David W. Reduker

Factors Influencing Excystation in Cryptosporidium Oocysts from Cattle

Fisheries and Oceans, Biological Station, St. Andrews, New Brunswick, Canada EOG 2X0. We thank P. C. F. Hurley for organizing the collection of swordfish, Joyce Taylor for acquireries and Oceans, Bi logical Station, St. Ans, New Brunswick, C n da EOG 2X0. ing literature on pennellids, W. B. Scott for reviewing the manuscript, and Brenda Fullerton for typing the manuscript. pennel ids, . B. Scot for re-

Functional Analysis of the Masticatory Apparatus in Two Species of Myotis

Jaw and skull morphologies were analyzed in two morphologically distinct species of bats, Myotis evotis (substrate gleaner) and Myotis volans (aerial insectivore). Force vector analyses were performed on five major muscle-bone lever systems to compare the relative adductive forces produced at three functional points (the incisors, canines, and protoconids of the lower second molar) during jaw closure. Relative force produced by contraction of a given adductor was approximated using a modified muscle mass value. Seven angles of dentary position were analyzed for changes in mechanical advantage due to changing muscle fiber orientation. Results of the analyses suggest that M. evotis has a more forceful bite than M. volans , but only as the upswinging dentary approaches closure. Moreover, M. evotis has the potential for a quicker jaw closure than M. volans and the ability to produce series of rapid nipping motions. The masticatory apparatus of M. volans appears well suited for capture of insect prey using a wide gape followed by a forceful closure. These results are interpreted in view of available ecological data concerning the foraging behavior of the two species.

DNA probes distinguish geographical isolates and identify a novel DNA molecule of Babesia bovis.

A genomic DNA library of Babesia bovis was screened to identify DNA probe candidates for direct detection of the parasite. Two sequences, Bo6 and Bo25, had the highest sensitivity and further analysis revealed unique characteristics of each of these. Neither sequence hybridized detectably to bovine DNA. Bo6 detected 100 pg of both a Mexican and an Australian isolate of B. bovis, but Bo6 also detected 1.0 ng of Babesia bigemina DNA under identical conditions. A unique characteristic of Bo6 is that it hybridizes to an apparent 7.4-kilobase DNA in undigested genomic DNA of both B. bovis and B. bigemina. The sequence is well conserved between the 2 geographic isolates of B. bovis, but it is apparently divergent in B. bigemina. Bo25 did not hybridize detectably to bovine or B. bigemina DNA. This sequence detected 100 pg of homologous B. bovis Mexican isolate DNA, but the sensitivity was reduced to 1 ng for the Australian isolate DNA. The restriction enzyme profile of the Bo25 sequence in genomic DNA differed markedly in the number, size, and intensity of bands between the 2 B. bovis geographic isolates tested. Thus, the Bo25 sequence can distinguish geographic isolates of B. bovis.

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.

FELINE SERUM ANTIBODY RESPONSES TO TOXOPLASMA GONDII AND CHARACTERIZATION OF TARGET ANTIGENS

The Toxoplasma gondii-specific target antigens for feline immunoglobulin M (IgM) and immunoglobulin G (IgG) immune responses were studied longitudinally using western blot immunoassay in 8 cats experimentally inoculated with T. gondii strain ME49. Multiple antigens were recognized by IgM and IgG during the course of infection. Dense bands were associated with 12 antigens in the IgM western blot immunoassay and 30 antigens in the IgG western blot immunoassay. Immunoglobulin M responses were maximal on week 4 postinoculation (PI) and were greatly diminished by week 20 PI. Immunoglobulin G responses were maximal on week 12 PI. On week 20 PI, the 19-kDa (6/8 samples), 26-kDa (8/8 samples), 28-kDa (8/8 samples), 31-kDa (7/8 samples), 35-kDa (6/8 samples), 51-kDa (6/8 samples), 55-kDa (7/8 samples), and 65-kDa (7/8 samples) antigens were recognized most commonly in the IgG western blot immunoassay. When the western blot immunoassay results were compared to enzyme-linked immunosorbent assay (ELISA) results, there was no clear advantage to the development of IgM-ELISA, IgG-ELISA, IgM western blot immunoassay, or IgG western blot immunoassay using a single antigen instead of multiple antigens as the detection system for the diagnosis of recent infection.

PROTEINS AND ANTIGENS OF MEROZOITES AND SPOROZOITES OF EIMERIA BOVIS (APICOMPLEXA)

Proteins and antigens of first-generation merozoites and sporozoites of Eimeria bovis were examined using standard sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), Western blotting and lactoperoxidase iodination procedures. SDS-PAGE gels revealed both common and unique protein bands in merozoite and sporozoite extracts, ranging in molecular weight (Mr) from 15,000 to 215,000. Nitrocellulose immunoblots of separated proteins, when probed with sera obtained from immunized calves, revealed numerous IgG-binding antigens of Mr 18,000 to 180,000 in merozoites and Mr 28,000 to approximately 118,000 in sporozoites. Although merozoite and sporozoite preparations each contained antigens of different molecular weights, 4 antigens had the same migratory distance in both preparations (Mr 58,000, 70,000, 83,000, 98,000). Of 3 types of immune sera used to probe immunoblots, serum taken from a calf that had been inoculated with oocysts of E. bovis and boosted 10 wk later by subcutaneous injection with 2 X 10(7) live merozoites emulsified in Freunds complete adjuvant consistently identified and reacted more intensely with more antigens of merozoites and sporozoites than the other immune sera tested. Autoradiographic analysis of radioiodinated parasites revealed major surface proteins on merozoites of between 15,000 and 18,000 Mr and 3 surface proteins on sporozoites of Mr 28,000, 77,000, and 183,000. All but the 183,000 protein elicited an IgG antibody response in the host.

EIMERIA FROM BATS OF THE WORLD. II. A NEW SPECIES IN TADARIDA FEMOROSACCA FROM SONORA, MEXICO

Between 1979 and 1980, 104 bats representing 13 species in 4 families were collected in California and New Mexico, U.S.A., and Baja California and Sonora, Mexico, and were examined for coccidia; only 3 (3%) had oocysts in their feces. Bats examined and their infection rates were: Molossidae: 0 of 12 Tadarida brasiliensis, 1 of 18 (6%) T. femorosacca; Natalidae: 0 of 1 Natalus stramineus; Phyllostomatidae: 0 of 1 Choeronycteris mexicana, 0 of 2 Leptonycteris sanborni, 0 of 1 Macrotus californicus; Vespertilionidae: 0 of 9 Antrozous pallidus, 0 of 28 Eptesicus fuscus, 0 of 1 Lasionycteris noctivagans, 0 of 3 Lasiurus borealis, 2 of 22 (9%) L. cinereus, 0 of 1 L. ega, 0 of 5 Pipistrellus hesperus. Sporulated oocysts were only found in T. femorosacca and these represent a new species, Eimeria tadarida n. sp. They are subspheroidal to ellipsoidal, 19 x 25 (16-23 x 20-30) microns; a micropyle is absent, and fragments within the oocyst may be oocyst residuum or multiple polar bodies. The oocyst wall, approximately 1.5 microns, is composed of a mammillated outer layer and smooth inner layer. Sporocysts are ovoidal, 8 x 12 (6-9 x 10-14) microns, and have a small Stieda body and a wide substieda body. This is only the 14th eimerian to be described from bats worldwide. Only unsporulated or partially sporulated oocysts of an eimerian were seen in 2 L. cinereus. These measured 28 x 25 (27-29 x 24-26) microns and had a mammillated outer oocyst wall.