Parelaphostrongylus andersoni (Nematoda: Protostrongylidae) in white-tailed deer from Michigan

Dorsal-spined larvae in fecal samples from free-ranging white-tailed deer (Odocoileus virginianus) in Michigan and Pennsylvania were used as a source of larvae to infect a hand-raised white-tailed deer fawn. The fawn receive 200 third-stage larvae and passed dorsal-spined larvae in feces 66 days later. Muscleworm (Parelaphostrongylus andersoni), and meningeal worm (Parelaphostrongylus tenuis) were recovered at necropsy. Two white-tailed deer and seven wapiti (Cervus elaphus) exposed to larvae of the source from Pennsylvania harbored only P. tenuis. This is the first report of P. andersoni in the midwestern United States and extends the known range of this muscleworm in free-ranging white-tailed deer. Concurrent infections of P. andersoni and P. tenuis have not been established previously in experimentally infected fawns.     

Muscleworms, Parelaphostrongylus andersoni (Nematoda: Protostrongylidae), discovered in Columbia white-tailed deer from Oregon and Washington: implications for biogeography and host associations

Parelaphostrongylus andersoni is considered a characteristic nematode infecting white-tailed deer (Odocoileus virginianus). Host and geographic distribution for this parasite, however, remain poorly defined in the region of western North America. Fecal samples collected from Columbia white-tailed deer (O. v. leucurus) in a restricted range endemic to Oregon and Washington, USA, were examined for dorsal-spined larvae characteristic of many protostrongylid nematodes. Multilocus DNA sequence data (internal transcribed spacer 2 and cytochrome c oxidase subunit 1) established the identity and a new record for P. andersoni in a subspecies of white-tailed deer previously unrecognized as hosts. Populations of P. andersoni are now recognized along the basin of the lower Columbia River in Oregon and Washington and from south-central Oregon on the North Umpqua River. Current data indicate a potentially broad zone of sympatry for P. andersoni and Parelaphostrongylus odocoilei in the western region of North America, although these elaphostrongylines seem to be segregated, respectively, in white-tailed deer or in black-tailed and mule deer (Odocoileus hemionus) at temperate latitudes. The geographic range for P. andersoni in white-tailed deer is extended substantially to the west of the currently defined limit in North America, and we confirm an apparently extensive range for this elpahostrongyline. These observations are explored in the broader context of host and geographic associations for P. andersoni and related elaphostrongylines in North American cervids.     

Efficacy of ivermectin against Parelaphostrongylus andersoni (Nematoda, Metastrongyloidea) in white-tailed deer (Odocoileus virginianus)

Ivermectin was injected subcutaneously at 200 and 400 micrograms/kg of body weight into seven white-tailed deer (Odocoileus virginianus) in an attempt to control the muscle nematode Parelaphostrongylus andersoni. Counts of first-stage larvae in feces dropped to zero at 17 to 18 days posttreatment. Larvae reappeared in feces 1.5 to 6 wk later in six deer. Four deer were treated again approximately 9 wk after the first treatment; larval counts dropped to zero in 12 to 18 days. Larvae reappeared in low numbers 45 to 55 days after the second treatment. Because deer were held indoors on cement and the prepatent period of these worms is approximately 2 mo, the reappearance of larvae was not due to reinfection by accidental ingestion of gastropod intermediate hosts. Results suggest that ivermectin at dosages of 200 or 400 micrograms/kg of body weight suppressed larval production by adult female nematodes for several weeks or destroyed first-stage larvae in the lungs.     

Serological diagnosis of Parelaphostrongylus tenuis infection in white-tailed deer and identification of a potentially unique parasite antigen

Serological diagnosis of Parelaphostrongylus tenuis infection should offer many advantages over the currently used method of fecal analysis that relies on a patent infection. Toward this end, we investigated the presence of P. tenuis-specific antibodies in experimentally infected white-tailed deer (WTD) and of unique P. tenuis antigens that may be exploited for serodiagnosis. WTD infected with 6, 20 or 100-150 P. tenuis third-stage larvae (L3) had anti-parasite antibodies from as early as 21 days postinoculation (dpi) until the end of the experiment (147 dpi). Peak anti-P. tenuis enzyme-linked immunosorbent assay (ELISA) titers in individual animals ranged from 1:70 to 1:5,700. Serum from infected WTD reacted with 5 distinct P. tenuis L3 antigens (105, 45, 37, 32, and 19 kDa) as detected by the immunoblotting technique. Serum from caribou infected with Parelaphostrongylus andersoni or Elaphostrongylus rangiferi reacted with all antigens except the 37-kDa antigen of L3, indicating that it may be unique to P. tenuis and can serve as a serodiagnostic antigen. The 37-kDa antigen appears to be present in the adult P. tenuis but not adult E. rangiferi or E. cervi. The development of an ELISA utilizing the unique antigen of P. tenuis should lead to a reliable diagnostic assay for P. tenuis infection in WTD.     

A search for Parelaphostrongylus andersoni in white-tailed deer from Maine.

Longissimus dorsi muscles from 42 white-tailed deer (Odocoileus virginianus) from Maine (USA) were examined for the Parelaphostrongylus andersoni. No adult nematodes were found. Prevalence based on the Poisson approximation of a binomial distribution could have been between 0 and 9% (95% C.I.). However, based on prevalence documented elsewhere (10 to 18%), it is unlikely that P. andersoni occurs in white-tailed deer in central Maine.     

Gastropod availability and habitat utilization by wapiti and white-tailed deer sympatric on range enzootic for meningeal worm

Gastropod occurrence and the utilization of habitat by sympatric populations of wapiti (Cervus elephus) and white-tailed deer (Odocoileus virginianus) on range enzootic for meningeal worm (Parelaphostrongylus tenuis) were studied on Cookson Hills Wildlife Management Area (WMA) in eastern Oklahoma. Visual observations and fecal pellet group transect data indicated that wapiti spent the majority of their time in open fields and meadows where we recovered the least numbers of gastropods. Although deer were frequently observed in open areas, visual sightings and transect data indicated that they spent more time in forested areas where we recovered the most gastropods. Gastropods harbored low numbers of P. tenuis larvae (0.00 to 0.06 larvae/gastropod) in all habitat types with the greatest recovery from red oak white oak-hickory forests (0.34 larvae/gastropod). Our results indicate that the reason a viable wapiti herd exists on Cookson Hills WMA in a P. tenuis enzootic area is at least partially because of the habitat preference by wapiti and the reduced availability of infected gastropods in the selected areas prefered by the wapiti. We were not able to detect any free-ranging wapiti that were shedding P. tenuis larvae nor were we able to detect past or sub-clinical infections with P. tenuis in wapiti.     

Experimental haemonchosis in white-tailed deer

White-tailed deer (Odocoileus virginianus) were successfully infected with Haemonchus contortus of sheep origin. Individual deer in each of three groups were inoculated with 0, 25,000, and 100,000 larvae respectively. Severity of infection was related to dose and signs of infection were most obvious in the heavily inoculated animals. Infected deer were weak, emaciated, and anemic, similar to the clinical response in sheep. Hemoglobin, packed cell volume, and total serum protein values for both infected groups were significantly lower than for the controls. Inhibition of larval growth was noted in both infected groups, but was most pronounced in the group which received 100,000 larvae. Inhibition of egg production was also noted in this group. The potential importance of H. cortortus in deer populations was discussed.     

Bot fly larvae (Cephenemyia spp., Oestridae) in mule deer (Odocoileus hemionus) from Utah

Ninety-nine mule deer (Odocoileus hemionus) from four Utah counties (Cache, Utah, Sanpete and Sevier) were examined for larvae of Cephenemyia spp. in 1985 and 1986. Numbers of first, second and third stage bot fly instars were related to age, sex, year and geographic location of the mule deer. Fawns and adult deer greater than or equal to 5.5 yr had a significantly (P less than or equal to 0.05) higher intensity (means = 37 and means = 68, respectively) of infection than the 1.5- and 2.5-yr-old age groups (means = 19 and means = 26, respectively). Infection by larvae was not significantly different between sexes. Infection was 100% in both years, but the mean intensity was significantly lower in 1986 (P less than 0.05). The decline may be related to differences in soil moisture between the 2 years. In 1985, 82% of the deer examined were infected with all three instars. Seventy-seven percent of all first instar larvae were observed in the trachea, usually in the fold immediately posterior to the epiglottis and corniculate cartilages. This new site of attachment for first instar larvae has not previously been recognized.     

Faecal egg output and herbage contamination with infective larvae of species of Ostertagia and Oesophagostomum from naturally infected farmed sika deer Cervus nippon in Lithuania

Faecal egg outputs and subsequent herbage larval contamination with third stage larvae (L3) of Ostertagia spp. and Oesophagostomum spp. from a herd of naturally infected sika deer Cervus nippon were examined in the same pasture in 2001/2002 in Lithuania. Sika deer were infected with Ostertagia circumcincta, O. kolchida, O. spiculoptera, Oesophagostomum radiatum, O. columbianum and O. venulosum. Faecal egg output in adult deer peaked in the spring during the periparturient period and also in late August, compared with a peak in egg output in calves during September to November. Herbage contamination with L3 of Ostertagia spp. peaked in June but larvae were not present on pastures from the end of September. Hence the highest risk of infection was in early born calves grazed on pastures in July. Infective larvae of Oesophagostomum spp. did not survive during the winter, but the nematodes were reintroduced onto the pastures by adult deer in the spring.     

The potential for false-positive diagnosis of protostrongyliasis by extraction of larvae from feces

The potential of protostrongylid first-stage larvae (L1) to survive passage through the alimentary canal of non-infected mammals was investigated. Parelaphostrongylus tenuis L1 were collected from feces of an experimentally infected white-tailed deer (Odocoileus virginianus). We utilized two red deer (Cervus elaphus) and four laboratory rats (Rattus norvegicus) which were each fed the L1 of P. tenuis. Larvae were recovered, intact and alive, from the fecal samples of all six animals. Larvae of P. tenuis, and probably of other related species, can survive passage through the alimentary canal of uninfected mammals and they can be collected from feces using the Baermann technique and other related larval extraction methods. Rain water was found to be successful in the dispersal of P. tenuis L1 from the feces of infected animals. These findings raise the possibility of ingestion of L1 and their subsequent passage, by uninfected animals. This potential for false-positive diagnosis of infection in live animals necessitates accurate interpretation of a host's infection-status. Such findings reinforce the need for a reliable method of diagnosing infections in live animals.     

Prevalence of Parelaphostrongylus tenuis in white-tailed deer in northern New York.

The prevalence and distribution of "brainworm" (Parelaphostrongylus tenuis) were examined in northern New York (USA) from 1986 to 1989. Sixty nine (46%) of 151 white-tailed deer (Odocoileus virginianus) heads examined, contained adult P. tenuis. The proportion of infected individuals was not significantly different between males and females. Prevalence was significantly greater in the adult age class as compared to the juvenile age class (P less than 0.01). Deer pellet samples were examined for prevalence of P. tenuis-like larvae. Pellet samples in New York had an overall prevalence of 60%. The effects of precipitation and host density on prevalence of P. tenuis in deer was not significant.     

Parelaphostrongylus odocoilei: Life cycle in experimentally infected cervids including the mule deer, Odocoileus h. hemionus

The life cycle of a metastrongyloid nematode, Parelaphostrongylus odocoilei, was successfully completed in three members of the Cervidae: mule deer (Odocoileus h. hemionus), black-tailed deer (O. h. columbianus), and moose (Alces alces andersoni). The terrestrial gastropod, Triodopsis multilineata, was the experimental intermediate host. White-tailed deer (O. virginianus dacotensis) were refractory to infection. The prepatent period of P. odocoilei was significantly shorter in mule deer (X = 53 days) than in the black-tailed deer or moose. There was an inverse relationship between the size of the infective inoculum and the duration of the prepatent period of P. odocoilei in mule deer, but not in black-tailed deer. The duration and intensity of larval production of P. odocoilei were higher in mule deer than in the other hosts. Peak larval production in the feces (approximately 14,000 larvae/g) of mule deer was in excess of previous reports for elaphostrongyline nematodes, regardless of the size of the infective inoculum. Daily larval production, estimated at 3 to 4 × 10⁶ larvae/day, was six times higher than estimates for other elaphostrongylines. The duration of patency was not clearly established, but three mule deer and one black-tailed deer passed larvae for 12, 18, 24, and 18 months, respectively. On the basis of the reduced prepatent period and increased length and intensity of larval production, O. h. hemionus is considered the primary host of P. odocoilei.     

Heterologous transmission with Dictyocaulus capreolus from roe deer (Capreolus capreolus) to cattle (Bos taurus)

Eight Swedish Red Breed cattle, about 2 months old, were experimentally infected with a Swedish isolate of Dictyocaulus viviparus (Dviv-Se) from cattle and D. capreolus from roe deer. The aims were to determine whether the roe deer lungworm is infective to cattle or if it can induce seroconversion in cattle against D. viviparus as measured with an ELISA. Four calves which were given 500 Dviv-Se infective larvae (L3) each by larval dosing for two successive days developed patent infection between days 23 and 25 post-inoculation (PI). Larval output varied among the calves and during the patent period. However, maximum recovery occurred between 28 and 56 days PI with peak shedding on day 37 PI. Shedding ceased at day 58 PI and adult worms were recovered from one calf at necropsy (day 67 PI). No immature worms were recovered from the lungs at necropsy. Seroconversion was detected on days 35-42 PI. One Dviv-Se infected calf became seronegative on day 67 PI whereas the other calves still remained seropositive during this period. Prepatency and patency periods of D. viviparus and serological findings in this study basically conform to previous studies. Each calf that was infected with 400 L3 of D. capreolus for two successive days, and about 800 L3 of the same species about 8 weeks later, did not develop to patency based on faecal and post-mortem examinations. Consequently, under the conditions of this study, D. capreolus was not infective to cattle. Two of the four calves that were infected with L3 from roe deer were challenged with L3 cultured from faeces of the Dviv-Se-infected calves. This infection did not develop to patency. Whether this was due to cross-protection as a result of the prior priming with L3 from roe deer is not clear. However, if it is so, it opens up the possibility of using D. capreolus L3 for preventing bovine dictyocauliasis.     

Identification of dorsal-spined larvae from free-ranging wapiti (Cervus elaphus) in southwestern Manitoba, Canada

Dorsal-spined first-stage larvae recovered from feces of free-ranging wapiti (Cervus elaphus) were passaged through snails (Triodopsis multilineata) and two hand-raised white-tailed deer fawns (Odocoileus virginianus). A total of 74 adult Parelaphostrongylus tenuis were recovered from the fawns; no other protostrongylid nematodes were recovered. The study indicates that wapiti may be infected with natural infections of meningeal worm and pass larvae suitable for transmission to gastropod intermediate hosts. Wapiti from areas endemic with P. tenuis should not be translocated to areas currently free of the parasite.     

Persistent Ehrlichia chaffeensis infection in white-tailed deer

Four white-tailed deer (Odocoileus virginianus) were inoculated intravenously with a deer-origin isolate (15B-WTD-GA) of Ehrlichia chaffeensis. The course of infection was monitored using indirect fluorescent antibody (IFA), polymerase chain reaction (PCR), and culture over a 9 m period. All deer became rickettsemic within 24 days post inoculation (DPI), and all developed antibody titers >1:64 to E. chaffeensis by 17 DPI. Titers in all deer fell below 1:64 during 87 to 143 DPI. One deer exhibited a second period of seropositivity (peak titer of 1:256) from 207 to 271 DPI but was culture and PCR negative during this period. Rickettsemia was confirmed by reisolation of E. chaffeensis as late as 73 to 108 DPI in three deer. Positive PCR results were obtained from femur bone marrow of one deer and from rumenal lymph node of another (leer at 278 DPI. None of the deer developed clinical signs, hematologic abnormalities, or gross or microscopic lesions attributable to E. chaffeensis. Two uninoculated control deer were negative on all tests through 90 DPI at which time they were removed from the study. Herein we confirm that white-tailed deer become persistently infected with E. chaffeensis, have initial rickettsemias of several weeks duration and may experience recrudescence of rickettsemia, which reaffirm the importance of deer in the epidemiology of E. chaffeensis.     

Persistence of Trichinella spiralisin Rat Carcasses Experimentally Mixed in Different Feed

Trichinella spiralis infected rat carcasses were incubated for 6 weeks in several animal feeds to assess how long Trichinella can present a risk for an outbreak in contaminated feeds. In groups of 6, 24 infected target rats were placed in silage, grained barley, propionic acid-preserved feed, and also into simulated pasture conditions. Test environments were sampled after one-, 2-, 4-, and 6-week-incubations. Trichinella larvae were recovered by digestion, and their infectivity was evaluated in rats. A two-week incubation reduced the number of recovered larvae, but still after 6 weeks low numbers were isolated from all feeds except from the experimental group simulating pasture conditions. After 2 weeks storage, the larvae were infective in all storage environments. However, up to 4 weeks, they survived only in the propionic acid-fermented feed and there in small numbers with reduced reproductive capability. This indicates the possibility of farm animals to get infection from rats or other infected material being hazardously mixed with hay or other feed. If silage is stored for at least one month before use, however, the risk from this forage appears to be minimized.     

Potential for cross-transmission of Dictyocaulus viviparus between cattle and white-tailed deer

Development of an in vitro culture system for infectious Dictyocaulus viviparus larvae made it possible to study the potential cross-transmission of D. viviparus between white-tailed deer (Odocoileus virginianus) and cattle (Bos taurus). Between 26 September 1995-29 February 1996, six parasite-free bull calves were individually inoculated with 15 to 50 infective third stage larvae (L3)/kg of body weight cultured from adult D. viviparus collected from white-tailed deer. Three bull calves were simultaneously inoculated with 45 L3/kg of body weight recovered from cattle either by the Baermann technique or by in vitro culture as above. All three calves inoculated with the homologous cattle strain became patently infected while all six calves inoculated with the heterologous deer strain remained negative for the presence of D. viviparus in the feces and in the lungs upon necropsy.     

Meningeal worm is a long-lived parasitic nematode in white-tailed deer

A natural infection of the meningeal worm, Parelaphostrongylus tenuis, persisted for at least 3.7 yr in a white-tailed deer (Odocoileus virginianus). The deer was 5-7 yr old and was shedding dorsal-spined nematode larvae at the time of quarantine. Larvae were extracted from all fecal samples collected up to 730 days post-quarantine (dpq) and thereafter only at 862 dpq and at necropsy (1,350 dpq). Live adults of P. tenuis, one male and one female, were recovered from the cranium at necropsy. Parelaphostrongylus tenuis infections are long lived and latent periods may be extended. Our findings reaffirm the need for reliable antemortem diagnosis to identify non-patent P. tenuis infections to prevent inadvertent introduction of infected animals to non-endemic areas.     

Trichinella spiralis: antifecundity and antinewborn larvae immunity in swine

The development of antifecundity and antinewborn larvae immunity in swine infected with Trichinella spiralis was investigated. In primary infections, adult female worm fecundity dropped sharply after 3 weeks, although adults could be recovered from the small intestine for at least 7 weeks after infection. In challenge infections of pigs infected previously, adult female worm fecundity was depressed up to 51% and the adults were expelled within 3 weeks. Since immune pigs are almost completely resistant to the secondary establishment of muscle larvae, this suggested the existence of immune effector mechanisms also acting on the newborn larvae. This was supported by observations, using an indirect fluorescent antibody assay, that pig antibody bound to the surface of the newborn larvae. Passive transfer of immune pig serum resulted in a large reduction in muscle larvae burden in both infected pig and rat recipients. Adult female worm fecundity in such immune serum recipients was reduced only by 20% and worm survival in the intestine was unaffected. These results indicate that immunity to the newborn larvae, in addition to antifecundity effects, are responsible for the high levels of acquired resistance to T. spiralis in swine.     

Mortality of fallow deer (Dama dama) experimentally-infected with meningeal worm, Parelaphostrongylus tenuis.

Six fallow deer (Dama dama) fawns died after receiving 25 to 150 infective larvae of Parelaphostrongylus tenuis. Fawns given higher doses usually died sooner (6 to 23 days) than those given lower doses (54 to 67 days). Early deaths were associated with severe acute peritonitis resulting from perforation of the intestinal wall; later deaths were associated with paralysis and inability to rise. Numerous adult P. tenuis were found within neural tissues of the brain and spinal cord in the three fawns with paralysis. One white-tailed deer (Odocoileus virginianus) exposed to infective larvae from the same source survived infection without exhibiting clinical signs and began passing larvae in feces 88 days post-exposure. At the doses used in this study, meningeal worm caused fatal infections in fallow deer. Results are compared to published observations of fallow deer naturally-infected with P. tenuis.