Abstract: Gastric or cutaneous habronemosis caused by Habronema microstoma Creplin, 1849 and Habronema muscae Carter, 1865 is a parasitic disease of equids transmitted by muscid flies. There is a paucity of information on the epidemiology of this disease, which is mainly due to limitations with diagnosis in the live animal and with the identification of the parasites in the intermediate hosts. To overcome such limitations, a molecular approach, based on the use of genetic markers in the second internal transcribed spacer (ITS-2) of ribosomal DNA, was established for the two species of Habronema. Characterisation of the ITS-2 revealed sequence lengths and G+C contents of 296 bp and 29.5% for H. microstoma, and of 334 bp and 35.9% for H. muscae, respectively. Exploiting the sequence difference (approximately 40%) between the two species of nematode, primers were designed and tested by the polymerase chain reaction (PCR) for their specificity using a panel of control DNA samples from common equid endoparasites, and from host tissues, faeces or muscid flies. Effective amplification from each of the two species of Habronema was achieved from as little as 10 pg of genomic DNA. Hence, this molecular approach allows the specific identification and differentiation of the DNA from H. microstoma and H. muscae, and could thus provide a molecular tool for the specific detection of Habronema DNA (irrespective of developmental stage) from faeces, skin and muscid fly samples. The establishment of this tool has important implications for the specific diagnosis of clinical cases of gastric and cutaneous habronemosis in equids, and for studying the ecology and epidemiology of the two species of Habronema.

Abstract: The present study evaluated the engorgement and drop-off rhythms of Anocentor nitens females feeding on horses. Drop-off rhythm was evaluated at 6h-intervals (06:00, 12:00, 18:00, and 00:00 h) on horses held in stalls or in a pasture. A new method of marking feeding female ticks (the bowknot technique) was developed to evaluate ticks on horses in pasture that attached to different parts of the horse's body. This technique was highly successful, indicating no significant interference on tick engorgement rate or final tick weight, length and reproductive capability. Horses held in the pasture during the summer produced only 28.2% of the tick detachment during the daylight period from 06:00 to 18:00 h. In contrast, 53.4% of the ticks detached during this same 12 h-period during the winter. This difference was probably related to the longer scotoperiod during the winter. Different drop-off rhythms were observed for females attached to different anatomical parts of the horse's body. For example, ticks attached to the ears, perineum, and tail showed similar drop-off patterns, but were different from ticks attached to mane, rump and other body parts. The idiosoma length of the feeding female ticks was individually measured every 6 h until the engorged female detached naturally. The engorgement rate (increase in millimeters of the body length per hour) was evaluated during the last 96 h of parasitism. The highest engorgement rates were observed during the last 24 h of parasitism (approximately 0.16 mm/h), which were four-fold higher than the engorgement rates of the previous 3 days ( approximately 0.04 mm/h), demonstrating that these lower and higher values corresponded to the slow and rapid feeding phases reported elsewhere. Based on these data, the 6 mm idiosoma length was estimated as the minimal length that would correspond to the time point (i.e. 24 h before detachment) during which ticks would undergo the rapid feeding phase and detach as fully engorged females. When this 6 mm length was tested to estimate the number of engorged females detaching from horses in a period of 24 h, the estimated accuracy varied from 58.5 to 97.7% (mean: 73.3%).

Abstract: Once a month, from June 1992 to May 1993, collections of tabanids on horse were conducted in the Nhecolandia, Pantanal State of Mato Grosso do Sul, Brazil. Tabanid catches using hand nets were conducted from sunrise to sunset at grassland and cerradao (dense savanna) habitats. A total of 3,442 tabanids from 21 species,12 genera, and 3 subfamilies were collected. Although species abundance varied seasonally depending on habitat, no habitat specificity was observed for the most abundant species. In the grassland, 1,625 (47.2%) tabanids belonging to 19 species were collected, while 1,817 (52.8%) tabanids from 17 species were caught in the cerradao. The number of tabanid species varied from 7 during winter (July/August) to 15 in the spring (October). Tabanus importunus (56%) was the most abundant species, followed by T. occidentalis (8.2%), and T. claripennis (8.1%). The tabanid peak, in October, coincided with the beginning of the rainy season. The population peak of most species, including those with higher vector potential, suggests that the rainy season can be considered as the period of potentially higher risk of mechanical transmission of pathogens by tabanids to horses in the region.