Experimental Demonstration of Reservoir Competence of the White-Footed Mouse, Peromyscus leucopus (Rodentia: Cricetidae), for the Lyme Disease Spirochete, Borrelia mayonii (Spirochaetales: Spirochaetaceae).

The white-footed mouse, Peromyscus leucopus (Rafinesque), is a reservoir for the Lyme disease spirochete Borrelia burgdorferi sensu stricto in the eastern half of the United States, where the blacklegged tick, Ixodes scapularis Say (Acari: Ixodidae), is the primary vector. In the Midwest, an additional Lyme disease spirochete, Borrelia mayonii, was recorded from naturally infected I. scapularis and P. leucopus. However, an experimental demonstration of reservoir competence was lacking for a natural tick host. We therefore experimentally infected P. leucopus with B. mayonii via I. scapularis nymphal bites and then fed uninfected larvae on the mice to demonstrate spirochete acquisition and passage to resulting nymphs. Of 23 mice fed on by B. mayonii-infected nymphs, 21 (91%) developed active infections. The infection prevalence for nymphs fed as larvae on these infected mice 4 wk post-infection ranged from 56 to 98%, and the overall infection prevalence for 842 nymphs across all 21 P. leucopus was 75% (95% confidence interval, 72-77%). To assess duration of infectivity, 10 of the P. leucopus were reinfested with uninfected larval ticks 12 wk after the mice were infected. The overall infection prevalence for 480 nymphs across all 10 P. leucopus at the 12-wk time point was 26% (95% confidence interval, 23-31%), when compared with 76% (95% confidence interval, 71-79%) for 474 nymphs from the same subset of 10 mice at the 4-wk time point. We conclude that P. leucopus is susceptible to infection with B. mayonii via bite by I. scapularis nymphs and an efficient reservoir for this Lyme disease spirochete.

Genetic evidence of enzootic leishmaniasis in a stray canine and Texas mouse from sites in west and central Texas.

We detected Leishmania mexicana in skin biopsies taken from a stray canine (Canis familiaris) and Texas mouse (Peromyscus attwateri) at two ecologically disparate sites in west and central Texas using polymerase chain reaction (PCR). A single PCR-positive dog was identified from a sample of 96 stray canines and was collected in a peri-urban area in El Paso County, Texas. The PCR-positive P. attwateri was trapped at a wildlife reserve in Mason County, Texas, from a convenience sample of 20 sylvatic mammals of different species. To our knowledge, this represents the first description of L. mexicana in west Texas and extends the known geographic range of the parasite to an area that includes the arid Chihuahuan Desert. Our finding of L. mexicana in P. attwateri represents a new host record and is the first description of the parasite in a wild peromyscid rodent in the United States.

Cytokine mRNA expression in Peromyscus yucatanicus (Rodentia: Cricetidae) infected by Leishmania (Leishmania) mexicana.

Peromyscus yucatanicus, the main reservoir of Leishmania (Leishmania) mexicana in the Yucatan peninsula of Mexico, reproduces clinical and histological pictures of LCL in human as well as subclinical infection. Thus, we used this rodent as a novel experimental model. In this work, we analyzed cytokine mRNA expression in P. yucatanicus infected with L. (L.) mexicana. Animals were inoculated with either 2.5×10(6) or 1×10(2) promastigotes and cytokine expressions were analyzed by real-time RT-PCR in skin at 4 and 12weeks post-infection (wpi). Independently of the parasite inoculum none of the infected rodents had clinical signs of LCL at 4wpi and all expressed high IFN-γ mRNA. All P. yucatanicus inoculated with 2.5×10(6) promastigotes developed signs of LCL at 12wpi while the mice inoculated with 1×10(2) remained subclinical. At that time, both IFN-γ and IL-10 were expressed in P. yucatanicus with clinical and subclinical infections. Expressions of TNF-α and IL-4 were significantly higher in clinical animals (2.5×10(6)) compared with subclinical ones (1×10(2)). High TGF-ß expression was observed in P. yucatanicus with clinical signs when compared with healthy animals. Results suggested that the clinical course of L. (L.) mexicana infection in P. yucatanicus was associated with a specific local pattern of cytokine production at 12wpi.

Nitric oxide production by Peromyscus yucatanicus (Rodentia) infected with Leishmania (Leishmania) mexicana.

Peromyscus yucatanicus (Rodentia: Cricetidae) is a primary reservoir of Leishmania (Leishmania) mexicana (Kinetoplastida: Trypanosomatidae). Nitric oxide (NO) generally plays a crucial role in the containment and elimination of Leishmania. The aim of this study was to determine the amount of NO produced by P. yucatanicus infected with L. (L.) mexicana. Subclinical and clinical infections were established in P. yucatanicus through inoculation with 1 x 10 2 and 2.5 x 10 6 promastigotes, respectively. Peritoneal macrophages were cultured alone or co-cultured with lymphocytes with or without soluble Leishmania antigen. The level of NO production was determined using the Griess reaction. The amount of NO produced was significantly higher (p ≤ 0.0001) in co-cultured macrophages and lymphocytes than in macrophages cultured alone. No differences in NO production were found between P. yucatanicus with subclinical L. (L.) mexicana infections and animals with clinical infections. These results support the hypothesis that the immunological mechanisms of NO production in P. yucatanicus are similar to those described in mouse models of leishmaniasis and, despite NO production, P. yucatanicus is unable to clear the parasite infection.

Nitric oxide production by Peromyscus yucatanicus (Rodentia) infected with Leishmania (Leishmania) mexicana

Peromyscus yucatanicus (Rodentia: Cricetidae) is a primary reservoir of Leishmania (Leishmania) mexicana (Kinetoplastida: Trypanosomatidae). Nitric oxide (NO) generally plays a crucial role in the containment and elimination of Leishmania. The aim of this study was to determine the amount of NO produced by P. yucatanicus infected with L. (L.) mexicana. Subclinical and clinical infections were established in P. yucatanicus through inoculation with 1 x 10 2 and 2.5 x 10 6 promastigotes, respectively. Peritoneal macrophages were cultured alone or co-cultured with lymphocytes with or without soluble Leishmania antigen. The level of NO production was determined using the Griess reaction. The amount of NO produced was significantly higher (p ≤ 0.0001) in co-cultured macrophages and lymphocytes than in macrophages cultured alone. No differences in NO production were found between P. yucatanicus with subclinical L. (L.) mexicana infections and animals with clinical infections. These results support the hypothesis that the immunological mechanisms of NO production in P. yucatanicus are similar to those described in mouse models of leishmaniasis and, despite NO production, P. yucatanicus is unable to clear the parasite infection.

Five new Mexican species of the flea genus Strepsylla Traub, 1950 (Siphonaptera: Ctenophthalmidae: Neopsyllinae: Phalacropsyllini) with a phylogenetic analysis.

The genus Strepsylla Traub is redescribed and compared with the other genera of the tribe Phalacropsyllini. Thirteen recognized species are distributed in Mexico, Guatemala, Honduras, Costa Rica, and Panama, where they parasitize small rodents, especially mice of the genus Peromyscus. Five new species are reported and described (Strepsylla ixtlanensis n. sp., Strepsylla longicauda n. sp., Strepsylla queretana n. sp., Strepsylla tezontli n. sp., and Strepsylla vargasi n. sp.). Keys for the genera of the tribe and for the species of Strepsylla are included, and relevant notes for each species are given. A cladistic analysis of Strepsylla led to the recognition of 2 clades: one includes S. mina, S. villai, and S. tezontli, which are distributed along the Trans-Mexico Volcanic Belt; and the other includes the remaining species, which have a northern Mexico distribution.

A new dicrocoeliid (Digenea: Dicrocoeliinae) parasite of rodents from Tlaxcala, Mexico.

Caballerolecythus ibunami n. gen., n. sp. is described from the intestine of 2 species of rodents (Liomys irroratus [Gray, 1868] and Peromyscus difficilis [Allen, 1891]) from Piñonal, El Carmen Tequexquitla, Tlaxcala state, Mexico. These specimens represent a new genus and a new species of Dicrocoeliinae by possession of an extremely short ceca (ending at equatorial level of ovary); vitelline follicles that begin posterior to these structures, without overlapping; and a body that is long and slender (length/width ratio 1:17-21). To the best of our knowledge, this combination of characters has not been reported in other genera of this subfamily (Athesmia Looss, 1899; Unilaterilecithum Oshmarin in Skrjabin and Evranova, 1952; and Pseudathesmia Travassos, 1942).

Polyplax guatemalensis sp. n. (Phthiraptera: Anoplura), a new sucking louse from Peromyscus grandis, a montane cloud forest rodent from Guatemala.

The adult male and female of Polyplax guatemalensis sp. n. are described from the sigmodontine murid rodent Peromyscus grandis Goodwin collected in the Reserva de Biosfera, Sierra de las Minas, Guatemala, at an elevation of 2,200 m. The new species extends the number of known native species of Polyplax in the New World to four with none of them recorded south of Panama. Polyplax guatemalensis is morphologically most closely related to Polyplax auricularis which parasitises a cluster of closely related New World sigmodontine rodents from Canada to Panama. These two species can be distinguished from all other known species of Polyplax by the presence of partially overlapping, subtriangular, anterior abdominal plates in both sexes. Polyplax guatemalensis can be separated from P. auricularis by the abundant tergal abdominal setae and longer pseudopenis in males, and by the presence of one fewer anterior abdominal, subtriangular tergite and sternite in females.

[2 new species of Stilestrongylus (Nematoda: Heligmonellidae) parasites of Peromyscus (Rodentia: Cricetidae) from Mexico]. / Dos nuevas especies de Stilestrongylus (Nematoda: Heligmonellidae) parásitos de Peromyscus (Rodentia: Cricetidae) de México.

Stilestrongylus peromysci n. sp. collected from Peromyscus difficilis (Hidalgo state, México), differs from other species in the genus in number of the spines (30) in the synlophe (both sexes) and because the eighth ray arises from the root of the ninth ray; S. hidalguensis n. sp. parasitised Peromyscus sp. and differs from all other congeneric species in the presence of 24 spines in the male synlophe and in the arrangement of the bursal rays (2-2-1 in the right lobe and 2-3 in the left lobe). A key to the species of Stilestrongylus is provided.

Dynamics of plague in a Gunnison's prairie dog colony complex from New Mexico.

A plague (Yersinia pestis) epizootic spread through Gunnison's prairie dogs (Cynomys gunnisoni), and possibly other rodent species, in the Moreno Valley in north-central New Mexico between winter 1984-1985 and autumn 1987. We observed the progress of the epizootic and subsequent population recovery at four prairie dog towns within the valley during this period. At two towns (Midlake and Val Verde) the prairie dogs were marked prior to the epizootic. At two additional towns (Vega and South Entrance) prairie dogs were marked following the epizootic. In 1988, a second epizootic occurred at Vega. One hundred thirty-nine serum samples were collected from prairie dogs and other rodents and 1,750 fleas were collected from animals and burrows. Fleas infected with Y. pestis were collected from prairie dogs, deer mice (Peromyscus maniculatus), and thirteen-lined ground squirrels (Spermophilus tridecemlineatus). Prairie dog fleas included Oropsylla hirsuta, O. labis and O. tuberculata, deermouse associated fleas were Aetheca wagneri and Rhadinopsylla sectilis, and Oropsylla bacchi was associated with thirteen-lined ground squirrels. All of the above flea species were collected from prairie dog burrows. All rodent species shared some flea species. Thirteen-lined ground squirrels disappeared shortly before plague was identified in prairie dogs at Midlake. Meadow voles were rare following the epizootic at Vega in 1986, became abundant in 1987, and disappeared at the time of the second prairie dog epizootic in summer 1988. Although we collected serum from Gunnison's prairie dogs, thirteen-lined ground squirrels, deer mice, and meadow voles (Microtus pennsylvanicus), we identified elevated serum titers against Y. pestis only in Gunnison's prairie dogs. Prairie dog mortality at all towns affected by plague was in excess of 99%. Serum antibody titers indicate that more than 40% of the few prairie dogs left to establish colonies following epizootics survived plague infection.

Parasites of mammals on the Sevilleta National Wildlife Refuge, Socorro, New Mexico: Cuterebra austeni and C. neomexicana (Diptera:Oestridae) from Neotoma and Peromyscus (Rodentia:Muridae), 1991-1994.

In total, 6,486 rodents representing 3 families (Muridae, Heteromyidae, and Sciuridae) and 24 species were trapped May through August of 1991 through 1994. Of these, only the white-throated woodrat. Neotoma albigula Hartley; piñon mouse, Peromyscus truei Shufeldt; and white-footed mouse, P. leucopus Rafinesque, were infested with Cuterebra Clark larvae. Of the 594 N. albigula that were captured 103 (17.3%) were infested with 139 Cuterebra larvae with all infestations occurring in the throat region. N. albigula infestations were observed in 4 of 5 habitats sampled. The highest prevalence of infestation occurred during May-June (27.2%) versus July-August (9.1%) and in males (25.2%) versus females (18.3%). Prevalence of infestation was not significantly different between animals from the mark-release webs versus removal webs or adults versus juveniles. Also, there was no correlation between relative density of N. albigula and prevalence of infestation. Fifteen adults were reared from puparia and identified as C. austeni Sabrosky. Of 716 P. truei captured, 22 (3.1%) were infested with a total of 25 Cuterebra larvae with all infestations occurring in the scapular region. Although P. truei were captured in all 5 habitats, they were only infested in the piñon-juniper habitat: the highest prevalence of infestation occurred during July-August (10.9%) versus May-June (3.3%). Prevalence of infestation was not significantly different between animals from mark-release webs versus removal webs, males versus females, or adults versus juveniles. As in the N. albigula, there was no correlation between relative density of P. truei and prevalence of infestation. The adult reared from a puparium was identified as C. neomexicana Sabrosky. Of the 310 P. leucopus captured, only 3 (1%) were infested with Cuterebra larvae.

Leishmaniasis in Texas: prevalence and seasonal transmission of Leishmania mexicana in Neotoma micropus.

The annual prevalence of Leishmania mexicana in Neotoma micropus from 16 southern Texas localities was determined by screening 192 N. micropus using in vitro culture. Eight woodrats also were screened using the polymerase chain reaction (PCR). Forty-six Sigmodon hispidus from four localities were also tested. Seasonal transmission was investigated through trap-recapture studies at three localities and analysis of infections in juveniles. Fourteen N. micropus from four localities were culture-positive for L. mexicana, indicating an annual non-zero prevalence at these localities of 5.6-27%. Four additional infections at two foci were detected only by the PCR. Transmission occurred in the fall in each case in which season of transmission could be determined. No S. hispidus were positive. These data support the hypothesis that N. micropus is a reservoir of L. mexicana in Texas.

Prevalence and control of babesiosis in the Americas.

This review presents up-to-date information on the distribution and control measures of babesiosis in Latin America. Bovine babesiosis caused by Babesia bovis and B. bigemia will be emphasized. The disease is endemic in most countries and poses a serious economic burden on livestock production in the region (U.S. $1365 million/year, FAO, 1989). Of the estimated 250 million cattle in Central and South America, approximately 175 million (70%) are in tick-infested regions. Humid, tropical and subtropical areas favor development of the main vector, the one-host tick Boophilus microplus. In many regions bovine babesiosis is enzootically stable as a consequence of a balanced host-parasite relationship. However, Latin America offers a wide range of epidemiologic conditions that are influenced by variations from tropical to cool climates and by susceptible purebred cattle that are regularly imported to upgrade local stocks. The control measures employed in most countries for babesiosis essentially rely on chemotherapy, use of acaricides for B. microplus, and to a lesser degree, on immunization methods. In general, these measures are expensive, time consuming, and in many cases, provide limited success. Finally, the zoonotic potential of babesiosis will be addressed, with special emphasis on the situation in the United States. Even though bovine babesiosis has long been eradicated from the U.S.A., human babesiosis is endemic in the northeastern region of the country.

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.