Introduction There are about 700 species of sand flies of which only 70 species have been incriminated as vector

so far. They bite human and transmit disease to them. The sand flies belong to the subfamily Phlebotominae of the family psychodidae. Sand flies are found mainly in tropics with a few species also found in the temperate regions. Adult flies are mainly called sandflies.

ClASSIFICATION OF SANDFLY. Phylum: Arthropoda. Class: Insecta Order: Diptera Family: Psycodidae. Genus: Phlebotomus Species: (a) P. papatasii (b) P. Sergenti (c) P. argentipes

Taxonomy The family Psychodidae includes six subfamilies (Duck- house, 1973), only two of which include species of known public health and veterinary importance: the Psychodinae and Phlebotominae. Adults of Phlebotominae (Fig. 9.2A) have relatively long mouthparts, long antennal segments, and long legs and hold the wings sloping upward (troughlike) when at rest. The mandibles are well developed. The sub- family includes about 600 species distributed globally be- tween about 50~ and 40 ~ S. About 380 species occur in the New World, including 14 in the United States and Canada. There are no records from Alaska or Hawaii. Canadian records include only British Columbia, Alberta, and Ontario. The New World genus Lutzomyia and the Old World genus Phlebotomus include species of public health and veterinary importance. Members of the New World genera Brumptomyia and Warileya and the Old World genus Sergentomyia are zoophilic and rarely bite humans. The gcnus Lutzomyia was rcviewed by Young and Duncan (1994), and the genus Phlebotomus was reviewed by Lewis (1982). The Phlebotominac of North America, north of Mexico, were reviewed by Young and Perkins (1984). Somc authors treat thc Phlebotominae as a separatc family, the Phlebotomidae, and some treat the subgenus Psychodopygus of Lutzomyia as a separate genus. There is evidence of cryptic species in some taxa. Many new approaches are being applied to problems of classification and identification; these include electron micrography, karyotyping, nucleic acid probes, isoen- zyme electrophoresis, polymerase chain reaction assay, and analysis of cuticular hydrocarbons.

Confusing common names When people talk about sand flies it can be very confusing because there are several different bloodsucking insects around the world called sand flies. This is why scientists use latin names for species to avoid confusion. Although the English name of sand fly is adequate for phlebotomines living in Middle Eastern deserts it is inappropriate for those in other areas such as humid Neotropical forests. Furthermore, both black flies (Simuliidae) and biting midges (Ceratopogonidae) are also known as sand flies in various parts of the world. The latter are the ones most likely to be biting people on beaches, particularly members of the genus Culicoides. This confusion extends even to the Encyclopaedia Britannica, which states (wrongly!) that sand flies have aquatic larvae (that) live in the intertidal zone of coastal beaches, in mud, or in wet organic debris. Identification All phlebotomine sand flies can be distinguished by the lay person by the following characteristics: Size Sand flies are smaller than mosquitoes but larger than midges, with a body length of 2-3mm

Color All sand flies are brownish in daylight but their bodies are densely covered in oily hairs which give the insects a whitish appearance when illuminated (see Fig. 1). This explains some of their common names, e.g. manta blanca (white mantle) in Ecuador, palomilla (little dove) in Colombia and asa branca (white wing) in Brazil. V-shaped wings This is perhaps the most distinctive feature of the group. Phlebotomines at rest hold their wings in a raised V (Fig. 2). The wings are never closed or laid flat across the body. This feature explains another vernacular name in Brazil, cangalinha or little yoke. Flight Phlebotomines have a weak, direct flight and once on the host progress by a series of small hops. They do not hover round a host and as such are often not recognised as a biting nuisance. Noise The wingbeat frequency of phlebotomines is inaudible to the human ear. They thus do not produce a buzzing or whining noise before biting, which again reduces the perceived nuisance to man. Nocturnal habit Phlebotomines are crepuscular or nocturnal biters, although they may bite during the day if disturbed from their resting sites or when deep shade or clouds produce low light levels. Painful bite Phlebotomines are pool feeders or telmophages which suck blood from a small wound they make in the skin of the host. Their bite is therefore relatively painful, and has been likened to a drop of hot oil or a cigarette burn. In Colombia the insects are sometimes known as quemadores (burners) or pringadores (stingers) GENERAL CHARACTERISTICS The body of a sand fly is divided into 3 parts 1. Head, 2. Thorax, 3. Abdomen. HEAD : The head bears a pairs of long slender and hairy antennae, palpi and proboscis. The antennae are composed of small bead like segments.

THORAX : The thorax bears a pair of legs and three pairs of wings. The legs are long & slender. The wings are upright, lancelets in shape and densely hairy. The second longitudinal vein on the wings branches twice. ABDOMEN : The abdomen has ten segments & is covered with hair. In the female, the tip of the abdomen is rounded. In the male there are claspers, which are conspicuous and attached to the last abdominal segment.

MORPHOLOGY Eggs of the Phlebotominae are about 400 #m long, elon- gate, dark brown, and shiny, with fine surface markings. Markings useful in classification and identification of species include irregular patterns, polygons, ellipses, ridges, pits, and mountain- or volcano-like features (Feliciangeli et al., 1993). Mature larvae of the Phlebotominae are elongate, leg- less, and up to 5 mm in length (Fig 9.2B). The head, thorax, and abdomen bear numerous, prominent, clavate setae that are used in classification and identification. The well-developed head bears eyespots, short antennae, and heavy, toothed mandibles, which oppose a heavy, platelike, serrate labium. The thoracic and abdominal seg- ments are secondarily divided into annuli, with two an- nuli comprising the thoracic and first abdominal segments and three comprising abdominal segments 2-7. Abdom- inal segments 1-8 each bear a medioventral proleg, or pseudopodium. There are two pairs of spiracles: an ante- rior pair on the prothorax and a posterior pair on the greatly reduced abdominal segment 9, which bears two or four long, conspicuous caudal setae adjacent to the spira- cles. Larvae are whitish, with a dark head and dark caudal setae. Phlebotomine pupae attach to substrates in an erect position with the exuviae of the last larval instar attached at the caudal end. They can be distinguished from pupae of the Psychodinae by the clavate body setae and long caudal setae of the larval exuviae. Appendages of the head and thorax are visible and closely appressed to the body. Phlebotomine adults (Fig. 9.2A) are usually less than 5 mm long, densely hairy, and grayish, brownish, or yel- lowish in color. The head is small and hypognathous, eyes conspicuously dark, and ocelli absent. The long, slender, 12- to 16-segmented antennae are similar in males and females. The segments are closely covered with short se- tae, and each segment has a whorl of long setae. The thorax is strongly humped. The wings are large, broadly ovate to elliptical or pointed, densely hairy, and without cross veins beyond the basal area. The abdomen is six- to eight-segmented. The male genitalia are large and con- spicuous. The mouthparts form a short probocis, bearing long, recurred, five-segmented palps with scattered setae. The mouthparts of the female consist of six broad, knife- like stylets (labrum, paired mandibles and maxillae, and hypopharynx), which are held within the fleshy labium when not in use. The mandibles and maxillae are toothed distally. The mandibles cut the skin with scissors-like and sawing movements while the maxillary teeth engage the sides of the wound and hold the mouthparts in place. Blood is taken from a subcutaneous pool produced by injury to the vessels. The food canal is formed by appo- sition of the labrum above and the hypopharynx, which contains the salivary duct, below. Males lack mandibles and do not bite, but some have been observed to take blood from wounds made by the females.

LIFECYCLE

The sand flies prefer to breed in dark places, rich in organic matter and moisture. Its difficult to study the lifecycle of sand flies because the larvae are tiny and dont live in well defined-places, like mosquito larvae. The entire life cycle takes 20 to 40 days except in diapering speaks. The life cycle of sand fly is characterized by complete metamorphosis, having 4 developmental stages egg, larva, pupae & adult. EGG : The eggs are elongated oval, dark brown in color with polygonal sculpturing over the chorion. The eggs are approximately 0.31 mm long and 0.10 mm wide. The female generally lays eggs in the damp dark places in the cattle sheds & poultry. The eggs vary from 40-60 in numbers. Eggs hatch within 1-2 weeks. LARVAE : The larva is maggot like structure, having large head, thorax and abdomen and two long bristle on last abdominal segment. There are 4 larval instars. The mature larvae is 3-6 mm long.Larva feed on decaying organic matter and become a pupa in about 2 weeks.

PUPAE : Pupae are obtect, measuring approximately 2.6 mm in length looking much like a butterfly chrysalis. They are glued to the substrate by the terminal abdominal segments consisting of the terminal pseudopodium of the ninth abdominal segment and the entire exuvium of the fourth instar larva The pupa stage lasts for about 1 week. ADULT : Adults are small, silvery-brownish, long-legged flies with narrow bodies. The wings are less than 3 mm long, and are held erect above the body. They are nocturnally active insects with weak, direct flight capability, typically not dispersing more than a half kilometer (Morrison et al. 1993). Adults shelter during the day in dark, humid places such as tree holes, animal burrows or under rocks. The average life of a sandfly is about 2 weeks. BEHAVIOR AND ECOLOGY Phlebotomine adults are found in and around the breeding sites where suitable resting places are available. Resting sites include forest litter, tree trunks and tree hollows, leaves of plants, caves, excavations, burrows and nests, livestock pens, buildings, cracks and crevices of rocks, masonry and ruins, and other dark, humid, pro- tected sites. Neotropical forest species may be found in forest litter (e.g., L. trapidoi), on understory plants (e.g., L. pessoana), on the trunks of trees (e.g., L. trinidadensis), or in the forest canopy (e.g., L. rorotaensis). The Asian P. caucasicus is commonly found in animal burrows. Certain species are more or less peridomestic, including L. longipalpis, L. verrucarum, P. argentipes, and P. papa- tasi. In southeastern Asia, P. argentipes is found in dark corners of houses and behind hanging clothing and pictures. Sand flies have a hesitating, running motion and fly readily. Their flight is noiseless, consisting of short, dis- crete hops, often only a few centimeters long, and slow, steady flights of longer duration. Flight is inhibited by wind and rain; P. orientalis does not fly at wind speeds above 15 kph. The flight ranges of neotropical forest species are usually less than 200 m, but some species migrate daily between the forest floor and canopy. Flight ranges of P. argentipes and P. oriental# may be 500 m or more, and those of L. longipalpis and P. caucasicus may be 1000 m or more. Longer records include a female P. ariasi recaptured 2.2 km from a release point in France and a male P. perniciosus captured on the island of Jersey (UK) off the coast of France, 25 km from the near- est source on the mainland. Adult sand flies feed on nectar, fruit juices, sap, honey dew, and other sugar sources. Females of P. papatasi are known to pierce the stems and leaves of plants to ob- tain sap. Female sand flies feed on blood in addition to sugars. Blood feeding is limited to areas of exposed skin such as the ears, eyelids, nose, feet, and tail. Males of L. longipalpis fly to the hosts of blood-feeding females, establish territories at blood-feeding sites on the host, and defend territories against other males. Males emit a ter- penoid pheromone from glands on the abdomen to at- tract females, which then feed and mate within the males' territories. Mating is preceded by elaborate courtship behavior. The biting activity of most species of sand flies occurs during twilight or darkness. Some, however, bite freely in daylight, including several medically important vec- tor species. L. panamensis, L. pessoana, L. sanguinaria, and L. trapidoi of the tropical lowlands of Panama bite only at temperatures above 20~ while L. verrucarum of the cool mountain valleys of Peru bites at tempera- tures as low as 10~ Some species are endophilic (e.g., L. verrucarum and P. papatasi), and others are exophilic (e.g., L. trapidoi and P. perniciosus). Most sand flies have broad host ranges, but some are narrowly restrictive. L. gomezi is known to feed on birds and on five different orders of mammals, while L. vexator feeds exclusively on lizards and L. vespertilionis feeds exclusively on bats. P. papatasi is anthropophilic through- out its range. P. argentipes is anthropophilic in

some areas but zoophilic, feeding preferentially on cattle, in others. Light-trap collections in Panama indicate the presence of a primarily anthropophilic association represented by L. gomezi, L. panamensis, and L. dysponeta and a pri- marily zoophilic association represented by L. carpenteri, L. triramula and L. camposi. In some locations, the anthropophilic and zoophilic associations alternate by season. Phlebotomine species often exhibit characteristic sea- sonal and biotopic patterns. In tropical areas, populations of most species increase during or shortly after the rainy season. In India, populations of P. argentipes and P. pa- patasi decrease in the dry season and increase during the monsoon season. In Africa, population densities of P. duboscqi and P. martini vary seasonally with those of the rodents on which they feed. Sand fly populations in Panama are strongly correlated with the degree of deve- lopment of the forest cover, increasing from grassy to secondary forest biotopes and from secondary to mature forest biotopes. Similar situations exist in Kenya, where populations of S. bedfordi and S. antennata increase from thickets to open-canopy forests and from open-canopy forests to closed-canopy forest. In Nigeria, populations of S. bedfordi and S. antennata increase from the open plains to more heavily vegetated habitats.

PUBLIC HEALTH IMPORTANCE Sand flies can be annoying biting pests in places where they are abundant. They may bite or probe several times before and after feeding, each time causing a sharp, pricking sensation. Residents of Peru have difficulty sleeping in areas that are highly infested with L. verrucarum. In one study, the mean biting rate was estimated to be 20-50 bites per person per night. One in- dividual received an estimated 300 bites in a single night. Other highly anthropophilic species are L. diabolica in the United States; L. gomezi, L. olmeca, L. panamensis, L. pessoana, L. sanguinaria, L. trapidoi, and L. ylephiletor in Central America; L. wellcomei in Brazil; and Phleboto- mus sergenti and P. papatasi in the Old World. The initial bites received by an individual typically in- duce sensitization, resulting in immediate or delayed skin reactions to subsequent bites. The reaction to the bite of P. papatasi is a pink or red papule about 2-3 mm in diameter and 0.5 mm high, which remains prominent for 4-5 days before gradually disappearing. Moderate to severe itching usually occurs. Individuals that become hypersensitive often develop hives, with pronounced swelling of the eyelids and lips if those sites are bitten. Prolonged exposure to sand fly bites results in eventual desensitization. Chronically exposed individuals living in areas with high sand fly populations may exhibit no reac- tion to their bites. Many species of Lutzomyia and Phlebotomus are vectors of viral, bacterial, and protozoan pathogens of humans (Table I). Zoophilic species, including species of Brump- tomyia, Warileya, and Sergentomyia, as well as Lutzomyia and Phlebotomus, may be involved in the maintenance of zoonotic diseases. DISEASE TRANSMITTED Phlebotomus argentipes Phlebotomus papatasi Phlebotomus sergenti S. Punjabensis Kala azar Sandfly fever Oriental sore Sandfly fever

VECTOR FOR LEISHMANIASIS L. donovani L.tropica L. braziliensis : visceral leishmaniasis : cutaneous leishmaniasis : mucocutaneous Visceral Leishmaniasis Visceral leishmaniasis is also known as kala-azar (from the Hindi for "black fever") and dumdum fever (from Dum Dum, the location of a former British arsenal near Calcutta). It is a chronic systemic disease that begins with an inconspicuous cutaneous lesion at the site of inoc- ulation by the bite of an infective sand fly. From this site the parasites are distributed through the body in the bloodstream, producing chronic fever, wasting, marked hepatosplenomegaly (especially splenomegaly), pancy- topenia, and hypergammaglobulinemia. Incomplete syn- dromes are common, and early clinical manifestations are variable. Untreated visceral leishmaniasis is usually fatal. Leishmania tropica, however, may produce inapparent or subclinical visceral infections, a condition called vis- cerotropic leishmaniasis. The intrinsic incubation period is usually 2 to 4 months. Cutaneous lesions may appear after apparent recovery or cure and may persist for up to 20 years in the absence of treatment. Such lesions are known as post-kala-azar cutaneous leishmaniasis or der- mal leishmanoid. Known and suspected vectors and reservoirs of visceral leishmaniasis are shown in Table I. In India, Nepal, and Bangladesh, humans and sand flies are the only known hosts. Transmission of L. donovani by P. argentipes in India is a classic example of an epidemic disease. An epi- demic in Assam State in the 1890s depopulated whole villages and decimated populations over large areas. Sub- sequently, large epidemics occurred in Assam State, Bihar State, and the Bengal region. A resurgence of visceral leishmaniasis occurred in the 1970s when the World Health Organization program of spraying homes with dichlorodiphenyltrichloroethane (DDT) for eradication of malaria was discontinued. In Bihar State, 100,000 cases were reported in 1977. Cutaneous Leishmaniasis Cutaneous leishmaniasis begins with a macule that de- velops into a nodular lesion at the site of inoculation by the bite of an infective sand fly. The intrinsic incu- bation period may be a week to many months. Multi- ple lesions may develop at sites of multiple bites. Lesions may remain nodular or develop into sharply demarcated, granulomatous ulcers that discharge necrotic material (Fig. 9.8). New areas of the body become involved by extension of the primary lesions or by metastasis via the blood or lymph (disseminated cutaneous leishmaniasis). Several clinical manifestations of cutaneous leishmani- asis have acquired specific common names: In the Old World, the condition characterized by single or multiple cutaneous ulcers due to L. tropica or L. major has been called oriental sore, tropical sore, Aleppo boil, Baghdad boil, Jericho boil, or Delhi boil. The form of lesion caused by L. tropica in the Mediterranean region and eastward to India has been called the classical form or dry form, and the form of lesion caused by L. major in Central Asia has been called the wet form. In Central America, the condition characterized

by single or multiple ulcers on the face or ears due to L. mexicana is known as chiclero ulcer. In French Guiana, the condition characterized by moderate ulcers due to L. amazonensis or L. guyanensis is known as pian bois. In Peru and Ecuador, the condition characterized by numerous, small, benign lesions due to L. peruviana is lmown as uta. In South America, mu- cocutaneous leishmaniasis due to L. amazonensis or L. braziliensis is known as espundia. Cutaneous leishmaniasis may be self-limiting or chronic. Known and suspected vectors and reservoirs of cuta- neous leishmaniasis are shown in Table I. Transmission of L. major by P. caucasicus in semiarid regions of cen- tral Asia is a classic example of an endemic zoonosis. In this area, P. caucasicus breeds in burrows of gerbils (Rhombomys opimus) and ground squirrels (Sperm0phil0p- sis leptodactylus) and transmits the agents of cutaneous leishmaniasis from animal to animal and from animals to humans. When gerbils were eradicated from the vicinity of a construction camp in Turkestan, leishmaniasis disap- peared from the construction workers. Cutaneous leishmaniasis due to L. mexicana occurs rarely, but widely, in south-central Texas (Bexar, Cameron, Gonzales, Uvalde, and Wells counties) and the adjoining states of Coahuila, Nuevo Leon, and Tamaulipas in Mexico. Typical and diffuse forms have been reported. Lutzomyia anthophora is believed to trans- mit the disease among woodrats ( Neotoma micropus), and L. diabolica is suspected of transmission from woodrats to humans. Seropositive coyotes and an infected cat have been found in southern Texas. mucocutaneous Lesions of the mucous membranes of the nose, mouth, and pharynx ( mucocutaneous or nasopharyngeal leishma- niasis) may develop after the primary lesion has healed or in the absence of a recognized primary lesion (Fig. 9.9). Species with known potential to produce mucocutaneous infections are indicated in Table I. The term diffuse cu- taneous leishmaniasis is applied to a progressive, anergic, nonulcerative condition due to defective cell-mediated immunity. Mucocutaneous leishrnaniasis persists for many years and ultimately may be fatal. Chiclero ulcer and dif- fuse cutaneous leishmaniasis are also chronic, and dif- fuse cutaneous leishmaniasis is resistant to treatment. In- fections of L. tropica may recur at or near the site of the healed ulcer after apparent cure, a condition clini- cally known as leishmaniasis recidivans or chronic relaps- ing leishmaniasis. Sldn lesions of leishmaniasis are prone to secondary infections by bacteria and fungi, and infes- tation by fly larvae (myiasis) may occur. Disfiguring scars remain after healing. CONTROL SPRAY INSECTICIDES : Measures to control adult sand flies (Lutzomyia and Phlebotomus) include the use of insecticides (currently mostly pyrethroids) for residual spraying of dwellings and animal shelters, space-spraying, insecticide-treated nets, impregnated dog-collars and personal protection through application of repellents/insecticides to skin or fabrics. RESERVOIR CONTROL : Certain species of mammals are important reservoirs of Leishmania and by eliminating the reservoir species living near human habitation, disease rates can sometimes be decreased. For instance, rodenticides have been used against the great gerbil, Rhombomys opimus, in Central Asia. Insecticide spraying of larval habitat is usually normally not possible because very little is known about where the larvae occur

Sanitation: Removal of shrubs and vegetation, filling of cracks and crevices in the wall & floor and distance of cattle sheds and poultry from human habitations. Personal protection : DDT , DIMP are one of the most important methods of preventing bites from sand flies. A sandfly net is useful too. Methods for investigating suspected breeding sites of phlebotomine sand flies include the direct examination of soil and litter, extraction by Berlese funnel, wet sieving or flotation, and emergence trapping. Surveillance and col- lection methods for adult sand flies include trapping and aspiration from resting sites, humans, and bait animals. Effective trap designs include light traps, bait traps, sticky traps, and flight traps. Smoke, insect repellent spray, or a twig or stick can be used to flush sand flies from inacces- sible resting sites for collection. Insect repellents and protective clothing are effective personal protectants. Sand flies do not bite through cloth- ing; however, L. verrucarum is reported to crawl beneath clothing and bed sheets to reach the skin. Long sleeves, trousers, and socks should be worn in areas where sand flies are active. Head nets, gloves, repellent-treated net jackets and hoods, and repellent lotions and sprays pro- vide effective protection for the face, hands, arms, and other exposed areas of the body. Outdoor sleeping areas should be high, open, breezy, dry, and as far from potential breeding sites as possible. Bed nets should be used when sleeping in areas where sand flies are present. Standard 16- and 18-mesh bed nets, head nets, window screens, and screen doors do not ex- clude sand flies, but both standard and large-mesh screens and nets are effective when treated with contact insec- ticides or repellents. Fine-mesh nets and screens are lit- tle used because they impede circulation of the air. Sand flies will bite through untreated nets in contact with the skin. Ultraviolet electrocutor traps have been recommended for control of Phlebotomus papatasi. Indoor flight activity can be reduced by use of electric fans. Sand flies do not usually fly into the upper stories of buildings. The use of insecticides can be helpful in reducing sand fly numbers. Natural and synthetic pyrethroid and thio- cyanate aerosols provide effective control indoors when applied twice daily, at night and in the early morning. Smoke from "mosquito coils" containing pyrethroids is also effective. Organochlorine, organophosphate, carba- mate, and pyrethroid insecticides are effective for resi- dual control of adult sand flies; however, resistance to organochlorine insecticides has been reported in P. papatasi. Insecticides should be applied to inside walls and ceilings, to the inside and outside of screens and doors, and to 1-2 feet of the outer wall around window and door casements and along foundations. Wall hang- ings impregnated with pyrethroids have been used for sand fly control in homes in Kenya. Area treatments may be needed out-of-doors. Area treatments should be di- rected toward potential breeding and resting sites, such as outcroppings, fences, walls, buildings, caves, burrows, and tree trunks. Another approach for effective control is the elimina- tion of breeding sites and resting sites of sand flies. Breed- ing and resting sites of P. papatasi in and around homes can be destroyed by filling cracks and crevices of walls, ceilings, and floors and by clearing and rolling, tamping, or paving outdoor areas. In Italy, transmission of leish- manial agents by P. perfiliewi has been reduced by locat- ing piles of farm manure where the sand flies breed at safe distances from homes. In Kenya, the transmission of leishmanial agents by P. martini has been reduced by destroying all termite mounds, in which this species breeds, within 20 m of homes. In Panama and French Guiana, leishmaniasis has been reduced by eliminating breeding and resting sites of sand

flies by deforestation. Control of animals that serve as reservoirs for leishma- nial organisms may be feasible in some areas. Elimina- tion of dogs is an effective preventive measure where dogs have been implicated as reservoirs. Reduction of feral dog and jackal populations by disposal of offal from slaughter houses and poultry farms has significantly reduced the in- cidence of visceral leishmaniasis in Iraq. P. papatasi has been controlled in Jordan and Russia by digging, plow- ing, or flooding gerbil burrows. The use of parasites and predators in sand fly control has not been demonstrated. The bacteria Bacillus sphae- ricus and B. thuringiensis var. israelensis ( Bti, or serotype H12), and certain gregarine protozoa and tylenchid nematodes, are potential agents. The immunization of humans by inoculation of liv- ing Leishmania tropica, called leishmanization, has been employed for control of cutaneous leishmaniasis in Iran, Israel, and Russia.

CONCLUTION