Development of the large American liver fluke, Fascioloides magna, in white-tailed deer, cattle, and sheep.

The comparative development of Fascioloides magna in white-tailed deer, cattle and sheep has been studied. Flukes were recovered from 72% of 32 deer administered 40 to 500 metacercariae, from 82% of 11 cattle administered 10 to 500 metacercariae, and from 53% of 15 sheep administered 8 to 200 metacercariae. The percentage recovery of the flukes administered as metacercariae was 4.1% of 6,130 in deer, 5.7% of 2,510 in cattle, and 4.7% of 1,213 in sheep. Flukes were recovered only from livers of infected deer, while in cattle, 1 fluke was also found in the lungs of each of 2 animals. In sheep, all but 10 flukes were recovered from the livers; 6 were found in the lungs and 4 in the abdominal cavities. The black iron porphyrin pigment associated with F. magna infection was found to be most widespread in cattle and sheep, but was also a pathognomonic feature in deer. Growth of the fluke was similar in all 3 host species tested, but eggs were passed only from deer, the normal definitive host. In cattle, the eggs were retained in the liver, and F. magna was lethal to sheep before its own maturity was attained. In cattle and deer, flukes matured approximately 7 months after exposure, but immature migrating flukes were found 12 months after infection and apparently can remain in this retarded state for an undetermined period of time.     

Domestic sheep as a rare definitive host of the large American liver fluke Fascioloides magna

Two domestic sheep were inoculated orally with 250 metacercariae of Fascioloides magna. One sheep died 4 mo after inoculation and 22 immature F. magna were recovered. The second sheep remained healthy and was killed 8 mo after inoculation. Ten mature F. magna were recovered from hepatic parenchyma, and eggs of F. magna were observed in areas of the liver where flukes resided and from feces. Although F. magna is almost always lethal to sheep, this sheep was a rare definitive host.     

Fascioloides magna in white-tailed deer (Odocoileus virginianus): observations on the pairing tendency

Fascioloides magna (Bassi 1875) was recovered from 227 of 312 (73%) white-tailed deer (Odocoileus virginianus) in southern Texas between 1971 and 1975. Deer from age classes more than 1 year old had infection rates of from 64 to 84%. Few dead flukes were found. Flukes occurred exclusively in the livers of infected animals and were paired in 256 of 301 (85%) fibrous hepatic capsules. The prevalence of immature flukes with an average of one immature per infected liver was similar in all age classes, suggesting a relationship between fluke pairing and maturation.     

Serologic survey for transmissible gastroenteritis virus neutralizing antibodies in selected feral and domestic swine sera in the southern United States

Serum samples collected from feral and domestic swine (Sus scrofa) in Florida and feral swine in Georgia and Texas were assayed by plaque reduction for their virus neutralizing (VN) antibodies against the porcine transmissible gastroenteritis virus (TGE). None of 560 samples collected from feral swine contained VN antibodies for TGE virus, but experimentally infected feral swine seroconverted. None of 665 samples from domestic swine contained TGE-VN antibodies. These results indicate feral swine are not a significant reservoir for TGE virus in southern states, but are capable of becoming infected and developing VN antibodies against TGE.     

Liver enzymes and blood metabolites in a population of free-ranging red deer (Cervus elaphus) naturally infected with Fascioloides magna

We investigated the effects of Fascioloides magna infection on the serum biochemistry values of the naturally infected red deer population in eastern Croatia. The investigation was performed on 47 red deer with F. magna infection confirmed patho-anatomically in 27 animals (57.4%). Fibrous capsules and migratory lesions were found in 14 deer while only fibrous capsules without migratory lesions were found in 13 deer. In 13 deer both immature and mature flukes were found, in 5 deer only immature flukes were found and in 9 deer only mature flukes were found. Fascioloides magna infected deer with fibrous capsules and migratory lesions had significantly higher values for lactate dehydrogenase (LDH), glutamate dehydrogenase (GLDH) and globulin, and lower values for albumin/globulin ratio and glucose compared to uninfected deer. Fascioloides magna infected deer with fibrous capsules without the presence of migratory lesions had higher values for alanine aminotransferase (ALT) and globulin, and lower values for albumin/globulin ratio and glucose, than the uninfected deer. The number of immature flukes was positively correlated with values of aspartate aminotransferase (AST), γ-glutamyltransferase (GGT), LDH, GLDH, urea and triglycerides. The number of migratory lesions was positively correlated with GGT, GLDH, globulin and urea values. The creatinine value was positively correlated with the number of mature flukes. The trial showed that F. magna infection causes significant changes in serum biochemistry. Moreover, these changes do not completely resemble changes following F. hepatica infection. Further investigation of changes in liver enzymes and other serum metabolites in controlled, experimentally induced fascioloidosis in red deer is needed to better understand the pathogenesis of F. magna.     

DNA-based identification of a hepatic trematode in an elk calf

Liver fluke infection was identified as a probable cause of clinical disease in an approximately 6-mo-old elk (Cervus elaphus) in coastal Oregon. Clinical pathology and necropsy findings are described. The alcohol-fixed flukes that were submitted for identification were similar in size to Fasciola hepatica, but their shape resembled Fascioloides magna in that they lacked a distinctive anterior cone. A few structures consistent with the eggs of F. magna were observed in liver lesions, suggesting that at least some of the worms were sexually maturing. Due to difficulties in morphologic identification associated with improper fixation technique, DNA analysis was used to compare small subunit (SSU) and internal transcribed spacer 2 ribosomal RNA gene sequences of the recovered parasites with those of F. hepatica and F. magna, confirming these small, but sexually mature flukes were F. magna. This is the first publication of the SSU gene sequence for F. magna. Phylogenetic analysis showed that it is related to, but is an outlier, to the genus Fasciola. Due to the high mortality rate associated with this disease outbreak, the overall significance of trematodiasis in the herd is unclear.     

Fasciola hepatica: site of resistance to reinfection in cattle

The effective site of resistance to reinfection of cattle with Fasciola hepatica was examined by recovery of challenge flukes from either the liver or body cavity. Calves infected 18 or 26 weeks previously with F. hepatica showed levels of resistance to reinfection of 56 and 94%, respectively, as assessed by recovery of flukes from the liver 15-16 weeks after challenge. Plasma glutamate dehydrogenase (EC 1.4.1.3; GLDH) enzyme activity estimations revealed only a marginal increase in these latter resistant calves compared with previously naive controls, indicating minimal liver damage as a result of migrating flukes. By comparison, when immature challenge flukes were recovered from the body cavity 4 or 14 days after infection of corresponding previously infected or naive calves, there was no significant difference in numbers. It appears, therefore, that, in cattle, resistance mechanisms are effective against challenge flukes at or soon after penetration of the liver.     

Efficacy of triclabendazole against natural infections of Fascioloides magna in wapiti.

Fourteen free-ranging adult wapiti (Cervus elaphus nelsoni) were captured in Banff National Park, Alberta (Canada) and held in captivity near Edmonton. A 24% suspension of triclabendazole at doses of 30 to 100 mg/kg body weight was drenched into the rumen of eight females and four males. Two male wapiti were used as untreated controls. Animals were killed and examined at 4 (n = 3), 6 (n = 4), or 8 (n = 4) wk after treatment. Efficacy was 90% against immature Fascioloides magna collected 4 wk after treatment and 98% against adult flukes collected 4, 6 or 8 wk after treatment. All 32 flukes recovered from control wapiti were active and apparently healthy. Treatment at 50 to 60 mg/kg is recommended against F. magna in wapiti. A protocol for treating infected wapiti is outlined.     

Potential sites of virus latency associated with indigenous pseudorabies viruses in feral swine

Free-ranging feral swine (Sus scrofa) are known to be present in at least 32 states of the USA and are continuously expanding their range. Infection with pseudorabies virus (PRV) occurs in feral swine and the primary route of transmission in free-living conditions seems to be venereal. Between 1995 and 1999, naturally infected feral swine and experimentally infected hybrid progeny of feral and domestic swine, were kept in isolation and evaluated for occurrence of latent PRV indigenous to feral swine in sacral and trigeminal ganglia and tonsil. Sacral ganglia were shown, by polymerase chain reaction (PCR) amplification of the thymidine kinase (TK) gene of PRV, to be the most frequent sites of latency of PRV. Nine (56%) of 16 sacral ganglia, seven (44%) of 16 trigeminal ganglia, and five (39%) of 13 tonsils from naturally infected feral swine were positive for PCR amplification of TK sequences of PRV. These tissues were negative for PRV when viral isolation was attempted in Vero cells. DNA sequencing of cloned TK fragments from the sacral ganglia of two feral swine, showed only one nucleotide difference between the two fragments and extensive sequence homology to fragment sequences from various domestic swine PRV strains from China, Northern Ireland, and the USA. The hybrid feral domestic swine, experimentally inoculated with an indigenous feral swine PRV isolate by either the genital or respiratory route, acquired the infection but showed no clinical signs of pseudorabies. Virus inoculated into either the genital or respiratory tract could, at times, be isolated from both these sites. The most common latency sites were the sacral ganglia, regardless of the route and dose of infection in these experimentally infected hybrids. Nine of 10 sacral ganglia, six of 10 trigeminal ganglia, and three of 10 tonsils were positive for PCR amplification of TK sequences. No virus was isolated from these tissues in Vero cells. The demonstration of the sacral ganglia as the most common sites of latency of pseudorabies viruses indigenous to feral swine, supports the hypothesis that these viruses are primarily transmitted venereally, and not by the respiratory route as is common in domestic swine, in which the trigeminal ganglia are the predominant sites of virus latency.     

Effects of six fasciolicides against Fascioloides magna In white-tailed deer

Thirty-three white-tailed deer (Odocoileus virginianus) of various ages, both sexes, and in good physical condition were captured for anthelmintic evaluation of six compounds against the large American liver fluke, Fascioloides magna. Based on fluke mortality, hexachlorophene administered at the rate of 12 to 26 mg/kg of body weight was lethal to 5 of 10 mature flukes in seven deer. Nitroxynil at 11 to 24 mg/kg inhibited egg production, but did not kill mature flukes in eight deer. Rafoxanide at 12 to 25 mg/kg killed 6 of 8 (75%) immature flukes in eight deer, but was not effective against 17 mature flukes. Clioxanide at 16 to 38 mg/kg, diamphenethide at 255 to 280 mg/kg, and hexachloroethane at 463 to 629 mg/kg were not effective against F. magna in four, two and four deer, respectively. There was no indication that treatment with fasciolicides at the higher dose rates was more efficacious than at the lower dose rates. Detection of fluke eggs in the feces was a reliable method for diagnosing the presence of mature F. magna in deer prior to treatment, but was not reliable for measuring fasciolicidal activity of all compounds tested.     

Venereal transmission of pseudorabies viruses indigenous to feral swine

Between 1995 and 1998, we designed a series of studies in which we attempted to determine the main routes of transmission involved in the natural infection of pseudorabies virus (PRV) indigenous to free-ranging feral swine (Sus scrofa). Naturally infected feral sows transmitted the infection to uninfected feral boars, with which they had been commingled for a 6-wk period. Pseudorabies virus was isolated from boar preputial swabs, but not from nasal swabs. Three of the same PRV-infected feral sows did not transmit the infection to domestic boars during a 16 wk commingling period, despite the fact that they became pregnant. Feral boars, naturally infected with PRV transmitted the virus to domestic gilts while penned together during 6 wk. Pseudorabies virus was isolated from vaginal swabs, but not from nasal swabs of gilts, after 2 and 3 wk of commingling. When the same infected boars were commingled with either feral or domestic boars for 13 wk, PRV transmission did not occur. None of the exposed boars developed neutralizing antibodies or yielded virus from their preputial or nasal swabs. Our results indicate that PRV indigenous to feral swine is preferentially transmitted to feral or domestic swine of the opposite sex by the venereal route. This mode of transmission differs from that seen in the natural transmission of PRV prevalent in domestic swine, where contaminated secretions, excretions and aerosols are responsible for the spread of the virus. Based on these results, we feel that as long as feral swine do not come into direct contact with domestic swine, PRV-infected feral swine probably pose only a limited risk to the success of the National Pseudorabies Eradication Program. The fact that PRV is usually transmitted from feral to domestic swine at the time of mating would indicate that the isolation of domestic herds by the use of a "double fence," should be adequate protection against reinfection with PRV.     

Molecular epidemiology of Brucella abortus isolates from cattle, elk, and bison in the United States, 1998 to 2011

A variable-number tandem repeat (VNTR) protocol targeting 10 loci in the Brucella abortus genome was used to assess genetic diversity among 366 field isolates recovered from cattle, bison, and elk in the Greater Yellowstone Area (GYA) and Texas during 1998 to 2011. Minimum spanning tree (MST) and unweighted-pair group method with arithmetic mean (UPGMA) analyses of VNTR data identified 237 different VNTR types, among which 14 prominent clusters of isolates could be identified. Cattle isolates from Texas segregated into three clusters: one comprised of field isolates from 1998 to 2005, one comprised of vaccination-associated infections, and one associated with an outbreak in Starr County in January 2011. An isolate obtained from a feral sow trapped on property adjacent to the Starr County herd in May 2011 clustered with the cattle isolates, suggesting a role for feral swine as B. abortus reservoirs in Starr County. Isolates from a 2005 cattle outbreak in Wyoming displayed VNTR-10 profiles matching those of strains recovered from Wyoming and Idaho elk. Additionally, isolates associated with cattle outbreaks in Idaho in 2002, Montana in 2008 and 2011, and Wyoming in 2010 primarily clustered with isolates recovered from GYA elk. This study indicates that elk play a predominant role in the transmission of B. abortus to cattle located in the GYA.     

Epidemiological Survey on the Infection of Intestinal Flukes in Residents of Muan-gun, Jeollanam-do, the Republic of Korea

Infection status of intestinal flukes was investigated in residents of Muan-gun, Jeollanam-do, the Republic of Korea. Total 1,257 fecal samples of residents were examined by formalin-ether sedimentation technique and Kato-Katz thick smear method. Helminth eggs were detected from 95 (7.6%) residents, and eggs of heterophyid flukes and Clonorchis sinensis were found from 62 (4.9%) and 40 (3.2%) cases, respectively. The larger heterophyid eggs, somewhat dark-brown in color and 37.7 × 21.5 µm in average size, and found in 32 (2.6%) out of 62 egg positive cases of heterophyid flukes. To confirm the adult flukes, we performed worm recovery from 12 cases after praziquantel treatment and purgation with MgSO₄. A total of 1,281 adult flukes, assigned to 7 species, were recovered from 9 cooperative cases. Heterophyes nocens (total 981 specimens) was collected from 9 cases, Stictodora fuscata (80) from 7, Gymnophalloides seoi (75) from 5, Pygidiopsis summa (140) from 3, Stellantchasmus falcatus (3) from 2, and Stictodora lari and Acanthotrema felis (each 1 worm) from 1 case each. The intrauterine eggs of S. fuscata collected from the recovered worm were identical with the larger heterophyid eggs detected in the stool examination. By the present study, it was confirmed that A. felis is a new intestinal fluke infecting humans, and residents in Muan-gun, Jeollanam-do are infected with variable species of intestinal trematodes.     

Visceral helminth communities of an insular population of feral swine

Nine species of helminths, all nematodes, were recovered from the viscera of 48 feral swine (Sus scrofa) from Cumberland Island, Georgia. Both the overdispersed frequency distributions and the abundances of the four common species of helminths (Stephanurus dentatus, Metastrongylus apri, M. pudendotectus and Gongylonema pulchrum) did not vary significantly across the main and interactive effects of host sex and/or seasons. Whether or not the present low population densities of feral swine on Cumberland Island has influenced the pattern of fluctuations in abundances of helminth species across seasons as often observed in helminth communities from other hosts was not resolved. The apparent recent decline in prevalences and abundances, and the loss of certain species from the helminth communities of feral swine on the island may be explained partially by the decreasing transmission potentials of direct life cycle species caused by a recent marked reduction of numbers of individuals in the host population. Conversely, the apparent increased prevalence and abundance of three species of helminths (S. dentatus, M. apri and M. pudendotectus) may be related to their common utilization of earthworms as paratenic or intermediate hosts. Gongylonema pulchrum was the only helminth in which abundances seemed to remain unchanged. This was the only species that was not strictly host specific to feral swine. We found no evidence that helminth infections were responsible for morbidity or mortality in this feral swine population.     

Trichinella spiralis in an agricultural ecosystem. III. Epidemiological investigations of Trichinella spiralis in resident wild and feral animals

As part of a larger epidemiological study examining the transmission of Trichinella spiralis in an agricultural ecosystem, resident wild and feral animals were trapped to determine the extent of their involvement in the natural, on-farm cycling of the parasite among swine. During a 21-mo-study, seven of 15 skunks (Mephitis mephitis), one of three opossums (Didelphis virginiana), two of two feral domestic cats and a raccoon (Procyon lotor) were found to be infected, while five shrews (Blarina brevicauda) and 18 deer mice (Peromyscus spp.) were uninfected. Most of the former hosts probably became infected by scavenging dead infected swine or rats (Rattus norvegicus). However, infections obtained through predation of living rats, particularly with regard to the cats, cannot be excluded. Our observations do not suggest that there was transmission of T. spiralis from the wild animals to swine. Therefore, transmission of T. spiralis appeared to occur only from the farm's swine and rats to the associated wild and feral animals.     

Brucella abortus in wildlife on selected cattle farms in Alabama

Two studies of brucellosis in wildlife on farms where the brucellosis infection prevalence in cattle was known are reported. On a research farm, 233 feral animals of 22 mammalian species and 12 of seven avian species were trapped during three time periods. Sixty were studied before cattle were introduced, 128 were studied while 501 cattle infected with Brucella abortus were calving and aborting, and 60 specimens were collected 20 mo after the last infected cow calved. Selected tissues from 229 wild animals were cultured and sera from 138 were examined using the brucellosis card, standard tube agglutination (STA), 2-mercaptoethanol (2-ME) and rivanol (RIV) tests. Brucella abortus was not recovered from any animals sampled prior to cattle being introduced and all sera collected were negative. Brucella abortus was isolated from four opossums (Didelphis virginiana) and one raccoon (Procyon lotor) in the group of animals trapped during the calving period. Three serums were tested and had STA titers ranging from 1:100 to 1:200. Of 68 sera only one had antibodies. Brucella were not isolated from 59 animals trapped after the calving period and only one of 42 serums had antibodies. On regional cattle farms, 243 wild animals were trapped. Brucellae were not isolated from 223 animals which were cultured. No serums had significant titers. The data from this study suggest opossums and raccoons can be infected from cattle but are unlikely to maintain the infection.     

Hyperparasitism of trichodinid ciliates on monogenean gill flukes of two marine fish

Two unusual cases of hyperparasitism of trichodinid ciliates on monogenean gill flukes are described from southern Israel (Red Sea). The first case occurred in cultured European sea bass Dicentrarchus labrax infected by Diplectanum aequans, while the second was observed in a feral devil firefish Pterois miles infected by Haliotrema sp. In both cases, the trichodinids heavily co-infested the host fish gills. The flukes were completely coated by the ciliates, which gave them a cobblestone appearance, but no damage to their tegument was apparent. Both cases are most likely a result of accidental hyperparasitism, brought about by perturbed environmental conditions.     

Isolation of Mycobacterium avium subsp paratuberculosis (Map) from feral cats on a dairy farm with Map-infected cattle

Paratuberculosis is an economically important disease of dairy cattle caused by Mycobacterium avium subsp. paratuberculosis (Map). The role of nonruminant, nondomestic animals in the epidemiology of paratuberculosis in cattle is unclear. To examine nonruminant, nondomestic animals for the presence of Map, 25 feral cats, nine mice (species unknown), eight rabbits (Sylvilagus floridanus), six raccoons (Procyon lotor), and three opossums (Didelphis virginiana) were collected from a mid-western dairy with known Map-infected cattle. Mycobacterium avium subsp. paratuberculosis was isolated from the mesenteric lymph node from seven of 25 (28%) feral cats. Ileum was culture-positive for three of these seven cats, and an isolation of Map was also made from the ileum of one of nine (11%) mice. Tissue samples from other species were negative as determined by Map culture; microscopic lesions consistent with paratuberculosis were not seen in any animal. Restriction fragment polymorphism analysis of isolates from cats and dairy cattle suggest interspecies transmission. The means by which interspecies transmission occurred may be through ingestion of Map-contaminated feces or waste milk or through ingestion of Map-infected prey. Shedding of Map from infected cats was not evaluated. The epidemiologic role of Map-infected feral cats on dairy farms requires further investigation.     

Experimental Fascioloides magna infections of mule deer (Odocoileus hemionus hemionus).

Six mule deer (Odocoileus hemionus hemionus) and one white-tailed deer (Odocoileus virginianus), approximately 5-mo-old, each were inoculated orally with 500 metacercariae of Fascioloides magna. All mule deer died from liver fluke infection between 69 and 134 days (mean = 114, SE = 9.9) after inoculation. Between 38 and 326 immature F. magna (mean = 102, SE = 45.5) were recovered from each deer at necropsy. Flukes were present in livers, lungs, and free in pleural and peritoneal spaces. Infection was characterized by necrotizing hepatitis, fibrosing peritonitis and pleuritis, and hematin pigment accumulation in liver, lung, and many other internal organs. Eggs of F. magna first were detected in feces of the white-tailed deer 28 wk after inoculation, and weekly thereafter until the healthy deer was euthanized at 31 wk. At necropsy, 205 F. magna, including 12 encapsulated mature and 193 nonencapsulated immature flukes were recovered from liver, lungs, and free in abdominal and thoracic spaces of the white-tailed deer. Based on these results, F. magna may be fatal to mule deer within 5 mo of infection. Like domestic sheep and goats, mule deer may be highly susceptible to infection, and it is unlikely mule deer can survive infection with large numbers of F. magna.     

Helminth parasites of intermingling axis deer, wild swine and domestic cattle from the island of Molokai, Hawaii

Helminth infections of axis deer (Cervus axis), wild swine (Sus scrofa) and domestic cattle (Bos taurus) were studied among intermingling herds on the Puu-O-Hoku Ranch, Molokai, Hawaii. Twenty-four species of helminths were collected from the 10 deer, 10 swine and 10 cattle. Capillaria bovis, Cooperia punctata, Haemonchus contortus and Trichostrongylus axei infected both axis deer and cattle, whereas Gongylonema pulchrum infected both axis deer and wild swine. None of the species of helminths occurred in both wild swine and cattle nor was any species found in all three hosts. Wild swine and domestic cattle supported separate and distinct helminth communities. In contrast, the helminth community of axis deer appeared to be derived from the helminth communities of cattle and wild swine and consisted only of those species capable of parasitizing either a broad range of ruminants or many mammalian taxa.