Natural infection of red deer with bovine tuberculosis

Six bovine tuberculosis-free red deer hinds were introduced in October 1993 to a 1.8 ha enclosure, within a larger field study site known to contain tuberculous possums, and kept there for 9 months. A Mycobacterium bovis-infected possum was found in the vicinity of the deer enclosure 3 weeks after the introduction. Subsequently, a further eleven infected possums were found in the area. The deer were monitored by repeated composite antibody detection ELISA and lymphocyte transformation assays for tuberculosis, interpreted in parallel, by skin testing and by routine culturing of samples collected from potential excretion sites. Lymphocyte transformation assay evidence of M. bovis infection in four hinds was first observed 4 months after introduction. One other hind became bovine tuberculin lymphocyte transformation assay positive in the 5th month. Positive or equivocal bovine reactivity remained evident at most test episodes. A comparative cervical skin test performed in July 1994, shortly before slaughter, was positive in these five hinds. Mycobacterium bovis was recovered off swabs from the oropharyngeal tonsils of two hinds during routine sampling. Detailed necropsy of the six deer revealed a single typical tuberculous lesion in only one, but culturing of various tissue specimens ascertained that the five blood test and comparative cervical skin test-positive animals were all infected. Mycobacterium bovis was cultured from the oropharyngeal tonsils of four and medial retropharyngeal lymph nodes of two of the deer with no typical gross lesions. Six additional tuberculosis-free hinds were introduced to the enclosure in April 1994 and kept there for 12 months. Four of these animals showed a positive lymphocyte transformation assay response to M. bovis after 9 weeks, but no significant reactivity thereafter. Concurrent observational studies suggest that five of the first six deer probably became infected through close inspection and investigation of the tuberculous possums, although the possibility of deer-to-deer transmission cannot be totally excluded. The likely deer-possum contact, and thus exposure to M. bovis, was related to the curiosity and social ranking of the hinds. The second group appear to have had transient exposure to M. bovis, possibly caused by direct contact with the infected hinds introduced earlier. This group never showed any curiosity toward, or interaction with, possums during the periods of observation.     

Phylogenetic analysis of Theileria sp. from sika deer, Cervus nippon, in Japan

The 18S rRNA genes of Theileria species detected in sika deer, Cervus nippon centralis in Yamaguchi and Cervus nippon yesoensis in Hokkaido, were analyzed. The percent identities of the nucleotide sequences of Theileria from Cervus nippon centralis and Cervus nippon yesoensis were more than 99%. The percent identities of the Theileria sp. from sika deer and Theileria sergenti, Theileria buffeli and Theileria cervi were 97, 96 and 95%, respectively. Phylogenetic analysis of the gene sequences also revealed that Theileria sp. detected from sika deer comprise a clade that is clearly distinct from the clade comprised of Theileria from cattle.     

Mycobacterium avium subsp. paratuberculosis infection in wildlife on three deer farms with a history of Johne's disease

Abstract

AIM: To determine the prevalence of Mycobacterium avium subsp. paratuberculosis (Map) infection in wildlife, in pastoral landscapes with a recent history of clinical Johne's disease in livestock.

METHODS: A total of 449 wild mammals and birds from three farms in the South Island of New Zealand with recent histories of clinical Johne's disease in their deer herds were trapped and examined for gross pathological changes in the gastrointestinal tract. Additionally, individual mesenteric lymph nodes from 380 mammals, and segments of gastrointestinal tract from 32 birds were excised, homogenised and cultured for viable Map bacilli. The prevalence of Map infection was then calculated for the various species. Faecal samples from those mammals which had culture-positive tissues were further cultured for the presence of Map.

RESULTS: Gross pathological changes were identified in the gastrointestinal tract of four brushtail possums, one cat, six ferrets, 12 hares, six hedgehogs, three rabbits, one stoat, and one paradise shelduck. Infection with Map in the gastrointestinal tract was confirmed in only three of these cases, one each of brushtail possums, hares and hedgehogs. In contrast, Map infection in the absence of gross pathological changes was frequently recorded in enteric tract tissues of mammals and birds. Among mammals, Map infection was recorded in 18/73 (25%) brushtail possums, 4/23 (17%) cats, 15/42 (36%) hedgehogs and 29/113 (26%) rabbits. Among birds, intestinal tract tissue Map infection was recorded in 3/17 (18%) paradise shelducks. Among 64 of the 74 mammals which had Map culture-positive tissues, 38% (n=5) of hedgehogs and 11% (n=3) of rabbits also had culture-positive faecal samples.

CONCLUSIONS: This study is the first to identify that Map infection can be prevalent in wildlife in New Zealand. There was a high prevalence of Map infection among both scavenging and grazing wild animals. Both mammals and birds are capable of harbouring viable Map organisms in their gastrointestinal tract; further, viable Map was excreted into the environment via faeces by hedgehogs and rabbits.

CLINICAL RELEVANCE: Previous studies overseas have postulated a role of wildlife as reservoirs of Map infection and possible vectors of Johne's disease to livestock. Here, brushtail possums, hedgehogs and rabbits and in particular were identified as potential wildlife hosts for Map infection in NewZealand. This suggests that several wildlife species could contribute to the persistence of Map infection within a wildlife/livestock complex, and potentially, perhaps more importantly, to the spread of infection between farms.     

甘肃马鹿导入东北马鹿杂交改良试验观察

以东北马鹿为父本．甘肃肃南鹿场人工驯养的甘肃马鹿为母本．进行导血杂交改良试验。结果表明：甘肃马鹿导入东北马鹿杂交改良．其杂交后代初生重、断奶重、周岁重和2～3周岁公鹿产茸量均显著高于本地马鹿。     

Evidence of Mycobacterium avium subsp. paratuberculosis infection in Dutch farmed red deer

Paratuberculosis is a chronic disease in ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). Most economic losses due to MAP occur in the dairy industry. However, the infection is not restricted to cattle, but also occurs in other ruminants, such as sheep, goat, and deer. Although deer are of minimal economic importance in The Netherlands, they may constitute a source of infection for the dairy industry. This pilot study was conducted to estimate the prevalence of Johne's disease in farmed red deer in The Netherlands. Serum and faecal samples were collected from 140 animals, originating from 8 different farms. Four of the farms had animals that tested positive for Johne's disease. The within-herd MAP seroprevalence varied between 4.8% and 21.2%. In conclusion, this pilot study provides evidence of MAP infection in the Dutch farmed deer population, and thus there might be a risk of MAP transmission between farmed red deer and dairy cattle.     

Osteochondrosis associated with copper deficiency in young farmed red deer and wapiti x red deer hybrids

Osteochondrosis is reported in association with copper deficiency in young red deer and wapiti X red deer hybrids on eight deer farms throughout New Zealand. On two farms, more than 30% of fawns were affected. Affected animals were lame, often had one or more swollen joints, and in some cases had an abnormal "bunny-hopping" gait or "cow-hocked" stance. Lesions were most common in the carpal, tarsal, stifle and hip joints, and were usually bilateral. Defects in articular cartilage ranged from loose flaps to complete separation with exposure of subchondral bone and the presence of cartilage fragments within the joint space. In advanced cases, the joints had features of degenerative arthropathy. Bilateral epiphyseolysis of the femoral head was observed in some severely lame deer. All deer with osteochondrosis had low serum and/or liver copper concentrations.     

Osteochondrosis associated with copper deficiency in young farmed red deer and wapiti x red deer hybrids

Osteochondrosis is reported in association with copper deficiency in young red deer and wapiti x red deer hybrids on eight deer farms throughout New Zealand. On two farms, more than 30% of fawns were affected. Affected animals were lame, often had one or more swollen joints, and in some cases had an abnormal “bunny-hopping” gait or “cow-hocked” stance. Lesions were most common in the carpal, tarsal, stifle and hip joints, and were usually bilateral. Defects in articular cartilage ranged from loose flaps to complete separation with exposure of subchondral bone and the presence of cartilage fragments within the joint space. In advanced cases, the joints had features of degenerative arthropathy. Bilateral epiphyseolysis of the femoral head was observed in some severely lame deer. All deer with osteochondrosis had low serum and/or liver copper concentrations.     

Onchocerca sp. in an imported Zangersheide gelding causing suspensory ligament desmitis

A 5-year-old imported Zangersheide gelding was evaluated for SC swellings over both forelimbs and lameness localized to the distal metacarpus. Ultrasound examination of the SC masses was compatible with verminous granulomas. Linear hyperechoic foci were present within the suspensory ligament branches of both forelimbs, suggestive of ligamentous parasitic infiltrates. A diagnosis of onchocerciasis was confirmed on biopsy of a SC mass. The gelding was treated with ivermectin and a tapering course of PO dexamethasone but was eventually euthanized. Necropsy confirmed the presence of SC eosinophilic granulomas and degenerative suspensory ligament desmitis, both with intralesional nematodes. Given the location and appearance of the nematode, a diagnosis of Onchocerca sp., most likely O. reticulata, was made. Onchocerciasis should be included as a differential diagnosis for multifocal suspensory ligament desmitis with these sonographic characteristics when paired with SC masses in imported European Warmbloods.     

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.     

BTV infection in wild ruminants, with emphasis on red deer: a review

The distribution of bluetongue virus has changed, possibly related to climate change. Vaccination of domestic ruminants is taking place throughout Europe to control BT expansion. The high density of wild red deer (Cervus elaphus) in some European regions has raised concerns about the potential role that unvaccinated European wild ungulates might play in maintaining or spreading the virus. Most species of wild ruminants are susceptible to BTV infection, although frequently asymptomatically. The red deer population density in Europe is similar to that of domestic livestock in some areas, and red deer could account for a significant percentage of the BTV-infection susceptible ruminant population in certain regions. High serum antibody prevalence has been found in red deer, and BTV RNA (BTV-1, BTV-4 and BTV-8) has been repeatedly detected in naturally infected European red deer by means of RT-PCR. Moreover, red deer may carry the virus asymptomatically for long periods. Epidemiological studies suggest that there are more BT cases in domestic ungulates in those areas where red deer are present. Vector and host density and environmental factors are implicated in the spatial distribution of BT. As in domestic ruminants, BTV transmission among wild ruminants depends almost exclusively on Culicoides vectors, mainly C. imicola but also members of the C. obsoletus and C. pulicaris complex. However, BTV transmission from red deer to the vector remains to be demonstrated. Transplacental, oral, and mechanical transmissions are also suspected. Thus, wild red deer contribute to the still unclear epidemiology of BTV in Europe, and could complicate BTV control in domestic ruminants. However, further research at the wildlife host-vector-pathogen interface and regarding the epidemiology of BT and BT vectors in wildlife habitats is needed to confirm this hypothesis. Moreover, red deer could be used as BT sentinels. Serum and spleen tissue of calves sampled from late autumn onwards should be the target samples when establishing a BTV surveillance program.     

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.     

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.     

Artificial infection of the tick Rhipicephalus appendiculatus with Theileria parva.

Methods for infecting Rhipicephalus appendiculatus ticks with Theileria parva by injection and by artificial feeding were confirmed and compared. The injection method proved simpler and at best as effective and suggested improvements are described.     

Experiments on transmission of an unidentified Theileria sp. to small ruminants with Haemaphysalis qinghaiensis and Hyalomma anatolicum anatolicum

Experiments on the transmission of an unidentified Theileria sp. infective for small ruminants by Haemaphysalis qinghaiensis and Hyalomma anatolicum anatolicum were carried out. Three Theileria-free batches of adult, larvae, and nymphs of laboratory reared H. qinghaiensis and Hy. a. anatolicum ticks were infected by feeding them on sheep infected with Theileria sp. The Theileria sp. was originally isolated from adult ticks of H. qinghaiensis, by inoculation of blood stabilates or tick transmission. H. qinghaiensis has been shown to be capable of transmitting the Theileria sp. infective for small ruminants transstadially to sheep and goats. The nymphs developed from the larvae engorged on the sheep infected with the parasite transmitted the pathogen to splenectomized sheep with prepatent periods of 30, 31 days, respectively; but the subsequent adult ticks of H. qinghaiensis derived from the nymphs did not transmit the pathogen to sheep. However, adults developed from the nymphs engorged on the sheep infected with the parasite transmitted the pathogen to sheep with prepatent periods of 24-27 days. The larvae, nymphs and adult ticks derived from female H. qinghaiensis ticks engorged on infected sheep were not able to transmit the parasite transovarially. The same experiments were done with Hy. a. anatolicum, but examination for presence of piroplasma of Theileria sp. from all animals were negative, demonstrating that Hy. a. anatolicum could not transmit the organism to sheep or goats.     

Molecular detection of Theileria sp. in ticks and naturally infected sheep

Seven healthy sheep and 10 sheep diagnosed with piroplasmosis based on clinical signs were tested for the presence of babesiae and theileriae. Using the molecular techniques, two species of theileriae were detected and characterized. Theileria ovis was present mostly in healthy sheep and in Rhipicephalus ticks collected from infected sheep. Theileria sp. OT3 parasite was detected mostly in ill animals which represent additional evidence to the possible pathogenic nature of Theileria sp. OT3. The presence of babesiae in sheep or in ticks was not determined. The results of this study showed that ovine piroplasmosis due to Theileria is present in Southern Croatia. It was concluded that clinical diagnosis of ovine piroplasmosis should be confirmed by molecular analysis in order to identify the species of piroplasm, to select the appropriate treatment and to exclude the threat for public health.