Susceptibility of Amblyomma americanum to natural and experimental infections with Theileria cervi

One hundred fifty Amblyomma americanum were examined between March and September 1986 from Cookson Hills Wildlife Refuge in eastern Oklahoma (USA). Of these ticks, 11% (17 of 150) were infected with Theileria cervi. Field-collected nymphal ticks had an 8% (3 of 37) prevalence of infection averaging 1.0 infected acini/nymph. Female ticks had a 16% prevalence of infection averaging 1.6 infected acini/female; T. cervi was not observed in salivary glands of field-collected male ticks. When laboratory reared A. americanum nymphs were allowed to feed on experimental white-tailed deer (Odocoileus virginianus) with varying T. cervi parasitemias (less than 1, 2, 6 and greater than 20%), only ticks which fed on deer with parasitemias greater than 1% became infected. Although prevalence and intensity of infection varied in the infected ticks, there was no significant difference in prevalence of infection between males and females. However, females did acquire significantly greater intensities than males. The data from these studies confirm that T. cervi overwinters in A. americanum and suggests that the prevalence, intensity and abundance of infection of T. cervi in ticks is influenced by the parasitemia of the deer host. Furthermore, fawns may play a more important role in the epidemiology of T. cervi transmission than do adult deer because of the coordination between tick activity patterns and deer fawning.     

Two Theileria cervi SSU RRNA gene sequence types found in isolates from white-tailed deer and elk in North America.

Two Theileria cervi SSU rRNA gene sequence Types, F and G, from white-tailed deer (Odocoileus virginianus) and elk (Cervus elaphus canadensis) isolates in North America were confirmed. Previously, nucleotide sequencing through a single variable (V4) region showed the presence of SSU rRNA gene Types F and G in T. cervi isolates from white-tailed deer and an elk. In this study, both sequence types were found in four T. cervi isolates (two from deer and two from elk). Microheterogeneity only appeared in the Type G gene, resulting in Subtypes G1, G2 and G3. Subtype G1 was found in two elk and one white-tailed deer T. cervi isolate; Subtypes G2 and G3 were found in a white-tailed deer T. cervi isolate. The Type F SSU rRNA genes were identical in nucleotide sequence in both elk and white-tailed deer T. cervi isolates. The high degree of conservation in the Type F variable regions may be exploited to design specific oligonucleotide primers for parasite detection by the polymerase chain reaction in cervine or tick hosts.     

First report of Elaphostrongylus cervi in Spanish red deer Cervus elaphus hispanicus

Elaphostrongylus cervi Cameron, 1931 was identified in six Cervus elaphus hispanicus sampled in Cuenca, central Spain. A total of 23 adult worms were found in the central nervous system with a mean of 3.8. Although E. cervi is reported to be widespread in cervids, this is the first time it has been recorded in Spanish red deer.     

The Prevalence of Elaphostrongylus Cervi Infection in Moose (Alces alces) in Southern Norway

The prevalence of Elaphostrongylus cervi infection in moose shot during the hunting season 1983 in 47 different municipalities in Southern Norway was studied. E. cervi seemed to occur endemically in the population, as infection was found in 35 % of 1,982 moose examined. Diagnosis was made on the basis of the presence of eggs and first-stage larvae (L1) and associated histopathological lesions in the lung, and on the presence of L1 in the faeces. The prevalence varied considerably between different municipalities. No relationship was found between the prevalence of E. cervi and the population density of moose. The highest prevalence was found in municipalities where red deer harbouring E. cervi were also present. The prevalence among males was higher than among females, and that among yearlings higher than among calves and adults. In no case was infection found in animals 51/2 years of age or older. Carcass weight in infected adult moose was significantly lower than in non-infected adults.     

Aspects of the life cycle and pathogenesis of Elaphostrongylus cervi in red deer (Cervus elaphus)

Aspects of the migratory life cycle and pathogenesis of Elaphostrongylus cervi were studied in red deer (Cervus elaphus) using 2 farmed calves experimentally infected with 450 third-stage larvae killed 40 and 45 days postinfection and using 3 wild calves and 3 wild yearlings with natural infections killed during autumn hunting. A full necropsy was carried out on the experimental calves, but only the head, eviscerated carcass, and lungs were examined from the naturally infected animals. Histological examination included extensive studies of the central nervous system (CNS), spinal nerve roots, and lungs. The experimental calves had prepatent infections, with many immature adult nematodes in the CNS, whereas the wild calves showed CNS lesions indicating a very recent E. cervi infection. The yearlings had patent infections, with many mature E. cervi in their skeletal muscles, reflecting acquisition of infection during the previous summer. Our findings showed that E. cervi develop to the adult stage in the CNS (subarachnoid spaces) and subsequently migrate into the skeletal muscles, where the mature nematodes live in reproductive pairs and groups. In the nervous system, the nematode caused encephalomyelitis, focal encephalomalacia and gliosis, meningitis, radiculitis, ganglionitis, and perineuritis.     

Identification of antigens with potential for immunodiagnosis of Parelaphostrongylus tenuis and Elaphostrongylus cervi infections in red deer (Cervus elaphus elaphus)

Red deer (Cervus elaphus elaphus) were infected experimentally with Parelaphostrongylus tenuis in New Brunswick, Canada, and with Elaphostrongylus cervi in New Zealand. Excretory-secretory (E-S) antigens from adult P. tenuis were evaluated for their serodiagnostic potential in identifying P. tenuis and heterologous E. cervi infections in a Western blot. The antigen recognition profile of sera from animals infected with P. tenuis varied between individuals and with duration of infections, whereas that of pooled sera from animals infected with E. cervi showed less variation. A single molecule of 42-43 kDa was recognized consistently by sera from all animals infected with either P. tenuis or E. cervi. Sera from unexposed control deer and from those with other heterologous nematode infections did not consistently identify this antigen. Serorecognition of the 42-43-kDa antigen by deer infected with P. tenuis resulted in a sensitivity of 99% and a specificity of 85% (> or =1 mo postinfection). Although antibody to this antigen waned with time, the persistence of recognition up to 34 mo postinfection with P. tenuis exemplifies its diagnostic value. The sensitivity and specificity of diagnosis using this molecule were each 100% for identifying deer infected with E. cervi (> or =3 mo postinfection). Two other molecules from E-S of adult P. tenuis, 26-28 and 10-12 kDa, were also diagnostic, although their recognition was not persistent throughout infections. These 2 molecules may prove useful in combination with the 42-43-kDa antigen to help identify all infected animals during all phases of infections. This research represents the first conclusive identification of antigens with real potential for reliable antemortem immunodiagnosis of both P. tenuis infections and heterologous E. cervi infections.     

Failure to detect infection in fallow deer (Cervus dama) exposed to Theileria cervi from white-tailed deer

A frozen stabilate was produced from Theileria cervi sporozoites in salivary glands of adult Amblyomma americanum. The stabilate was inoculated into three fallow deer (Cervus dama) and two white-tailed deer (Odocoileus virginianus). Following inoculation, the white-tailed deer developed parasitemias as determined by blood smear examination at 11 and 13 days postexposure. Repeat examination of blood from the three fallow deer for 30 days postexposure failed to reveal observable piro-plasms. These findings indicate that fallow deer are not as susceptible to the Theileria cervi found in white-tailed deer from North America. Thus, there are some questions regarding the taxonomic position of this organism.     

Elaeophorosis in red deer from Spain

Elaeophorosis, caused by Elaeophora elaphi, was observed in red deer (Cervus elaphus) from Toledo Province (Spain) for the first time. Adult specimens of Elaeophora elaphi were found in the hepatic vessels of nine of 151 red deer between October 1994 and September 1995; intensity of infection was two to 18 nematodes per host. Adult nematodes were only found during the period from fall through early spring. No differences were present between sex or age groups. Parasites were not found in a limited sample from fallow deer (Dama dama). Blood samples were negative for the presence of microfilariae.     

Elaphostrongylus cervi in a population of red deer (Cervus elaphus) and evidence of cerebrospinal nematodiasis in small ruminants in the province of Varese, Italy

Thirty-one faecal samples were collected from red deer in the northern area of Varese, in the Italian region of Lombardy, between August and October 2008. The animals had either been hunted or accidently killed. Examination for internal parasites showed a prevalence of 45.2% for Elaphostrongylus cervi larvae and species identification was confirmed by polymerase chain reaction (PCR). Ninety-seven faecal samples were also collected from two goat flocks grazing in the same area between December 2007 and May 2008. These showed a prevalence of 74.7% for lungworms. Furthermore, the central nervous systems from five goats and one sheep from this area with a history of neurologically related lameness were examined. Histopathology confirmed E. cervi cerebro-spinal nematodiasis in five cases out of six. This study demonstrates E. cervi transmission from wild to domestic ruminants when the animals graze in the same area, and the possible occurrence of clinical disease in infected goats and sheep associated with high prevalence in deer.     

Abomasal parasites in wild sympatric cervids, red deer, Cervus elaphus and fallow deer, Dama dama, from three localities across central and western Spain: relationship to host density and park management

A survey of abomasal parasites in cervids from Central Spain was conducted at 3 sites, Quintos de Mora (Toledo), Malué?ez de Arriba (Cáceres), and La Herguijuela (Cáceres). Commonly occurring helminths belonged to 3 polymorphic species of the Ostertagiinae: Spiculopteragia asymmetricalS. quadrispiculata, Ostertagia leptospicularis/O. kolchida, and O. drozdzi/O. ryjikovi. Trichostrongylus axei was found in very few cases. Ostertagia drozdzi/O. ryjikovi and the minor male morphotype, S. quadrispiculata, are reported for the first time in red deer from Spain. The 3 ostertagiine species are also reported for the first time in fallow deer from Spain. These 3 species of Ostertagiinae are primarily parasites of cervids, and nematode species characteristic of domestic ruminants were not present. Prevalence of infection by gastrointestinal parasites in cervids was high, ranging from 97.5 to 100%, across the 3 areas sampled. Mean intensity of infection and abundance showed a positive relationship to the population density of red deer. Helminth burdens were higher in fallow deer than in the sympatric red deer and may reflect the gregarious social structure and different foraging patterns of fallow deer.     

Theileriosis in a white-tailed deer (Odocoileus virginianus) fawn

A white-tailed deer (Odocoileus virginianus) fawn was collected in Missouri (USA) and submitted for diagnostic evaluation. Necropsy and histologic examination revealed severe Amblyomma americanum infestation, pronounced icterus, and marked hemosiderin deposition in the liver and spleen. Whole blood evaluation revealed a normocytic normochromic anemia and a piroplasm parasitemia of approximately 70%. The piroplasm was identified as Theileria cervi by polymerase chain reaction and sequencing of the V4 variable region of the 18S rRNA gene from a paraffin-embedded section of lung. Although T. cervi parasitemias have been commonly reported in healthy white-tailed deer, the severe parasitemia in this fawn may have contributed to overt clinical disease, perhaps as part of a combined malnutrition and parasitemia syndrome.     

Onchocercosis in red deer (Cervus elaphus) from Spain.

Onchocercosis, caused by Onchocerca flexuosa, was observed in red deer (Cervus elaphus) from Spain for the first time. Adult specimens of P. flexuosa were found in nodules in subcutaneous tissues in 42 of 125 (33%) red deer between October 1994 and September 1995; intensity of infection +/- SD was 3.93 +/- 5.26 nodules per infected host. A clear seasonal pattern in the distribution of nodules was observed, with higher values of prevalence and intensity in fall and winter in contrast to spring and summer. Significant differences were found among age groups in prevalence, but not in the mean intensity. No differences in infection were apparent between male and female adult red deer.     

Detection and identification of Theileria infection in sika deer ( Cervus nippon ) in China

The sika deer ( Cervus nippon ) is a first-grade state-protected animal in China and designated a threatened species by the World Conservation Union. To detect hemoparasite infection of sika deer, blood samples were collected from 24 animals in the Hubei Province Deer Center. Genomic DNA was extracted, and the V4 hypervariable region encoding 18S rRNA was analyzed by reverse line blot hybridization assay. PCR products hybridized with Babesia / Theileria genus-specific probes but failed to hybridize with any of the Babesia or Theileria species-specific probes, suggesting the presence of a novel, or variant, species. Here 18S rRNA and internal transcribed spacer (ITS) genes were amplified, cloned, and sequenced from 7 isolates. Alignment and BlastN of the cloned sequences revealed high similarities to the homologous 18S rRNA genes and ITS genes of Theileria cervi (AY735122), Theileria sp. CNY1A (AB012194), and Theileria sp. ex Yamaguchi (AF529272). Phylogenetic analysis based on the 18S rRNA gene and ITS sequences showed that all cloned sequences were grouped within the Theileria clade. Phylogeny based on the 18S rRNA gene divided the organisms into 2 groups. Group 1 was closest to Theileria sp. ex Yamaguchi (AF529272), and group 2 was distinct from all other identified Theileria and Babesia species. These results suggest the existence of Theileria sp. infection in sika deer in China. To our knowledge, this is the first report of cervine Theileria sp. in China.     

Assessment of domestic goats as a patent host of Elaphostrongylus cervi

A study was undertaken to determine whether domestic goats can serve as patent hosts of Elaphostrongylus cervi under natural or experimental conditions. Three-hundred and two fecal samples from 124 domestic goats raised outdoors in New Zealand, where E. cervi is enzootic, were tested for nematode larvae by the Baermann method. All samples were negative for E. cervi dorsal-spined larvae. Twenty juvenile male Nubian and Saanen goats obtained locally were assigned randomly to 5 dosage groups and were orally administered 5, 15, 35, 65, or 125 third-stage larvae of E. cervi, respectively. Two yearling female red deer (Cervus elaphus elaphus) each received 35 or 65 third-stage larvae as positive controls, and 2 uninoculated juvenile male goats served as negative controls. Fecal Baermann testing of pooled samples from the inoculated goats was conducted weekly for the first 80 days postinoculation (DPI) and daily thereafter until 250 DPI. No dorsal-spined larvae were recovered. One goat that had received 15 third-stage larvae displayed a mild transient posterior ataxia suggestive of cerebrospinal elaphostrongylosis. Gross postmortem examination did not reveal any direct evidence of nematodes in any of the goats, and only a few minor lesions were present. Histologically, these lesions were consistent with a parasite etiology. Histological evaluation of grossly normal lumbar and sacral spinal cord from 2 goats that had each received 125 third-stage larvae revealed eosinophilic meningoencephalitis and leukomyelitis, respectively, suggestive of the presence of parasites in the central nervous system. The 2 positive control red deer became patent with dorsal-spined larvae consistent with E. cervi at 131 DPI. These findings suggest that goats, at least those breeds utilized in this study, are not suitable patent hosts for E. cervi.     

Mandibular osteomyelitis in red deer (Cervus elaphus hispanicus) and fallow deer (Dama dama): occurrence and associated factors in free-living populations in southern Spain

The prevalence of mandibular osteomyelitis, which results in a condition called lumpy jaw, and factors associated with its occurrence were investigated in syntopic free-living populations of red deer (Cervus elaphus hispanicus) and fallow deer (Dama dama) in Spain. The study material consisted of 3,586 mandibles from 2,548 red deer and 1,038 fallow deer shot during sport hunting, herd management culls, and programs for population control between 1988 and 1997 (period 1) and 2002 and 2009 (period 2) in eastern Sierra Morena, southern Spain. Disease prevalence ranged from 0.36% to 10.91% among age groups. Older animals were significantly more likely to be affected than younger ones. Red deer stags had higher prevalence than other groups. There was a significantly higher prevalence in period 1, probably associated with differences in climatic and population conditions. High population densities of female red deer contributed significantly to occurrence of disease. Intensive herd management and poor environmental conditions were considered risk factors that increased susceptibility to disease. The study of this affliction could be useful for monitoring general herd welfare and ecologic changes in Mediterranean ecosystems.     

Foot infections associated with Arcanobacterium pyogenes in free-living fallow deer (Dama dama)

We describe foot infection associated with Arcanobacterium pyogenes in three adult male free-living fallow deer (Dama dama) from Sueve Regional Hunting Reserve (Principality of Asturias, Spain). Affected fallow deer were culled in November 1997 and 1998 during the hunting season. Necropsy, radiography, and microbiologic analysis were carried out for each animal. Unilateral swelling of one extremity at the coronary band was observed in all three cases. Areas of bone loss, severe periosteal reaction, and soft tissue swelling were seen on radiography. Lead fragments were observed in one fallow deer. Seven bacterial species were isolated, but only Arcanobacterium pyogenes was routinely found. Weather conditions in the area (mild temperatures and high humidity), the land (alternating pasture land and rock), the animal population density (both fallow deer and domestic herds of cows, horses, sheep, and goats, live side by side in the same areas), and hunting activities could be related to the frequency of these infections.     

Bovine virus diarrhea virus in free-living deer from Denmark

Free-living deer are suggested as a possible source of infection of cattle with bovine virus diarrhea (BVD) virus. To examine this hypothesis blood samples from 476 free-living deer were collected during two different periods and tested for BVD virus and antibody in Denmark. In 1995-96, 207 animals were tested. These included 149 roe deer (Capreolus capreolus), 29 fallow deer (Dama dama), 20 red deer (Cervus elaphus) and one sika deer (Cervus sika). For the remaining eight animals no species information was available. In 1998-99, 269 animals were tested including 212 roe deer and 57 red deer. The animals were selected from areas with a relatively high prevalence of cattle herds with a BVD persistent infection status in 1997 and 1998. All 207 samples from 1995-96 were found antibody-negative except two samples from red deer. Only 158 of the 207 samples were tested for virus and were all found negative. Of the 269 samples from 1998-99 all but one were antibody negative. The positive sample was from a red deer. All samples were virus-negative. It appears that BVD infection does not occur in roe deer in Denmark. The presence of antibody in a few red deer from various districts in Jutland probably results from cattle to deer transmission, rather than spread among deer. Hence, the possibility of free-living deer as a source of infection for cattle in Denmark seems to be remote.     

Hypodermosis of red deer in Spain

From February 1998 to January 1999, 106 red deer in the Autonomic Organism National Park "Quintos de Mora" (Toledo, central Spain) were evaluated to determine the prevalence and dynamics of infection with Hypoderma sp. by detection of subcutaneous larvae. Between six and 13 deer shot in selective hunting were examined monthly. Hypoderma sp. larvae were detected throughout the period except in June, July and August of 1998. Excluding the period during which no subcutaneous larvae were detected, the number of animals sampled was 80 (52 males and 28 females), belonging to three age classes: 12 calves (<1-yr-old), 19 yearlings (1-yr-old), and 49 adults (2- to 10-yr-old). All the third instar (L3) collected were identified as H. actaeon. Total prevalence during the period of larval detection was 61%. Prevalence in yearling and adult deer shot during the official hunting season was 89%. Monthly prevalence increased from September to January and decreased from February to May. In September and October, a small percentage of larvae were classified as first instar (L1). The rest of larvae collected between September and December were second instar (L2). Third instar (L3) predominated in January and February and was the only stage collected from March to May. Intensity ranged from 1 to 145 larvae. Intensities were >100 larvae in 6% of animals. Possible relationships of intensity or prevalence of infection with sex or age of the animals were evaluated. Significant differences in prevalence were observed among different host age classes. Prevalence was higher in yearlings (84%) than in adults (63%) and lowest in calves (17%).     

Seasonal changes in prevalence and intensity of Hypoderma actaeon in Cervus elaphus from central Spain

Hypodermosis in red deer, Cervus elaphus Linnaeus (Artilodactyla: Cervidae), caused by the warble fly, Hypoderma actaeon Latreille (Diptera: Oestridae), was studied at Quintos de Mora (Toledo, Spain) over a one-year period between October 1994 and September 1995. One hundred and twenty-five red deer were examined for the presence of warble fly larvae. The prevalence of warbles was 44.8% and the intensity of infection was 38.29 (SD +/- 61.32) warbles/deer infected. Clear seasonal variations were observed in prevalence and intensity, with the highest values in the autumn and winter. Statistically significant differences were observed in prevalence and intensity among age groups. In contrast, no differences were found in prevalence and intensity between males and females.