Following the 1993 hantavirus pulmonary syndrome (HPS) epidemic in the southwestern United States, mammalogists and epidemiologists instituted long-term studies to monitor population density and prevalence of infection in rodents which constitute the reservoir for Sin Nombre virus (SNV). In this study, field techniques used in sampling small mammals for SNV infection were evaluated to determine if trapping and handling protocols were having significant effects on future trapability or mortality of animals. We compared rodent mark-recapture control plots, on which all rodents were simply measured, marked, and released on site, with experimental plots on which all animals were anesthetized with methoxyflurane, sampled for blood and saliva, measured, marked, and released. Blood samples were obtained from anesthetized animals on the experimental plots via a retro-orbital sinus puncture using a heparinized capillary tube. Dacron tipped oral swabs were used to collect buccal cells and saliva from the rodent's oral cavity. Field data were collected monthly from August 1994 to August 1996 at two sites in New Mexico (USA). Analyses were based on 3,661 captures of 1,513 individuals representing 21 species from three rodent families (Rodentia: Muridae, Heteromyidae, Sciuridae) and two species of rabbits (Lagomorpha: Leporidae). Overall, for most murid rodents (including five Peromyscus spp., Neotoma albigula, and Onychomys leucogaster) and one rabbit species (Sylvilagus floridanus), the handling/bleeding procedures had no significant effects on recapture rates or mortality. In contrast, several species of heteromyids (Dipodomys ordii and Perognathus flavus), one murid (Reithrodontomys megalotis) and one leporid (S. auduboni) suffered higher mortality rates, and heteromyid kangaroo rats (D. ordii and D. merriami) exhibited lower trapability as a result of the anesthesia and sampling procedures. In view of the overall non-significant influence of the sampling procedures on murid rodents, the anesthesia and blood/saliva sampling protocols described herein appear to be appropriate for hantavirus research, and may serve as a model for environmental monitoring of other zoonotic agents and their reservoirs.

The objective of this study was to develop a bait for delivering an oral rabies vaccine to free-ranging coyotes (Canis latrans) in southern Texas. Captive trials were conducted from January to April, 1994, to determine bait preferences and behavioral responses of coyotes (n = 42) to selected baits and attractants. Baits were hollow rectangular cubes made of polymer dog food or fish meal. Attractants had sweet (watermelon), fruity (raspberry), sulfurous (synthetic WU), and lard (beef lard) fragrances. Captive coyotes did not exhibit a preference for either bait bases or attractants; however, coyotes chewed dog food baits 1.6 times more than fish meal baits. Average proximity of coyotes eliciting a response to baits was 2.2 ± 1.3 m (x̄ ± SE). Captive coyotes readily accepted dog food baits containing 2 ml of liquid rhodamine B, a biological marker. Rhodamine B staining of the oropharyngeal region was evident in each captive coyote. Results from the field evaluation of baits and attractants were consistent with that of the captive trials. Of 2,070 bait station-nights conducted from February to April, 1994, coyotes comprised the greatest single species visitation and uptake rates with 31% and 28%, respectively. Bait uptake rates of free-ranging coyotes did not differ among bait-attractant combinations. Coyotes took baits 93% of the time they encountered a bait, regardless of bait type.

This study sought to develop a baiting strategy to deliver an oral rabies vaccine to free-ranging coyotes (Canis latrans) in southern Texas. To determine bait longevity, dog foodlard baits were placed (n = 50) on- and off-roads during July 1994 and January 1995. Coyote visitation and uptake rates did not differ between on-road and off-road placement of baits. To evaluate bait stations as possible visual cues, baits were placed out both with (n = 50) arid without (n = 50) bait stations. A visual cue of a bait station did not affect coyote response to baits. Bait longevity was shorter during July (≤4 days) than January because of consumption of baits by imported fire ants (Solenopsis invicta). The effect of two different bait densities on coyote acceptance rates was determined on six 93.5 km² study areas in southern Texas. Three study areas received a bait density of 19 baits/km² (50 baits/mi²) and the remaining three study areas received 58 baits/km² (150 baits/mi²). Coyote bait uptake rates, based upon the proportion of coyotes marked with either tetracycline hydrochloride or rhodamine B or both, were 83% (n = 99 coyotes) and 87% (n = 101 coyotes) for the 19 and 58 baits/km2 densities, respectively. Bait uptake rates did not differ (P > 0.54) between the two bait densities. Rodents and rabbits, which were fed baits containing tetracycline hydrochloride and a simulated oral rabies vaccine sachet containing rhodamine B, did consume the bait but not the rhodamine B sachet. These animals then were killed and fed to captive coyotes (n = 9). Canine teeth were extracted from coyotes and processed for tetracycline determination. Each coyote tested negative for tetracycline. Therefore, it was unlikely that coyote bait consumption rates were overestimated because of coyotes secondarily marking themselves by ingesting prey items that consumed baits.

Thirty-five free-ranging healthy spiny-tailed iguanas (31 Ctenosaura bakeri, 4 C. similis) and 14 green iguanas (Iguana iguana rhinolopha) were caught and held in captivity for 2 days. Blood was collected from all animals and their sera were evaluated for antibody titres against reptilian reoviruses, reptilian paramyxoviruses, and avian paramyxovirus-1 (PMV-1). Cloacal and pharyngeal swabs also were collected and examined for viral content by incubation on chicken embryo fibroblasts (CEF) and terrapene heart cells (TH-1). No virus was isolated from the pharyngeal and cloacal swabs on CEF and TH-1. Twenty-three (47%) of 49 sera samples tested positive for reptilian reoviruses by virus neutralization tests. Twenty (41%) of 49 samples had antibodies against one reptilian PMV isolate by virus neutralization tests and 3 (9%) of 34 by hemagglutination inhibition tests. No antibodies were detected against the other PMV isolate of reptilian origin nor against avian PMV-1. This is the first description of serum antibodies against reptilian reoviruses and PMV in wild iguanas.

Serum samples from 21 of 36 Eskimo harvested bowhead whales (Balaena mysticetus) were positive by virus neutralization (50% endpoint titer ≥1:28 and/or 100% endpoint titer ≥1:20) for antibodies to at least one virus serotype from the calicivirus family, vesicular exanthema of swine virus (VESV) and San Miguel sea lion virus (SMSV). Many animals were positive to more than one serotype when using the Spearman-Karber (S-K) method for calculating antibody titers. The most common serotype detected was VESV F55 with 6 of 36 (17%) by the Monto and Bryan (MB) titer calculation method, and 17 of 36 (47%) by the S-K titer calculation method. Vesicular exanthema of swine virus 1934B antibody was detected in 3 of 36 (8%) and 5 of 36 (14%) whales using the MB and S-K methods, respectively. Vesicular exanthema of swine virus J56 antibody was detected in 3 of 36 (8%) by the S-K method only. All whales <8.5 m (estimated yearlings, n = 6) were seronegative for VESV J56 and 1934B while 10% and 17% of the whales >8.5 m were positive, respectively. Whales assumed to be sexually mature (>13 m) had a higher prevalence of antibody to VESV 1934B and SMSV 8 than those <13 m. Gender had an effect on seroprevalence of antibody to VESV 1934B as titers ≥1:28 (S-K method) occurred in 18% of the females and 7% of the males. Antibody to other serotypes (SMSV 8 and 12) occurred less frequently (<6%) at an antibody titer ≥1:28 by the S-K method. All 36 whale sera were negative for antibody to VESV-A48, B51, C52, D53, E54, G55, H54, 155, and K54; Tillamook calicivirus, and dolphin morbillivirus; and SMSV-1, 2, 4, 5, 6, 7, 9, 10, 11, and 13 by the S-K method.

The 5′ untranslated region (5′ UTR) of cytopathogenic pestiviruses isolated from two seronegative roe deer (Capreolus capreolus) in northern Germany was partially sequenced and compared with those of 28 other pestiviruses. Due to the occurrence within a narrow location and the complete identity of the sequenced fragments from both roe deer isolates (SH9 and SH11) they seem to belong to the same bovine virus diarrhea virus (BVDV) strain called SH9/11. This strain is highly homologous (up to 93% identity) to “classical” BVDV strains. However, SH9/11 has characteristic variations in its 5′ UTR distinct from all other pestiviruses analyzed in this study. Strain SH9/11 is more similar to BVDV group I than to group II, although it is clearly separated from all other cattle isolates tested. In monoclonal antibody (mAb)-typing studies, isolate SH9 reacted with one pestivirus-specific mAb (C16), with two BVDV specific mAbs (N2B12 and D5), and with one mAb (f48) raised against the E2 protein of classical swine fever virus out of a panel of 13 mAbs. The separate position of strain SH9/11 again was demonstrated by the unique reaction pattern of isolate SH9 when compared with other mAb f48-positive BVDV and BDV strains. All these results indicate that distinct BVDV strains might exist among freeranging roe deer in Germany.

In order to determine the importance of wild felids in the epidemiology of Bartonella spp. infection, 136 Nobuto strips or serum samples from free-ranging mountain lions (Felis concolor) and bobcats (Felis rufus) captured in California (USA) between 1985 and 1996 were tested for B. henselae antibodies (titer ≥ 1:64) using an immunofluorescence test. Similarly, 124 serum samples from 114 captive wild cats representing 26 species or subspecies collected between 1991 and 1995 were retrieved from the serum banks of four California zoological parks. Fifty-three percent (33/62) of the bobcats, 35% (26/74) of the mountain lions, and 30% (34/114) of the captive wild felids (genera Acinonyx, Panthera and Felis) had B. henselae antibodies. In captive wild felids, prevalence varied widely among the species, but seropositivity was more likely to occur in the genus Felis than in the genus Acinonyx or Panthera. Prevalence was evenly distributed between sexes, except for free-ranging mountain lions. Antibody prevalence ranged from 25% in 0- to 2-yr-old captive felids to 35% in cats ≥9-yr-old, but the highest antibody' titers were observed in cats <5-yr-old.

In a captive breeding center near Rome (Italy), cases of embryonic and neonatal death were recorded during the breeding seasons in the European eagle owl (Bubo bubo), peregrine falcon (Falco peregrinus), buzzard (Buteo buteo), and lanner falcon (Falco biarmicus). Salmonella havana and S. virchow were isolated. Three pulli, clinically infected with S. havana, were successfully treated with enrofioxacin. From two groups of healthy 3- to 4-wk-old eagle owls, Salmonella sp. group 61 (61:r:-) and S. havana were collected. A strain of S. paratyphi B was detected in a pharyngeal swab and a fecal sample from an adult female goshawk (Accipiter gentilis), affected with pharyngeal trichomoniasis. A S. hadar strain was collected from a healthy 1-yr-old female eagle owl and S. livingstone was isolated from a 1-mo-old female peregrine, dead of an acute respiratory syndrome. Lesions of fibrinous polyserositis and multivisceral congestion were observed. From frozen 1-day-old chicks, on which adult and young raptors were fed, S. havana and S. livingstone isolates with similar biochemical and drug susceptibility patterns to those isolated from raptors were identified. A surveillance program on infectious diseases reduced embryonic and neonatal death rates in the following breeding seasons.

Fecal and cloacal swabs or feces of wild mammalian, avian and reptilian species, either farmed or free-ranging, and of racing pigeons (Columba livia) kept in lofts were cultured for Salmonella spp., Campylobacter spp., and Yersinia spp. Of 291 free-ranging mammals tested, 6 (2%) and 1 (<1%) yielded positive cultures of Salmonella spp. and Campylobacter spp., respectively. Salmonella newport was the predominant serotype isolated and the opossum (Didelphis marsupialis insularis) had the significantly highest prevalence (29%) of Salmonella spp. infection compared to other species such as deer (Mazama americana trinitatis), lappe (Agouti paca), tattoo (Dasypus novemcinctus), agouti (Dasyprocta leporina), and wild hog (Tayassu tajacu). Among 14 species of farmed wildlife studied, 13 (7%) and 10 (5%) of 184 fecal or cloacal samples tested were positive for Salmonella spp. and Campylobacter spp., respectively. Salmonella javiana accounted for 50% of the Salmonella spp. isolates and C. jejuni represented 90% of the Campylobacter spp. cultured. Only 1 (1%) of 124 cloacal swabs of free-flying avian species yielded Salmonella spp. compared to 21(17%) samples positive for Campylobacter spp. Of 171 racing pigeons which originated from 8 fanciers, 8 (5%) yielded Salmonella spp. all of which were serotype typhimurium while only 1 (1%) was positive for Campylobacter spp. Seven (88%) of 8 Salmonella spp. isolates were recovered from one fancier. Yersinia spp. was not cultured from any of the above samples. Although the prevalences of Salmonella spp. and Campylobacter spp. in wildlife in Trinidad are low, the practice of wildlife farming and the increased consumption of meat from wildlife may increase the health risk to human consumers.

To improve assessment of cellular immune responses in seals, northern elephant seal (Mirounga angustirostris) interleukin 2 (IL-2) has been characterized. The gene was cloned and sequenced from a 658 base pair (bp) cDNA generated from total RNA by reverse transcription-polymerase chain reaction (RT-PCR). The sequence encoded a 154 amino acid (aa) polypeptide that included a 20 aa putative signal peptide. Seal IL-2 was found to share considerable identity with published sequences. Nucleotide sequence analysis of phocine (seal) IL-2 with canine, feline, human, trichechine (manatee), bovine and murine sequences demonstrated 93, 92, 86, 82, 78 and 71% identity, respectively. Analysis of the derived amino acid sequences demonstrated 88, 89, 78, 71, 66 and 60% identity, respectively. Interleukin-2 sequence identities appear to reflect evolutionary proximity among the analyzed species, and importantly, those residues identified as critical to IL-2 biological activity and receptor binding are largely conserved. To examine the kinetics of IL-2 mRNA expression, northern elephant seal lymphocytes were stimulated with the mitogen concanavalin A (Con A), and RNA was collected at several time points thereafter. The RT-PCR demonstrated that seal IL-2 mRNA expression peaks in the first 8 hr following Con A stimulation. Lastly, genomic DNA from northern elephant seal, harbour seal (Phoca vitulina) and California sea lion (Zalophus californianus) was used as template to identify and clone genomic IL-2. Partial sequence of the genomic clones demonstrated nearly complete identity among the three species. Sequence identity indicates that probes constructed from the northern elephant seal IL-2 gene will be effective in assessing IL-2 in other pinniped species.

Pathologic examination of green turtles (Chelonia mydas) from the Hawaiian Islands (USA) was performed to determine the primary cause of mortality. Lesions were associated with fibropapillomatosis (FP) and/or spirorchidiasis (SP) in 16 of 17 green turtles examined. Gross lesions included moderate to severe emaciation, lobulated fibropapillomas of different size classes, serous atrophy of fat, and edema of subcutaneous tissue and muscle. Anasarca, hydropericardium and pulmonary edema were common findings. The neoplastic lesions observed in the gastrointestinal tract, lungs, liver, and kidneys of 29% of turtles examined were histologically characterized as fibromas. A generalized thickening and hardening of major vessels and thrombosis with partial or complete lumen occlusion were observed in turtles with FP and SP. Histologically, lymphoplasmocytic endarteritis was observed in vessels of turtles with both conditions. Multifocal granulomas were associated with trematode ova in the parenchyrna of most organs of all turtles with FP and SP. Spirorchidiasis and FP were considered the primary causes of mortality in the turtles examined. Further studies should focus on the pathogenic interaction of both conditions and their synergism as debilitating and fatal diseases in this threatened species.

During winter 1994–95, four and three sperm whales (Physeter niacrocephalus) were stranded along the Belgian and the Dutch coasts, respectively. Necropsies and tissue samplings were collected 24 hrs post mortem. Lesions on several whales included round and linear skin scars, ventral skin abrasions, acute skin ulcers, acute ulcerative stomatitides, acute to chronic external otitides, and passive visceral congestion. In addition, these sperm whales appeared to be debilitated with severe weight deficit, had blubber thickness reduction, the absence of abdominal fat, and the intestinal tracts were almost empty. Three categories of lesions and their possible relation with the stranding were evaluated. Cutaneous scars observed on the seven whales appeared to have no relation with the stranding. The poor body condition and acute integument ulcerative lesions were present before the stranding. Ventral skin abrasions and visceral passive congestion were caused by the strandings. Absence of food in the alimentary tracts, evidence of weight loss and blubber thickness reduction were compatible with an extended presence of the sperm whales in the North Sea, where adequate food is not available. This might lead to progressive weakness, predisposing the animals to secondary pathogens such as viral diseases. Finally, the coastal configuration of the southern North Sea makes it a trap for sperm whales which have entered the area during their wanderings.

The ways in which comprehensive condition profiles, incorporating morphometric, histologic, physiologic, and diet quality indices, responded to changes in density of a white-tailed deer (Odocoileus virginianus) population were examined. Changes in these condition indices were monitored in a northeastern Oklahoma deer herd as density declined from peaks of 80 and 72 deer/km² in 1989 and 1990 (high-density) to lows of 39 and 41 deer/km₂ in 1991 and 1992 (reduced-density), respectively. Compared to a reference population (6 deer/km₂), deer sampled during high-density exhibited classic signs of nutritional stress such as low body and visceral organ masses (except elevated adrenal gland mass), low fecal nitrogen levels, reduced concentrations of serum albumin, elevated serum creatinine concentrations, and a high prevalence of parasitic infections. Although density declined by one half over the 4-yr study, gross indices of condition (in particular body mass and size) remained largely unchanged. However, selected organ masses, serum albumin and non-protein nitrogen constituents, and fecal nitrogen indices reflected improvements in nutritional status with reductions in density. Many commonly used indices of deer condition (fat reserves, hematocrit, total serum protein, and blood urea nitrogen) were not responsive to fluctuations in density.

Recent research has demonstrated the potential of pregnancy diagnosis in elk (Cerlvus elaphus nelsoni) using immunoassays of fecal steroid concentration. However, multiple sampies are required to insure accurate results, limiting its utility for free-ranging animals. We attempted to develop an accurate one-sample pregnancy diagnosis using 153 fecal samples that were collected from free-ranging, radio-collared, adult female elk in Yellowstone National Park (Wyoming, USA) and from captive elk maintained at the Starkey Research Facility (La Grande, Oregon, USA) February through April 1992 and 1997. The pregnancy status of each animal was diagnosed using serum pregnancy-specific protein B (PSPB) assays providing fecal samples from 38 nonpregnant and 115 pregnant animals. Fecal radioimmunoassay (RIA) indicated that mean (± SD) progestagens (P₄) were elevated significantly in pregnant (2.96 ± 1.49 μg/gm) compared to nonpregnant (0.43 ± 0.26 μ/gm) individuals. Confidence intervals (1.96 ± SE) for the two groups were widely separated (nonpregnant 0.34–0.51, pregnant 2.69–3.24) with little overlap in the range of concentrations measured for each group (nonpregnant 0.09–0.98, pregnant 0.90–8.29). These results indicate that fecal progestagens RIA provides a reliable method of noninvasive pregnancy diagnosis using single fecal samples collected from elk during late gestation. However, independent validation of the suggested discrimination criteria should be performed before routine application.

Trypanosoma cruzi frequently infects wild mammals in the southern United States but little is known about the effect of the parasite on reservoir hosts such as the raccoon (Procyonlotor). To investigate this issue, 30 raccoons trapped on St. Catherine's Island (Georgia, USA) during September, 1994 were tested for T. cruzi infection by examination of wet mounts of fresh blood and by culturing blood in liver infusion tryptose medium. Thirteen animals (43%) were found to be infected with T. cruzi. Heart tissues from 10 of the infected raccoons and 4 uninfected raccoons were fixed, sectioned, stained and examined for the presence of parasites and evidence of tissue damage. One T. cruzi pseudocyst was found in cardiac tissue from the left ventricle of a female raccoon. In addition, Sarcocystis sp. sarcocysts and schizonts of Hepatozoon sp. were observed in heart tissue from seven of the T. cruzi-infected raccoons. Mild, multifocal and interstitial inflammation was observed in the heart tissues of all 10 of the infected animals. No evidence of T. cruzi pseudocysts or tissue damage was observed in heart tissue from C3H/HeJ mice infected with culture forms of the parasites isolated from raccoons. Our findings suggest that the T. cruzi parasites isolated from raccoons in Georgia are not pathogenic to this host or C3H/HeJ mice and may be of low virulence.

Because conservation biologists have postulated that infectious diseases may have potentiated the endangerment of the Attwater's prairie chicken (Tympanuchus cupido attwateri), free-living prairie chickens were surveyed from all remaining populations for helminthic endoparasites and antibody against the etiological agents of nine infectious diseases. Samples from 4 of 27 adult males were positive for anti-Pasteurella multocida antibody. All other serologic tests were negative (n = 19). We identified Dispharynx nasuta, a parasite previously associated with disease in other grouse from North America, in one of three adult Attwater's prairie chickens examined. Evidence of Tmichostrongylus cramae was found for eight of nine suitable samples, which represents the first report of this parasite in prairie grouse. The mean intensity of T. cramae in Attwater's prairie chicken was 1,019.3 (Range = 3-1,906; n = 3). Further work is needed to determine whether P. multocida, T. cramae, or D. nasuta are detrimental to Attwater's prairie chicken populations. If so, conservation biologists could reduce the prevalence and incidence of these parasites and potentially gain more time to address the habitat conditions thought to be the ultimate cause of population declines

12-yr old Springhaas (Pedetes capensis; Pedetidae, Rodentia) imported into the USA from the Republic of South Africa was severely debilitated as the result of abberant migrations of adult Filaria versterae (Filarioidea: Filariidae) normally confined to the subcutaneous tissues of this host in Africa. Some nematodes invaded the peritoneal and thoracic cavities, the pericardial sac and the lungs. Invasion of the lungs resulted in life-threatening respiratory distress necessitating euthanasia. Filana versterae is redescribed.

Parasitological data were used to test the hypothesis that the wood mouse (Apodemus sylvaticus) population of Fair Isle, Shetland, originated from the British Isles rather than Scandinavia, as is usually argued. This study was based on the assumption that the mice were likely to share most of their parasite fauna with conspecifics from their ancestral home. The ecto- and endo-parasites of wood mice on Fair Isle between 18 June to 10 July 1987 and 17 to 26 August 1991 were identified and compared with those reported from conspecifics in the two putative source areas. All eight species of metazoan parasites that infected A. sylvaticus on Fair Isle were common to mice of the British Isles, whereas just one parasite on Fair Isle, a mite, had been recorded from Scandinavia. This lends support to the hypothesis that the mice originated from Britain rather than Scandinavia.

Gnathostorna doloresi, a nematode parasite producing ulcerative and/or granulomatous lesions in the gastric wall of domestic pigs and wild boars (Sus scrofa leucomystax) throughout southeastern Asia, is an important zoonosis causing cutaneous larva migrans in humans. Patients infected with G. doloresi have been found yearly in the vicinity of Miyazaki Prefecture, Japan. Adult nematodes were found in 31 of 32 wild boars captured at two different geographic locations in the prefecture, a location in the high mountains and one at sea level. The parasite-free animal had gastric ilesions typical of a previous infection with this parasite. The nematode intensities were higher in wild boars caught in the mountains than those caught at near sea level. The extremely high prevalence of G. doloresi in wild boars indicates that the life cycle of this parasite is well established in this area of Japan.

Larvae of Trichinella sp. were found in two of 208 red foxes (Vulpes vulpes) and one of 125 coyotes (Canis latrans) obtained from trappers from Prince Edward Island (Canada) in 1995 and 1996. A polymerase chain reaction based DNA biotyping method revealed the larvae to be isolates of Trichinella spiralis. This is the first verified identification of T spiralis in sylvatic hosts from Canada.

From May 1990 to February 1996, 55 black bears (Ursus americanus floridanus) (38 males, 17 females) from 21 counties in Florida (USA), were examined for Dermodex ursi(Acari: Demodecidae). The sample included cubs <1-yr-old (n = 12), juveniles (1 to <3 yr, n = 20), and adults (3 to 12 yr, n = 23). Demodex ursi was collected from four bears by deep scrapings of 15 sites on the head and forelimb of each bear. All four bears with demodicosis had crusty, scaly skin lesions, and moderate alopecia. No D. ursi were seen in any of the 750 scrapings from the other 51 bears, nor did any of those bears have signs of demodi cosis. Demodicosis was found only in bears from or near the Ocala population in northcentral Florida.

Thin blood smears of 75 wood storks (Mycteria americana) from Georgia (USA) were made during the summers of 1994–96 and examined for blood parasites. Haemoproteus crnmenium was found in one of 71 juveniles and in two adults from a sample of two subadults and two adults. Intensity of infection in the juvenile and in each of the two adults was 11, 3, and 2 parasites/5,000 erythrocytes, respectively. This is the first record of H. crumenium in the wood stork from Georgia and the second published record of H. crumenium infecting this host in North America. Additionally, one juvenile was infected with a microfilarid.

Forty-three wild rats (Rattus norvegicus) trapped in Milan (Italy) were examined for gastric spiral bacteria and to evaluate associated histological lesions. Spiral bacteria were histologically detected in the stomach of 10 rats (23%). The morphological features of these microorganisms, observed using both light and electron microscopy, correspond to those of Helicobactem heilmannii. Mild gastritis characterized by focal lymphoplasmacytic infiltration of the lamina propria was observed in six rats with gastric spiral bacteria. Our findings suggest a causal role of these bacteria in inducing a gastric inflammatory response.

From 1993 to 1995, 100 blood samples and 97 spleen samples of free-ranging rabbits (Oryctolagus cuniculus) from northern Germany were tested for prevalence of different bovine viral diarrhea virus (BVDV) antibodies and BVDV antigen, respectively. Forty sera (40%) were positive for antibodies to one or more of three cytopathogenic BVDV strains (NADL, Grub 313/83, and roe deer isolate SH9/11). Microneutralization test (NT) titers ranged from 5 to 13. Moreover, 13 of 35 NT positive sera also were positive by enzymelinked immunosorbent assay for detection of anti-BVDV antibodies. Therefore, we assume that the rabbits either had passively acquired antibodies or had experienced natural infection with BVDV. Furthermore, 97 spleen samples were used for attempts to isolate BVDV and tested by reverse transcription polymerase chain reaction for viral nucleic acid using vanous protocols. Bovine viral diarrhea virus was not identified in these samples.

Archived serum samples from 111 Peromyscus leucopus, collected 1984–88 in Baltimore City (Maryland, USA), were analyzed by indirect immunofluorescence assay. Sera from two (2%) individuals contained antibodies reactive to Ehrlichia equi and the human granulocytic ehrlichiosis (HGE) agent. This suggests that the HGE agent or an antigenically related organism has been present in rodent populations in this locality for more than a decade, and was present in the region at least 12 yr before a case of human disease was recognized.

A cryopreservation method and improved isolation techniques for detection of Pasteurella multocida from wetland samples were developed. Wetland water samples were collected in the field, diluted in dimethyl sulfoxide (DMSO, final concentration 10%), and frozen at −180 C in a liquid nitrogen vapor shipper. Frozen samples were transported to the laboratory where they were subsequently thawed and processed in Pasteurella multocida selective broth (PMSB) to isolate P. multocida. This method allowed for consistent isolation of 2 to 18 organisms/ml from water seeded with known concentrations of P. multocida. The method compared favorably with the standard mouse inoculation method and allowed for preservation of the samples until they could be processed in the laboratory.