Enzyme variation of Eimeria arizonensis from Peromyscus truei and P. boylii

Cricetid rodents, Peromyscus truei and P. boylii, were inoculated with sporulated oocysts of Eimeria arizonensis collected from wild P. truei maintained in the lab. In P. truei the prepatent period was 4-5 days, the patent period was 9-11 days, and sporulated oocysts were 21.5 x 25.0 (20-23 x 24-26) microns with sporocysts 7.7 x 12.0 (6-8 x 10-13) microns. In P. boylii the prepatent period was 6-7 days, the patent period was 8-9 days, and sporulated oocysts were 20.1 x 23.2 (18-22 x 21-24) microns with sporocysts 6.8 x 10.0 (5-8 x 9-12) microns. Sporulated oocysts from both host species were used in direct side-by-side comparison of isozyme banding patterns using protein electrophoresis. The parasite has polytypic loci for leucine aminopeptidase (LAP), lactate dehydrogenase (LDH), and 6-phosphogluconate dehydrogenase (6-PGD). In oocysts from P. truei, LAP showed one band with fast migration and LDH and 6-PGD each showed two bands, one with fast and one with slow migration. In oocysts from P. boylii, LAP and LDH each had one band with slow migration and 6-PGD had one band with moderate migration. Oocysts of E. arizonensis collected from P. boylii were used to inoculate P. truei. The prepatent and patent periods, structural measurements, and isozyme banding patterns of the resultant oocysts were the same as those from P. truei when inoculated with oocysts from P. truei.     

Cross-transmission studies with Eimeria arizonensis, E. arizonensis-like oocysts and Eimeria langebarteli: host specificity at the genus and species level within the Muridae

Cross-transmission experiments were done using sporulated oocysts of Eimeria arizonensis from Peromyscus truei and Peromyscus maniculatus, and oocysts of 2 putative species that resemble E. arizonensis, i.e., Eimeria albigulae from Neotoma albigula, and Eimeria onychomysis from Onychomys leucogaster. Oocysts of each species were inoculated into representatives of P. maniculatus and the latter 2 rodent species. Other experiments were conducted wherein oocysts of Eimeria langebarteli from Peromyscus leucopus were given to P. truei and P. maniculatus. Oocysts of E. arizonensis from P. truei and P. maniculatus could be transmitted only to P. maniculatus; likewise, oocysts of E. albigulae and E. onychomysis produced patent infections only in N. albigula and O. leucogaster, respectively. Oocysts of E. langebarteli from P. leucopus could be transmitted to P. truei, but not P. maniculatus. These results indicate that E. arizonensis, and the morphologically similar E. albigulae and E. onychomysis, are distinct species that are not transmissible between the genera of their respective hosts (Peromyscus, Neotoma, Onychomys), and that some isolates of E. langebarteli, reported from 6 species of Peromyscus and Reithrodontomys megalotis, may not always be infective to P. maniculatus.     

Hammondia pardalis sp. n. (Sarcocystidae) from the Ocelot, Felis pardalis, and Experimental Infection of Other Felines

Synopsis.
Hammondia pardals sp. n. (Eimeriorina: Sarcocystidae) from Panama Canal Zone is described as an obligate heteroxenous coccidian, with felids as the final host and laboratory mice as the experimental intermediate host. Ovoid oocysts. measuring 40.8 (36–46) × 28.5 (25–35) m. are shed unsporulated. Oocysts were infective only for the intermediate host. the laboratory mouse, Mus musculus , and the intracellular cysts were infective only for felids. Attempted passage of tissue cysts from mouse to mouse was unsuccessful.
Mice fed 5 × 10⁴ sporulated oocysts were found to harbor small intracellular cysts, 13–16 × 10–15 m, in the mesenteric lymph nodes, lungs, and intestinal submucosa 15 days postinfection. The meronts in these early cysts were stubby and measured 3 × 6 m. The prepatent period in the felids was 5 to 8 days and the patent period 5–13 days. Experimental infections of definitive hosts were successful with 6/6 domestic laboratory-reared kittens, Felis catus ; 5/5 ocelots, F. pardalis ; and 1/1 jaguarundi, F. yagouaroundi. None of the exposed raccoons, Procyon lotor , shed oocysts.     

Eimeria species (Apicomplexa: Eimeriidae) infecting Peromyscus rodents in the southwestern United States and northern Mexico with description of a new species

Of 198 deermice (Peromyscus spp) collected from various localities in the southwestern United States and northern Mexico, 106 (54%) had eimerian oocysts in their feces when examined. These included 50 of 106 (47%) Peromyscus truei, 34 of 54 (63%) Peromyscus maniculatus, 4 of 17 (24%) Peromyscus leucopus, and 18 of 21 (86%) Peromyscus eremicus. The following Eimeria were identified from infected mice: Eimeria arizonensis and Eimeria langebarteli from P. truei; E. arizonensis, Eimeria peromysci, and Eimeria delicata from P. maniculatus; E. arizonensis and Eimeria lachrymalis n. sp. from P. eremicus; and E. langebarteli from P. leucopus. Of the 106 Peromyscus found positive for Eimeria, 97 (91.5%) harbored only a single eimerian species at the time of examination. Sporulated oocysts of E. lachrymalis n. sp. were ellipsoid, 27-35 X 17-21 (30.8 +/- 1.7 X 19.1-0.9) micron, possessed a smooth wall and one polar granule, but lacked a micropyle and an oocyst residuum. Sporocysts were teardrop-shaped, 9-13 X 6-10 (10.9 +/- 0.9 X 7.9 +/- 0.5) micron, and had a Stieda body and sporocyst residuum, but no substieda body. Prepatent periods in experimental infections were 3-6 days after inoculation (DAI) for E. arizonensis (hosts: P. eremicus, P. maniculatus, P. truei); 4-5 DAI for E. peromysci (host: P. maniculatus); 6-9 DAI for E. langebarteli (hosts: P. truei, P. leucopus); and 8-10 DAI for E. lachrymalis (host: P. eremicus). Patency in these infections lasted 6-11 days for E. arizonensis, 5-10 days for E. peromysci, 14-40+ days for E. langebarteli, and 19-50+ days for E. lachrymalis. Eimeria lachrymalis appears to produce occult infections in P. eremicus that can be reactivated upon inoculation of the host with E. arizonensis.     

Dorisiella arizonensis n.sp., a Coccidium from the Desert Woodrat, Neotoma lepida

SUMMARY. Dorisiella arizonensis n. sp. (Sporozoa: Eimeriidae) is described from oocysts in the feces of a desert woodrat, Neotoma lepida , from Grand Canyon National Park, Arizona. The oocysts are spherical to subspherical, measure 20.8–21.8 × 20.8–22.9 μ, with a mean of 21.0 × 21.8 μ, and have a two-layered wall. They contain 1 to 3 refractile granules but no residual body. The two lemon-shaped sporocysts have a Stieda body, 8 sporozoites, and a few to many scattered, round, clear residual granules or bodies.     

Infection of llamas with stored Eimeria macusaniensis oocysts obtained from guanaco and alpaca feces

Oocysts obtained from a guanaco and an alpaca with natural infections were identified as Eimeria macusaniensis and evaluated for host specificity and infectivity over time. In 3 separate trials conducted over 4 yr, 4 adult llamas were fed 500-5,000 sporulated oocysts obtained from guanaco feces stored under laboratory conditions for 41-84 mo. Infections with prepatent periods of 36-41 days and patent periods of 38-55 days developed in 4/4 llamas. In a fourth trial, 3 adult llamas and 1 alpaca were each fed 1,000 sporulated E. macusaniensis oocysts obtained from alpaca feces stored in the laboratory for 3 mo. Infections with prepatent periods of 33-34 days and patent periods of 14-20 days developed in 3/3 llamas. Infection in the alpaca had a prepatent period of 58 days and a patent period of 1 day. Clinical signs associated with infection, if any, were minimal and included increased fecal mucus and occasional soft feces. These results provide evidence that E. macusaniensis is a single species transmissible amongst alpacas, llamas, and guanacos and that oocysts of this species can remain infective for many years.     

Eimeria scholtysecki n. sp. from Ord's Kangaroo Rat Dipodomys ordii*

SYNOPSIS. Eimeria scholtysecki n. sp. is described from Ord's kangaroo rat Dipodomys ordii. The sporulated oocysts are broadly ovoid to ellipsoid, averaging 24.6 by 19.6 μ. A polar granule is present. A micropyle and oocyst residuum are absent. The ovoid sporocysts average 12.1 by 8.0 μ, and have small, flattened Stieda bodies. The distinctive sporocyst residuum is composed of coarse granules. The mean prepatent period was 8.2 days. Five inoculated rats apparently became reinfected and discharged oocysts for 30 days or more.     

Fixing coccidian oocysts is not an adequate solution to the problem of preserving protozoan type material

Fresh (36 days old) sporulated oocysts of Eimeria nieschulzi were divided into 7 groups. Control oocysts were maintained at 23 C in 2% aqueous (w/v) K2Cr2O7. The 6 experimental groups were mixed with either Bouin's solution, 10% aqueous (v/v) buffered formalin, Karnovsky's solution, glutaraldehyde, paraformaldehyde, or 70% aqueous (v/v) ethanol (EtOH). After 115 days, oocysts from all 7 groups were examined under oil immersion to determine the effect of fixation on their structural integrity. The parameters examined were lengths and widths of oocysts and sporocysts, percent sporulation (%S), and percent crenation (%C) of oocysts and sporocysts. The highest destruction (%S and %C) occurred in oocysts exposed to glutaraldehyde and Karnovsky's fixatives where 100% of both oocysts and sporocysts crenated and only 8% and 48%, respectively, remained sporulated. Of the oocysts in paraformaldehyde, 93% remained sporulated, but 95%of these oocysts and 100% of the sporocyst crenated. In Bouin's solution, 75% of the oocysts were intact structurally, but of these, only 60% were still sporulated with 70% of their sporocysts crenated. Oocysts preserved in 70% EtOH were 80% intact and 70% remained sporulated, but nearly 60% of their sporocysts collapsed even though the oocyst walls were intact. Oocysts preserved in 10% buffered formalin maintained structural integrity but had lower numbers of sporulated oocysts (84%) and greater numbers of crenated oocysts (18%) than control oocysts maintained in the dichromate solution (95% and 0%, respectively).     

Infectivity of preserved Cryptosporidium parvum oocysts for immunosuppressed adult mice

Abstract The present study was undertaken to determine the infectivity of Cryptosporidium parvum oocysts for immunosup-pressed adult C57BL/6N mice after the oocysts had been stored from 1–48 months at 4°C in 2.5% potassium dichromate. All mice inoculated with oocysts 1–18 months old developed patent infections, while mice inoculated with older oocysts remained uninfected. The prepatent period was extended from 2 to 6 or 7 days as the storage time for oocysts increased. The finding that C. parvum oocysts remain infective for mice for at least 18 months offers important economic and time-saving advantages for investigators who frequently require large numbers of oocysts that must be painstakingly purified from calf manure.     

Coccidia from the Tiger Salamander, Ambystoma tigrinum, in Northeastern Colorado and in Northern New Mexico

SYNOPSIS. Oocysts of Eimeria ambystomae Saxe, 1955, Eimeria microcapi sp. n., and Eimeria urodela sp. n. are described from the tiger salamander, Ambystoma tigrinum , collected in Colorado and New Mexico. The oocysts of E. ambystomae are ellipsoid, 29.8 × 17.3 (24–38 × 15–25) μm, and the sporocysts lanceolate, 22.6 × 5.4 (16–27 × 5–7) μm. Oocyst and sporocyst residua are present, but not a polar granule and a micropyle. The oocysts and sporocysts of E. microcapi are ellipsoid, measuring respectively 38.1 × 25.3 (35-41 × 23-26) μm and 18.1 × 7.4 (16-19 × 6–8) μm. Oocyst and sporocyst residua, a micropyle (mean 3 μm), and a distinct micropyle cap (2 μm high) are present, but not a polar granule. The oocysts of E. urodela are spheroid, 22.2 (14-26) μm, and the sporocysts lanceolate, 16.3 × 5.8 (12-19 × 4-7) μm. Oocyst and sporocyst residua are present, but a polar granule and a micropyle are absent.     

Cross-transmission studies with Eimeria arizonensis-like oocysts (Apicomplexa) in New World rodents of the Genera baiomys, Neotoma, Onychomys, Peromyscus, and Reithrodontomys (Muridae).

Cross-transmission experiments were performed using oocysts of an Eimeria arizonensis-like coccidian from Peromyscus leucopus and Peromyscus truei, an E. arizonensis-like coccidian from Reithrodontomys fulvescens, Eimeria baiomysis and Eimeria taylori from Baiomys taylori, Eimeria albigulae from Neotoma albigula, and Eimeria onychomysis from Onychomys spp., between representatives of the above host genera. The E. arizonensis-like coccidian from R. fulvescens infected Reithrodontomys megalotis, Reithrodontomys montanus, and Peromyscus leucopus. Oocysts of E. arizonensis from P. leucopus could be transmitted to both P. leucopus and R. megalotus. Oocysts of E. baiomysis and E. taylori infected only B. taylori. Oocysts of E. arizonensis from P. truei infected P. truei but not Neotoma mexicana or Onychomys leucogaster. Oocysts of E. albigulae from N. albigula were infective for N. mexicana but not for P. truei or O. leucogaster. Oocysts of E. onychomysis from Onychomys spp. infected O. leucogaster but not N. mexicana or P. truei. These results demonstrate that Peromyscus and Reithrodontomys, genera known to be related very closely evolutionarily, are capable of sharing E. arizonensis, whereas morphologically similar coccidians (E. albigulae, E. baiomysis, and E. onychomysis) from more distantly related hosts, are probably distinct and more stenoxenous. This also is the first report of coccidians infecting species of Reithrodontomys.     

The Sporulated Oocysts of Eimeria illinoisensis n. sp. and of Other Species of Eimeria of the Ox

SYNOPSIS. The sporulated oocysts of 12 species of Eimeria occurring in the ox Bos taurus in the United States are described and differentiated. They are E. alabamensis, E. auburnensis, E. bovis, E. brasiliensis, E. bukidnonensis, E. canadensis, E. cylindrica, E. ellipsoidalis, E. illinoisensis n. sp., E. subspherica, E. wyomingensis and E. zuernii. Two other species, not yet found in North America, which are recognized as valid are E. pellita and E. thianethi. The sporulated oocysts of E. illinoisensis n. sp. are ellipsoidal or slightly ovoid, 24–29 by 19–22 μ with a mean of 26.3 by 20.7 μ; their sporocysts are 13–16 by 6–7 μ with a mean of 15.3 by 6.5 μ. This species was found in 3 cattle from one farm in Illinois.     

Sarcocystis leporum in cottontail rabbits and its transmission to carnivores.

Muscle from Sarcocystis-infected cottontail rabbits (Sylvilagus floridanus) was fed to coccidia-free cats (Felis domestica) and dogs (Canis familiaris). Only cats became infected and shed sporocysts in their feces. The prepatent period ranged from 10 to 25 days and the patent period from 3 to 46 days. Sporocysts were fully sporulated when shed. They contained 4 sporozoites and a coarse granular residuum and averaged 9.4 by 13.6 micron (N=55). Doses of 200-75,000 sporocysts were orally administered to 5 domestic rabbits (Oryctolagus cuniculus). Domestic rabbits did not become infected, suggesting a strict host specificity for the intermediate host S. floridanus.     

Isospora vulpina Nieschulz and Bos, 1933: Description,and Transmission from the Fox (Vulpes vulpes) to the Dog*†

SYNOPSIS. Oocysts of Isospora vulpina were found in silver foxes (Vulpes vulpes) on a fox farm in Wisconsin. They were 29.7 (25-38) × 24.3 (21-32) μm. The sporocysts were 17.7 (15–23) × 13 (11–16) μm. Five coccidia-free puppies were inoculated with 22,000–42,000 oocysts each of I. vulpina from the fox: a patent infection resulted after 6-7 days. The infection was then transferred from 1 of these dogs to another coccidia-free puppy. After a 7-day prepatent period the puppy passed oocysts for 7 days.     

Excystation of the Sporozoites of Eimeria tenella in Apparently Unbroken Oocysts in the Chicken

SYNOPSIS. Chickens experimentally fed sporulated oocysts of Eimeria tenella were necropsied 5–300 minutes later to study excystation, especially in apparently unbroken oocysts. In birds killed 75–125 minutes after inoculation, there was evidence of excystation in some oocysts recovered from the small and large intestines. Altho not visibly broken, the shells of about 10% of the ones studied were wrinkled or otherwise deformed; many contained active sporozoites, some inside and some outside the sporocysts. During examinations, numerous sporozoites emerged from the sporocysts. Altho evidence was not obtained that excystation from unbroken oocysts was occurring inside the birds, the fact that it occurred in some oocysts during examinations may indicate that it could be taking place. Evidence of excystation was not seen in oocysts with shells that were not visibly altered.     

Eimeria lancasterensis Joseph, 1969 and E. confusa Joseph, 1969 from the Grey Squirrel Sciurus carolinensis*

SYNOPSIS. Forty grey squirrels Sciurus carolinensis were examined for coccidia during a 2-year period. Eimeria lancasterensis was found in all of them. The ellipsoidal oocysts of this species averaged 24.6 by 14.6 μ. They had no micropyle or oocyst residuum. A polar body was present. The sporocysts averaged 14.1 by 8.4 μ. The endogenous phases of the parasite were found in the epithelial cells of the villi thru the entire length of the small intestine. E. confusa was found in one of 40 squirrels. The oocysts of this species were subspherical, occasionally ellipsoidal or rarely spherical; they averaged 33.2 by 26.7 μ. Oocyst residuum and micropyle were absent. Polar granules ranged in number from 0–5. The sporocysts averaged 19.6 by 12.1 μ. The prepatent period for this species was 7–8 days and the patent period 6–13 days. E. ontarioensis was found in 3 of the 40 squirrels.     

Eimeria procyonis sp. n., an Isospora sp., and a Redescription of E. nuttalli Yakimoff and Matikaschwili, 1932 (Protozoa: Eimeriidae) from the American Raccoon (Procyon lotor)

SYNOPSIS. Thirty-two of 48 raccoons examined were infected with a previously undescribed species of Eimeria which is herein named E. procyonis. Of the 32 infected animals, 10 also harbored E. nuttalli and 1 had Isospora sp. oocysts.
The ellipsoid to ovoid oocysts of E. procyonis measured 23.4 × 18.0 (16–29 × 13–24) μm; its sporocysts measured 12.1 × 9.3 (11.5–15 × 7–10) μm, each containing a slightly flattened substiedal body. The sporocyst residuum consisted of numerous scattered granules each ∼1 μm in diameter. The oocyst wall was double-layered. The outer layer appeared rough and pitted, measuring 1.5 μm, except at the micropyle where it was 1 μm thick.
The oocysts of the Isospora sp. measured 16.8 × 13.7 (16–18.5 × 12.5–15.5) μm. The wall consisted of a single layer ∼0.5 μm thick. The sporocysts measured 11.2 × 9.1 (9.5–11.5 × 8–10) μm, and each contained 4 elongate sporozoites. The oocysts of E. nuttalli measured 17.5 × 13.6 (12-21 × 11-15) μm, with a smooth single-layered wall approximately 0.7 μm thick. The sporocysts measured 12.2 × 7.1 (9-13 × 5.5–11) μm. Each sporocyst had a thin, dark, Stieda body and the sporocyst residuum consisted of many fine granules.     

Coccidian parasites (Apicomplexa: Eimeriidae) from insectivores. IV. Four new species in Talpa europaea from England

Moles from England were examined for coccidian oocysts and all 64 Talpa europaea were infected; of 64 infected hosts, 56 (88%) had multiple infections representing two to six coccidian species when examined. Oocysts in 31 of the 64 samples remainedunsporulated. Three eimerians and one isosporan were studied from the 33 fecal samples that had sporulated oocysts and these are described as new species; Cyclospora talpae Pellérdy & Tanyi, 1968, and Isospora sofiae (Golemansky, 1978) Levine & Ivens, 1979, are redescribed; and Cyclospora sp., similar to C. talpae, is discussed. Sporulated oocysts of C. talpae are ellipsoidal, 14.3 X 9.6 (12-19 X 6-13) microns with sporocysts ovoid, 9.4 X 5.7 (6-13 X 4-8) microns; it was found in 21 of the 33 (63.6%) sporulated samples. Sporulated oocysts of Cyclospora sp. are subspheroidal to ellipsoidal, 12.5 X 8.9 (10-14 X 6-12) microns with sporocysts ovoid, 8.6 X 5.3 (6-10 X 4-6) microns; it was found in 21 of the 33 (63.6%) sporulated samples. Sporulated oocysts of Eimeria avonensis n. sp. are elongate-ellipsoidal, 15.0 X 9.6 (13-20 X 7-12) microns with sporocysts ovoid, 6.6 X 3.6 (5-9 X 3-7) microns; it was found in 15 of the 33 (45.5%) sporulated samples. Sporulated oocysts of Eimeria berea n. sp. are subspheroidal, 12.1 X 10.5 (10-15 X 8-14) microns with sporocysts ovoid, 6.3 X 3.9 (5-10 X 2-5) microns; it was found in 8 of the 33 (24.2%) sporulated samples.(ABSTRACT TRUNCATED AT 250 WORDS)     

Molecular phylogenetics of the Peromyscus boylii species group (Rodentia: muridae) based on mitochondrial cytochrome b sequences

Variation in the mitochondrial cytochrome b gene (1143 bp) was examined to estimate the phylogenetic relationships of taxa within the Peromyscus boylii species group. In addition, phylogenetic relationships among the aztecus, boylii, and truei species groups were addressed. Maximum-likelihood, neighbor-joining, and maximum-parsimony (weighted and equally weighted) analyses produced similar topologies with P. boylii, P. beatae, P. simulus, P. stephani, P. madrensis, P. levipes, and three undescribed taxa from western Mexico forming a monophyletic unit. At least two of the undescribed taxa from western Mexico potentially represent species. Members of the P. aztecus species group formed a clade separate from the P. boylii group and should be recognized as a distinct species group. P. sagax, P. polius, and P. pectoralis, formerly placed in the P. boylii species group, generally formed an unresolved polytomy with the P. truei, P. aztecus, and P. boylii species groups. P. attwateri formed a sister taxon relationship with members of the P. truei species group (P. difficilis and P. nasutus) and should be considered a member of this group. Members of the P. truei species group did not form a monophyletic unit, indicating that this species group is not monophyletic and may be composed of two assemblages.     

Eimeria ornata N. Sp. (Apicomplexa: Eimeriidae) from the Ornate Box Turtle, Terrapene ornata ornata (Reptilia: Testudines), in Texas

Eimeria ornata n. sp. is described from the feces of 6/16 (37.5%) ornate box turtles, Terrapene ornata ornata , in northcentral Texas. Endogenously sporulated oocysts are ellipsoid 17.9 × 15.7(16-21 × 14-18) μm, with a thin, single-layered wall; shape index 1.14 (1.0-1.3). A micropyle is absent but a polar granule was present in one third of the oocysts. An oocyst residuum was present, consisting of numerous small globules situated either in a distinct mass or scattered within the oocyst. The sporocysts are elongate, 11.1 × 5.4 (9-13 × 5-6) μm, with an indistinct Stieda body at 1 pole. A sporocyst residuum is present, consisting either as a compact mass or as scattered granules. The sporozoites are elongate, 9.5 × 2.0 (8-12 × 2) μm, in situ, with spherical anterior and posterior refractile bodies. The new species is distinguished from the similar Eimeria carri Ernst & Forrester, 1973, from eastern box turtles, T. Carolina , by slight differences in oocyst morphology and endogenous sporulation.