Introduction

Species of subfamily Phlebotominae (Diptera, Psychodidae), commonly known as sand flies, feed on the blood of vertebrates, including humans. Some species are known to transmit agents of disease, most notably Leishmania species that cause visceral or cutaneous Leishmaniasis. Due to the role of sand flies as disease vectors, it is important for leishmaniasis surveillance programs to recognize them and understand their natural history, particularly factors which can influence the transmission dynamics of these diseases. Often little is known about the phlebotomine fauna in geographic regions where no leishmaniasis cases have been reported. In Mexico, most studies of these flies have been conducted in the southern and southeastern states, in areas where the greatest numbers of human leishmaniasis cases in Mexico are recorded.

According to the Mexican Health Secretariat (CENAPRECE, 2013, C. Guzmán, pers. com.), human leishmaniases have been recorded in all south and southeastern states, in the costal slope states of the Pacific toward Sinaloa (^{Ochoa-Díaz et al., 2012}, ^{Salazar-Mejía et al., 2010}) and in the Gulf of Mexico toward Tamaulipas, with a few cases recorded in the High Plateau states, specifically in Durango (^{Pérez-Vega et al., 2009}), Coahuila (^{Díaz-Nájera, 1971}) and Nuevo León (^{Simpson et al., 1968}). (^{Sánchez-Tejeda et al., 2001}) recorded about 326 cases of human local cutaneous leishmaniasis from 1987 to 1994 in Nayarit (currently a total of approximately 1,200 cases), but in neighboring southern states (Colima and Michoacán) there are almost no reports of cases, except one of diffuse cutaneous leishmaniasis in the Balsas River Basin of Michoacán. This scarcity of cases may be the reason for the low interest in the study of phlebotomine fauna in Michoacán and the paucity of records in that state for this group.

Eight species of Phlebotominae have previously been recorded in Michoacán based on few specimens collected on three occasions: in the year of 1935 by J. Ortega recording six species, in 1944 by the same collector recording one previously known species, and in 1951-1952 by A. Díaz-Nájera which found two additional species (Table 1) (^{Vargas & Díaz-Nájera, 1953}); for complete collection data of species see (^{Godinez-Alvarez & Ibáñez-Bernal, 2010}). All historical records are from the warmer, lower altitude (below 1000 m asl) Tierra Caliente area of Michoacán. No attempts have previously been made to collect phlebotomine sand flies from the highlands of Michoacán.

Table 1: Phlebotominae sandfly species recorded in Michoacán, Mexico, including the new records reported in this work. 

In this work we present new records of phlebotomine sand fly species collected in the High Plateau region of Michoacán (1500-1700 m asl) in the municipality of Sahuayo. Species listed in this paper can be identified using the comprehensive taxonomic keys of (^{Ibáñez-Bernal, 2005a}, ^2005b); however, in this work we include a taxonomic key to males and females of the ten species currently known from Michoacán.

Material and methods

Collections were made in the Universidad de la Ciénega (20° 00' 50.05" N- 102° 44' 40.73" W, about 1700 m asl), in the Municipality of Sahuayo, state of Michoacán, Mexico, in June 5 and 12, and August 6, 2014, using a CDC miniature light trap additionally baited with Octanol.

The municipality of Sahuayo is located in the northeast portion of Michoacán de Ocampo and about 215 km from Morelia, the capital of the state, south of Chapala Lake. Climate is temperate, with temperature between 10.4° to 26.0 ºC, with rainy summer, recording about 709.0 mm of annual precipitation. Landscape is grassland type, dominated by mesquite (Prosopis spp.), linaloe (Bursera spp.) and nopal (Opuntia spp.) (^{INAFED, 2010}).

Phlebotomine specimens were preserved in 70% ethanol and subsequently cleared, dissected and permanently mounted on microscope slides following the procedure outlined by (^{Ibáñez-Bernal, 2005a}). Observations were made using a Nikon Eclipse 50i compound microscope equipped with phase contrast and species identified using the keys of (^{Young & Duncan, 1994}, ^{Ibáñez-Bernal, 2005a}, ^2005b, and ^{Galati, 2003}). Abbreviations for genera and subgenera follow the proposal of (^{Marcondes, 2007}). All specimens are deposited in the Psychodidae Collection of Instituto de Ecología, A. C.

Results

A total of 63 phlebotomine specimens were obtained at the municipality of Sahuayo, belonging to three subtribes, four genera, four subgenera and four species, according to the classification proposed by (^{Galati, 2003}).

Tribe Phlebotomini Rondani, 1840

Subtribe Sergentomyiina Artemiev, 1991

Micropygomyia (Coquillettimyia) vindicator (Dampf, 1944)

Phlebotomus vindicator (Dampf, 1944): 248 (♀). Type locality: Mexico, Morelos, Cuautla; (Dampf, 1947): 205 (♂).

Diagnosis. Ascoids simple and long, reaching or extending beyond apex of flagellomere; palpal segment 5 longer than 3+4 segments. Male: Gonocoxite with a basal bipartite tubercle, the proximal covered with small spicules and the distal with ca. 8 long setae; gonostylus with a pair of spiniform proximal setae located at about the same level, one median spiniform seta inserted near the proximal setae and distant from the two apical spiniform setae, without subapical fine seta; paramere narrow at middle, with the apical portion broader apex rounded pointed and with a triangular small projection in the ventral edge near the apex, basally with a dorsal spinose rounded lobe; lateral lobe thin; ejaculatory ducts with lanceolate apex. Female: Cibarium with four radiated triangular horizontal teeth, an irregular row of small vertical teeth, and complete arch; spermathecal individual ducts shorter than 4 times the length of the furca stem; spermatheca broader than long, spherical, without annulations, narrowest than spermathecal common duct (^{Ibáñez-Bernal, 2003}).

Material examined. Mexico: Michoacán, Sahuayo, Universidad de La Ciénega, 5-vi-2014, 14 ♂♂, 15 ♀♀; 12-vi-2014, 4 ♂♂, 2 ♀♀; 6-viii-2014, 3 ♂♂, 2 ♀♀.

Distribution. This species is so far only known from Mexico, with records from the following states: Distrito Federal, Guerrero, Morelos, Oaxaca, San Luis Potosí (^{Ibáñez-Bernal, 2003}), and now Michoacán.

Remarks. (^{Young & Duncan, 1994}) suggested that this species probably bites reptiles, and since then, no additional information on natural history of this species has been recorded.

Subtribe Lutzomyiina (Abonnenc & Leger, 1976)

Lutzomyia (Tricholateralis) cruciata (Coquillett, 1907)

Flebotomus cruciatus Coquillett, 1907: 102. Type locality: Guatemala, Alta Vera Paz, Trece Aguas, Cacao.

Diagnosis. Antennal ascoids simple; palpus segment 5 long, longer than segments 3+4. Scutum, pronotum and paratergite pigmented and contrasting with the rest of the pale pleura. Male: gonocoxite with basal tuft of setae with alveoli morular arrangement; gonostylus as long or longer than 0.5 the length of gonocoxite, without subterminal fine seta and four spiniform setae present; paramere simple, apex rounded and without isolated setae near the middle on dorsal margin; ejaculatory ducts simple; lateral lobe thin. Female: Cibarium with four evenly disposed horizontal teeth, an irregular row of small vertical teeth, and without patches of lateral teeth; spermathecal ducts thin, no more than 0.3 as spermathecal diameter, spermathecal common duct shortest than spermatheca, and individual spermathecal duct at least 4.0 times the length of spermatheca; spermathecal pyriform, with globular apical portion differentiated from the basal annuli (^{Ibáñez-Bernal, 1999}).

Material examined. Mexico: Michoacán, Sahuayo, Universidad de La Ciénega, 12-vi-2014, 3 ♀♀.

Distribution. USA (Florida, Georgia) (^{Young & Perkins, 1984}); Mexico (Campeche, Chiapas, Guerrero, Hidalgo, Jalisco, Michoacán [see Table 1], Morelos, Nayarit, Nuevo León, Oaxaca, Puebla, Quintana Roo, San Luis Potosí, Tabasco, Tamaulipas, Veracruz, Yucatán) (^{Ibáñez-Bernal, 1999}), Belize, Guatemala, Honduras, El Salvador, Nicaragua, Costa Rica, Panama (^{Martins et al., 1978}; ^{Young & Duncan, 1994}).

Remarks. (^{Ibáñez-Bernal, 1999}, ^2001b) indicated Lutzomyia cruciata as a highly anthropophilic phlebotomine species with a wide distribution in Mexico. The distribution of this species corresponds with some of the reported cases of human local cutaneous leishmaniasis in the country. Recently, (^{Pech-May et al., 2010}) found natural infection of this species with Leishmania parasites, therefore its presence in that region represents a risk for human leishmaniasis which deserves to be considered by surveillance programs.

Dampfomyia (Dampfomyia) anthophora (Addis, 1945)

Phlebotomus anthophorus (Addis, 1945): 119 (♂, ♀). Type locality: USA: Texas, Uvalde Co., Uvalde.

Diagnosis. Palpus segment 5 longer than segments 3+4. Male: gonocoxite without basal setae tuft; gonostylus with one apical and one preapical spiniform strong setae and one thin setae at middle; paramere complex, with a short capitate superior branch which has point-curved setae along its margin and other group of setae at base of the apical region; ejaculatory ducts with no strong modified apices; lateral lobe thin. Female: Cibarium with two laminar horizontal teeth, a row of about 6 vertical teeth, and lateral teeth patches, arch complete; spermathecae amphora-like surrounded by many globular sacs that as a whole has a morular aspect. Females of Da. anthophora and Da. dodgei (^{Vargas & Díaz-Nájera, 1953}) are indistinguishable, but here associated by the presence of Da. anthophora males.

Material examined. Mexico: Michoacán, Sahuayo, Universidad de La Ciénega, 6-viii-2014, 1 ♂, 6 ♀♀; 12-vi-2014, 1 ♂, 2 ♀♀.

Distribution. Dampfomyia anthophora is known from USA (Texas), and Mexico (Guerrero, Michoacán [see Table 1 for municipalities of Michoacán records], Morelos, and Nuevo León) (^{Young & Perkins, 1984}; ^{Ibáñez-Bernal, 2001a}; Ibáñez-Bernal & Godinez-Alvarez, 2010).

Remarks. This species is not anthropophilic but has been associated with the transmission of Leishmania mexicana to Neotoma rodents in Texas (^{Young & Duncan, 1994}).

Subtribe Psychodopygina (Galati, 1995)

Psathyromyia (Forattiniella) texana (^{Dampf, 1938})

Phlebotomus texanus (^{Dampf, 1938}): 119. Type locality: USA, Texas, Bexar Co., San Antonio.

Diagnosis. Antennal ascoids with short proximal spur; palpus with segment 5 as long as 3+4; wing with apex of vein R₁ before middle of vein R₂. Male: gonocoxite with some non-caducous fine setae scattered at the middle; gonostylus with 4 spiniform setae, the preapical about the middle between the distal and the two proximal setae or nearest to the proximal setae, and without preapical fine seta; paramere simple; ejaculatory ducts with simple apex. Female: cibarium with about 12 horizontal small triangular teeth, one row of about 6 large vertical teeth and other row below of about four less differentiated teeth; arch complete but less visible at middle; spermathecae spherical smooth, without differentiated wide neck at the union with the individual spermathecal duct; individual spermathecal ducts as long as the furca stem and nearly inexistent common duct (^{Ibáñez-Bernal, 2002}).

Material examined. Mexico: Michoacán, Sahuayo, Universidad de La Ciénega, 5-vi-2014, 1 ♂, 6 ♀♀; 12-vi-2014, 1 ♀; 6-viii-2014, 1 ♂, 1 ♀.

Distribution. U. S. A. (Texas) (^{Young & Perkins, 1984}), Mexico (Chiapas [^{Mickery-Pacheco et al., 2012}], Coahuila, Guerrero, Jalisco, Morelos, Nayarit, Oaxaca, San Luis Potosí, Tamaulipas, Veracruz) (^{Ibáñez-Bernal, 2002}).

Remarks. Psathyromyia texana is associated with armadillo borrows and has been collected also in Atta sp. nests (^{Young & Perkins, 1984}).

Taxonomic key for male and female identification of the currently known species of Michoacán

This key is compiled based on portions of those from ^{Young & Duncan (1994)}, ^{Galati (2003)}, and ^{Ibáñez-Bernal (2005a}, ^2005b). It is intended to facilitate the rapid identification of taxa currently recorded only in Michoacán (Table 1). For this reason, it is not necessarily useful for other Mexican states or for other species that may be present but unrecorded in Michoacán. Moreover, the key does not reflect the complete set of characteristics typically used for differentiating supraspecific taxa.