Genetic relatedness of a rarely isolated Salmonella: Salmonella enterica serotype Niakhar from NARMS animal isolates.

BACKGROUND: In the United States, Salmonella enterica serotype Niakhar is infrequently isolated. Between 1997 and 2000, the animal arm of the National Antimicrobial Resistance Monitoring System-Enteric Bacteria (NARMS) assayed a total of 22,383 Salmonella isolates from various animal sources (swine, cattle, chickens, turkeys, cats, horses, exotics and dogs) for antimicrobial susceptibility. Isolates originated from diagnostic and non-diagnostic submissions. OBJECTIVES: To study the phenotypic and genotypic characteristics of Salmonella Niakhar. METHODS AND RESULTS: Only five (0.02%) of the 22,383 isolates were identified as Salmonella Niakhar. Antimicrobial resistance testing indicated that three isolates were pan-susceptible, one isolate was resistant to ampicillin and one isolate was resistant to ampicillin, chloramphenicol, ciprofloxacin, kanamycin, nalidixic acid, streptomycin, sulfamethoxazole, tetracycline and trimethoprim/sulfamethoxazole. RAPD-PCR analysis, PFGE and ribotyping indicated that two pan-susceptible isolates were genetically similar, whereas the three remaining isolates were genetically different. The one Salmonella Niakhar isolate that was multiresistant harboured a class I integron, intI1 and two large plasmids. CONCLUSIONS: This study represents the first report of a ciprofloxacin-resistant Salmonella isolate from the animal arm of NARMS.

Presence of Campylobacter jejuni in various organs one hour, one day, and one week following oral or intracloacal inoculations of broiler chicks.

Day-old broiler chicks (n=30) were obtained from a commercial hatchery and inoculated, either orally or intracloacally, with a characterized strain of Campylobacter jejuni. At 1 hr, 1 day, and 1 wk after inoculation, broilers (n = 5) from the orally and intracloacally inoculated groups along with control birds (n=4) were humanely killed by cervical dislocation. The broilers from the control and treatment groups were aseptically opened, and the thymus, spleen, liver/gallbladder, bursa of Fabricius, and ceca were aseptically removed and individually analyzed for C. jejuni. Overall, C. jejuni was isolated after oral inoculation from 13% (10/ 75), 17% (13/75), and 28% (14/50) of the 1-hr, 1-day, and 1-wk samples, respectively. Campylobacter jejuni was isolated from 10% (4/ 40), 8% (3/40), 10% (4/40), 25% (10/40), and 40% (16/40) of the thymus, spleen, liver/gallbladder, bursa of Fabricius, and ceca samples, respectively. Following the intracloacal route of inoculation, C. jejuni was recovered from 32% (24/75), 8% (6/75), and 16% (8/50) of the 1-hr, 1-day, and 1-wk samples, respectively. Campylobacter jejuni was isolated from 5% (2/40), 5% (2/40), 5% (2/40), 45% (18/40), and 40% (16/40) of the thymus, spleen, liver/gallbladder, bursa of Fabricius, and ceca samples, respectively, for all sampling periods. Campylobacter spp. were not recovered from sample sites examined from the control broilers from trial one, trial two, or trial three samples examined after 1 hr and 1 day. However, one control sample was positive from the 1-wk sampling from repetition three; therefore, those data were omitted. The rapid movement of Campylobacter to internal organs following both oral and intracloacal inoculation may be significant, particularly if it persists in these organs as reservoirs throughout the 65-wk life cycle of breeding birds.

Pulmonary hypertension syndrome in young chickens challenged with frozen and autoclaved cultures of Enterococcus faecalis.

Enterococcus faecalis, when administered in a growth medium or sterile saline, will cause pulmonary hypertension syndrome (PHS) in chickens. The objective of this study was to determine if frozen and/or autoclaved cultures of E. faecalis retain ability to evoke PHS. In Trial 1, chicks were inoculated with 3.6 x 10(7) E. faecalis (IA) in tryptic soy broth (TSB) from either a live culture or one that had been autoclaved (120 degrees C for 20 min). Controls received TSB. Autoclaved and live cultures produced the same degree of PHS in a majority of the birds. Trial 2 used the same protocol, except a frozen (-70 degrees C for 60 min) culture of E. faecalis was compared with the control. The results agreed with those of Trial 1, i.e., the frozen culture also produced PHS. Trial 3 was conducted to determine if E. faecalis caused PHS by producing and releasing some unknown substance into the supernatant. Incidence of PHS was based on percentage of birds exhibiting ascites fluid at 24 hr after challenge. Controls received sterile, frozen, or autoclaved TSB. As compared with controls, those birds that received challenge with E. faecalis alone, supernatant alone, and E. faecalis plus supernatant from live cultures exhibited similar incidence of ascites, whereas birds that received E. faecalis plus supernatant and supernatant alone from cultures that had been either frozen or autoclaved exhibited elevated incidence of ascites as compared with controls. Also, with frozen and autoclaved cultures, those birds that received only pelleted E. faecalis exhibited incidence of ascites that did not differ from controls. Apparently, E. faecalis produces PHS in chicks by producing and releasing an unknown toxin.

Bacteria in heart and lungs of young chicks.

AIMS: The purpose of these experiments was to determine whether the heart and lungs of young chicks harboured bacteria. METHODS AND RESULTS: Samples of the heart and lungs were aseptically removed from chicks on scheduled sampling days. Experiment 1 showed that of the 360 birds evaluated during the late embryonic and early post-hatching periods, only 10.8% harboured bacteria in the heart, lungs, and heart and lungs simultaneously. Experiment 2 suggested that bacteria in these organs were transient. Twenty-three bacterial species were found in the hearts whereas 30 were found in the lungs. Experiment 3 showed that only 1.4% of embryos harboured bacteria in the yolk, albumen, heart and lungs whereas 12.9% of the embryos had bacteria in the air cell. CONCLUSIONS: During the post-hatching period, there was a higher incidence of bacterial isolation in the heart and lungs, whilst during the embryonic development period, there was a lower incidence of bacterial isolation from these two organs. Results suggested that the heart and lungs do not have a residual bacterial flora; rather, opportunistic bacteria occasionally pass through these tissues. SIGNIFICANCE AND IMPACT OF THE STUDY: These experiments proved that bacteria could be isolated in the heart and lungs of healthy chicks reared from E17 to 3 weeks of age.

Morphological changes in heart and lungs of broilers experiencing pulmonary hypertension syndrome caused by Enterococcus faecalis.

In a previous report, a method of identification of birds experiencing early symptoms of pulmonary hypertension syndrome (PHS) caused by challenge with Enterococcus faecalis was delineated. This method involved subjective heart scores based on visual observation of a cavity on the external surface of the right ventricular wall (RVW), as well as tonicity and thickness of this wall. Accuracy in identifying birds 48 h postchallenge with E. faecalis was acceptable. However, this method did not attempt to offer other morphological or physiological characteristics for further understanding the etiology of PHS. In the present study, three trials were conducted to establish morphological characteristics of the heart from birds challenged with E. faecalis. In Trials 1 and 2, discrepancies were found in heart length (HL) and thickness of the RVW. In Trial 3, the dry weight of the right ventricle (RV) increased after challenge with E. faecalis, as was the ratio of the mass of the RV to the mass of the total ventricle (TV). Histopathological evidence of hearts and especially lungs of birds challenged with E. faecalis were suggestive of PHS. Results indicated that RV, RV:TV ratio, and histopatholgical evaluation of heart and lungs are complementary to diagnosis of PHS.

Biochemical and immunological changes in chickens experiencing pulmonary hypertension syndrome caused by Enterococcus faecalis.

Recent studies have proven that Enterococcus faecalis (1.5 x 10(7) live bacteria from a tryptic broth culture given s.c. or intra-abdominally (IA) to 5-wk-old broilers) caused pulmonary hypertension syndrome (PHS) in 97% of the birds within 48 h. Definitive diagnosis of PHS was made at necropsy by observing a cavity on the surface of the right ventricular wall and by increased ratio of left ventricular weight to total ventricular weight. A nonlethal method of diagnosing PHS would enhance the study of PHS and alert production poultrymen to the onset of ascites (waterbelly), which is the cuLminating event of PHS. In the present study, serum hemoglobin, glucose, protein, cholesterol, aspartate amino transferase (AST) and creatine kinase-MB (myocardial in origin) enzymes, differential leukocyte numbers, and specific antibody levels against Ent. faecalis were evaluated as nonlethal diagnostic indicators of PHS. Decreases in serum protein and cholesterol of 3 and 10%, respectively, plus increases in percentages of basophils and monocytes of 18 and 40%, respectively, appear to indicate that PHS has been initiated. An agglutinating antibody, specific against Ent. faecalis, but not against other closely related bacteria, has been developed. Presence of this antibody in a bird means that the bird has previously encountered Ent. faecalis. Thus, this antibody may become a diagnostic for PHS in fast-growing chickens.

Pulmonary hypertension syndrome in broilers caused by Enterococcus faecalis.

A field strain of Enterococcus faecalis was administered to broiler chicks at doses of 0, 3 x 10(6), 1.5 x 10(7), and 2 x 10(7) bacteria/bird either intra-abdominally or intravenously. In trials 1 to 3, birds were reared communally in a broiler house on pine shaving litter. In trial 4, challenged and control birds were maintained in separate isolation rooms in metal cages with raised wire floors. Challenged birds exhibited a characteristic cavity or depression in the external wall of the right ventricle. A subjective scoring system was devised to quantify challenge effects by assigning each heart a score of 1 to 4. The average number of birds, over all trials and over all dose levels, exhibiting the ventricular cavity was 93%. This value in controls was 5%. The average heart score for challenged birds was 3.1, and that for controls was 0.20. Heart scores of challenged and control chicks were not different in birds reared communally or in separate isolation rooms. Additionally, both routes of administration were equally effective. Results suggest that challenge with E. faecalis caused pulmonary hypertension.

Stress and nutritional quality of broilers.

Broiler chicks were reared in environmental chambers. All birds were started under ideal conditions, i.e., 30.6 C with 35% RH. Beginning at Day 36, half of the chicks were maintained at 24 C and 35% RH. The other half were subjected to a cyclic temperature-RH regime that approximated a typical August day in central Mississippi (heat treatment). Half of each of the described groups received implants of osmotic pumps that released adrenocorticotropin (ACTH) at 8 IU/kg BW/d for 7 d. The remaining birds received placebo pumps. The main effects of ACTH and heat treatments were similar. Both treatments caused reductions in BW, carcass weight (CW), carcass protein (CP), and muscle calorie (C) content. ACTH, but not heat, reduced carcass moisture (M). Carcass fat and ash, however, were not affected. Most changes were not reversed after 1 wk of recovery. Although visible signs of pale, soft, exudative muscle (PSE) were present, "white" areas of muscle were absent. The decreased meat yield and detrimental changes in meat quality suggest that stress, whether induced hormonally or by exposure to over-heating, caused losses that were as severe as those associated with PSE under field conditions.