Faecal bacteria of wild ruminants and the alpine marmot

Faecal samples from 60 red deer (Cervus elaphus), 13 roe deer (Capreolus capreolus), 7 chamois (Rupicapra rupicapra), 41 alpine marmot (Marmota marmota) and soils mixed with deer faeces from the Stelvio National Park were examined forCampylobacter sp. andSalmonella sp. with negative results.The same material, especially deer faeces, was a habitat highly suitable forYersinia sp.:Y. enterocolitica (two biotypes) was isolated twice,Y. kristen-senii (two serotypes) was isolated 19 times,Y. frederiksenii andY. intermedia were isolated once.Antibiotic-resistantEscherichia coli were isolated from 16 specimens from wild ruminants, one from marmot and two from feeding places.This article was presented as a paper at the 8th International Symposium Enteric Infections and their Control of the World Association of Veterinary Microbiologists, Immunologists and Specialists in Infectious Diseases (Perth, Western Australia, 20 August 1983).     

Faecal survey of deer for Yersinia pseudotuberculosis and Salmonella sp

The results of a national survey of faeces from clinically normal deer for Salmonella sp. and Yersinia pseudotuberculosis are reported. Five isolates of Y. pseudotuberculosis and none of Salmonella sp. were made from 3810 faeces representing 122 farms.     

The identity and prevalence of coccidia species in sheep and cattle in New Zealand

The results of a national survey of faeces from clinically normal deer for Salmonella sp. and Yersinia pseudotuberculosis are reported. Five isolates of Y. pseudotuberculosis and none of Salmonella sp. were made from 3810 faeces representing 122 farms.     

The isolation of Yersinia sp. from feral and farmed deer faeces

Faecal samples from clinically normal farmed red deer, wapiti, fallow deer; and feral red deer and white tail deer were examined for members of the genus Yersinia. From 922 samples 176 strains of Y.enterocolitica, 56 strains of Y.frederiksenii, 29 strains of Y.kristensenii, eight strains of Y.intermedia, and seven strains of Y.pseudotuberculosis were isolated. High isolation rates of Yersinia sp. were recorded from some farms. Two herds had isolation rates of 33.3% and 36.8%. Sixteen strains of Yersinia sp. in addition to strains of Y.psuedotuberculosis were found to be Hela cell invasive. The majority of these strains were confined to a single herd and represented Y.enterocolitica biotypes I, II and III, Y.intermedia, Y. fredericksenii, and Y.kristensenii.     

Yersinia pseudotuberculosis recovered from the faeces of clinically healthy deer

Extract

Sir:— A recent paper by Henderson and Hemmingsen(1) reported the recovery of Yersinia pseudotuberculosis from the faeces of 5 of 3810 (0.13%) clinically healthy deer in a national survey conducted on 122 farms. The isolates were obtained from animals on four (3.27%) different farms. The authors concluded on the basis of these results and unpublished data that Y. pseudotuberculosis was only rarely recovered from faeces of healthy deer.     

Isolation and characterization of Babesia pecorum sp. nov. from farmed red deer (Cervus elaphus)

The diversity of Babesia species infecting cervids in parts of central and southern Spain was analyzed by collecting blood from farmed red deer (Cervus elaphus). Babesia sp. was isolated in vitro from two red deer herds in Cádiz and Ciudad Real. The number of Babesia sp. carriers differed between the two herds: 36/77 in Cádiz and 1/35 in Ciudad Real. Hyalomma lusitanicum was the most prevalent tick species identified on the Cádiz farm vegetation and on sampled animals, and is therefore a candidate vector. The molecular characteristics of 21 isolates were determined by complete (8 isolates) or partial (13 isolates) 18S rRNA gene sequencing. The sequences were highly similar (over 99.4% identity) and 6 sequence types were identified at the level of one herd only, demonstrating a rather high genetic diversity. They formed a monophyletic clade, and members of the three main sequence types shared a similar morphology and the same erythrocyte susceptibility pattern. This clade also included Babesia sp. Xinjiang isolated from sheep in China and Babesia sp. identified in giraffe in South Africa, with identities higher than 98.3% and statistically relevant phylogenetic support. None of the biological properties analyzed for both Babesia from red deer and Babesia sp. Xinjiang allowed their differentiation (ability to develop in vitro in erythrocytes from cattle and sheep, as well as in erythrocytes from different cervids, unsuccessful infection of calves). We propose the Babesia isolated from red deer as a new species named B. pecorum. Whether Babesia sp. Xinjiang and the Babesia characterized in South Africa belong to the same species is debated.

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-014-0078-7) contains supplementary material, which is available to authorized users.     

Yersiniosis in Farmed Deer

Samples from 77 chital (Axis axis), 42 fallow (Dama dama), 26 red (Cervus elaphus), 7 rusa (Cervus timorensis) and 1 sambar deer (Cervus unicolor) were examined. Yersinia pseudotuberculosis infection was diagnosed as the cause of death in 6 (23%) of the red and 23 (30%) of the chital deer. Yersiniosis was the most common infectious cause of death diagnosed. Affected deer were usually found moribund or dead, often with faecal staining of the perineum. Gross pathology in chital included a fibrinous enterocolitis, enlarged congested mesenteric lymph nodes and multiple pale foci through the liver. Gross changes in red deer were limited to intense congestion of the intestinal mucosa and enlargement and congestion of mesenteric lymph nodes. Microscopic intestinal changes in both species consisted of microabscessation or diffuse suppurative inflammation of the intestinal mucosa with numerous bacterial colonies in the lamina propria. Multifocal suppurative mesenteric lymphadenitis was a common finding. Multifocal suppurative or non-suppurative hepatitis was frequently present in the liver of chital but was uncommon in the red deer. Yersiniosis occurred during the cooler months from June to November, with younger age classes most commonly affected. Y. pseudotuberculosis serotypes I, II and III were isolated in the ratio 17:3:0 in the chital deer and 1:1:2 in red deer. The clinical, epidemiological and bacteriological features are similar to those documented previously by New Zealand workers. The increased susceptibility to disease of red deer and chital compared to fallow deer and perhaps other species has not previously been documented.     

Larvae of Elaphostrongylus cervi in the deer of Scotland

Protostrongylid larvae were recovered from the faeces or lungs of red deer (Cervus elaphus), roe deer (Capreolus capreolus) and reindeer (Rangifer tarandus) in Scotland during 1981. Typical protostrongylid first-stage larvae were also recovered from possible intermediate hosts, the grey field slug (Agriolimax reticulata) and the white-soled slug (Arion fasciatus). All these protostrongylid larvae were microscopically identical to those of the nematode Elaphostrongylus cervi. Despite careful search, adult E cervi were not found, but it is concluded that infection with E cervi is widespread in Scottish deer.     

Prevalence of Cryptosporidium and Giardia in roe deer (Capreolus capreolus) and wild boars (Sus scrofa) in Galicia (NW, Spain)

Faecal samples from 224 roe deer (Capreolus capreolus) and 381 wild boars (Sus scrofa) shot during the 2008-2009 hunting season (August-January) in Galicia (NW Spain) were examined to determine the presence and intensity of infection by Cryptosporidium and Giardia. Analysis of a single sample from each of the roe deer revealed that the prevalence of cryptosporidiosis and giardiosis was 1.3% and 5.3% respectively. The prevalence of Giardia infection was significantly higher in juvenile female roe deer than in adult females, but no other significant differences were found in relation to age and sex. In wild boars, the prevalence of cryptosporidiosis and giardiosis was 7.6% and 1.3% respectively. The prevalence of Cryptosporidium infection was significantly higher in juvenile male wild boars than in adult males, but no other significant differences were found in relation to age or sex. In both groups of wild animals, the number of Cryptosporidium oocysts per gram of faeces (OPG) ranged from 5 to 200 and the number of Giardia cysts per gram of faeces (CPG) was between 5 and 47; there were no significant differences between the two groups with respect to number of infections. This is the first large study of Cryptosporidium and Giardia in roe deer and wild boars in hunting areas in Spain and the results demonstrate a low, but widespread prevalence of Cryptosporidium and Giardia in these animals.     

A model for study of lungworm (Dictyocaulus sp.) and gastrointestinal nematode infection in young red deer (Cervus elaphus)

A model of sub-clinical parasitism in young red deer, using concurrent trickle infections of lungworm (Dictyocaulus sp.) and mixed gastro-intestinal (GI) nematodes of deer-origin was evaluated. 20 parasite-free deer calves were artificially reared indoors from 4 days of age. A further five calves were naturally reared on pasture with their dams, treated with anthelmintic and brought indoors at 3-4 months. At 4-4.5 months of age they were individually housed and allocated to five groups (n=5). Groups were dosed 3 x per week, for 9 weeks with 0, 100 and 500, 200 and 1000 (2 groups), 400 and 2000 infective larvae of lungworm and mixed GI nematodes, respectively, cultured from deer faeces. Liveweight and voluntary feed intake measurements and faecal and blood samples were taken weekly. In the fourth week following cessation of trickle infection, deer were euthanased and lung and GI nematodes recovered. Both lungworm and GI nematode infections became patent at Week 4 of infection. Maximum group arithmetic mean faecal egg counts were 100-190 epg. Maximum group arithmetic mean faecal lungworm larval counts were 58-123 lpg. Group arithmetic mean nematode counts at slaughter ranged from 439-806 for GI nematodes and 31-73 for lungworm, respectively. Despite low nematode counts, reduced liveweight gain, voluntary feed intake and serum albumin concentration, elevated serum pepsinogen, gastrin and globulin concentrations and elevated peripheral eosinophil counts and slight haemoconcentration, but no clinical signs, were observed. The reduction in liveweight gain was related to the reduction in voluntary feed intake (r2=0.83; p<0.088). Naturally-reared deer had similar liveweight gains, voluntary feed intake and nematode counts to artificially-reared deer. Thus, methods of infection to produce concurrent sub-clinical lungworm and GI nematode burdens for study of sub-clinical parasitism in young deer have been defined.     

Fatal yersiniosis in farmed deer caused by Yersinia pseudotuberculosis serotype O:3 encoding a mannosyltransferase-like protein WbyK.

Sudden death of 9 deer occurred in a large enclosed deer farm with approximately 400 heads of cervids. Fatal yersiniosis was diagnosed in 2 deer that were submitted for laboratory diagnosis. Histopathologically, the disease was characterized by multifocal pulmonary hemorrhage and mild interstitial pneumonia, marked diffuse cholangiohepatitis, minimal myocarditis with mild myocardial degeneration, and mild multifocal suppurative cystic colitis. Yersinia pseudotuberculosis was isolated from the lungs and colon of the affected animals. The isolates were PCR-positive for genes virF, inv, yopB, and yopH, which are essential for invasion and colonization of host intestine and lung. The isolates reacted with polyclonal antibodies against serotype O:3 antigen. The O-genotyping patterns of the isolates were identical with each other, but different from those of the 21 O-genotypes (or serotypes) reported previously. In addition to the O-antigen genes possessed by classical serotype O:3, a gene (wbyK) encoding a mannosyltransferase-like protein was detected in these isolates. The wbyK gene of the isolates showed 94% of DNA sequence homology with the wbyK gene harbored by Y. pseudotuberculosis O:1b. On the basis of pathology, bacteriology, and serology, the authors concluded that the acute deaths of these deer were caused by Y. pseudotuberculosis infection. Molecular characterization of the isolate revealed a genetic heterogeneity in the O-antigen gene cluster of Y. pseudotuberculosis serotype O:3.     

Deer as a potential wildlife reservoir for Parachlamydia species

Wildlife populations represent an important reservoir for emerging pathogens and trans-boundary livestock diseases. However, detailed information relating to the occurrence of endemic pathogens such as those of the order Chlamydiales in such populations is lacking. During the hunting season of 2008, 863 samples (including blood, conjunctival swabs, internal organs and faeces) were collected in the Eastern Swiss Alps from 99 free-living red deer (Cervus elaphus) and 64 free-living roe deer (Capreolus capreolus) and tested using ELISA, PCR and immunohistochemistry for members of the family Chlamydiaceae and the genus Parachlamydia. Parachlamydia spp. were detected in the conjunctival swabs, faeces and internal organs of both species of deer (2.4% positive, with a further 29.5% inconclusive). The very low occurrence of Chlamydiaceae (2.5%) was in line with serological data (0.7% seroprevalence for Chlamydia abortus). Further investigations are required to elucidate the zoonotic potential, pathogenicity, and distribution of Parachlamydia spp. in wild ruminants.     

Effects of grazing undrenched weaner deer on chicory or perennial ryegrass/white clover pasture on the viability of gastrointestinal nematodes and lungworms

This study determined the in vitro effects on the viability of internal parasites of grazing undrenched weaner deer on either chicory (Cichorium intybus) or perennial ryegrass (Lolium perenne)/white clover (Trifolium repens) pasture. One experiment investigated the hatching and development of gastrointestinal nematode eggs and larvae, and the development and motility of L1 lungworm (Dictyocaulus eckerti) larvae, and a second experiment used larval migration inhibition assays to test the viability of L1 lungworm larvae extracted from the faeces of weaner deer grazed on either chicory or pasture when they were incubated with rumen and abomasal fluids from fistulated deer also grazing on chicory or pasture. The incubations were undertaken with and without added condensed tannins purified from chicory and with or without polyethylene glycol (PEG) to bind the tannins. Chicory had no effect on the hatching and development of gastrointestinal nematode eggs and larvae. Grazing chicory reduced the number of lungworm larvae developing to the L3 stage, and L1 lungworm larvae from the faeces of chicory-grazed deer were less viable in rumen and abomasal fluid than larvae from pasture-grazed animals. Abomasal fluid was significantly (P < 0.001) less inhibitory to the migration of L1 lungworms than rumen fluid. When the larvae were incubated in rumen and abomasal fluids from chicory-grazed deer, their passage through sieves was significantly (P < 0.001) reduced in comparison with when they were incubated in the fluids from pasture-grazed deer Adding condensed tannins to rumen fluid increased the inhibition of the migration of L1 lungworm larvae but PEG removed this inhibition; this effect was not observed with abomasal fluid.     

Experimental in vitro transmission of Babesia sp. (EU1) by Ixodes ricinus

Babesia sp. (EU1), first characterized in 2003, has been implicated in human cases of babesiosis in Italy, Austria and Germany. It has been identified in roe deer and in its suspected tick vector, Ixodes ricinus, in several European countries. The aim of the present study was to validate the competence of I. ricinus as a vector of Babesia sp. (EU1) via experimental infections. For this purpose, a parasite strain isolated from roe deer was cloned in sheep erythrocytes. After experimental infections, parasite DNA was successfully amplified by PCR in both eggs and larvae originating from infected I. ricinus females and in the salivary glands of females exposed to Babesia sp. (EU1) as nymphs. We also demonstrate that infected females were able to transmit parasite DNA during a new blood meal. Together with previous epidemiological studies, these results validate I. ricinus as a competent vector for Babesia sp. (EU1).     

Prevalence of gastrointestinal bacterial pathogens in a population of zoo animals

Faecal prevalence of gastrointestinal bacterial pathogens, including Campylobacter, Escherichia coli O157:H7, Salmonella, Shigella, Yersinia, as well as Arcobacter, were examined in 317 faecal specimens from 44 animal species in Belfast Zoological Gardens, during July-September 2006. Thermophilic campylobacters including Campylobacter jejuni, Campylobacter coli and Campylobacter lari, were the most frequently isolated pathogens, where members of this genus were isolated from 11 animal species (11 of 44; 25%). Yersinia spp. were isolated from seven animal species (seven of 44; 15.9%) and included, Yersinia enterocolitica (five of seven isolates; 71.4%) and one isolate each of Yersinia frederiksenii and Yersinia kristensenii. Only one isolate of Salmonella was obtained throughout the entire study, which was an isolate of Salmonella dublin (O 1,9,12: H g, p), originating from tiger faeces after enrichment. None of the animal species found in public contact areas of the zoo were positive for any gastrointestinal bacterial pathogens. Also, water from the lake in the centre of the grounds, was examined for the same bacterial pathogens and was found to contain C. jejuni. This study is the first report on the isolation of a number of important bacterial pathogens from a variety of novel host species, C. jejuni from the red kangaroo (Macropus rufus), C. lari from a maned wolf (Chrysocyon brachyurus), Y. kristensenii from a vicugna (Vicugna vicugna) and Y. enterocolitica from a maned wolf and red panda (Ailurus fulgens). In conclusion, this study demonstrated that the faeces of animals in public contact areas of the zoo were not positive for the bacterial gastrointestinal pathogens examined. This is reassuring for the public health of visitors, particularly children, who enjoy this educational and recreational resource.     

Experimental transmission of sheep-associated malignant catarrhal fever from sheep to Japanese deer (Cervus nippon) and cattle

The assumption that sheep carry ovine herpesvirus-2 (OvHV-2), the causative agent of sheep-associated malignant catarrhal fever (SA-MCF), is widely accepted, albeit OvHV-2 has not been isolated. We attempted experimental contact transmission of MCF from Japanese sheep persistently infected with OvHV-2 to Japanese deer (Cervus nippon) and cattle. In Experiment 1, a deer was kept in close quarters with an infected ewe. In Experiment 2, a second deer was kept with the same ewe. In Experiment 3, two cows were each kept with two infected wethers. In Experiment 1, the deer developed clinical signs at 138 days after first contact and then died. OvHV-2 genes by polymerase chain reaction (PCR) and fluorescent antibodies to Alcelaphine herpesvirus-1 were detected in the affected deer. Moreover, sequences of PCR products (423bp), obtained by amplification of materials from the sheep and from the affected deer, coincided. These results clearly confirmed that the sheep was a carrier of OvHV-2, and that this virus had induced SA-MCF in a deer. In other experiments, no OvHV-2 infection occurred in deer and cattle during the 6-18 months periods of contact, though viral genes were detected in the nasal swabs and white blood cells of the sheep. To our knowledge, this is the first report on successful experimental transmission of MCF from OvHV-2-infected sheep to deer.     

Gamma herpesvirus carrier status of captive artiodactyls

Between 1998 and 2000, 103 individuals of 19 species of the order Artiodactyla at Whipsnade Wild Animal Park were tested for evidence of infection with gamma herpesviruses in order to distinguish between species which are susceptible to malignant catarrhal fever (MCF), caused by alcelaphine herpesvirus-1 (AlHV-1) of wildebeest (Connochaetes sp.) or ovine herpesvirus-2 (OvHV-2) of domestic sheep, and species which carry related viruses sub-clinically. Gamma herpesvirus DNA was detected in the known, or suspected, carrier species: roan antelope (Hippotragus equinus), scimitar-horned oryx (Oryx dammah), gemsbok (Oryx gazella), musk ox (Ovibos muschatus) and mouflon (Ovis musimon). In six other species: lowland anoa (Bubalus depressicornis) yak (Bos grunniens), sitatunga (Tragelaphus spekei), greater kudu (Tragelaphus strepsiceros), waterbuck (Kobus ellipsiprymnus) and Nile lechwe (Kobus megaceros), DNA was present in some newborn calves and over 30% of adults, strongly suggesting a carrier state. In contrast five Père David's deer (Elaphurus davidianus) and two swamp deer (Cervus duvauceli) died of MCF during the study. A virus isolated from scimitar-horned oryx calves produced cytopathic effects in scimitar-horned oryx kidney cell-culture and caused MCF in a rabbit.     

A preliminary study of Salmonella, verocytotoxigenic Escherichia coli/Escherichia coli O157 and Campylobacter on four mixed farms

The aims of this study were to investigate the incidence of Salmonella, verocytotoxigenic Escherichia coli (VTEC)/Escherichia coli O157 and Campylobacter on four mixed farms and to characterize the isolates in terms of a range of virulence factors. Eighty-nine composite (five different samples from the same animal species combined) faecal [cattle (24), pigs (14), sheep (4), poultry (4), horses (7), deer (4), dogs (9), rodents (2) and wild birds (20)] samples, 16 composite soil samples plus 35 individual water samples were screened using culture-based, immunomagnetic separation and molecular methods. Salmonella was detected in bovine faeces, cattle and poultry house water. Salmonella serotypes/phage types included Dublin, Kiel and Typhimurium DT193, and most isolates were spvC, invA and rck positive. The pefA and rck genes were found exclusively in the non-Typhimurium strains, while Salmonella Dublin and Salmonella Kiel strains carried Salmonella genomic island I marker(s). VTEC/E. coli O157 were found in deer and dog faeces only. The E. coli O157 isolate was an enteroinvasive E. coli, while the VTEC isolate was untypable but carried the vt1, eaeA, hlyA, tir and eptD genes. This article reports the first confirmed carriage of E. coli O157 in Irish deer. Campylobacter species were not detected over the course of this study. It was concluded that [1] Salmonella, VTEC and Campylobacter have low (<5%) prevalence or are absent on the farms in this study; [2] water was an important source of bacterial pathogens; [3] both dogs and deer may act as a source of pathogenic E. coli and [4] key virulence and resistance determinants are widespread in farm Salmonella strains. This study highlights the need to control water as a source of pathogens and suggests that the domestic pets and deer should be considered in any farm risk assessment.     

The effect of a single disinfection of a farm on cryptosporidiosis infection in calves

The disinfection of a farm with Dikonit (active substance: sodium dichlorocyanuran) exerted no significant influence on the course of the spreading of cryptosporidiosis infections in newborn calves. The oocysts of Cryptosporidium sp., isolated from the faeces of a spontaneously infected calf and left in a 2.5% disinfecting solution of Dikonit under laboratory conditions for four hours, did not lose their viability. Laboratory mice experimentally infected with these "disinfected" oocysts, began to produce oocysts of Cryptosporidium sp. the sixth day from infection. The findings of different developmental stages of this protozoan obtained during the histological examination of the intestinal tissue of test mice are also considered as evidence of successful experimental infection. The cryptosporidium-free period which lasted only 14 days from disinfection was mainly due to thorough mechanical cleaning of the area where the calves were kept after birth. This is also proved by the results of the examination of old residues of calf faeces scraped from the floors: only individual individual oocysts of Cryptosporidium sp. were found in these samples.     

The studies on the aetiology of diarrhoea in neonatal calves and determination of virulence gene markers of Escherichia coli strains by multiplex PCR

The purpose of this study was to determine aetiological agents of diarrhoea in neonatal calves and to investigate virulence gene markers of Escherichia coli strains isolated from calves by multiplex polymerase chain reaction (PCR). Eighty-two diarrhoeic calves and 18 healthy calves were used as subjects. Faeces were taken from the rectums of all the calves and were subjected to bacterial culture. Antigen enzyme-linked immunosorbent assay (ELISA) was performed to detect rotavirus, coronavirus and E. coli K99 in faeces of all the calves. A multiplex PCR was used to characterize E. coli strains in all the calves. Escherichia coli was isolated from 37 faeces samples, Enterococcus ssp. was isolated from 22 faeces samples and Salmonella was isolated from one faeces sample in diarrhoeic calves. Furthermore, only E. coli was isolated from all 18 faeces samples of healthy calves. Of the 37 E. coli isolated from diarrhoeic calves, K99 (18.9%), F41 (18.9%), heat-stable enterotoxin a (STa) (18.9%), Shiga toxin 1 (Stx1; 13.5%) and Shiga toxin 2 (Stx2; 5.4%) and intimin (8.1%) genes were identified by multiplex PCR. Of the 18 E. coli isolated from healthy calves, K99 (16.6%) and intimin (55.5%) genes were identified by PCR. A total of 15 rotavirus, 11 coronavirus and 11 E. coli K99 were detected in diarrhoeic calves by the antigen ELISA. As a result, this study shows that rotavirus, coronavirus, E. coli and Enterococcus ssp. were determined to play a role in the aetiology of diarrhoea in the neonatal calves. K99, F41, STa, Stx1 and Stx2 were found as the most common virulence gene markers of E. coli strains isolated from calves with diarrhoea. Multiplex PCR may be useful for characterization of E. coli isolated from calves.