circulating GH levels during juvenile period. The increase in body weight was essentially linear until 8 months of age, whereas plasma GH concentrations exhibited bimodal peaks for the meanwhile. The results suggest that ghrelin may not play internal cueing in GH secretion in juvenile beagle dogs.     

Occurrence of a new microsporidium in the skeletal muscle of the flying fish Cypselurus pinnatibarbatus japonicus (Exocoetidae) from Yakushima, Japan

A new microsporidium was observed in the flying fish Cypselurus pinnatibarbatus japonicus (Franz) (Exocoetidae) from Yakushima, Japan. Visual examination revealed the microsporidium to form white elongate nodules in the host's trunk muscle. Monomorphic spores were ovoid to pyriform in shape, with average dimensions of 4.1 x 2.2 microm and possessing a polar tube describing 13-15 coils. Histological observations showed that each parasite focus of infection was encapsulated by a host-produced fibrous membrane. The presence of sporophorous vesicles was not clearly determined. Ribosomal DNA sequence analyses showed the microsporidium to be discrete from other known fish muscle-infecting species and to be most closely related to a clade comprising the Pleistophoridae and Glugea spp. The parasite is provisionally placed as Microsporidium cypselurus sp. n.     

Ultrastructural study on the epithelial responses against attachment of indigenous bacteria to epithelial membranes in peyer's patches of rat small intestine

The ultrastructure of epithelial responses against the membrane adhesion of indigenous bacteria was investigated in the follicle-associated epithelium (FAE) of rat small intestine. The most frequent adherence of the various morphological types of bacteria to the epithelial membranes was found at the apex of the FAE. The attachment sites were deeply invaginated, and their bottoms were deformed into a sharp cone shape. Four layers with different electron densities were formed just beneath the apical membranes by microfilaments which surrounded the invaginations. The electron density of each layer was gradually decreased as being apart from the invaginations. The extremities of some bacteria in the invaginations were deformed into sharpened shapes. The cell walls of the extremities of the bacteria were occasionally dissolved in the invaginations, and their cytoplasms were slightly swollen with low electron densities. In some invaginations, the attached bacteria were eliminated to leave their fragments such as filamentous debris and a part of cell walls. Finally these remnants disappeared completely. When the bacterial colonies existed in the middle region of the FAE, the attachment of bacteria resulted in the engulfment of bacteria by M cells. The degenerated bacteria whose cytoplasmic matrices were separated into high electron dense materials and cleared materials were occasionally engulfed by ordinary microvillous columnar epithelial cells or goblet cells throughout the FAE. These findings suggest that the epithelial cells reject the attachment of live indigenous bacteria and that the M cells absorb indigenous bacteria in rat Peyer's patches.     

Effect of dietary copper on intestinal mucosa enzyme activity, morphology, and turnover rates in weanling pigs.

Twenty-four pigs from four litters weaned at 21 d of age (6.6 kg of BW) were used to evaluate the influence of 250 ppm of dietary Cu on intestinal mucosa glucose-6-phosphatase (GP), alkaline phosphatase (AP), and adenosine triphosphatase (ATPase) activity; mucosal morphology; and the turnover rate of the intestinal mucosa throughout the gastrointestinal tract. Pigs were allotted into four pens of six pigs each based on sex, litter, and weight. Pens were then assigned to one of two treatments: 1) corn-soybean meal-whey diet with no antimicrobials (CO), or 2) CO + 250 ppm of Cu. Pigs were fed twice daily an amount approximately equal to ad libitum intake for 14 d. On d 14, pigs were injected i.p. with [3H]thymidine (50 microCi/kg of BW) 10 h after the morning meal. One pig from each pen was euthanatized at 1, 6, 12, 20, 32, and 44 h postinjection, and intestinal tissue was collected from the duodenum, two jejunum sites (upper and lower), ileum, cecum, and colon. The activity of GP and AP in the lower jejunum tended to decrease in pigs fed Cu (P less than .11, P less than .08, respectively). The ATPase activity was not affected by treatment (P greater than .10). Crypt death, villus height, or epithelial cell size (P greater than .10) were not affected by feeding Cu. Migration rate of epithelial cells up the villus was also not affected by treatment (P greater than .10).(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of zeolite a or clinoptilolite in diets of growing swine

Growth, nutrient balance, plasma ammonia levels and urinary p-cresol excretion were evaluated in growing pigs fed diets containing various levels of zeolite A or clinoptilolite. In one growth trial, crossbred pigs averaging 25 kg initial body weight were assigned to diets containing no zeolite, .3% zeolite A or .5% clinoptilolite for a 6-wk growing phase trial. Average daily gain (ADG), average daily feed intake (ADF) and feed/gain (F/G) were unaffected by supplementation of either zeolite in the diet, but metabolizable energy (ME) utilization was improved by feeding diets containing either zeolite. A second growth trial utilized the same crossbred pigs, which averaged 65 kg initial body weight, and were assigned to diets containing no zeolite, 1% zeolite A or 5% clinoptilolite for an 8-wk finishing phase trial. Average daily gain, ADF, and ME utilization were unaffected by feeding either zeolite diet, while F/G was increased in pigs fed the diet containing 5% clinoptilolite. In two nutrient balance trials, 16 crossbred pigs averaging 7.5 kg in initial body weight were fed diets containing 0, 1, 2 or 3% zeolite A in one trial and 16 crossbred pigs averaging 7.0 kg initial body weight were fed diets containing 0, 2.5, 5.0 or 7.5% clinoptilolite in a second trial. In both trials, digestible energy, ME, N-corrected ME and ME corrected for N balance and zeolite levels were linearly reduced as increasing amounts of either zeolite were fed. Daily fecal N increased and apparent digestibility of N was linearly reduced by feeding increasing amounts of zeolite A or clinoptilolite. Biological value of protein was improved linearly as higher levels of zeolite A were fed, indicating that there may be some ammonia binding to zeolite A in the gastrointestinal tract. Net protein utilization was reduced by feeding increasing levels of clinoptilolite in the diet. Calcium, P, Mg, Na, K and Fe retentions were linearly reduced by feeding increasing amounts of zeolite A in the diet, while increasing levels of clinoptilolite caused only P retention to be linearly reduced. Both free and conjugated forms of urinary p-cresol were linearly reduced by feeding increasing levels of clinoptilolite. Plasma ammonia levels were reduced at subsequent bleedings after a meal and by increasing levels of clinoptilolite.     

Ruminal and plasma concentrations of 3-methylindole associated with tryptophan-induced pulmonary edema and emphysema in cattle.

Five Hereford cows were given an intraruminal dose of L-tryptophan (0.35 g/kg of body weight), and 2 cows were used as controls. Of the 5 treated cows, 3 developed clinical signs of interstitial pul monary edema, and emphysema and severe pulmonary lesions were seen at necropsy after 96 hours. Another cow developed moderate clinical signs and pulmonary lesions, and the remaining cow had few clinical signs and mild pulmonary lesions. The severity of clinical signs in each cow was related to the severity of pulmonary lesions at necropsy. The 3-methylindole (3MI) was present in ruminal fluid and plasma within 6 hours after administration of tryptophan, and the concentrations increased to 3.0 and 9.0 mug/ml within 12 to 24 hours. Severity of pulmonary lesions was related to maximal concentration and duration of 3MI in the plasma. At necropsy, gross lesions were characterized by diffuse, pulmonary edema and interstital emphysema; and the lungs were dark red, firm, and heavier than normal. Predominant microscopic changes included accumulation of proteinaceous residue, hypertrophy and hyperplasia of alveolar lining epithelium, thickening of alveolar septums, and emphysematous thickening of interstitial tissues. These changes were similar to previously reported 3MI-induced pulmonary lesions. The presence of 3MI in ruminal fluid and plasma after administration of tryptophan and the relationship between concentration of 3MI and severity of clinical signs indicate that 3MI is the principal metabolite of ruminal fermentation which leads to the development of acute pulmonary edema and emphysema in cattle given tryptophan.